Fertiliser (Inorganic, Organic or Mixed) (Control) Order, 1985

Essential Commodities Act, 1955

Published vide Notification Gazette of India, Extraordinary, Part 2, Section 3(1), dated 25th September, 1985, (w. 53).

Last Updated 11th April, 2023 [act2219]

G.S.R. 758(E), dated 25th September, 1985. - In exercise of the powers conferred by Section 3 of the Essential Commodities Act, 1955 (10 of 1955), the Central Government hereby makes the following Order, namely -

1. Short title and commencement. - (1) This Order may be called the Fertiliser [(Inorganic, Organic or Mixed)] (Control) Order, 1985.

(2) It shall come into force on the date of its publication in the Official Gazette.

2. Definitions. - In this Order, unless the context otherwise requires, -

(a) "Act" means the Essential Commodities Act, 1955 (10 of 1955) ;

[(ab) "biostimulant" means a substance or microorganism or a combination of both whose primary function when applied to plants, seeds or rhizosphere is to stimulate physiological processes in plants and to enhance its nutrient uptake, growth, yield, nutrition efficiency, crop quality and tolerance to stress, regardless of its nutrient content, but does not include pesticides or plant growth regulators which are regulated under the Insecticide Act, 1968 (46 of 1968);]

(b) "Certificate of source" means a certificate given by a State Government, Commodity Board, manufacturer, pool handling agency or, as the case may be, wholesale dealer indicating therein the source from which fertilizer for purpose of sale is obtained ;

(c) "Commodity Board" means the Coffee Board constituted under Section 4 of the Coffee Act, 1942 (7 of 1942), or the Rubber Board constituted under Section 4 of the Rubber Act, 1947 (24 of 1947), the Tea Board constituted under Section 4 of the Tea Act, 1953 (29 of 1953), or, as the case may be, the Cardamom Board constituted under Section 4 of the Cardamom Act, 1965 (42 of 1965) ;

(d) "compound or complex fertilizer" means a fertilizer containing two or more [* * *] nutrients during the production of which chemical reaction takes place ;

(e) "controller" means the person appointed as Controller of Fertilizers by the Central Government and includes any other person empowered by the Central Government to exercise or perform all or of the powers, or as the case may be, functions of the Controller under this Order ;

[(ee) "Customised Fertiliser" means a granular multi nutrient carrier which contains Primary , Secondary and /or micro nutrient forms, both from inorganic and/or organic sources, manufactured through a systematic process of Fusion blend granulation, [or precision blend technology,] formulated on the basis of soil fertility data and include 100% water soluble specialty fertilizer as customized combination products;]

(f) "dealer" means a person carrying on the business of selling fertilizers, whether wholesale or retail, [or industrial use] and includes a manufacturer and a pool handling agency carrying on such business and the agents of such person, manufacturer or pool handling agency ;

(g) [* * *].

[(h) "fertiliser" means any essential substance, either in straight or mixed form and derived from either inorganic, organic or mixed sources, that is used or intended to be used to provide essential plant nutrients or beneficial elements or both for the soil or for the crop or makes essential plant nutrients available to the plants either directly or by biological processor by both in the soil or plant as notified from time to time by Central Government and specified in the schedules appended to this order or as may be notified by the State Governments [and includes a biostimulant].

Explanation. - For the purpose of Fertiliser, -

(i) "the essential plant nutrients" include Primary Nutrients (Nitrogen, Phosphorous and Potassium), Secondary Nutrients (Calcium, Magnesium and Sulphur) and Micro Nutrients (Zinc, Manganese, Copper, Iron, Boron and Molybdenum);

(ii) "Beneficial element" means any element as notified by the Central Government from time to time.]

(i) "Form" means a form appended to this order ;

[(i) "industrial dealer" means a dealer who sells fertilisers for industrial purposes.

(ii) "industrial purpose" means the use of fertilizer of purpose other than fertilisation of soil and increasing productivity of crops;]

[(j) "grade" means the nutrient contents in the fertilizers, expressed in percentage;]

(k) "granulated mixture" means a mixture of fertilizers [* * *] made by intimately mixing two or more fertilizer with or without inert material and granulating them together, without involving any chemical reaction ;

[(kk) "importer" means a person who imports fertilizer in accordance with the Export and Import Policy of the Central Government, as amended from time to time;]

(l) "inspector" means an Inspector of Fertilizers appointed under Clause 27;

(m) "manufacturer, [importer]" means a person who produces fertilizers or mixtures of fertilizers [* * *] and the expression 'manufacture, [importer]' with its grammatical variations shall be construed accordingly ;

[(ma) "Marketer" means such fertiliser companies who sell, offer for sale, carry on the business of selling of fertiliser manufactured by other fertiliser company;]

[(mb) the mixture of micronutrient are those which are made by mixing of micronutrient notified in Schedule I under the sub-heading "1(g) MICRONUTRIENTS" of said Order.]

[(n) "mixture of fertilizers" means a mixture of fertilizer made by physically mixing two or more fertilizers ; with or without inert material in physical or granular form and includes a mixture of NPK fertilizers, a mixture of micronutrient fertilizers and a mixture of NPK with micronutrient fertilizers ;]

[(nn) "Notified Authority" means an authority appointed under clause 26A];

(o) "offer for sale" includes a reference to an intimation by a person of a proposal by him for the sale of any fertilizer, made by publication of a price list, by exposing the fertilizer for sale indicating the price, by furnishing of a quotation or otherwise howsoever ;

(p) "physical mixture" means a mixture of fertilizers [made by physically, mixing two or more] fertilizers with or without inert material necessary to make a required grade, without involving any chemical reaction ;

(q) "prescribed standard" means, -

(i) in relation to fertilizer included in column I of Part A of Schedule I, the standard set out in the corresponding entry in column 2, subject to the limits of permissible variation as specified in Part B of that Schedule ; and

(ii) in relation to a mixture of fertilizers, the standard set out in respect of that mixture under sub-clause (1) of Clause 13 by the Central Government, subject to the limits of permissible variation as specified in Part B of Schedule I;

[(iii)] in relation to a mixture [mixture of fertiliser] the standard set out in respect of that mixture under sub-clause (2) of Clause 13 by the State Government, subject to limits of permissible variation as specified in Part B of Schedule I.

[(vii) prescribed standard means in relation to Customised Fertilizers, standards set out in respect of Customised Fertilizers under clause 20B by the Central Government, subject to limits of permissible variation as specified in part B of Schedule –I];

[(viii) in relation to a biostimulant included in column (2) of Part-A of Schedule VI, the standard set out in the corresponding entry in column (3), subject to the limits of permissible variations as specified in Part-B of the said Schedule;]

(r) "pool handling agency" means an agency entrusted by the. Central Government with functions relating to handling and distribution of imported fertilizers ;

(s) "registering authority" means a registering authority appointed under Clause 26 ;

(t) "retail dealer" means a dealer who sells fertilizers to [to farmers or plantations for agricultural use such as for fertilisation of soil and increasing productivity of crops];

(u) "Schedule" means a Schedule appended to this order ;

(v) "special mixture of fertilizers" means any mixture of fertilizers prepared for experimental purposes in pursuance of a requisition made by any person (including a person engaged in the cultivation of tea, coffee or rubber) for sale to that person in such quantity and within such period as may be specified in such requisition ; and

(w) "wholesale dealer" means a dealer who sells fertilizers otherwise than in retail [for agricultural use such as for fertilisation of soil and increasing productivity of crops].

II. Price Control

3. Fixation of prices of fertilizers. - (1) The Central Government may, with a view to regulating equitable distribution of fertilizers and making fertilizers available at fair prices, by notification in the Official Gazette, fix the maximum prices or rates at which any fertilizer may be sold by a dealer, manufacturer or a pool handling agency.

(2) The Central Government may having regard to the local conditions of any area, the period of storage of fertilizers and other relevant circumstances, fixed different prices or rates for fertilizers having different periods of storage or for different areas or for different classes of consumers.

(3) No dealer, manufacturer or pool handling agency shall sell or offer for sale any fertilizers at a price exceeding the maximum price or rate fixed under this clause.

4. Display of stock position and price list of fertilizers. - Every dealer, who makes or offers to make a retail sale of any fertilizers, shall prominently display in his place of business. -

(a) the quantities of opening stock of different fertilizers held by him on each day ;

Explanation. - The actual stocks at any point of time during the day may be different from that of the displayed opening stocks to the extent of sale and receipt of such fertilizers up to the time of inspection during that day.

(b) a list of prices or rates of such fertilizers fixed under Clause 3 and for the time being in force.

5. Issue of cash/credit memorandum. - Every dealer shall issue a cash or credit memorandum to a purchaser of a fertilizer in [Form M].

III. Control On Distribution Of Fermizers By Manufacturer

6. Allocation of fertilizers to various States. - The Central Government may, with a view to securing equitable distribution and availability of fertilizers to the farmers in time, by notification in the Official Gazette, direct any manufacturer to sell the fertilizers produced by him in such quantities and in such State or States and within such period as may be specified in the said notification.

[Chapter IV

Authorisation or Registration of Dealers]

[7. Registration of Industrial dealers and authorisation of other dealers. - No person shall sell, offer for sale or carry on the business of selling of fertilizer at any place as wholesale dealer or retail dealer except under and in accordance with clause 8:

Provided that a State Government may, if it considers it necessary or expedient, by notification in the Official Gazette, exempt from the provisions of this clause any person selling fertilizer to farmers in such areas and subject to such conditions as may be specified in that notification.]

8. Application for intimation or registration. - (1) Every person intending to sell or offer for sale or carrying on the business of selling of fertilizer as Industrial Dealer shall obtain a certificate of registration from the controller by making an application in Form A together with the fee prescribed under clause 36 and a Certificate of source [in Form O].

(2) Every person including a manufacturer, an importer, a pool handling agency, wholesaler and a retail dealer intending to sell or offer for sale or carrying on the business of selling of fertilizer shall make a Memorandum of Intimation to the Notified Authority, in Form A1 duly filled in, in duplicate, together with the fee prescribed under clause 36 and certificate of source in Form O.

(3) On receipt of a Memorandum of Intimation, complete in all respects, the Notified Authority shall issue an acknowledgement of receipt in Form A2 and its shall be deemed to be an authorisation letter granted and the concerned person as authorised dealer for the purposes of this Order:

Provided that certificate of registration granted before the commencement of the Fertiliser (Control) (Amendment) Order, 2003, shall be deemed to be an authorization letter granted under the provisions of this Order:

Provided Further that where the applicant is a State Government, a manufacturer or an importer or a pool-handling agency, it shall not be necessary for it or him to submit Form O:

Provided also that a separate Memorandum of Intimation shall be submitted by an applicant for wholesale business or retail dealership, as the case may be:

Provided also that where fertilizers are obtained for sale from different sources, a certificate of source from each such source shall be furnished in Form O:

[Provided also that a wholesale dealer, excepting a manufacturer, a pool handling agency or a State Government, shall not issue the certificate of source of another wholesale dealer. A wholesale dealer shall issue a certificate of source only to a retail dealer:]

[Provided also that where the manufacturer of organic fertilizer is a State Government or municipality, it shall not be necessary for it to obtain the authorisation letter:

Provided also that where the manufacturer of vermi-compost, other than a State Government or municipality, has annual production capacity less than 50 metric tonne, it shall not be necessary for him to obtain the authorisation letter.]

[(4) No authorisation letter shall be granted to any applicant for retail dealership, unless the applicant possess the certificate course of fifteen days from any State Agriuclture University or Krishi Vigyan Kendras or National Institute of Agricultural Extension Management (MANAGE) or National Institute of Rural Development and Panchayati Raj (NIDPR) or Fetiliser Association of India or any other approved Government Institute:

Provided that a person in possession of Bachelor of Science in Agriculture or chemistry or Diploma in Agriculture Science from a recognised University or Institute or equivalent course having one of the subject on fertiliser or agri inputs, as notified by the State Government shall not be required to possess separate certificate course:

Provided further that a dealer who has been granted authorisation letter before commencement of the Fertiliser (Inorganic, Organic or Mixed) (Control) Fourth Amendment Order, 2018 shall not be required to possess the qualification at the time of renewal of their authorisation letter:

Provided also that the said qualification shall not be applicable for renewal of the authorisation letter of the registered Agricultural Cooperative Societies and State Marketing Federations subject to condition that such Society or Federation shall engage a person who possesses the qualification under this clause.]

9. Grant or refusal of certificate of registration. - The registering authority [or, as the case may be, the controller] shall grant a certificate of registration in Form B within thirty days of the receipt of application to any person who applies for it under CI 8 :

Provided that no certificate of registration shall be granted to a person. -

(a) if his previous certificate of registration is under suspension ; or

(b) if his previous certificate of registration has been cancelled within a period of one year immediately precedent the date of application ; or

(c) if he has been convicted of an offence under the Act, or any Order made thereunder within three years immediately preceding the date of making the application ; or

(d) if he fails to enclose with the application a certificate of source ; or

(e) if the application is incomplete in any respect ; or

[(f) if he makes an application for obtaining the certificate of registration for industrial dealer and, excepting if he is a manufacturer or pool handling agency, holds a valid certificate of registration for wholesale dealer or retail dealer or both, and as the case may be, the vice versa.]

10. Period of validity of certificate of registration. - Every certificate of registration granted under Clause [as case may be renewed under Clause 11] shall, unless suspended or cancelled, be valid for a maximum period of three years, from the date of issue. [Notwithstanding anything contained in the said clause, the letter of Authorisation granted to the manufacture of City Compost issued under clause 8, unless suspended or cancelled is valid in perpetuity.]

11. Renewal of certificate of registration. - (1) Every holder of a certificate of registration desiring to renew the certificate of registration granted under Clause 9 shall, before the date of expiry of such certificate of registration make an application for renewal to the registering authority [or, as the case may be, the controller"] in Form C, in duplicate, together with the fees prescribed under Clause 36 for such renewal and a certificate of source as required under Clause 8.

(2) On receipt of such application, together with such fee and certificate of source, the registering authority ["or, as the case may be, the controller"] may renew the certificate of registration :

Provided that a certificate of registration shall not be renewed, if the holder of the certificate of registration did not sell any fertilizer during the period of one year immediately preceding the date of expiry of the period of validity of the certificate of registration sought to be renewed.

(3) If any application for renewal is not made before the expiry of the period of validity of the certificate of registration but is made within one month from the date of such expiry, the certificate of registration may be renewed on payment of such additional fee as may be prescribed by the State Government, ["or, as the case may be, the controller"] in addition to the fee for renewal of the certificate of registration.

(4) Where the application for renewal is made within the time specified in sub - clause (1) or sub - clause (3) the application shall be deemed to have held a valid certificate of registration, until such date as the registering authority ["or, as the case may be, the controller"] passed orders on the application for renewal.

(5) If an application for renewal of a certificate of registration is not made within one month of the date of expiry of the period of validity of the certificate of registration, the certificate of registration shall be deemed to have lapsed on the date on which its validity expired and any business carried on after that date shall be deemed to have been carried on in contravention of Clause 7.

V. Manufacture Of Mixtures Of Fertilizers

12. Restriction on preparation of mixtures of fertilizers. - No person shall carry on the business of preparing any mixture of fertilizers [* * *] or special mixture of fertilizers except under and in accordance with the terms and conditions of a certificate of manufacture granted to him under Clause [15 to 16].

[13. Standards of mixtures of Fertilizers]. - (1) Subject to the other provisions of this order no person shall manufacture any mixture of fertilizers unless such mixture conforms to the standards set out in the notification to the issued by the Central Government in the Official Gazette.

[(2) Subject to the other provisions of this Order, no person shall manufacture any mixture of fertilisers unless such mixture conforms to the standards set out in the notification to be issued by the State Government in the Official Gazette ;

Explanation. - For the purposes of this sub - clause, mixture of fertilisers shall not include liquid fertilisers and 100% water soluble fertilisers, containing N.P.K.].

(3) Notwithstanding anything contained in his Order, those units which are already manufacturing [mixture of fertilisers] as on the date of publication of this order, for which standards have not yet been specified under 13(2), may continue to manufacture for sale, sell, offer for sale, stock or exhibit for sale or distribute such mixtures of micro - nutrient fertilizers for a period of [upto 27th July, 1992] from such date or till the standards are prescribed by the State Governments whichever is earlier subject to the condition that before the expiry of said period, they will follow the standards prescribed by State Government and obtain Certificate of Manufacture under Clause 15.

(4) No Certificate of Manufacture shall be granted in respect of any mixture of fertilizers or [* * *] which does not conform to the standards set out in the notification referred in sub - clause (1) or (2) ;

(5) Nothing in this clause shall apply to special mixtures of fertilizers.

14. Application for certificate of manufacture of mixtures of fertilizers. - (1) Every person desiring to obtain a certificate of manufacture for preparation of any mixture of fertilizers or special mixture of fertilizers [* * *] shall possess such qualification as may be prescribed for this purpose by the State Government or shall employ a person possessing such qualification for the preparation of such mixture [and possess the minimum laboratory facility as specified, in Clause 21 - A of this order.]

(2) An applicant for a certificate of manufacture for preparation of mixture of fertilizers or special mixture of fertilizers [* * *] shall make an application to the registering authority -

(a) if he is an applicant for a certificate of manufacture for any mixture of fertilizers, [* * *] in Form D, in duplicate, together with the fee prescribed therefor under Clause 36, or,

(b) if he is an applicant for a certificate of manufacture for any special mixture, in Form E, in duplicate, together with the fee prescribed therefor under the said Clause 36 and an attested copy of the requisition of the purchaser.

15. Grant or refusal of certificate of manufacture for preparation of mixtures of fertilizers [* **]. - (1) The registering authority may, for reasons to be recorded in writing, refuse to grant a certificate of manufacture in respect of any mixture of fertilizers, or special mixture of fertilizers [* * *] and shall furnish to the applicant a copy of the order so passed.

(2) Where an application for a certificate of manufacture for mixture of fertilizers [* * *] is not refused under sub - clause (1), the registering authority shall grant a certificate of manufacture in Form F and where an application for a certificate of manufacture for a special mixture is not refused under that sub - clause, such authority shall grant a certificate of manufacture to the applicant in Form "G".

16. Conditions for grant of certificate of manufacture in respect of special mixture of fertilizers and period of validity of such certificate. - (1) No certificate of manufacture in respect of any special mixture of fertilizers shall be granted to an applicant unless he holds a valid certificate of manufacture under this Order for any mixture of fertilizers.

(2) Every certificate of manufacture granted in respect of any special mixture of fertilizer's shall be valid for a period of three months from the date of its issue ;

Provided that the registering authority may, if it is satisfied that it is necessary so to do extend the said period to such further period or periods as it may deem fit, so however, that the total period or periods so extended shall not exceed six months.

17. Period of validity of certificate of manufacture for preparation of mixtures of fertilizers. - Every certificate of manufacture granted under Clause 15 for preparation of a mixture of fertilizers shall, unless suspended or cancelled, be valid for a period of three years from the date of issue.

18. Renewal of certificate of manufacture for preparation of mixtures of fertilizers [* * *]. - (1) Every holder of a certificate of manufacture for preparation of a mixture of fertilizers' [* * *] desiring to renew the certificate, shall, before the date of expiry of the said certificate of manufacture make an application to the registering authority in Form D in duplicate, together with the fee prescribed for this purpose under Clause 36.

(2) On receipt of an application for renewal as provided in sub - clause (1), and keeping in view the performance of the applicant and other relevant circumstance, the registering authority may, if he so decides, renew the certificate of registration by endorsement on Form F and in case the certificate of registration is not renewed, the registering authority shall record in writing his reasons for not renewing the certificate of registration.

(3) If an application for renewal is not made before the expiry of the certificate of registration but is made within one month from the date of expiry of the certificate of registration the certificate of registration may be renewed on payment of such additional fee as may be prescribed by the State Government for this purpose.

(4) Where the application for renewal is made within the time specified in sub - clause (1) or sub - clause (3), the applicant shall be deemed to have held a valid certificate of registration until such date as the registering authority passes order on the application for renewal.

(5) If an application for renewal of a certificate of manufacture is not made within the period stipulated under sub - clause (1) or, as the case may be, under sub - clause (3), the certificate of manufacture shall be deemed to have expired immediately on the expiry of its validity period, and any business carried on after that date shall be deemed to have been carried on in contravention of Clause 12.

VI. Restrictions On Manufacture, Sale, Etc. of Fertilizers

19. Restriction on manufacture, sale and distribution of fertilizers. - [***] No person shall himself or by any other person on his behalf -

(a) manufacture for sale, sell, offer for sale, stock or exhibit for sale or distribute any fertilizer which is not of prescribed standard ;

(b) manufacture for sale, sell, offer for sale, stock or exhibit for sale, or distribute any mixture of fertilizers [mixture of fertilizers] which is not of prescribed standard [(subject to such limits of permissible variation as may be specified from time to time by the Central Government)] or special mixture of fertilizers which [* * *] does not conform to the particulars specified in the certificate of manufacture granted to him under this Order in respect of such special mixture ;

(i) any fertilizer the container whereof is not packed and marked in the manner laid down in this Order ;

(ii) any fertilizer which is in limitation or a substitute for another fertilizer under the name of which it is sold ;

(iii) any fertilizer which is adulterated ;

Explanation. - A fertilizer shall be deemed to be adulterated, if it contains any substance the addition of which is likely to eliminate or decrease its nutrient contents or make the fertilizer not conforming to the prescribed standard.

(iv) any fertilizer the label or container whereof bears the name of any individual, firm or company purporting to be manufacturer of the fertilizer which individual, firm or company is fictitious or does not exist ;

(v) any fertilizer, the label or container whereof anything accompanying therewith bears any statement which makes a false claim for the fertilizer or which is false or misleading in any material particular ;

(vi) any substance as a fertilizer which substance is not, in fact, a fertilizer ; or

(vii) any fertilizer without exhibiting the minimum guaranteed percentage by weight of plan nutrient.

[19A. In cases where the samples were drawn from the dealers out of original sound bags (without any mark of tempering) and are found non-standard, then in such circumstances both dealer and manufacturer shall be made party for filling the case in the concerned court under the Act and proceedings under Clause 31 of this Order.]

20. Specifications in respect of imported fertilizers. - Notwithstanding anything contained in this Order, the Central Government may, by an order published in the Official Gazette, fix separate specifications in respect of imported fertilizers.

[20A. Specification in respect of provisional fertilizer. - Notwithstanding anything contained in this Order, the Central Government may, by order published in the Official Gazette, notify specifications, valid for a period not exceeding three years, in respect of fertilizers to be manufactured by any manufacturing unit for conducting commercial trials.]

[20B. Specification in respect of Customised Fertiliser. -

(i) Notwithstanding anything contained in this Order, the Central Government may by notification in the Official Gazette notify the General specification of customised fertiliser.

(ii) No person shall manufacture any grade of Customised Fertiliser unless such customised fertiliser conforms to the standards set out in the notification to be issued by the Central Government in the Official Gazette under sub-clause (i):

Provided that the grades of customized fertiliser, which the company will manufacture, must be based on the soil fertility data maintained by the Ministry of Agriculture and Farmers Welfare or State Governments:

Provided further that in case, the data for a district for which the company intending to formulate the grade is not available or still under process by the State Government, then the company shall use the scientific data obtained from soil testing results generated by testing in their own laboratories.

(iii) No person, except with the prior permission of Controller shall, manufacture any particular grade of customized fertilizer formulated as per the General specification notified under sub-clause (i).

(iv) Every person, desirous of obtaining a specific product approval of any particular grade of customised fertilizer, shall make an application in form Q in duplicate to the Controller of Fertiliser, Government of India.

(v) On receipt of application under clause (ii), the Controller shall by order in writing either grant or refuse to grant the permission, in respect of manufacturing of any particular grade of customised fertiliser and shall within three months from the date of receipt of application shall furnish a copy of order so passed to the applicant:

Provided that on completion of three years or earlier, manufacturing company of customised fertiliser shall again submit an application for approval for manufacturing of the said grade:

Provided further that the permission for manufacture and sale of Customised Fertiliser shall be granted to only such Fertiliser Companies whose annual production of fertilisers other than CFs is 5.00 lakh metric tonne:

Provided also that such manufacturing companies, having annual production of 5 LMT of fertilizer other than CFs, can set up manufacturing units of CF either on their own or through subsidiaries or joint venture through a minimum stake of 51% in such joint ventures.]

[20C. Specifications in respect of biostimulants. - (1) Notwithstanding anything contained in this Order, the specifications of biostimulants, including its name, active ingredients or where it is not possible to indicate its active ingredients, then its chemical composition, name of the crop to which it may be applied, its benefits and the method of analysis for determination of its active ingredients or chemical composition shall be specified by the Central Government in Schedule VI.

(2) The biostimulants specified in Schedule VI shall be classified under any of the following categories, namely:-

(a) botanical extracts, including seaweed extracts;

(b) bio-chemicals;

(d) vitamins;

(e) cell free microbial products;

(f) antioxidants;

(g) anti-transpirants;

(h) humic and fulvic acid and their derivatives

(3) Notwithstanding anything contained in this order, every manufacturer or importer of a biostimulant shall make an application to the Controller in Form G along with the following data relating to such product for specifying it as a biostimulant in Schedule VI, namely;-

A. Chemistry:

1. Source (natural extracts of plant/microbe/animal / synthetic)

2. Product Specification (with analysis from Good Laboratory Practice(GLP) or National Accreditation Board for Testing and Current Central Legislation/Lucknow Law Times Calibration Laboratory (NABL) accredited laboratory. Physical and Chemical properties of active ingredients and adjuvants, if any.

3. Method of analysis conforming to the specifications.

4. Shelf-life;

B. Bio-efficacy Trials:

1. Agronomic Bio-efficiency trials shall be conducted at National Agricultural Research System, including Indian Council of Agricultural Research, State Agricultural Universities.

2. Bio-efficacy trials shall be conducted at minimum three different doses for one season at three agroecological locations:

C. Toxicity: Name of the Institute (where the manufacturer of biostimulant obtained the certificate) The test report along with recommendation of following five basic acute toxicity tests (i) to (v) and four Eco toxicity tests (vi) to (ix) of GLP accredited laboratory shall be submitted along with the application, namely:-

(i) Acute oral (Rat)

(ii) Acute dermal (Rat)

(iii) Acute Inhalation (Rat)

(iv) Primary skin Irritation (Rabbit)

(v) Eye irritation (Rabbit)

(vi) Toxicity to bird

(vii) Toxicity to Fish (Freshwater)

(viii) Toxicity to honeybee

(ix) Toxicity to earthworm

Note: - Long-term chronic studies as in case of pesticides are not needed to be conducted;

D. Heavy metal analysis report;

E. The sample of the product along with an Affidavit by the manufacturer or importer on nonjudicial stamp paper of rupees ten stating that the product is not laced with pesticide beyond permissible limit of 0.01 ppm:

Provided that in case of a biostimulant having natural origin or other, the requirement of necessary data shall be as decided by the Central Biostimulants Committee as per the guidelines formulated by it in this regard:

Provided further that every manufacturer or importer of a biostimulant shall, notwithstanding that such biostimulant has been specified in Schedule VI, be required to make an application under this sub-clause.

(4) Notwithstanding anything contained in this order, any person manufacturing [or importing] a biostimulant as on the date of publication of this order for which no standards have been specified, may, subject to making an application for grant of provisional registration under sub-clause (5), [continue to manufacture or import] for sale, sell, offer for sale, stock or exhibit such biostimulant for a period of [three years] from the date of publication of this order.

(5) The manufacturer [or importer] referred to in sub-clause (4) shall, within a period of six months from the date of publication of this order, submit an application to the Controller for grant of provisional registration in Form G-1 along with-

(a) details of the product, its specifications, label and such other particulars as may be required;

(b) a certificate issued by the State Government in Form G-2 to the effect that-

(d) no incident has come to its notice about harmful effect of the product.

(6) On receipt of an application under sub-clause (5) for provisional registration, the Controller shall, by an order in writing, grant a provisional certificate of registration of biostimulant in Form G-3.

(7) No person shall manufacture or import any biostimulant unless such biostimulant is included in schedule VI.

(8) The Controller shall, by notification in the Official Gazette, publish the name of the manufacturer or importer of the biostimulant included in Schedule VI.

(9) No biostimulant shall contain heavy metals content over and above the following maximum limit prescribed for various metals, namely,-

In mg/kg (maximum)
Cadmium (as Cd)	5.00
Chromium (VI) (as Cr)	50.00
Copper (as Cu)	300.00
Zinc (as Zn)	1000.00
Lead (as Pb)	100.00
Arsenic (as As203)	10.0.

(10) No biostimulant shall contain any pesticide beyond the permissible limit of 0.01 ppm.]

21. Manufacturers/pool handling agencies to comply with certain requirements in regard to packing and making, etc. - Every manufacturer, import and pool handling agency shall in regard to packing and marking of containers of [fertiliser, non-edible de-oiled cake fertiliser or biostimulant] comply with the following requirements, namely: -

[(a) Every container in which any fertiliser is packed shall conspicuously be superscribed with the word "FERTILISER" and shall bear only such particulars and unless otherwise required under any law nothing else, as may from time to time, be [specified by the Controller in this behalf;

Provided that in case of containers the gross weight of which is 5 kg. or less, no such printing of superscription and other particular shall be necessary if such superscription and other particulars are printed on a separate label which is securely affixed to such container.]]

[(aa) Every container in which any Bio-fertilizer or Organic fertilizer is packed shall conspicuously be superscribed with the words BIO-FERTILIZERS/ORGANIC FERTILIZERS [OR NON-EDIBLE DE-OILED CAKE FERTILIZER, as the case may be] and shall bear only such particulars and unless otherwise required under any law nothing else, as may from time to time, be specified by the Controller in this behalf.]

(b) every container shall be so packed and sealed that the contents thereof cannot be tampered with without breaking the seal :

Provided that where fertilizer manufactured in India are packed in bags stitched in hand, such bags shall bear lead seals, so that the contents thereof cannot be tampered with without breaking the seals.

Provided further that lead sealing shall not be necessary, -

(i) if such bags are machine stitched in such a manner that contents thereof cannot be tampered with without a visible break in the stitching ; and

(ii) in the case of fertilizers imported from abroad and packed in bags stitched in hand, in such a manner that the contents thereof cannot be tampered with without visible break in the stitching.

[Provided also that in case fertilizer bags are in cut, torn or damaged condition during transportation or mishandling during loading or unloading operation, the manufacturer of such fertilizer may, under intimation to the State Government and the Central Government, repack the fertilizer in new bags or re-standardize the quantity in terms of declared weight.]

[(c) Every fertiliser bag in which any fertiliser packed for sale shall be of such weight and size as may be specified by the Central Government from time to time in this behalf.]

[(d) every container in which any biostimulant is packed shall be conspicuously superscribed with the word "Biostimulanf ' and shall bear only such particulars as the Central Government may, by notification in the Official Gazette, specify in this behalf, unless any other particulars are required under any other law for the time being in force.]

[21A. Manufacturers to comply with certain requirements for laboratory facilities. - Every manufacture shall, in order to ensure quality of their product, possess the minimum laboratory facility, as may be specified from time to time, by the Controller].

22. Bulk sale of fertilizers. - Notwithstanding anything contained in the Order, -

(a) a retail dealer may retain at any time one bag or container of each variety of fertilizer in an open and unsealed condition for the purpose of sale ;

(b) a manufacturer may sell the fertilizer manufactured by him in bulk to a manufacturer of mixture of fertilizers, compound/complex fertilizers or special mixture of fertilizers ; and

(c) the Central Government may by notification published in the Official Gazette in this behalf authorise a manufacturer to sell any fertilizer manufactured by him in bulk also direct to farmers for such period as may be specified in that notification:

Provided that a certificate indicating the minimum guaranteed percentage of plant nutrients is issued by the manufacturer to each former at the time of such sale.

23. Disposal of non-standard fertilizers. - (1) Notwithstanding anything contained in this Order, a person may sell, offer for sale, stock or exhibit for sale or distribute any fertilizer which, not being an adulterated fertilizer, does not conform to the prescribed standard (hereinafter in this Order referred to as non - standard fertilizer) subject to the conditions that, -

(a) the container of such non - standard fertilizer is conspicuously superscribed in red colour with the words "non - standard" and also with the sign "X", and

(b) an application for the disposal of non - standard fertilizers in Form H is submitted to the registering authority to grant a certificate of authorisation for sale of such fertilizers and a certificate of authorisation with regard to their disposal and price is obtained in Form I ;

(c) such non - standard fertilizer shall be sold only to the manufacturers of mixtures of fertilizers or special mixtures of fertilizers or research farms of Government or universities of such bodies.

(2) The price per unit of the non - standard fertilizer shall be fixed by the registering authority after satisfying itself that the sample taken is a representative one, and after considering the nutrient contents in the sample determined on the basis of a chemical analysis of the non - standard fertilizer.

(3) The Central Government may, by notification in the Official Gazette and subject to the conditions, if any, laid down in that notification, and subject to guidelines issued in this regard by the Central Government exempt such pool handling agencies, as it deems fit, from complying with conditions laid down in paragraphs (a) and (b) of the sub-clause (1).

[(4) Where any fertiliser imported by the Central Government is found to be of non-standard and the Central Government decides that the fertilizer cannot be permitted for direct use in agriculture, it may permit the use of such fertiliser by manufacturers of complex fertilisers, mixture of fertiliser or special mixture of fertiliser to be sold at such price as may be fixed by the Central Government.

(5) If a manufacturer or importer, defects or has reasonable doubt about the standard of the fertiliser manufactured or imported by him, and dispatched for sale has deteriorated in quality during transit due to natural calamity and is not of the prescribed standards, he may, within fifteen days from the dated of dispatch from factory or port, apply with detailed justifications to the Central Government for obtaining permission for reprocessing the same in a factory to meet the prescribed standards and the Central Government may, after considering the facts, permit the re-processing of such fertilizer on the terms and conditions as may be notified by the Central Government in this behalf:

Provided that no such application for permission to reprocess the fertilizer by the manufacturer or importer shall be accepted by the Central Government after the expiry of the said period of fifteen days.]

[(6) Disposal of Biofertilizers (non-standard or expired). - Notwithstanding anything contained in this (Order), every manufacturer shall recycle the carrier based biofertiliser after sterilisation of the stock lying at manufacture's site, is not of prescribed standard (hereinafter referred as nonstandard), or can dispose solid or liquid biofertiliser or both by spraying or by sprinkling over the compost piles or windrows; in case of liquid biofertiliser it shall be destroyed by autoclaved first and then spread or sprinkled on the soil in nearby fields.]

[23A. Reprocessing of fertiliser damaged during storage. - (1) Notwithstanding anything contained in this order, where the stock of fertiliser damaged during its storage, the manufacturer or the importer of such fertiliser may make an application for grant of certificate of reprocessing in Form I-1 along with prescribed fee to the Notified Authority for reprocessing such fertiliser in the manufacturing unit of such manufacturer or the unit mentioned by the importer in his application.

(2) The Notified Authority may, on being satisfied, allow the application made under sub-clause (1) and grant the certificate of reprocessing in Form -I-2, subject to the following conditions, namely: -

(a) the container of such damaged material shall be conspicuously superscribed in red color with the word "Damaged" and also with sign "X".

(b) the damaged fertiliser shall be moved from the godown within one week from the date of grant of certificate of reprocessing.]

[23B. Disposal of Damaged and Expired stock of Bio-fertilisers/Organic fertilizers. - (1) Notwithstanding anything contained in this order where the stock of biofertiliser damaged during storage or expired or does not conform to the prescribed standard at dealer end or during transit from manufacturing unit to dealer end, such material shall be taken back by manufacturer or returned by dealer by intimating to the Notified authority within seven days, in case where the material is damaged during transit or in case where the material get expired, the manufacturer shall intimate the authority within seven days from the date of expiry for transacting the material for its disposal as per the procedure prescribed in sub-clause (6) of Clause 26:

Provided that the material of such damaged material shall be conspicuously superscribed in red color with word damaged and also with sign "X";

(2) Notwithstanding anything contained in this order where the stock of organic fertiliser damaged during storage or during transit and does not conform to the prescribed standard, such material shall be taken by manufacturer for reprocessing it by mixing with fresh biomass or in case where it is non-standard due to heavy metal content then the manufacturer immediately with draw the material for reprocessing by mixing with fresh biomass or such material shall be used in gardens, golf courses etc or sent to landfills. If stock is at dealers end, it shall be taken back or returned to the manufacturer for disposal as mentioned above.

(3) The manufacturer shall intimate to the Notified authority within seven days in case where the material is damaged during transit or, within seven days in case the material damaged or not of prescribed standard during storage at any dealer level or in storage godown:

Provided that such damaged material shall be conspicuously superscribed in red color with word damaged and also with sign "X".]

24. Manufacturers/pool handling agencies to appoint officers responsible with compliance of the order. - Every manufacturing organisation [importer] and pool handling agency shall appoint in that organisation and in consultation with the Central Government, an officer, who shall be responsible for compliance with the provisions of this Order.

25. Restriction on sale/use of fertilizers. - (1) No person shall, except with the prior permission of the Central Government and subject to such terms and conditions as may be imposed by such Government, shall or use fertilizer, for purposes other than fertilization of soils and increasing productivity of crops :

Provided that the price of fertilizers permitted for sale for industrial use shall be no profit no loss price, excluding all subsidies at the production, import, handling or on sale for agricultural consumers :

Provided further that wherever customs or excise duties are chargeable, these may be added to the price so fixed :

Provided also that in the case of non - standard fertilizers, reductions shall be made from the no profit no loss price, indicated above, proportionate to the loss of nutrient contents.

[(2) Notwithstanding anything contained in sub - clause (1), no prior permission for use of fertiliser for industrial purposes shall be necessary when the fertiliser for such purpose is purchased from the industrial dealer possessing a valid certificate of registration granted under Clause 9.

(3) Any person possessing a valid certificate of registration for industrial leader, unless such person is a State Government, a manufacturer or a pool handling agency, shall not carry on the business of selling fertilisers for agricultural purposes, including a wholesale dealer or a retail dealer. However, in case of a State Government, a manufacturer of a pool handling agency possessing a valid certificate of registration of sale of fertiliser for industrial use and also for sale of fertiliser for agriculture use, whether in wholesale or retails or both, shall not carry on the business of selling fertilisers both for industrial use and agricultural use in the same premises.]

VII. Enforcement Authorities

26. Appointment of registering authority. - The State Government may, by notification in the Official Gazette, appoint such number of persons, as it thinks necessary, to be registering authorities for the purpose of this Order, [excepting for grant or renewal of certificate of registration for industrial dealer] and may, in any such notification, define the limits of local area within which each such registering authority shall exercise his jurisdiction.

26A. Notified Authority. - The State Government may, by notification in the Official Gazette, appoint such number of persons, as it thinks necessary, to be Notified Authorities for the purpose of this Order and define the local limits within which each such Notified Authority shall exercise his jurisdiction.

27. Appointment of inspectors. - The State Government, or the Central Government may, by notification in the Official Gazette, appoint such number of persons, as it thinks necessary, to be inspectors of fertilizers for the purpose of this Order, and may, in any such notification, define the limits of local area within which each such inspector shall exercise his jurisdiction.

[27A. Qualifications for appointment of fertilizer inspectors. - No person shall be eligible for appointment as fertilizer inspector under this order, unless he possesses the following qualifications, namely : -

(1) graduate in Agriculture or Science with Chemistry as one of the subjects, from a recognised University, and

(2) training or experience in the quality control of fertilizers and working in the State or Central Department of Agriculture.]

27B. Qualifications for appointment of fertiliser Inspectors for Biofertiliser and Organic Fertiliser. - No person shall be eligible for appointment as inspector of biofertiliser and Organic fertilizer under this Order unless he may possess the following qualifications, namely: -

(1) Graduate in agriculture or science with chemistry/microbiology as one of the subject; and

(2) Training or experience in the field of quality control of biofertilisers/organic fertilizers.

28. Powers of Inspectors. - (1) An Inspector may, with a view to securing compliance with this Order, -

(a) require any manufacturer, pool handling agency, wholesale dealer or retail dealer to give any information in his possession with respect to the manufacture, storage and disposal of any fertilizer manufactured or, in any manner handled by him;

(b) draw samples of any fertilizer in accordance with the precedure of drawal of samples laid down in Schedule II :

Provided that the Inspector shall prepare the sampling details in duplicate in Form J, and hand over one copy of the same to the dealer or his representative from whom the sample has been drawn ;

[(ba) draw samples of any Bio-fertilizers in accordance with the procedure of drawal of samples laid down in Schedule III:

[Provided that the inspector shall prepare the sampling details in duplicate in Form J and hand over one copy of the same to the dealer or his representative from whom the sample has been drawn;]

(bb) draw samples of any Organic fertilizers in accordance with procedure of drawal of samples laid down in Schedule IV:]

[Provided that the inspector shall prepare the sampling details in duplicate in Form J and hand over one copy of the same to the dealer or his representative from whom the sample has been drawn;]

[(bc) draw samples of any non-edible de-oiled vake fertilizer in accordance with procedure of drawl of samples laid down in Schedule V:]

[Provided that the inspector shall prepare the sampling details in duplicate in Form J and hand over one copy of the same to the dealer or his representative from whom the sample has been drawn;]

[(bd) draw samples of any biostimulants in accordance with the procedure of drawal of samples laid down in Schedule VI;

Provided that the Inspector shall prepare the sampling details in duplicate in Form-J and handover one copy of the same to the dealer or his representative from whom the sample has been drawn;]

(c) enter upon and search any premises where any fertilizer is manufactured or stored or exhibited for sale, if he has reason to believe that any fertilizer has been or is being manufactured, sold, offered for sale, stored, exhibited for sale or distributed contrary to the provisions of this Order ;

(d) seize or detain any fertilizer in respect of which he has reason to believe that a contravention of this Order has been or is being or is about to be committed ;

(e) seize any books of accounts or documents relating to manufacture, storage or sale of fertilizers, etc, in respect of which he has reason to believe that any contravention of this Order has been or is being or is about to be committed :

Provided that the inspector shall give a receipt for such fertilizers or books of accounts or documents so seized to the person from whom the same have been seized :

Provided further that the books of accounts or documents so seized shall be returned to the person from whom they were seized after copies thereof or extracts therefrom as certified by such person, have been taken.

[Provided also that the Inspector shall give the stop sale notice in writing, to the person whose stocks have been detained and initiate appropriate actions as per the provisions of this Order within a period of twenty - one days. If no action has been initiated by the Inspector within the said period of twenty - one days from the date of issue of the said notice, the notice of stop sale shall be deemed to have been revoked.]

(2) Subject to the proviso to paragraphs (d) and (e) of sub - clause (1), the provisions of the Criminal Procedure Code, 1973 (2 of 1974), relating to search and seizure shall, so far as may be, apply to searches and seizures under this clause.

(3) Where any fertilizer is seized by an Inspector under this clause he shall forth with report the fact of such seizure to the Collector whereupon the provisions of Sections 6 - A, 6 - B, 6 - C, 6 - D and 6 - E of the Act, shall apply to the custody, disposal and confiscation of such fertilizers.

(4) Every person, if so required by an inspector, shall be bound to afford all necessary facilities to him for the purpose of enabling him to exercise his powers under sub - clause (1).

[28A. Appointment of officer for keeping the sample in custody. - The State Government may, by notification in Official Gazette, appoints an officer not below the rank of Joint Director of Agriculture Department for keeping the samples in its custody and for sending the second part for analysis to National test House and the third part for referee analysis.

28B. Sample drawn by the Inspector. - (1) Out of three parts made by the Inspector drawn under sub-clause (6) of Clause 28, the first part shall be sent to any of the State Government notified lab by the state Inspector under intimation to the officer notified under Clause 28-A and the second part and third part shall be kept in custody of officer nominated by the State Government.

(2) The company or dealer, from whom sample is drawn may request to such authority within one week from the date of drawal of sample, to send the second part of the sample to any of the National Test House Laboratories on payment of fees as provided by the Central Government from time to time and in case, there is variation in the results of the first and second analysis, then the said authority invariably shall send the third part of the sample to Central Fertiliser Quality Control and Training Institute, Faridabad for final referee analysis.

(3) In case where the sample is drawn by the Central Government Inspector then out of three parts made by the Inspector, the first part shall be sent to any of the Regional Quality Control Laboratory at Mumbai, Kalyani or Chennai and , the second part and third part shall be kept in custody of incharge of the Regional Fertiliser Quality Control Laboratory at Chennai, Kalyani or Mumbai as the case may be.

(4) The company or dealer, from whom sample is drawn may request within one week to such authority to send the second part of the sample to any of the National Test House Laboratories on payment of fees as provided by the Central Government from time to time and in case, there is variation in the result of the first and second analysis, then such authority invariably send the third part of the sample to Central Fertiliser Quality Control and Training Institute, Faridabad for final referee analysis.

(5) The referee analysis report received from the laboratory shall be treated as final.

(6) In case, where the appeal of non-standard fertiliser under sub-clause (3) of Clause 32 and sub-clause (3) of Clause 32-A have not been considered then the report of the regional fertiliser Quality Control Laboratory or the notified Fertiliser Testing Laboratory, as the case may be shall be treated as final.]

VIII. Analysis Of Samples

29. Laboratory for analysis. - [(1) A fertilizer sample drawn by an Inspector, shall be analysed in accordance with the instructions contained in Schedule II -

(a) in regional fertilizer control laboratories at Mumbai, Chennai or Kalyani (Kolkata) and the National Test House Laboratories at Mumbai, Chennai, Kolkatta, Ghaziabad and Jaipur as per sub-clause (2) of Clause 28-B; or

(b) in any laboratory notified for this purpose by the State Government with the prior approval of the Central Government and the National Test House Laboratories at Mumbai, Chennai, Kolkatta, Ghaziabad and Jaipur as per subclause (4) of Clause 28-B;]

[(1A) Bio-fertilizer samples drawn by an inspector shall be analysed in accordance with the instructions laid down in Schedule III in the [Regional centre of Organic farming at Bengaluru, bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula] or any other laboratory notified by Central or State Government.

(1B) [Organic fertilizer and non-edible de-oiled cake fertilizer samples] drawn by an inspector shall be analyzed in accordance with the instructions laid down in Schedule IV in the [Regional centre of Organic farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jablapur, Nagpur and Panchkula] or any other laboratory notified by Central or State Government.]

[(1C) A biostimulant sample drawn by an inspector shall be analysed in accordance with the procedure contained in Part C of Schedule VI in the Central Fertiliser Quality Control and Training Institute, Faridabad or its regional laboratories or in any other laboratory notified for this purpose by the State Government with prior approval of Central Government.]

[(2) Every laboratory referred to in sub-clause (1) shall in order to ensure accurate analysis, of fertiliser samples, possess, minimum equipment and other laboratory facilities, as may be specified from time to time by the Controller in this behalf.]

[29A. Qualifications for appointment of fertilizer analyst in the fertilizer quality control laboratories. - No person shall be eligible for appointment as fertilizer analyst or analysis of fertilizer samples in the laboratories notified under Clause 29 of the order, unless the possesses the following qualifications, namely :

(1) graduate in Agriculture or Science with Chemistry as one of the subject from a recognised University, and

(2) training in fertilizer quality control and analysis at Central Fertilizer Quality Control and Training Institute, Faridabad.

Provided that the fertilizer analysts appointed before the commencement of this order, who do not possess the requisite training, shall undergo prescribed training, within a period of three years, in the Central Fertilizer Quality Control and Training Institute. Faridabad, from the date of commencement of this order.]

[29AA. Regular training of Fertiliser Analysts. - Every Fertiliser Analyst shall undergo training after every three years in the Central Fertiliser Quality Control and Training Institute or any Regional Fertiliser Quality Control Laboratory at Mumbai, Kalyani or Chennai.]

[29AB. No person shall be eligible to be notified as analyst for analysis of sample of Biofertiliser, Organic fertiliser and non-edible de-oiled cake fertilizers in the laboratories notified under sub-clauses (1A) and (1B) of clause 29, unless the analyst possesses the following qualifications, namely: -

(a) Postgraduate degree in Agriculture Chemistry and Soil Science or Microbiology or Plant Pathology or Chemistry or Biotechnology or Horticulture or Bio-Engineering;

(b) training in analysis of Biofertiliser, Organic fertiliser and non-edible de-oiled cake fertilizers at National Centre of Organic Farming or at a recognized laboratory or Institute.

Provided that the analyst who has been appointed prior to the date of commencement of this Order, but does not possess the requisite training or experience in analysis of Biofertiliser, Organic fertiliser and non-edible de-oiled cake shall undergo prescribed training at the National Centre of Organic Farming or at a recognized lab or Institute within a period of three years from the date of such commencement.]

[29B. The Central Fertiliser Quality Control and Training Institute, Faridabad shall be designated as referee laboratory for the purpose of analysis of any sample of fertiliser under sub-clauses (1), (2) and (3) of Clause 28-B.]

[29C. Laboratories for referee analysis of Biofertiliser. - (1) [Regional centre of Organic Farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula] and every laboratory referred to in clause 29(1A) shall be designated as referee laboratory for the purpose of analysis of any sample of Bio-fertiliser:

Provided Further that in respect of any sample the analysis of which has been challenged, may be sent for referee analysis to any one of the other laboratories except those which are located in the State or where the first analysis has been done.

Provided further that [Regional centre of Organic Farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula] shall be considered as one group of laboratories and a sample first analysed by any one of them, shall not be sent for referee analysis to any other in that group, but only to any other laboratory notified by a State Government or Central Government.

(2) Notwithstanding anything contained in this Order, the Appellate Authority as specified in sub-clause (1) of clause 32A in case of sample collected by [Regional centre of Organic Farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula], as the case may be, shall decide and send, one of the two remaining samples, for reference analysis as provided under sub-clause (1).

29D. Laboratories for referee analysis of Organic fertiliser. - (1) [Regional centre of Organic Farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula] and every laboratory referred to in clause 29(1A) shall be designated as referee laboratory for the purpose of analysis of any sample of Organic fertiliser:

Provided that no such laboratory which carried out the first analysis of fertiliser sample shall be so designated in respect of that sample:

Provided further that in respect of any sample the analysis of which has been challenged, may be sent for referee analysis to any one of the other laboratories except those which are located in the State or where the first analysis has been done:

(2) Notwithstanding anything contained in this Order, the Appellate Authority as specified in sub-clause (1) of clause 32A in case of sample collected by the State Government laboratory, or the Controller, in case of sample collected by [Regional centre of Organic Farming at Bengaluru, Bhubaneswar, Ghaziabad, Imphal, Jabalpur, Nagpur and Panchkula], as the case may be, shall decide and send, one of the two remaining samples, for reference analysis as provided under sub-clause (1).]

30. Time limit for analysis and communication of result. - (1) Where sample of a fertilizer has been drawn, the same shall be despatched, along with a memorandum in Form K [and in case of organic fertilizers, bio-fertilizers and non-edible de-oiled cake fertilizers in [Form K]] to the laboratory for analysis within a period of [three working days] from the date of its drawal.

[(2) The laboratory shall analyse the sample and forward the analysis report in Form L, within fifteen days, and in case of the sample of organic, fertiliser, biofertiliser and non-edible de-oiled cake fertilisers, in Form L1, L2, L3, respectively, within thirty days, from the date receipt of the sample in the laboratory to the authority specified in the said memorandum.]

(3) The authority to whom the analysis report is sent under sub-clause (2) shall communicate the result of the analysis to the dealer/manufacturer/[importer]/pool handling agency from whom the sample was drawn within [seven days] from the date of receipt of the analysis report of the laboratory.

IX. Miscellaneous

31. Suspension/Cancellation of registration certificate. - (1) A registering authority [or, as the case may be, the controller] may, after giving the holder of a certificate of registration or a certificate of manufacture or any other certificate granted under this Order, an opportunity of being heard, suspend or cancel such certificate on any of the following grounds, namely :

(a) that such certificate has been obtained by misrepresentation as to material particulars.

(b) that any of the provisions of this Order or any of the terms and conditions of such certificate has been contravened or not fulfilled :

Provided that while cancelling the certificate the holder thereof may be allowed a period of 30 days to dispose of the balance stock of fertilizers if any, held by him :

Provided further that the stock of fertilizer lying with the holder after the expiry of the said 30 days shall be confiscated.

(2) Where the contravention alleged to have been committed by a person is such as would, on being proved, justify cancellation of the certificate of registration or, as the case may be, certificate of manufacture or any other certificate granted under this Order to such person the registering authority [or, as the case may be, the controller] may, without any notice, suspend such certificate, as an interim measure :

Provided that the registering authority [or, as the case may be, the controller] shall immediately furnish to the person details and the nature of contravention alleged to have been committed by such person and, after giving the person an opportunity of being heard, pass final orders either revoking the order of suspension or cancelling the certificate within fifteen days from the date of issue of the order of suspension :

Provided further that where no final order is passed within the period as specified above, the order of interim suspension shall be deemed to have been revoked without prejudice, however, to further action which the registering authority [or, as the case may be, the controller] may take against the holder of the certificate under sub-clause (1).

(3) Wherever a certificate is suspended or cancelled under this clause, the registering authority [or, as the case may be, the controller] shall record a brief statement of the reasons for such suspension or, as the case may be, cancellation and furnish a copy thereof to the person whose certificate has been suspended or cancelled.

[(4) Wherever the person alleged to have committed the contravention is an industrial dealer, the registering authority may also take action against the holder of such certificate of registration, under sub-clauses (1) and (2) :

Provided that where such certificate is suspended or cancelled the registering authority shall within a period of fifteen days from the date of issue of such order of suspension or cancellation, furnish to the Controller also, besides sending the same to the person whose certificate has been suspended or cancelled, a detailed report about the nature of contravention committed and a brief statement of the reasons for such suspension or, as the case may be, cancellation :

Provided further that the Controller shall, in case of the order for suspension passed by the registering authority, on receipt of the detailed report and after giving the person an opportunity of being heard, pass final order either revoking the order of suspension or cancelling the certificate of registration, within fifteen days from the date of receipt of the detailed report from the registering authority, falling which the order of interim suspension passed by the registering authority shall be deemed to have been revoked without prejudice, however, to further action which the Controller may take against the holder of certificate under sub - clause (1) :

Provided also that the order of cancellation passed by the registering authority shall remain effective as if it has been passed by the Controller till such time the Controller, on receipt of the detailed report from the registering authority, and if deemed necessary, after giving the person a fresh opportunity of being heard, pass the final order either revoking or confirming the order of cancellation.]

32. Appeal. - [Any person, excepting an industrial dealer or, as the case may be, the person desiring to obtain a certificate of registration for industrial dealer, aggrieved by]: -

(1) an order -

(a) refusing to grant, amend or renew a certificate of registration for sale of fertilizers ; or

(b) refusing to grant a certificate of manufacture for preparation of mixture of fertilizers or special mixture of fertilizers ; or

[*****]

[(3) in cases where the company or dealer has not applied for the second analysis due to some valid reason then the company or dealer may made an appeal to the Controller, as the case may be, within seven days from the date of receipt of analysis report of first part of the sample and the Controller shall decide on the grounds of the facts whether the matter qualifies to be considered for third analysis.]

32A. Appeal at the State Government level. - (1) The State Government shall, by notification in the Official Gazette, specify such authority as the Appellate authority before whom the appeals may be filed within 30 days from the date of the order appealed against by any person, except by an industrial dealer, aggrieved by any of the following Orders or action of registering authority or a Notified Authority, namely: -

(i) Refusing to grant a certificate of manufacture for preparation of mixture of fertilisers or special mixture of fertilizers;or

(ii) Suspending or canceling a certificate of manufacture; or

(iii) Suspending or canceling authorization letter or debarring from carrying on the business of selling of fertilizer, or

(iv) non-issuance of certificate of manufacture within the stipulated period; or

(v) non-issuance of amendment in authorization letter within the stipulated period.

[****]

[(3) In cases where the company or dealer has not applied for the second analysis due to some valid reason then the company or dealer may made an appeal in the Fertiliser (Inorganic, Organic or Mixed) (Control) Order, 1985 to the Appellate authority and the Appellate Authority shall decide on the grounds of the facts whether the matter qualifies to be considered for third analysis.]

33. Grant of duplicate copies of [authorization letter or Certificate of manufacture] certificate of registrations, etc. - Where [an authorisation letter or] a certificate of registration or a certificate of manufacture or any other certificate granted or, as the case may be, renewed under this Order is [lost or defaced, the Notified Authority], the registering authority [or, as the case may be, the Controller] may, on an application made in this behalf, together with the fee prescribed for this purpose under Clause 36, grant a duplicate copy of such certificate.

[34. Amendment of authorisation letter, certificate of manufacture and certificate of Registration. - The Notified Authority, registering authority or controller, as the case may be, may, on application being made by the holder of an authorization letter, a certificate of registration or certificate of manufacture, together with the fee prescribed for the purpose under clause 36, amend an entry in such authorization letter, certificate of Registration or certificate of manufacture, as the case may be.]

35. Maintenance of records and submission of returns, etc. - (1) The Controller may be an order made in writing direct the dealers, manufacturers, and pool handling agencies -

(a) to maintain such books of accounts, records, etc. relating to their business [in Form N] [or maintain digital stock register in the form which clearly exhibits the date wise stock position, opening balance, receipts during the day, sales during the day and closing stock]; and

(b) to submit to such authority, returns and statements in such form and containing such information relating to their business and within such time as may be specified in that order.

(2) Where a person holds certificates of registration for retail sale and wholesale sale of fertilizers, he shall maintain separate books of accounts for these two types of sales made by him.

[(3) Where a State Government, a manufacturer and a pool handling agency holds valid certificates of registration for sale of fertilisers in wholesale or retail or both and also for sale for industrial use, he shall maintain separate books of accounts for these two, or three types of sales made by him.]

[(4) Every importer shall inform the Director of Agriculture of the State in which he intends to discharge the imported fertiliser, under intimation to the Central Government, before the import is made or within a period of fifteen days after an indent for import as placed, the following details, namely : -

(i) name of fertiliser,

(ii) name of the country of importer,

(iii) name of manufacturer,

(iv) quantity to be imported,

(v) date of arrival of the consignment,

(vi) name of the discharge part,]

[(vii) name, designation of authorised or responsible person alongwith mobile number;

(viii) other information.]

36. Fees. - (1) The fees payable for grant, amendment or renewal of [an authorisation letter or] a certificate of registration or certificate of manufacture [or certificate of authorisation for sale of non-standard fertiliser or certificate for reprocessing of fertiliser damaged during storage] a duplicate of such certificates or renewal thereof under this Order shall be such as the State Government may, [***], from time to time, fix, subject to the maximum fees fixed for different purposes by the Central Government and different fees may be fixed for different purposes or for different classes of dealers or for different types of mixtures of fertilizer or special mixture.

[***]

(2) The authority to whom and the manner in which the fee fixed under sub-clause (1) shall be paid shall be such as may be specified by the State Government by notification in the official Gazette.

(3) Any fee paid under sub-clause (1) shall not be refundable unless the grant or renewal of any certificate of registration or certificate of manufacture, [Import] or duplicate copy of such certificate or renewal under this Order has been refused.

[(4) The fees payable for grant, amendment, renewal or duplicate copy of certificate of registration for industrial dealer and the authority to whom and the manner in which such fee shall be paid, shall be such as may be specified by the Controller, from time to time by notification in the Official Gazette.]

37. Service of orders and directions. - Any order or direction made or issued by the Controller or by any other authority under this order shall be served in the same manner as provided in sub - section (5) of Section 3 of the Act.

38. Advisory Committee. - (1) The Central Government may be notification in the Official Gazette and on such terms and conditions as may be specified in such notification, constitute a Committee called the Central Fertilizer Committee consisting of a Chairman and not more than ten other persons, having experience or knowledge in the field, who shall be members of the Committee, to advise the Central Government regarding -

(i) inclusion of a new fertilizer, under this Order ;

(ii) specifications of various fertilizers

(iii) grades/formulations of physical granulated mixtures of fertilizers that can be allowed to be prepared in a State ;

(iv) requirements of laboratory facilities in a manufacturing unit, including a unit manufacturing physical/granulated mixtures of fertilizers ;

(v) Methods of drawal and analysis of samples ;

(vi) any other matter referred by the Central Government to the Committee.

(2) The Committee may, subject to the previous approval of the Central Government, make bye-laws fixing the quorum and regulation its own procedure and the conduct of all business to be transacted by it.

(3) The Committee may co-opt such number of experts and for such purposes or periods as it may deem fit, but any expert so co-opted shall not have the right to vote.

(4) The Committee may appoint one or more sub-committees, consisting wholly of members of the Committee [***] or partly of the members of the Committee and partly of co-opted members, as it thinks fit, for the purpose of discharging such its functions as may be delegated to such sub-committee or sub-committees by the Central Fertilizer Committee.

[(5) The State Government may be notification in the Official Gazette and on such terms and conditions as may be specified in such notification, constitute a Committee called the State Fertilizer Committee consisting of a Chairman and not more than 5 other members, having experience or knowledge in the field, including a representative from State Agricultural University, the Fertilizer Industry and Indian Micro Fertilizers Manufacturers' Association to advise the State Government regarding the grades/formulations of [mixture of fertilizer] and their specifications].

[38A. Central Biostimulant Committee. - (1) The Central Government may, by notification in the Official Gazette, constitute a Committee to be called 'Central Biostimulant Committee' with the Agriculture Commissioner as its Chairperson and seven other members, subject to such terms and conditions, as may be specified therein.

(2) The Central Biostimulant Committee shall advise the Central Government on the following issues, namely: -

(i) of a new biostimulant;

(ii) specifications of various biostimulants;

(iii) methods of drawing of samples and its analysis;

(iv) minimum requirements of laboratory;

(v) method of testing of biostimulants;

(vi) any other matter referred to the Committee by the Central Government.

(3) The Committee may, subject to the previous approval of the Central Government, make bye-laws fixing the quorum and regulating its own procedure and the conduct of all business to be transacted by it.

(4) The committee may co-opt such number of experts and for such purposes or periods as it may deem fit, but any expert so co-opted shall not have the right to vote.

(5) The committee may appoint one or more sub-committees, consisting wholly of its members or partly of its members and co-opted members, as it thinks fit, for the purpose of discharging such for its functions as may be delegated to such sub-committee.

(6) The Central Biostimulants Committee shall frame guidelines laying down the requirements of toxicology testing and other related tests.]

39. Repeal and saving. - (1) The Fertilizers (Control) Order, 1957 is hereby repealed except as respects things done or omitted to be done under the said Order before the commencement of this Order.

(2) Notwithstanding such repeal, an order made by any authority, which is in force immediately before the commencement of this Order and which is consistent with this Order, shall continue in force and all appointments made, prices fixed, certificates granted and directions issued under repealed Order and in force immediately before such commencement shall likewise continue in force and be deemed to be made, fixed, granted or issued in pursuance of this order till revoked.

Schedule I

Refer clause 2(h) & (q)

[Part A]

Specifications of Fertilisers

1(a) Straight Nitrogenous Fertilisers

1- Ammonium Sulphate

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Ammoniacal nitrogen per cent by weight, minimum

20.5

(iii)

Sulphate Sulphur (as 5) per cent by weight, minimum

23-0

(iv)

Free acidity (as H₂SO₄) per cent by weight, maximum (0.04 for material obtained from by product ammonia and by-project gypsum)

0.025

(v)

Arsenic (as As₂O₃) per cent by weight, maximum

0.01

2. Urea (46% N)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight, (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Particle size-Minimum [80 per cent] of the material shall be retained between 1 mm and 26 mm 15 sieve

3. Ammonium Chloride

(i)

Moisture per cent by weight, maximum

2.0

(ii)

Ammoniacal nitrogen per cent by weight, minimum

25.0

(iii)

Chloride other than ammonium chloride (as NaCT) per cent by weight, maximum basis, maximum

2.0

4. Calcium Ammonium Nitrate (25% N)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total ammoniacal and nitrate nitrogen per cent by weight, minimum

25.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

12.5

(iv)

Calcium nitrate per cent by weight, maximum

0.5

(v)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

5. Urea Super Granulated

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Particle size-Minimum 80 per cent of the material shall be retained between 13.2 mm and 9.5 mm 15 sieve

6. Urea (Granular)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Particle size-Minimum 90 per cent of the material shall be retained between 4 mm and [2mm] 15 sieve

7. Urea Ammonium Nitrate (32%N) (liquid)

(i)

Total nitrogen (Urea, Ammoniacal and Nitrate) per cent by weight, minimum

32.0

(ii)

Urea nitrogen per cent weight, maximum

16.5

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.5

(iv)

Nitrate nitrogen per cent by weight, minimum

7.5

(v)

Specific gravity (at 15'C)

1.30-1.35

8. Neem Coated Urea

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight, (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Neem oil content soluble in [(binary mixture of n-hexane acetone (4.1)] per cent by weight, minimum

0.035

[(v)

the total melanin content in the oily residue, per cent by weight, minimum

1.0]

[Particle size-Minimum 80 per cent, of the material shall be retained between 1 mm and 2.8 mm 15 sieve]

[Foot Note: The term 'Neem oil' means the oil conforming to BIS standards (15 4765-1975-Rev.1).

Explanation: The total melacin content means consisting of at least three of the major melacin namely. Azadirachtin 'A' and 'B', nimbin, salanin, 6-desacety nimbin and 3-desicety salanin;]

9. Urea Briquettes

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Particle size-Minimum 90 per cent of the material shall be retained between 5.7 mm and 3.8 mm 15 sieve

10. Sulphur Coated Urea

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen per cent by weight (on dry basis), minium

37.0

(iii)

Elemental Sulphur (as 5) per cent by weight, minimum

17.0

(iv)

Biuret per cent by weight, maximum

1.5

(v)

Particle size-Minimum 90 per cent of the material shall be retained between 8 mm and 2 mm 15 sieve

11. Neem Coated Urea (Granular)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen, per cent by weight, (on dry basis), minimum

46.0

(iii)

Biuret per cent by weight, maximum

1.5

(iv)

Neem oil content soluble in [(binary mixture of n-hexane acetone (4:1)], per cent by weight, minimum

0.035

(v)

Particle size-Minimum 90 per cent of the material shall be retained between 2 mm and 4 mm 15 sieve

[(vi)

the total meliacin content in the oily residue, per cent by weight, minimum

1.0

Foot note: The term 'Neem oil' means the oil conforming to BIS standards (i5 4765-1975-Rev.1);

Explanation 1: The total melaicin content means consisting of at least three of the major meliacin Namely, Azadirachtin 'A' and 'B', nimbin, salanin, 6-desacety, nimbin and 3-desacety salanin.]

[12. Calcium Cyanamide

(i)

Moisture per cent, by weight, maximum

1.0

(ii)

Total Nitrogen per cent, by weight, minimum

19.5

(iii)

Cyanamide nitrogen per cent, by weight, minimum

15.0

(iv)

Nitrate nitrogen per cent by weight, maximum

1.5

(v)

Particle size-Not less than 90 per cent of the material shall pass through 4 mm sieve and be retained on 1 mm 15 sieve

Requirements-Every importer who intends to import Calcium Cyanamide into the country shall be required to follow the following requirements.-
(i) the importer shall print or cause to be printed on the bags containing fertilizers to the effect that the material is irritant to skin and eyes and the same shall be used by wearing hand gloves only:
(ii) the importer shall pack or cause to be packed hand gloves for use of farmers along with each container;
(iii) the importer shall provide or cause to be provided a leaflet indicating the following in the bags of fertilisers in which the material is packed;
(i) Avoid contact of the product with skin;
(ii) Wear protective clothing and gloves;
(iii) Wash hands and exposed skin after work and before meal;]

1(b). Straight Phosphorous Fertilisers

1. Single Superphosphate (16% F₂O₃ Powdered)

(i)

Moisture per cent by weight, maximum

12.0

(ii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

11.0

(v)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

4.0

2. Triple Superphosphate

(i)

Moisture per cent by weight, maximum

12.0

(ii)

Total phosphorus (asP₂O₃) per cent by weight, minimum

46.0

(iii)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

42.5

(iv)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

3.0

3. Rock Phosphate

(i)

Total phosphorous (as P₂O₃) per cent by weight, minimum

18.0

(ii)

Particle size-Minimum 90 per cent of the material shall pass through 0.15 mm 15 sieve and the balance 10 per cent of material shall pass through 0.25 mm 15 sieve.

4. Single Superphosphate (16% P₂O₃ Crenulated)

(i)

Moisture per cent by weight, maximum

5.0

(ii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

11.0

(v)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

4.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

5. Superphosphoric Acid (70% P₂O₃) Liquid)

(i)

Total phosphorous (as P₂O₃) per cent by weight, minimum

70.0

(ii)

Polyphosphate (as P₂O₃) per cent by weight, minimum

19.0

(iii)

Methanol insoluble matter, per cent by weight maximum

1.0

(iv)

Magnesium as MgO, per cent by weight, maximum

0.5

(v)

Specific gravity (at 24'C)

1.95-2.00

1(c) Straight Potassium Fertilisers

1. Potassium Chloride (Muriate of Potash)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Water soluble potassium content (as K₂O) per cent by weight, minimum

60.0

(iii)

Sodium as NaCI per cent by weight (on dry basis), maximum

3.5

(iv)

Particle size-Minimum 65 per cent of the material shall be retained between 0.25 mm and 1.7 mm 15 sieve

2. Potassium Sulphate

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Water soluble potassium (as K₂O) per cent by weight, minimum

50.0

(iii)

Sulphate Sulphur (as 5) per cent by weight, minimum

17.5

(iv)

Sodium as NaCI per cent by weight, maximum

2.0

(v)

Total chlorides (as CI) per cent by weight, maximum

2.5

3. Potassium Schoenite

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Water soluble potassium (as K₂O) per cent by weight, minimum

23.0

(iii)

Magnesium (as MgO) per cent by weight, maximum

11.0

(iv)

Sodium (as NaCI) per cent by weight, maximum

1.5

4. Potassium Chloride (Muriate of Potash) (Granular)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Water soluble potassium (as K₂O) per cent by weight, minimum

60.0

(iii)

Sodium (as NaCI) per cent by weight, maximum

3.5

(iv)

Magnesium (as MgCl₂) per cent by weight, maximum

1.0

(v)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

5. Potash Derived from Molasses

(i)

Moisture per cent by weight, maximum

5.0

(ii)

Water soluble potassium (as K₂O), per cent by weight, minimum

14.5

[6. Dehydrate Poly halite

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Water soluble potassium (as K₂O) per cent by weight, minimum

[12.5]

(iii)

Water soluble Calcium (as CaO) per cent by weight, minimum

[15.0]

(iv)

Water soluble Magnesium (as MgO) per cent by weight, minimum

[5.0]

(v)

Sulphate Sulphur (as S) per cent by weight, maximum

[17.5]

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Arsenic (as As) per cent by weight, maximum

0.01]

[7. Potassium Magnesium Sulphate (granular)

(i)

Moisture per cent, by weight, maximum

0.5

(ii)

Magnesium as MgO per cent, by weight, minimum

10.0

(iii)

Potash as K₂O per cent.by weight, minimum

30.0

(iv)

Sulphate Sulphur as S per cent, by weight, minimum

17.0

(v)

Total Chlorides per cent, by weight, maximum

2.5

(vi)

Particle Size 90% of the material shall be retained between 5 mm IS sieve and 2mm IS sieve]

1(d) Straight Sulphur Fertilisers

1. Sulphur 90% (Powder)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total Elemental sulphur (as S) per cent by weight, minimum

90.0

(Note: the product may contain any inter filler material such as Bentonite etc. up to the maximum extent of 10 per cent by weight)

2. Sulphur 90% (Granular)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Total elemental sulphur (as S) per cent by weight, minimum

90.0

(iii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

(Note: the product may contain any inert filler material such as Bentonite etc. up to the maximum extent of 10 per cent by weight)

1(e) N.P. Complex Fertilisers

1. Diammonium Phosphate (18-46-0)

(i)

Moisture per cent by weight, maximum

2.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

18.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

15.5

(iv)

Available phosphorus (as PxO₃) per cent by weight, minimum

16.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

39.5

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

2. Ammonium Phosphate Sulphate (16-20-0)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total ammoniacal nitrogen per cent by weight, minimum

16.0

(iii)

Available phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(iv)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

17.0

(v)

Sulphate Sulphur (as S) per cent by weight, minimum

13.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

3. Ammonium Phosphate Sulphate (20-20-0)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen (ammoniacal+urea) per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

18.0

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(v)

Water soluble phosphorus (as PxO₃) per cent by weight, minimum

17.0

(vi)

Sulphate Sulphur (as S) per cent by weight, minimum

13.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

4. Ammonium Phosphate Sulphate Nitrate (20-20-0)

(i)

Moisture per cent by weight maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

18.0

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

17.0

(vi)

sulphate Sulphur (as S) per cent by weight, minimum

13.0

(vii)

Particle size- Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

5. Nitro Phosphate (20-20-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

20.0

(iii)

Nitrogen in ammoniacal form per cent by weight, minimum

10.0

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

12.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

6. Urea Ammonium Phosphate (28-28-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

28.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

9.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight minimum

28.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

24.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve.

7. Urea Ammonium Phosphate (24-24-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and Urea) per cent by weight, minimum

24.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.5

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

24.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

20.5

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

8. Urea Ammonium Phosphate (20-20-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

6.5

(iv)

Available phosphorous (as P₂O₃) per cent weight by minimum

20.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

17.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

9. Mono Ammonium Phosphate (11-52-0)

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen all in ammoniacal form per cent by weight, minimum

11.0

(iii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

52.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

44.5

(v)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

10. Ammonium Nitrate Phosphate (23-23-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (Ammoniacal and Nitrate) per cent by weight, minimum

23.0

(iii)

Nitrogen in ammoniacal form per cent by weight, minimum

13.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

23.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

11. Ammonium Poly-phosphate (10-34-0)(Liquid)

(i)

Total nitrogen (all as ammoniacal nitrogen) per cent by weight, minimum

10.0

(ii)

Total Phosphorous (as P₂O₃) per cent by weight, minimum

34.0

(iii)

Poly-phosphorus (as P₂O₃) per cent by weight, minimum

22.0

(iv)

Specific gravity (at 27'C)

1.2-1.6

(v)

pH (5% solution)

5.8-6.2

12. Ammonium Phosphate (14-28-0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and Urea) per cent by weight, minimum

14.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.0

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

28.9

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

24.0

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

13. NPK 13-33:0:155

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Ammoniacal nitrogen per cent by weight, minimum

13.0

(iii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

33.0

(iv)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

28.5

(v)

Total elemental+sulphate sulphur (as S), per cent by weight, minimum

15.0

(vi)

Sulphate sulphur (as S), per cent by weight, minimum

7.5

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

14. Nitrophosphate (24-24-9)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

24.0

(iii)

Nitrogen in ammoniacal form per cent by weight, minimum

13.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

24.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

20.5

(vi)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

[15. Neem Coated Urea Ammonium Phosphate 28-28-0

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total Nitrogen (Ammoniacal& Urea) per cent, by weight, minimum

28.0

(iii)

Ammoniacal nitrogen percent by weight, minimum

9.0

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

28.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

24.0

(vi)

Neem oil content soluble in Benzene per cent by weight, minimum

1.035

(vii)

Particle size-Minimum 90 per cent, of the material shall be retained between 1 mm 15 sieve 4mm 15 sieve]

1(f). N.P.K. Complex Fertilisers

1. Nitrophosphate with Potash (15-15-15)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

15.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

4.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

15.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1mm and 4mm 15 sieve

2. N.P.K. (10-26-26)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight minimum

10.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

26.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

22.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

26.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

3. N.P.K. (12-32-16)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight minimum

12.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

9.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

32.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

27.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

16.0

(vii)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

4. N.P.K. (22-22-11)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

22.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

22.0

(v)

Water soluble phosphorous (asP₂O₃) per cent by weight, minimum

19.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

11.0

(vii)

particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve.

5. N.P.K. (14-35-14)

(i)

Moisture per cent by weight maximum

1.5

(ii)

Nitrogen in ammoniacal form per cent by weight, minimum

14.0

(iii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

35.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

30.0

(v)

Water soluble potassium (as K₂O) per cent by weight, minimum

14.0

(vi)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

6. N.P.K. (17-17-17)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

17.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

5.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight minimum

17.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

17.0

(vii)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

7. N.P.K. (14-28-14)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

14.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

6.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

28.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

24.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

14.0

(vii)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

8. N.P.K. (19-19-19)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

19.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

5.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

19.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

16.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

19.0

(vii)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

9. N.P.K. (17-17-17)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

17.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

17.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

13.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

17.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

10. N.P.K. (20-10-10)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

4.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

10.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

8.3

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

10.0

(vii)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

11. N.P.K. (15-15-15)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

12.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

15.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

13.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

15.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

12. N.P.K (15-15-15-9(51)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

12.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

15.0

(v)

Water soluble potassium (as P₂O₃) per cent by weight, minimum

13.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

15.0

(vii)

Sulphate Sulphur (as 5) per cent by weight, minimum

9.0

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

13. N.P.K. (12:11:18)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

12.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

11.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

8.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

18.0

(vii)

Magnesium (as Mg) per cent by weight, minimum

1.0

(viii)

Sulphate sulphur (as 5) per cent by weight, minimum

7.5

(ix)

Total chlorides (as CI) per cent by weight, maximum

1.0

(x)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

14. N.P.K. (16:16:16)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

16.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

16.0

(v)

Water soluble phosphorous (as P₂O₆) per cent by weight, minimum

12.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

16.0

(vii)

Particle size- Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

15. N.P.K. (9:25:25)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrogen) per cent by weight, minimum

9.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

6.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

25.0

(v)

Water soluble phosphorous (as P₂O₃ per cent by weight, minimum

21.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

25.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

16. Nitrophosphate with Potash (14-14-21)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

14.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

14.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

9.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

21.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

17. Nitrophosphate with Potash (21-06-13)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

21.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

10.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

6.0

(v)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

4.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

13.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

18. Nitrophosphate with Potash Grade II (15-15-15)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

15.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

10.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

15.0

(vii)

Sulphate Sulphur (as 5) per cent by weight, minimum

3.5

(viii)

Total chloride (as CI) per cent by weight, maximum

3.5

(ix)

Particle size-Minimum 90 per cent of the material shall be between 1 mm and 4 mm 15 sieve

19. Nitrophosphate with Potash Grade II (15-9-20)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and nitrate) per cent by weight, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

8.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

9.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

6.0

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

20.0

(vii)

Sulphate Sulphur (as 5) per cent by weight minimum

3.5

(viii)

Magnesium (as Mg) per cent by weight, minimum

0.5

(ix)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

1(g) Micro Nutrients

1. Zinc Sulphate Heptahydrate (XnSO₆7H₂O)

(i)

Matter insoluble in water per cent by weight, maximum

1.0

(ii)

Zine (as Zn) per cent by weight, minimum

21.0

(iii)

Sulphate sulphur (as 5) per cent by weight, minimum

10.0

(iv)

pH (5% solution) not less than

4.0

(v)

Lead (as Pb) per cent by weight, maximum

0.003

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Arsenic (as As) per cent by weight, maximum

0.01

2. Manganese Sulphate

(i)

Manganese (as Mn) content per cent by weight minimum

30.5

(ii)

Sulphate sulphur (as 5) per cent by weight, minimum

17.0

(iii)

Matter insoluble in water per cent by weight, maximum

1.0

(iv)

pH (5% solution) not less than

4.0

(v)

Lead (as Pb) per cent by weight, maximum

0.003

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Arsenic (as As) per cent by weight, maximum

0.01

3. Borax (Sodium Tetraborate) (Na₂B₂O₇, 10H₂O) for Soil Application

(i)

Content of boron as (B) per cent by weight, minimum

10.5

(ii)

Matter insoluble in water per cent by weight, maximum

1.0

(iii)

pH (3.8% solution)

9.0-9.5

(iv)

Lead (as Pb) per cent by weight, maximum

0.003

(v)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vi)

Arsenic (as As) per cent by weight, maximum

0.01

4. Copper Sulphate (CusO₄,5H₂O)

(i)

Copper (as Cu) per cent by weight, minimum

24.0

(ii)

Sulphate sulphur (as 5) per cent by weight, minimum

2.0

(iii)

Matter insoluble in water per cent by weight, maximum

1.0

(iv)

pH (5% solution) not less than

3.0

(v)

Lead (as Pb) per cent by weight, maximum

0.003

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Arsenic (as As) per cent by weight, maximum

0.01

5. Ferrous Sulphate (FeSo₄, 7H₂O)

(i)

Ferrous iron (as Fe) per cent by weight, minimum

19.0

(ii)

Sulphate sulphur (as 5) percent by weight, minimum

10.5

(iv)

Free acid (as H₂So₄) per cent by weight, maximum

1.0

(v)

Matter insoluble in water, per cent by weight, maximum

1.0

(vi)

pH (5% solution) not less than

3.5

(vii)

Lead (as Pb) per cent by weight, maximum

0.003

(viii)

Cadmium (as Cd) per cent by weight, maximum

0.00025

(ix)

Arsenic (as As) per cent by weight, maximum

0.01

6. Ammonium Molybdate (NH₄)₆ Mo₇O₂₄, 4H₂O

(i)

Molybdenum (as Mo) per cent by weight, minimum

52.0

(ii)

Matter insoluble in water per cent by weight, maximum

1.0

(iii)

Lead (as Pb) per cent by weight, maximum

0.0003

(iv)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(v)

Arsenic (as As) per cent by weight, maximum

0.01

7. Chelated Zinc an Zn-EDTA

(i)

Appearance-Free flowing crystalline or powder or Tablet

(ii)

Zinc content (Expressed as Zn) per cent by weight, minimum in the form of Zn-EDTA

12.0

(iii)

pH (5% solution)

6.0-6.5

(iv)

Lead (as Pb) per cent by weight, maximum

0.0003

(v)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vi)

Arsenic (as As) per cent by weight, maximum

0.01

8. Chelated Iron as Fe-EDTA

(i)

Appearance-Free flowing crystalline/powder

(ii)

Iron content (expressed as Fe), per cent by weight minimum in the form of Fe-EDTA

12.0

(iii)

pH (5% solution)

5.5-6.5

(iv)

Lead (as Pb) per cent by weight, maximum

0.003

(v)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vi)

Arsenic (as As) per cent by weight, maximum

0.01

9. Zinc Sulphate Mono-hydrate (ZnSO₄H₂O)

(i)

Free flowing powder form

(ii)

Zinc (as Zn) per cent by weight, minimum

33.0

(iii)

Sulphate sulphur (as 5) per cent by weight, minimum

15.0

(iv)

Matter insoluble in water per cent by weight, maximum

1.0

(v)

pH (5% solution) not less than

4.0

(vi)

Iron (as Fe) per cent by weight, maximum

1.0

(vii)

Lead (as Pb) per cent by weight, maximum

0.003

(viii)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(ix)

Arsenic (as As) per cent by weight, maximum

0.01

10. Magnesium Sulphate

(i)

Magnesium (as Mg) per cent by weight, minimum

9.5

(ii)

Sulphate Sulphur (as 5) per cent by weight, minimum

12.0

(iii)

Matter insoluble in water per cent by weight, maximum

1.0

(iv)

pH (5% solution)

5.0-8.0

(v)

Lead (as Pb) per cent by weight, maximum

0.003

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Arsenic (as As) per cent by weight, maximum

0.01

11. Boric Acid (H₃BO₃)

(i)

Boron (as B) per cent weight, minimum

17.0

(ii)

Matter insoluble in water per cent by weight, maximum

1.0

(iii)

Lead (as Pb) per cent by weight, maximum

0.003

(iv)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(v)

Arsenic (as As) per cent by weight, maximum

0.01

12. Di-Sodium Octa Borate Tetra Hydrate (Na₂B₈O₁₃-4H₂O)

(i)

Boron (as B) per cent by weight, minimum

20.0

(ii)

Matter insoluble in water per cent by weight, maximum

1.0

(iii)

Lead (as Pb) per cent by weight, maximum

0.003

(iv)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(v)

Arsenic (as As) per cent by weight, maximum

0.01

13. Di-Sodium Tetra Borate Penta Hydrate (Granular)

(i)

Boron (as B) per cent by weight, minimum

14.5

(ii)

Matter insoluble in water per cent by weight, maximum

1.0

(iii)

Lead (as Pb) per cent by weight, maximum

0.003

(iv)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(v)

Arsenic (as As) per cent by weight, maximum

0.01

(vi)

Particle size-Minimum 90% of the material shall be retained between 1.4 mm and 5 mm 15 sieve

14. Zinc Sulphate Monohydrate (Granular)

(i)

Zinc (as Zn) per cent by weight, minimum

33.0

(ii)

Sulphate-Sulphur (as 5) per cent by weight, minimum

15.0

(iii)

Iron (as Fe) per cent by weight, maximum

1.0

(iv)

Matter-insoluble in water per cent by weight, maximum

1.0

(v)

pH (5% solution) not less than

4.0

(vi)

Lead (as Pb) per cent by weight, maximum

0.003

(vii)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(viii)

Arsenic (as As) per cent by weight, maximum

0.01

(ix)

Particle size- Minimum 90 per cent of the material shall be retained between 4 mm and 2 mm 15 sieve

15. Zinc Oxide Suspension Concentrate (39.5% Zn)

(i)

Dense suspension concentrate of liquid Zinc

(ii)

Zinc (as Zn) per cent by weight, minimum

39.5

(iii)

pH (5% solution)

9.0+1

(iv)

Arsenic (as As) per cent by weight, maximum

0.001

(v)

Lead (as Pb) per cent by weight, maximum

0.003

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

(vii)

Specific gravity (at 15'C)

1.71-1.75

16. Di-Sodium Tetra Borate Penta Hydrate

(i)

Appearance

Free flowing crystalline

(ii)

Boron (as B) per cent by weight, minimum

14.5

(iii)

Matter insoluble in water per cent by weight, maximum

1.0

(iv)

Arsenic (as As) per cent by weight, maximum

0.001

(v)

Lead (as Pb) per cent by weight, maximum

0.001

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025

17. Anhydrous Boron (Na₂B₄O₇)

(i)

Boron (as B) per cent by weight, minimum

20.5

(ii)

Matter insoluble in water per cent by weight, maximum

0.1

(iii)

Arsenic (as As) per cent by weight, maximum

0.001

(iv)

Lead (as Pb) per cent by weight, maximum

0.001

(v)

Appearance

Free flowing form

[18. Concentrated Liquid Calcium 11%

(i)

Calcium (as Ca) per cent by weight, minimum

11.0

(ii)

9.5+/1.00

(iii)

Specific gravity (kg/1)

1.3-1.4]

[19. Zinc polyphosphate

(i)

Appearance

Free flowing

(ii)

Zinc (as Zn) per cent, by weight, minimum

16.0

(iii)

DTPA soluble Zinc (as Zn) per cent by weight, minimum

15.0

(iv)

Water soluble Zinc (as Zn) per cent by weight, maximum

1.0

(v)

Magnesium as Mg per cent, by weight, maximum

3.0

(vi)

Lead as Pb per cent by weight, maximum

0.003

(vii)

Arsenic as As per cent by weight, maximum

0.01

(viii)

Cadmium (as Cd) per cent by weight, maximum

0.0025

20. Chelated Zinc as Zn-HEDP

(i)

Appearance

free flowing crystalline

(ii)

Zinc (as Zn) per cent, by weight, minimum in the form of Zn-HEDP

17.0

(iii)

Lead (as Pb) per cent by weight, maximum

0.003

(iv)

Arsenic (as As) per cent by weight, maximum

0.01

(v)

Cadmium (as Cd) per cent by weight, maximum

0.0025

21. Chelated iron as Fe-HEDP

(i)

Appearance

free flowing crystalline

(ii)

Iron (as Fe) per cent by weight, minimum in the form of Fe-HEDP

17.0

(iii)

Lead (as Pb) per cent, by weight, maximum

0.003

(iv)

Arsenic (as As) per cent, by weight, maximum

0.01

(v)

Cadmium (as Cd) per cent by weight, maximum

0.0025

22. Colemanite

(i)

Boron as B per cent by weight, minimum

11.0

(ii)

Sodium (as NaCl) per cent, by weight, maximum

0.5

(iii)

Moisture per cent, by weight maximum

6.50

(iv)

Lead (as Pb) per cent by weight, maximum

0.003

(v)

Arsenic (as As) per cent by weight, maximum

0.01

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025]

[23. Boron ethanol amine

(i)

Boron (as B), per cent by weight, minimum

10.0

(ii)

9.5+1

(iii)

Specific gravity

1.3-1.4

(iv)

Lead (as Pb) per cent by weight, maximum

0.003

(v)

Arsenic (as As) per cent by weight, maximum

0.01

(vi)

Cadmium (as Cd) per cent by weight, maximum

0.0025]

[24. Magnesium Hydroxide and Zinc Phosphate

(i)

Magnesium as Mg per cent, by weight minimum

24.0

(ii)

Zinc as Zn per cent, by weight minimum

10.0

(iii)

pH (50 g/L)

8.5+/-1

(iv)

Available Phosphorus as P₂O₅, per cent, by weight minimum

2.5.]

[25. Manganese carbonate Suspension concentrate

Manganese per cent by weight, minimum

26.0

9+/-1

Specific gravity

1.8 - 1.9

Arsenic (as As) per cent by weight, maximum

0.001

Lead (as Pb) per cent by weight maximum

0.003

Cadmium (as Cd) per cent by weight, maximum

0.0025]

[Note: The "Micronutrient Mixture grades are made by physically mixing of above specified fertilizers or grades.]

1(h) Fortified Fertilisers

1. Boronated Single Superphosphate (16% P2O3 Powdered)

(i)

Moisture per cent by weight, maximum

12.0

(ii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

11.0

(v)

Boron (as B) per cent by weight

0.2-0.3

(vi)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

4.0

2. Zincated Urea

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Total nitrogen (in Urea form) per cent by weight, minimum

43.0

(iii)

Zinc (as Zn) per cent by weight, minimum

2.0

(iv)

Biuret, per cent by weight, maximum

1.5

(v)

Particle Size-Minimum 80 per cent of the material shall be retained between 1 mm and 2.8 mm 15 sieve

3. Zincated Phosphate (Suspension) - for Seed Treatment

(i)

Total Phosphorous (as P2O3) per cent by weight, minimum

3.0

(iii)

Total zinc (Zn) per cent by weight, minimum

17.5

(iv)

Lead (as Pb) per cent by weight, maximum

0.003

(v)

pH (5% solution)

8+1

4. Boronated NPK Complex (10:26:26)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

10.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

26.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

22.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

26.0

(vii)

Boron (as B) per cent by weight

0.2-0.3

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

5. Bomnated NPK Complex (12:32:16)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

12.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

9.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

32.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

27.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

16.0

(vii)

Boron (as B) per cent by weight

0.2-0.3

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

6. Boronated Diammonium Phosphate (18:46:0)

(i)

Moisture per cent by weight, maximum

2.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

18.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

15.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

46.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

41.0

(vi)

Boron (as B) per cent by weight

0.2-0.3

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

7. Zincated NPK Complex (10:26:26)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

10.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

26.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight minimum

22.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

26.0

(vii)

Zinc (as Zn) per cent by weight, minimum

0.5

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

8. Zincated NPK Complex (12:32:16)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

12.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

9.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

32.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

27.5

(vi)

Water soluble potassium (as K₂O) per cent by weight, minimum

16.0

(vii)

Zinc (as Zn) per cent by weight, minimum

0.5

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

9. Boronated Calcium Nitrate

(i)

Total nitrogen (ammoniacal and nitrate N) per cent by weight, minimum

14.5

(ii)

Nitrate nitrogen as N per cent by weight, minimum

13.5

(iii)

Water soluble calcium as per cent by weight minimum

17.0

(iv)

Boron (as B) per cent by weight

0.2-0.3

10. Boronated Nitrophosphate with Potash (15:15:15)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and Nitrate N) per cent by weight, minimum

15.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.5

(iv)

Available phosphorous (as P₂O) per cent by weight, minimum

15.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight,

4.0

(vi)

Water soluble potassium (as P₂O₃) per cent by weight, minimum

15.0

(vii)

Boron (as B) per cent by weight

0.2-0.3

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

11. Zincated DAP (18:46:0)

(i)

Moisture per cent by weight, maximum

2.5

(ii)

Total nitrogen (ammoniacal and urea) per cent by weight, minimum

18.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

15.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

46.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

39.5

(vi)

Zinc (as Zn) per cent by weight, minimum

0.5

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

12. Zincated SSP (16% P2O3)

(i)

Moisture per cent by weight, maximum

12.0

(ii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

11.0

(v)

Zinc (as Zn) per cent by weight, minimum

0.5

(vi)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

4.0

13. Bononated NPK Complex (24:24:0)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (ammoniacal and Nitrate) per cent by weight, minimum

24.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

13.5

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

24.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

20.5

(vi)

Boron as B per cent by weight

0.2-0.3

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve.

14. Boronated Single Super Phosphate (16% P₂O₃) Granular)

(i)

Moisture per cent by weight, maximum

5.0

(ii)

Available phosphorous (as P₂O₃) per cent by weight minimum

16.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

11.0

(v)

Boron (as B) per cent by weight

0.2-0.3

(vi)

Free phosphoric acid (as P₂O₃) per cent by weight, maximum

4.0

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

15. Zincated Bentonite Sulphur

(i)

Moisture per cent by weight, maximum

1.0

(ii)

Elemental Sulphur (as 5) per cent by weight, minimum

65.0

(iii)

Zinc (as Zn) per cent by weight, minimum

18.0

(iv)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

16. Zincated Urea Ammonium Phosphate 28-28-0

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen per cent by weight, minimum

28.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

9.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

28.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

25.2

(vi)

Zinc (as Zn) per cent by weight

0.5

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

17. Zincated NPK 14-35-14

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Ammoniacal nitrogen per cent by weight, minimum

14.0

(iii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

35.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

29.0

(v)

Water soluble potassium (as K₂O) per cent by weight, minimum

14.0

(vi)

Zinc (as Zn) per cent by weight

0.5

(vii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

18. Zincated Ammonium Phosphate Sulphate (20-20-0-13)

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total Nitrogen per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

18.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight minimum

20.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

17.0

(vi)

Sulphate Sulphur (as 5) per cent by weight, minimum

13.0

(vii)

Zinc (as Zn) per cent by weight

0.55

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 115 sieve

19. NP 24-24-0 fortified with Sulphur

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total Nitrogen (Ammoniacal and Urea) per cent by weight, minimum

24.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

7.5

(iv)

Nitrogen in the form of Urea, maximum

16.5

(v)

Available phosphorous (as P₂O₃) per cent by weight, minimum

24.0

(vi)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

20.0

(vii)

Total Sulphur per cent by weight, minimum

8.0

(viii)

Elemental Sulphur per cent by weight, minimum

4.0

(ix)

Sulphate Sulphur (as 5) per cent by weight, minimum

4.0

(x)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm 15 sieve

20. Boronated Ammonium Phosphate Sulphate

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (Ammoniacal and Urea) per cent by weight, minimum

20.0

(iii)

Ammoniacal nitrogen per cent by weight minimum

18.0

(iv)

Available phosphorous (as P₂O₃) per cent by weight, minimum

20.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

17.0

(vi)

Sulphate sulphur (as 5) per cent by weight, minimum

13.0

(vii)

Boron (as B) per cent by weight

0.2-0.3

(viii)

Particle size-Minimum 90 per cent of the material shall be retained between 1 mm and 3 mm 15 sieve

[21. NPK 14-35-14 fortified with Zinc and Boron

(i)

Moisture per cent by weight maximum

1.0

(ii)

Nitrogen in ammoniacal form per cent by weight minimum

14.0

(iii)

Available phosphorous (as P₂O₃) per cent by weight, minimum

35.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

29.0

(v)

Water soluble potash (as K₂O) per cent by weight, minimum

14.0

(vi)

Zinc (as Zn) per cent by weight, minimum

0.5

(vii)

Boron (as B) per cent by weight, minimum

0.3

(viii)

Particle size-Minimum 90 per cent of the material shall retain between 1 mm 15 sieve and 4 mm 15 sieve. Not more than 5 per cent shall be below 1 mm 15 sieve

22. SSP fortified with Zinc and Boron (Granular)

(i)

Moisture per cent by weight, maximum

5.0

(ii)

Available phosphorus (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

14.5

(iv)

Zinc (as Zn) per cent by weight, minimum

0.5

(v)

Boron (as B) per cent by weight, minimum

0.2

(vi)

Free phosphoric Acid (as P₂O₃) per cent by weight, maximum

4.0

(vii)

Particle size-Minimum 90 per cent of the material shall retain between through 1.0 mm 15 sieve 4 mm 15 sieve

[(viii)

Sulphate Sulphur (as 5) per cent, by weight, minimum

11.0]

23. Boronated Sulphur (Granular)

(i)

Moisture per cent by weight maximum

0.50

(ii)

Total elemental sulphur (as 5) per cent by weight, minimum

80.0

(iii)

Boron (as B) per cent by weight, minimum

1.20

(iv)

Particle size-Minimum 90 per cent of the material shall retain between through 1.0 mm 15 sieve 4 mm 15 sieve

(Note: the product may contain insert filler material as Bentonite etc. up to the extent of 14 per cent by weight, maximum]

[24. Urea Ammonium Phosphate 20-20-0 fortified with 13% Sulphur

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total Nitrogen in (ammoniacal and urea) per cent by weight, minimum

20.0

(iii)

Available phosphorus (as P₂O₃) per cent by weight, minimum

20.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

17.0

(v)

Elemental Sulphur (as 5) per cent by weight, minimum

6.5

(vi)

Sulphate Sulphur (as 5) per cent by weight, minimum

6.5

(vii)

Particle size-Minimum 90% of the material shall retain between 1 mm 15 sieve and 4 mm 15 sieve

25. Zincated SSP (Granular)

(i)

Moisture per cent by weight, maximum

5.0

(ii)

Available phosphorus (as P₂O₃) per cent by weight, minimum

16.0

(iii)

Water soluble phosphorus (as P₂O₃) per cent, by weight, minimum

14.5

(iv)

Free phosphoric acid per cent by weight, maximum

4.0

(v)

Sulphate Sulphur (as 5) per cent, by weight, minimum

11.0

(vi)

Zinc (as Zn) per cent by weight, minimum

0.5

(vii)

Particle size-Minimum 90% of the material shall retain between 4 mm 15 sieve and 1 mm 15 sieve]

[26. Elemental Sulphur fortified with Zinc Oxide in microgranule form

(i)

Elemental Sulphur as 5 per cent, by weight minimum

67.0

(ii)

Zinc as Zn per cent, by weight, minimum

14.0

(iii)

Moisture, per cent, by weight, maximum

2.0

(iv)

Particle size-80% of the material shall retain on 0.149 mm 15 sieve]

[27. NPK 8:21:21 fortified with Mg. S, Zn and B

(i)

Moisture per cent by weight, maximum

1.5

(ii)

Total nitrogen (Ammoniacal and Urea) per cent by weight, minimum

8.0

(iii)

Ammoniacal nitrogen per cent by weight, minimum

5.5

(iv)

Available phosphorus (as P₂O₃) per cent by weight, minimum

21.0

(v)

Water soluble phosphorus (as P₂O₃) per cent, by weight, minimum

18.0

(vi)

Water soluble Potassium (as K₂O) per cent, by weight, minimum

21.0

(vii)

Magnesium (as Mg), per cent by weight, minimum

1.20

(viii)

Total Sulphur (as 5) per cent, by weight, minimum

2.0

(ix)

Zinc (as Zn) per cent by weight minimum

1.0

(x)

Boron (as B) per cent by weight

0.2-0.3

(xi)

Particle size-Not less than 90 per cent of the material shall be retained between 1 mm 15 sieve and 4 mm 15 sieve

28. Diammonium Phosphate fortified with 4% Sulphur

(i)

Moisture, per cent, by weight, maximum

2.5

(ii)

Ammoniacal nitrogen per cent by weight, minimum

15.5

(iv)

Available Phosphorus (as P₂O₃) per cent, by weight, minimum

46.0

(v)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

41.0

(vi)

Total Sulphur (as 5) per cent by weight, minimum

4.0

(vii)

Elemental Sulphur (as 5) per cent by weight, minimum

3.0

(viii)

Particle size-Not less than 90 per cent of the material shall be retained between 1 mm 15 sieve and 4 mm 15 sieve]

[29. NPK 9:24:24 fortified with Mg S Zn B

(i)

Total nitrogen (ammoniacal and urea), per cent, by weight, minimum

9.0

(ii)

Ammoniacal nitrogen, per cent, by weight minimum

6.3

(iii)

Available phosphorous (as P₂O₃) per cent, by weight, minimum

24.0

(iv)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

20.5

(v)

Water soluble potassium (K₂O), per cent by weight, minimum

24.0

(vi)

Magnesium (as Mg),percent by weight, minimum

0.3

(vii)

Total Sulphur (S) per cent, by weight, minimum

2.1

(viii)

Total Zinc (Zn), per cent, by weight, minimum

0.6

(ix)

Total Boron (b), per cent by weight

0.2-0.3

(x)

Moisture per cent by weight, maximum

1.3

(xi)

Particle size- Minimum 90 per cent of the material shall be retained between 1 mm and 4 mm sieve

30. Boronated NP 28-28-0

(i)

Moisture per cent, by weight, maximum

1.5

(ii)

Total N (ammoniacal and nitrate) per cent, by weight, minimum

28.0

(iii)

Ammoniacal per cent,by weight, minimum

9.0

(iv)

Available phosphorous (P₂O₃) per cent, by weight, minimum

28.0

(v)

Water soluble phosphorous (P₂O₃) per cent by weight, minimum

24.0

(vi)

Boron (as B) per cent by weight

0.2

(vii)

Particle size-Min 90% of the material shall be retained between 1 mm and 4 mm 15 sieve]

[31. NPK 11-30-14 fortified with Magnesium, Sulphur, Zinc and Boron

(i)

Moisture per cent, by weight maximum

1.5

(ii)

Total Nitrogen (Ammonical + urea) per cent, by weight, minimum

11.0

(iii)

Ammoniacal nitrogen percent by weight, minimum

8.2

(iv)

Available Phosphorus (as P205) per cent, by weight, minimum

30.0

(v)

Water soluble phosphorus (as P205), per cent, by weight, minimum

25.5

(vi)

Water soluble potassium (as K20) per cent, by weight minimum

14.0

(vii)

Zinc (as Zn) per cent, by weight minimum

1.0

(viii)

Total Sulphur (as S) per cent, by weight, minimum

1.2

(ix)

Total boron (as B) per cent, by weight minimum

0.2-0.3

(x)

Magnesium (as Mg), per cent, by weight minimum

0.4

(xi)

Particle Size -90% of the material shall retain on 4 mm IS sieve and 1mm IS sieve.

32. SSP Fortified with Magnesium, Zinc and Boron (Granular)

Moisture percent by weight, maximum

5.0

Available phosphorous (as P205) percent by weight, minimum

16.0

Water soluble phosphorous (as P205) percent by weight, minimum

14.5

Magnesium (as Mg) percent by weight, minimum

0.5

Zinc (as Zn) percent by weight, minimum

0.5

Boron (as B) percent by weight, minimum

0.2

Sulphate Sulphur (as S) percent by weight, minimum

11.0

Free Phosphoric acid (as P205) percent by weight maximum

4.0

Particle size- 90 percent of the material shall retain between 4.0 mm IS sieve and 1 mm IS sieve

33. Ammonium Phosphate Sulphate (16-20-0) fortified with Magnesium

Moisture per cent, by weight, maximum

1.0

Total ammonical nitogen per cent, by weight, maximum

16.0

Available Phosphorus (as P205) per cent, by weight, minimum

20.0

Water soluble Phosphorus (as P205 per cent, by weight, minimum

17.0

Sulphate Sulphur (as S) per cent, by weight, minimum

13.0

Magnesium as (Mg) per cent, by weight, minimum

0.6

Particle Size - minimum 90% of the material shall be retained between 4 mm and 1 mmIS sieve.]

1(i) 100% Water Soluble Complex Fertilisers

1. Potassium Nitrate (13-0-45)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Total nitrogen (all in Nitrate form) per cent by weight, minimum

13.0

(iii)

Water soluble potassium (as K₂O) per cent by weight, minimum

45.0

(iv)

Sodium (as Na) per cent by weight, maximum

1.0

(v)

Total chloride (as CI) per cent by weight, maximum

1.5

(vi)

Matter insoluble in water, per cent by weight, maximum

0.5

2. Mono-Potassium Phosphate (0-52-34) (100% water soluble)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

52.0

(iii)

Water soluble potassium (as K₂O) per cent by weight, minimum

34.0

(iv)

Sodium (as NaCI) per cent by weight, maximum

0.5

3. Calcium Nitrate

(i)

Total nitrogen (Ammoniacal and Nitrate form) per cent by weight, minimum

15.5

(ii)

Nitrate nitrogen as N per cent by weight, minimum

14.5

(iii)

Water soluble calcium (as Ca) per cent by weight, minimum

18.5

(iv)

Matter insoluble in water per cent by weight, maximum

1.5

4. Potassium Magnesium Sulphate

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Water soluble potassium (as K₂O) per cent by weight, minimum

22.0

(iii)

Magnesium as MgO per cent by weight, minimum

18.0

(iv)

Sulphate Sulphur (as 5) per cent by weight, minimum

20.0

(v)

Total chloride (as CI) per cent by weight, maximum

2.5

(vi)

Sodium (as NaCI), per cent by weight, maximum

2.0

5. Mono Ammonium Phosphate 12:61:0 (100% water soluble)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Ammoniacal nitrogen per cent by weight, minimum

12.0

(iii)

Water soluble phosphorous (as P₂O₃) per cent by weight, minimum

61.0

(iv)

Sodium as NaCI per cent by weight, maximum

0.5

(v)

Matter insoluble in water per cent by weight, maximum

0.5

6. Urea Phosphate 17:44:0 (100% Water Soluble)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Total nitrogen (all in urea form) per cent by weight minimum

17.0

(iii)

Water soluble phosphorus (as P₂O₃) per cent by weight, minimum

44.0

(iv)

Matter insoluble in water per cent by weight, maximum

0.5

[***]

[7. Potassium Nitrate (prilled) (13-0-45) (soil application)

(i)

Moisture per cent by weight, maximum

0.5

(ii)

Total nitrogen (all in Nitrate form) per cent by weight, minimum

13.0

(iii)

Water soluble potassium (as K₂O) per cent by weight, minimum

45.0

(iv)

Sodium (as Na) per cent, by weight maximum

1.0

(v)

Total Chloride (as CI) per cent by weight, maximum

1.5

(vi)

Matter insoluble in water, per cent by weight, maximum

[1.5]

(vii)

Particle size 80 per cent of the material shall be retained between 1 mm and 2.8 mm 15 sieve]

[8. 24-24-0 100% Water Soluble Complex Fertiliser

S.No.	Parameter
(i)	Moisture per cent. by weight maximum	0.5
(ii)	Total Nitrogen (Ammoniacal and Nitrate) minimum	24.0
(iii)	Ammoniacal Nitrogen per cent. by weight minimum	14.0
(iv)	Nitrate Nitrogen per cent. by weight minimum	10.0
(v)	Water Soluble Phosphorus per cent. by weight minimum	24.0
(vi)	matter insoluble in water per cent. by weight maximum	0.5]

[9. Potassium Metaphosphate dimer (0-40-40)

(i)

Moisture per cent, by weight, maximum

0.5

(ii)

Water soluble phosphorus (as P205) per cent, by weight, minimum

40.0

(iii)

Water soluble Potassium (as K20) per cent by weight, minimum

40.0

(iv)

Matter insoluble per cent by weight maximum

0.5

(v)

Lead as Pb per cent, by weight maximum

0.0003

(vi)

Cadmium (as Cd) per cent by weight, minimum

0.0025

(vii)

Arsenic (as As) per cent, by weight, maximum

0.1]

[1(j) Beneficial Element Fertiliser

1. Ortho Silicic Acid (OSA) 2.0% WSL

Ortho Silicic Acid SI(OH)4, per cent, by weight, minimum /plant available Silicon (Sl) equivalent, per cent by weight, minimum

2.0
[0.6]

[***]

Matter insoluble in water percent by weight, maximum

0.5

Specific Gravity (g/m)

1.07-1.15

Sodium (as Na) per cent by weight, Maximum

0.10

Total Chloride (as C1) per cent by weight, Maximum

4.00]

[2. Diatomite Silicon

Moisture per cent. by weight maximum

12.0

ii.

pH, minimum

7.5

iii.

Plant available silicon (Si (OH)4 per cent. by weight minimum

0.08

iv.

Lead (as Pb) per cent by weight, maximum

0.003

Cadmium (as Cd) per cent by weight, maximum

0.0025

vi.

Arsenic (as As) per cent by weight, maximum

0.01

vii.

Particle Size : Not less than 80 per cent. of the material shall retain between 6 mm IS sieve and 2mm IS sieve.]

[3. Sodium Silicate (liquid)

Silicon as SiO₂ per cent, by weight minimum

23.8

ii.

Sodium as Na per cent, by weight minimum

6.0

iii.

Specific gravity

1.3]

[1(k) Liquid Fertiliser

1. NPK 8:8:8 (liquid) (for Sugar cane crop)

(i)

Total Nitrogen (ammoniacal+nitrate form) per cent by weight, minimum

8.0

(ii)

Nitrogen in ammoniacal form per cent by weight, minimum

3.0

(iii)

Available Phosphorous (as P₂O₃) per cent by weight, minimum

8.0

(iv)

Water soluble Potash (as K₂O) per cent by weight, minimum

8.0

(v)

Sulphate sulphur per cent by weight, minimum

2.0

(vi)

Water soluble Chlorine (as Cl) per cent by weight, maximum

0.5

(vii)

pH of 1 per cent solution

3.5+1.5

(viii)

Insoluble particle will not exceed 0.1 per cent by weight when filtered on muslim cloth

(ix)

Specify gravity

1.35

2. Calcium Di Hydrogen Phosphate (liquid)

(i)

Available Phosphorous (as P₂O₃), per cent by weight, minimum

222.0

(ii)

1.1+/-1

(iii)

Sp gravity (kg/1)

1.25-1.35]

[3. Chelated Zinc as Zinc-Glycine (liquid)

(i)

Zinc (as Zn), per cent. by weight , minimum

6.80

(ii)

pH (1 per cent. Distilled water

4.0-5.5

(iii)

Specific gravity

1.21-1.28

4. Chelated Calcium as Calcium-Glycine (liquid)

(i)

Calcium (as Ca), per cent. by weight, minimum

6.00

(ii)

pH (1 per cent. Distilled water)

5.5-7.0

(iii)

Specific gravity

1.15-1.22

5. Chelated Boron as Boron-Glycine (liquid)

(i)

Boron (as B), per cent. by weight ,minimum

5.0

(ii)

pH (1 per cent. Distilled water

8.0-9.0

(iii)

Specific gravity (g/ml)

1.10-1.23]

[6. NK 6:0:18 Fortified with Calcium, Magnesium & Boron (suspension)

(i)

Total nitrogen per cent. by weight, minimum

6.0

(ii)

Nitrate Nitrogen as N per cent by weight minimum

5.8

(iii)

Water Soluble potassium as K₂O per cent by weight minimum

18.0

(iv)

Water soluble Calcium (as CaO), per cent by weight, minimum

5.0

(v)

Water soluble Magnesium (as MgO), per cent by weight, minimum

2.0

(vi)

Boron as B

0.5-0.8

(vii)

pH (1 % Solution) at 200°C

8 to 9

7. NPK 11: 11:8 Fortified with Zinc & Boron (suspension)

(i)

Total nitrogen per cent. by weight minimum

11.0

(ii)

urea nitrogen, per cent by weight, minimum

7.2

(iii)

Ammonical nitrogen, per cent, by weight maximum

3.0

(iv)

Water soluble phosphorus (as P₂O₅), per cent. By weight minimum

11.0

(iii)

Water Soluble potassium (as K₂O) per cent. By weight minimum

8.0

(iv)

Zinc as Zn percent by weight minimum in the form of Zn-EDTA

0.7

(vi)

Boron as B

0.5-0.7

(vii)

pH (1 % Solution) at 200°C

7.0-8.0

8. Calcium Nitrate Fortified with Magnesium (suspension)

(i)

Total nitrogen per cent. By weight minimum

10.0

(ii)

Nitrate Nitrogen per cent by weight, maximum

8.5

(iii)

Water solube calcium as CaO per cent, by weight minimum

15.0

(iv)

Water solube magnesium as MgO per cent, by weight maximum

2.0

(v)

Total chloride as Cl per cent. by weight maximum

2.5

(vi)

pH (1% Solution) at 200 C

8.0 - 9.0]

9. [-]

10. [-]

[11. Fortified NP (7-21-0) Liquid
Total Nitrogen (as N), percent by weight, minimum	7.0
Water soluble Phosphorus (as P205), per cent by weight, minimum	21.0
Zinc (as Zn), percent by weight, minimum	0.2
Specific Gravity	1.29-1.33
pH	52-1.2
12. Fortified Calcium Suspension
Calcium (as Ca), percent by weight, minimum	21.0
Boron (as B), percent by weight, minimum	0.1
Zinc (as Zn), percent by weight, minimum	1.5
Specific Gravity	1.74-1.76
pH	8.5-10.0]

Part B

[Tolerance Limit in plant nutrients and physical parameters for various fertilisers

S.No.	Fertilisers	Tolerance limit
1	Straight Nitrogenous Fertilisers, Straight Phosphatic Fertilisers, Straight Potassic Fertilisers, Straight Sulphur Fertilisers, NP Complex Fertilisers, NPK Complex Fertilisers, Micro Nutrients, 100 percent Water Soluble Complex Fertilisers, 100 percent Water Soluble Mixture of fertilisers, Mixture of fertilisers	1.5 units for each nutrient
2.	For mixture of micro nutrients fertilisers	10 percent of each nutrient or 0.5 unit whichever is less subject to maximum of 7.5 percent of all the combined nutrients
[3.	For Customised fertilisers and fortified fertiliser
	Nutrient level% for primary nutrient (NPK) and secondary nutrient (S,Ca, Mg)	05. units for each nutrient
	For micro nutrients	10 percent of each nutrient or 0.5 unit whichever is less subject to maximum of 7.5 percent of all the combined nutrients]
4	Particle size	[7 units]
5	Moisture	1.3 units

Note: The tolerance limit besides total nutrient shall also be applicable on the forms of nutrient subject to the condition that it shall not exceed the value of tolerance limit applicable on the total nutrient.]

OLD LAW ▼

Schedule I

[See Clause 2(h)& (q)]

Part A

Specifications of fertilisers

Name of the Fertiliser	Specifications
1	2
1. (a) Straight Nitrogenous Fertilisers :
1. Ammonium Sulphate	(i) Moisture per cent by weight, maximum	1.0
	(ii) Ammoniacal nitrogen percent. by weight, minimum	20.6
	(iii) Free acidity (as H₂SO₄) per cent. by weight maximum (0.04 for material obtained from by product ammonia and by product gypsum)	0.025
	(iv) Arsenic as (A S₂O₃) per cent. by weight, maximum	0.01
2. Urea (46% N) (White free flowing)	(i) Moisture per cent. by weight maximum	1.00
	(ii) Total nitrogen, per cent. by weight, (on dry basis) minimum	46.00
	(iii) Biuret per cent. By' weight, maximum	1.5
	(iv) Particle size - 90 per cent. of the material shall pass through 2.8 mm IS sieve and not less than 80 per cent. by weight shall be retained on 1 mm IS sieve.	1.5
3. Urea (coated) (45% N) (White free flowing)	(i) Moisture per cent. by weight, maximum	0.5
	(ii) Total nitrogen per cent. by weight content with coating minimum	45.0
	(iii) Biuret per cent. by weight maximum	15
	(iv) Particle size - 90 per cent. of the material shall pass through 2.8 mm IS sieve and not less than 80 per cent. by weight shall be retained on i mm IS sieve.
4. Ammonium Chloride	(i) Moisture per cent. by weight, maximum	2.0
	(ii) Ammoniacal nitrogen per cent. by weight, minimum	25.0
	(iii) Chloride other than ammonium chloride (as NaCI) per cent. by weight (on dry basis) maximum	2.0
	(iv) [* * *]
5. Calcium Ammonium (Nitrate 25%N)	(i) Moisture percent. by weight, maximum	1.00
	(ii) Total ammoniacal and nitrate nitrogen	25.0
	(iii) Ammoniacal nitrogen per cent. by weight, minimum	12.5
	(iv) Calcium nitrate per cent. by weight.maximum	0.5
	(v) Particle size - 80 per cent. of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 10 per cent. shall be below 1 mm IS sieve.
6. Calcium Ammonium Nitrate (26% N)	(i) Moisture per cent, by weight, maximum	1.0
	(ii) Total ammoniacal and nitrate nitrogen per cent. by weight, minimum	26.0
	(iii) Ammoniacal nitrogen per cent. by weight, minimum	13.0
	(iv) Calcium nitrate per cent. by weight, maximum	0.5
	(v) Particle size - 90 per cent. of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 percent shall be below I lmm IS sieve.
7. Anhydrous Ammonia	(i) Ammonia per cent. by weight, minimum	99.0
	(ii) Water per cent. by weight, maximum	1.0
	(iii) Oil content by weight, maximum 20 ppm
[8. Urea Super Granulated.]	(i) Moisture, percent, by weight, maximum	1.00
	(ii) Total nitrogen, per cent. by weight, (on 46.00 dry basis) minimum	46.00
	(iii) Biuret per cent. by weight, maximum	1.5
	(iv) Particle size, 90 per cent. of the material shall pass through 13.2 mm IS sieve and not less than [80] per cent. by weight shall be retained on 9.5 mm IS sieve
[9. (Urea granular)]	(i) Moisture per cent. by weight, maximum	1.0
	(ii) Total nitrogen, per cent. by weight, minimum	1.5
	(iii) Biurate, per cent, by weight, maximum	1.5
	(iv) Particle size 90 per cent. of the material shall 15 pass through 4 mm IS sieve and be retained on 2 mm IS sieve. Not more than 5 per cent. shall be blow 2 mm IS sieve."	1.5
[12. Urea Briquettes	(i) Moisture per cent. by weight, maximum	1.0
	(ii) Total nitrogen per cent. by weight (on dry basis), minimum	46.0
	(iii) Biuret per cent. by weight, maximum	1.5
	(iv) Particle size: Not less than 90 per cent. of the materials shall pass through 5.7 mm IS Sieve, and not less than 80 per cent. by weight shall be retained on 3.8 mm IS sieve.]
1.(b) Straight Phosphatic fertilisers :
1. Single Super - phosphate (16% P₂O₅ powered)	(i) Moisture percent. by weight, maximum	12.0
	(ii).Free phosphoric acid (as P₂O₅) per cent. by weight, maximum	4.0
	(iii) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	16.0
[***	*	]
3. Triple Super - phosphate	(i) Moisture per cent. by weight, maximum	12.0
	(ii) Free phosphoric acid (as P₂O₅) per cent. by weight, maximum	3.0
	(iii) Total phosphates (as P₂O₅) per cent. by weight, minimum	46.0
	(iv) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	42.5
4. Bone meal, Raw.	(i) Moisture per cent. by weight, maximum	8.0
	(ii) Acid insoluble matter per cent. by weight, maximum	12.0
	(iii) Total phosphates (as P₂O₅) per cent. by weight, maximum	20.0
	(iv) 2 per cent. citric acid soluble phosphates (as P₂O₅) per cent. by weight, minimum	8.0
	(v) Nitrogen content of water insoluble portion per cent. by weight, minimum
	(vi) Particle size—The material shall pass wholly through 2.36 mm IS sieve of which not more than 30 per cent. shall be retained on 0.85 mm IS sieve.
5. Bone meal, Steamed	(i) Moisture per cent. by weight, maximum	7.0
	(ii) Total phosphates (as P₂O₅) per cent. by weight (on dry basis), minimum	22.0
	(iii) 2 per cent, citric acid soluble phosphates (as P₂O₅) per cent. by weight (on dry basis), minimum	16.0
	(iv) Particle size—Not less than 90 per cent. of the material shall pass through 1.18 mm IS sieve.	1.18
6. Rock - phosphate	(i) Particle size—minimum 90 per cent. of the material shall pass through 0.15 mm IS sieve and the balance 10 per cent. of material shall pass through 0.25 mm IS sieve.
	[(ii) Total phosphate (as P₂O₅) per cent. by weight, minimum]	18.0
[7. Single Super Phosphate (16% P₂O₅Granulated)]	(i) Moisture per cent, by weight, maximum	5.0
	(ii) Free phosphoric acid (as P₂O₅) per cent. by weight, maximum	4.0
	(iii) Water soluble phosphates (as P₂O₅) per cent. by weight, maximum
	(iv) Particle size - Not less than 90 per cent. of the material shall pass through 4 mm IS sieve. Not more than 5 per cent. shall pass through 1mm IS sieve.]
8. [* * *]
1. (c) Straight potassic fertilisers :
1. Potassium Chloride (Muriate of potash)	(i) Moisture per cent. by weight, maximum	0.5
	(ii) Water soluble potash content (as K₂O) per cent. by weight, minimum .	[60.0]
	(iii) Sodium as NaCI per cent. by weight (on dry basis), maximum
	(iv) Particle size - [minimum 95 per cent.] of the material shall pass through 1.7 mm IS sieve and be retained on 0.25 mm IS sieve.
2. Potassium Sulphate	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Potash content (as K₂O) per cent. by 6. [weight], minimum	[50.00]
	(iii) Total chlorides (as C1) as per cent. by weight (on dry basis), maximum
	(iv) Sodium as Nacl per cent. by weight (on dry basis), maximum	2.0
3. Potassium Schoenite	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Potash content (as K₂O) per cent. by weight (on dry basis), minimum	23.0
	(iii) Magnesium oxide (as MgO) per cent. by weight, maximum	10.0
	(iv) Total chlorides (as Cl) (on dry basis) per cent. by weight, maximum	2.5
	(v) Sodium (as NaCI) (on dry basis) per cent. by weight, maximum	1.5
[4. Potassium Chloride (Murriate of Potash (Grnular)]	(i) Moisture per cent. by weight, maximum 0.5
	(ii) Water soluble potash (at K20) per cent. by weight minimum 60.9
	(iii) Sodium (as NaCI) per cent. by weight maximum 3.5
	(iv) Magnesium (as MgCl₂) per cent. by weight maximum 1.0
	(v) Particle size - 90 per cent of the material shall pass through 3.35 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent. shall be blow 1 mm IS sieve.]
1.(d) N.P. Fertilisers : [* * *]
1. Diammonium Phosphate (18 - 46 - 0)	(i) Moisture per cent. by weight maximum	1.0
	(ii) Total nitrogen all in ammoniacal form per cent. by weight, minimum	18.0
	(iii) Total phosphates (as P₂O₅) per cent. by weight, minimum	46.0
	(iv) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum]	41.0
2. Diammonium Phosphate [* * *] (18 - 46 - 0)	(i) Moisture per cent. by weight maximum	1.5
	(i) Moisture per cent. by weight maximum	1.5
	(ii) Total nitrogen per cent. by weight, minimum	18.00
	(iii) Ammoniacal nitrogen form per cent by weight, minimum	15.5
	(iv) Total nitrogen in the form of urea per cent. by weight, maximum	2.5
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent. by weight, minimum
	(vi) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	41.0
	(vii) Particle size - 90 per cent. of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent. shall be below than 1 mm size.
3. Ammonium, phosphate Sulphate (16 - 20 - 0)	(i) Moisture per cent. by weight, maximum	1.0
	(ii) Total ammoniacal nitrogen per cent by weight, minimum	16.0
	(iii) Neutral ammonium citrate soluble Phosphates (as P₂O₅) per cent by weight, minimum	20.0
	(iv) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	19.5
	(v) Particle size - 90 per cent. of the material shall pass through 4 mm IS sieve and shall be retained on 1 mm IS sieve. Not more than 5 per cent. shall be below 1 mm IS sieve.
4. Ammonium phosphate Sulphate (20 - 20 - 0)	(i) Moisture per cent. by weight, maximum	1.0
	(ii) Total nitrogen per cent. by weight, minimum	20.0
	(iii) Ammoniacal nitrogen percent. by weight, minimum	18.0
	(iv) Nitrogen in the form of urea per cent. by weight, maximum	2.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent. by weight, minimum	20.0
	(vi) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	17.0
	(vii) Particle size - 90 percent. of the material shall pass through 4 mm IS sieve and shall be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
5. Ammonium phosphate Sulphate nitrate (20 - 20 - 0)	(i) Moisture per cent. by weight, maximum
	(ii) Total nitrogen per cent. by weight minimum	20.0
	(iii) Ammoniacal nitrogen per cent by weight, maximum	17.0
	(iv) Nitrate nitrogen per cent by weight maximum	3.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent. by weight, minimum	20.0
	(vi) Water soluble phosphates (as P₂O₅) per cent. by weight, minimum	17.0
	(vii) Particle size - 90 per cent. of the material shall be pass through 4 mm IS sieve. Not more than 5 per cent. shall be below 1 mm IS sieve.
6. Ammonium phosphate Sulphate (18 - 9 - 0)	(i) Moisture percent. by weight, maximum	1.0
	(ii) Ammoniacal nitrogen per cent by weight, minimum	18.0
	(iii) Nitrate ammonium citrate soluble phosphates (as P₂O₅) by weight minimum	9.0
	(iv) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	8.5
	(v) Particle size - - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent. shall be below 1 mm IS sieve.
7. Nitrogen Phosphate (20 - 20 - 0)	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Total nitrogen per cent. by weight, minimum	20.00
	(iii) Nitrogen in ammoniacal form per cent by weight, minimum	10.0
	(iv) Nitrogen in nitrate form per cent by weight, maximum	10.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum 1. Substitued. by S.O. 942(E), dated 10th December, 1993.	[12.0]
	(vii) Calcium Nitrate per cent by weight, maximum	1.0
	(viii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
8. Urea Ammonium Phosphate (28 - 28 - 0)	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Total nitrogen per cent by weight, minimum	28.0
	(iii) Ammoniacal nitrogen per cent by weight, minimum	9.0
	(iv) Neutral ammonium citrate soluble phosphate (as P₂O₅) per cent by weight, minimum
	(v) Water soluble phosphates (as P2O5) per cent. by weight, minimum	25.2
	(vi) Particle size - 90 per cent. of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
9. Urea Ammonium Phosphate (24 - 24 - 0)	(i) Moisture per cent by weight, maximum	15
	(ii) Total nitrogen per cent by weight, minimum	24.0
	(iii) Ammoniacal nitrogen per cent, by weight, minimum	7.5
	(iv) Nitrogen in the form of urea per cent by weight, maximum	16.5
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	24.0
	(vi) Water soluble phosphate (as P₂O₅) per cent by weight, minimum	20.4
	(vii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve (Note.—The products contain inert filler material such as sand or dolomite to the extent of 20% by weight, maximum	20.4
10. Urea Ammonium Phosphate (20 - 20 - 0)	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen per cent by weight, minimum	20.0
	(iii) Ammoniacal nitrogen per cent by weight, minimum	6.4
	(iv) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	20.0
	(v) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	17.0
	(vi) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve. (Note.—This product contains filler materials (inert soil) to the extent of 30 per cent by weight).
11. Mono Ammonium Phosphate (11 - 52 - 0)	(i) Moisture per cent by weight, maximum	1.0
	(ii) Total nitrogen all in ammoniacal for per cent weight, minimum	11.0
	(iii) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	52.0
	(iv) Water soluble phosphates (as P205) per cent by weight, minimum Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm sieve. Not more than 5 per cent shall be below 1 mm IS sieve.	44.2
[12. Nitrophosphate (23 - 23 - 0)]	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total Nitrogen per cent by weight, minimum	23.0
	(iii) Nitrogen in ammoniacal form per cent by weight, minimum	11.5
	(iv) Nitrogen in nitrate form per cent by weight, minimum	11.5
	(v) Neutral ammonium citrate soluble phosphate (as P₂O₅) per cent by weight, minimum	23.0
	(vi) Water soluble phosphate (as P₂O₅) per cent by weight, minimum	18.5
	(vii) Calcium nitrate per cent by weight, maximum	1.0
	(viii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve]
[13. Ammonium Nitrate Phosphate]	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen per cent by weight, minimum	23.0
	(iii) Nitrogen in ammoniacal form per cent by weight, minimum	13.0
	(iv) Nitrogen in nitrate form per cent by weight, maximum	10.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	23.0
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	20.5
	(vii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
[15. Ammonium Phosphates (14 - 28 - 0)]	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen per cent by weight, minimum	14.0
	(iii) Urea nitrogen per cent by weight, maximum	6.0
	(iv) Ammoniacal nitrogen per cent by weight, minimum	8.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	28.0
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	23.0
	(vii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5% shall be below 1 mm IS sieve.
1. (e) N.P.K. Fertiliser :
1. Nitrophosphate with Potash (15 - 15 - 15)	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen, minimum	15.0
	(iii) Ammoniacal nitrogen, per cent by weight, minimum	7.5
	(iv) Nitrate nitrogen, per cent by weight, maximum	7.5
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	4.0
	(vii) Water soluble Potash (as K₂O) per cent by weight, minimum	15.0
	(viii) Particle size - 90 per cent of the material shall pass through 4 mm IS sieve and be retained on 1 mm IS sieve.
	(ix) Calcium nitrate, per cent by weight, maximum
[2. N.P.K. (10 - 26 - 26)]	(i) Moisture per cent by weight, maximum	1.0
	(ii) Total nitrogen per cent by weight, minimum	10.0
	(iii) Ammoniacal nitrogen per cent by weight, minimum	7.0
	(iv) Nitrogen in the form of the urea per cent. by weight, maximum	3.0
	(v) Neutral ammonium citrate soluble phosphate (as P₂O₅) per cent by weight, minimum	26.0
	(vi) Water soluble potash (as K₂O) per cent by weight, minimum	26.0
	(vii) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	22.1
	(viii) Particle size—Particle size of the material will by such that 90 per cent of the material will be between 1 mm and 4 mm IS sieve and not more than 5 per cent will be below 1 mm size.
3. N.P.K. (12 - 36 - 16)	(i) Moisture per cent weight, maximum	1.0
	(ii) Total nitrogen per cent by weight, minimum	12.0
	(iii) Ammoniacal nitrogen per cent by weight, minimum	9.0
	(iv) Nitrogen in the form of urea per cent by weight, maximum	3.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	32.0
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	27.2
	(vii) Water soluble potash (as K₂O) per cent by weight, minimum	16.0
	(viii) Particle size—Particle size of the material will be such that 90 per cent of the material will be between 1 mm and 4 IS sieve and not more than 5 per cent will be below 1 mm size.]
4. N.P.K. (22 - 22 - 1)	(i) Moisture percent by weight, maximum	1.5
	(ii) Total nitrogen, per cent by weight, minimum	22.0
	(iii) Ammoniacal nitrogen, per cent by weight, minimum	7.0
	(iv) Urea nitrogen, per cent by weight maximum	15.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	22.0
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	18.7
	(vii) Water soluble potash (as K₂O) per cent by weight, minimum	11.0
	(viii) Particle size - 90 per cent of the material shall be pass through 4 mm IS sieve and be retained through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
5. N.P.K. (14 - 35 - 14)	(i) Moisture per cent by weight, maximum	1.0
	(ii) Nitrogen in ammoniacal form per cent. by weight. minimum '	[14.0]
	(iii) [* * *]
	(iv) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	35.0
	(v) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	29.0
	(vi) Water soluble potash (as K₂O) per cent by weight, minimum	14.0
	(vii) Particle size - 90 per cent of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
6. N.P.K.(17 - 17 - 17)	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen, per cent by weight, minimum	17.0
	(iii) Ammoniacal nitrogen, per cent by weight, minimum	5.0
	(iv) Urea nitrogen, per cent by weight, maximum	12.0
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	17.0
	(vi) Water soluble phosphates (as P₂O₅) per cent by weight, minimum	14.5
	(vii) Water soluble potash as (as K₂O) per cent by weight, minimum	17.0
	(viii) Particle size - 90 per cent of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
7. N.P.K.(14 - 28 - 14)	(i) Moisture per cent by weight, maximum	1.5
	(ii) Total nitrogen, per cent, by weight, minimum	14.0
	(iii) Ammoniacal nitrogen, per cent. by weight minimum	8.0
	(iv) Use nitrogen. per cent by weight maximum	6.0
	(v) Neutral ammonium citrate soluble phosphat (as P₂O₅) per cent by weight, minimum	28.0
	(vi) Water soluble phosphate (as P₂O₅) per cent. by weight, minimum	23.8
	(vii) Water soluble potash (as K₂O) per cent. by weight, minimum	14.0
	(viii) Particle size - 90 per cent. of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
8. N.P.K.(19 - 19 - 19)	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Total nitrogen, per cent. by weight, minimum	19.0
	(iii) Ammoniacal nitrogen, per cent by weight, minimum	5.6
	(iv) Urea nitrogen, per cent by weight, maximum	13.4
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	19.0
	(vi) Watersoluble phosphate (as P₂O₅) per cent by weight, minimum	16.2
	(vii) Water soluble potash (as K2O) per cent. by weight, minimum	19.0
	(viii) Particle size - 90 per cent. of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5 per cent shall be below 1 mm IS sieve.
[9. N.P.K. (17 - 17 - 17)]	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Total nitrogen, per cent. by weight, minimum	17.0
	(iii) Ammoniacal nitrogen per cent by weight, minimum	8.5
	(iv) Nitrate nitrogen, per cent by weight, maximum	8.5
	(v) Neutral ammonium citrate soluble phosphate (as P₂O₅) per cent by weight, minimum	17.0
	(vi) Water soluble phosphate (as P₂O₅) per cent. by weight, minimum	13.6
	(vii) Water soluble potash (as K₂O) per cent by weight, minimum	17.0
	(viii) Particle size - 80 per cent. of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 20 per cent shall be below 1 mm IS sieve.]
[10. N.P.K. (20 - 10 - 10)]	(i) Moisture per cent. by weight, maximum	1.5
	(ii) Total nitrogen, per cent. by weight, maximum	20.0
	(iii) Urea nitrogen per cent by weight, maximum	17.1
	(iv) Ammoniacal nitrogen, per cent by weight, minimum	2.9
	(v) Neutral ammonium citrate soluble phosphates (as P₂O₅) per cent by weight, minimum	10.0
	(vi) Water soluble phosphate (as P₂O₅) per cent by weight, minimum	8.5
	(vii) Water soluble potash (as K₂O) per cent. by weight minimum	10.0
	(viii) Particle size - 90% of the material shall be pass through 4 mm IS sieve and be retained on 1 mm IS sieve. Not more than 5% shall be below 1 mm IS sieve] ;
1. (f) Micronutrients
[1. Zinc Sulphate heptahydrate (ZnSO₄7H₂O)]	(i) Free flowing crystalline form
	(ii) Matter insoluble in water, per cent. by weight, maximum	1.0
	(iii) Zinc (as Zn), per cent. by weight, minimum	21.0
	(iv) Lead (as (Pb), per cent by weight, maximum	0.003
	(v) Copper (as Cu), per cent by weight, maximum	0.1
	(vi) Magnesium (as Mg), per cent. by weight, maximum	0.5
	(vii) pH	Not less than 4.0
2. Magnanese Sulphate	(i) Free flowing crystalline form
	(ii) Matter insoluble in water, per cent. by weight, maximum	1.2
	(iii) Manganese (as Zn), content per cent. by weight, minimum	30.5
	(iv) Lead (as Pb), per cent by weight, maximum	0.003
	(v) Copper (as Cu), per cent by weight, maximum	0.1
	(vi) Magnesium (as Mg), per cent. by weight, maximum	2.0
	(vii) pH	3.75 +0.25
[3. Borax (Sodium Tetraborate) (ii) Na₂ B₄O₇10H₂O)_ for soil application.]	(i) Content of Boron (as B) per cent. by weight, minimum	10.5
	Matter insoluble in water per cent. by weight, maximum	1.0
	(iii) pH	9.0 - 9.5
	(iv)Lead (as Pb) per cent. by weight, maximum	0.003
4. Solubor (Na₂B₄O₇5H₂ + Na₂ B₁₀ O₁₆ 10H₂ O)for foliar Sulphate	(i) Content of Boron (as B) per cent. by weight, minimum	19.0
	(ii) Matter insoluble in water per cent. by weight, maximum	1.0
	(iii) Lead (as Pb) per cent. by weight, maximum	0.003
5. Copper Sulphate (CuSO₂5H₂O)	(i)Copper (as Cu) per cent. by weight,minimum	24.0
	(ii) Matter insoluble in water per cent. by weight, maximum	1.0
	(iii) Soluble Iron & aluminium compounds (expressed as Fe), per cent. by weight, maximum	0.5
	(iv) Lead (as Pb) per cent. by weight, maximum	0.003
	(v) pH not less than	3.0
6. Ferrous Sulphate (FeSO₄7H₂O)	(i) Ferrous iron (as Fe), per cent. by weight, minimum	19.0
	(ii) Free Acid (as H₂SO₄) per cent. by weight maximum	1.0
	(iii) Ferric Iron (as Fe), per cent by weight maximum	0.5
	(iv) Matter insoluble in water, per cent. by weight, maximum	1.0
	(v) pH not less than	3.5
	(vi) Lead (as Pb) per cent. by weight, maximum
7. Ammonium Molybdate (HN₄)₆ Mo₇O₄4H₂O)	(i) Molybde num (as Mo) per cent. by weight, minimum	52.0
	(ii) Matter insoluble in water, per cent. by weight, maximum	1.0
	(iii) Lead (as Pb), per cent. by weight, maximum	0.003
8. Chelated Zinc as Zn - EDTA	(i) Appearance— Free flowing crystalline/powder
	(ii) Zinc content (Expressed as Zn), per cent. by weight, minimum in the form of Zn - EDTA	12.0
	(iii) Lead (as Pb), per cent. by weight maximum	0.003
	(iv) pH	6.0 - 6.5
9. Chetaled Iron as Fe - EDTA	(i) Appearance—Free flowing crystalline/ powder
	(ii) Iron content (expressed as Fe), per cent. by weight minimum in the form of Fe - EDTA	12.0
	(iii) Lead (as Pb) per cent by weight, maximum	0.003
	(iv) pH	5.5 - 6.5
[10. Zinc Sulphate Monohydrate (ZnSO₄ H₂O)]	[(i) Free flowing powder form]
	(ii) Matter insoluble in water, per cent. by weight, maximum	1.0
	(iii) Zinc (as Zn), per cent. by weight minimum	33.0
	(iv) Lead (as Pb), per cent. by weight maximum	0.003
	(v) Copper (as Cu), per cent, by weight maximum	0.1]
	(vi) Magnesium (as Mg), per cent. by weight maximum	0.5
	(vii) Iron (as Fe), per cent. by weight, maximum	0.5
	(viii) pH not less than	4.0]
[1] Magnesium Sulphate (i)	Free Flowing—crystalline form
	(ii) Matter insoluble in water, per cent. by weight, maximum	1.0
	(iii) Magnesium (as Mg), per cent. by weight, minimum	9.6
	(iv) Lead (as Pb), per cent. by weight, maximum
	(v) pH (5% solution)	5.0.8.0]
[1.(g) Fortified Fertilisers.
1. Boronated Single Super Phosphate (16% P₂O₅)	(i) Moisture per cent. by weight, maximum	12.0
1. Boronated Single Super Phosphate (16% P₂O₅)	(ii) Free phosphate acid (as P₂O₅) per cent. by weight, maximum	4.0
Powdered		4.0
	(iii) Water soluble phosphate (as P,05) per cent. by weight, minimum	16.0
	(iv) Boron (as B) per cent. by weight, maximum	0.18
2. Zincated Urea	(i) Moisture per cent. by weight, maximum	1.0
	(ii) Total nitrogen per cent. by weight (on dry basis), minimum	43.0
	(iii) Zinc (as Zn) per cent. by weight, minimum	2.0
	(iv) Biuret per cent. by weight, maximum	1.5
	(v) Particle size - 90 per cent of the material shall pass through 2.8 mm IS sieve and not less than 80 per cent. by weight shall be retained on 1 mm IS sieve.]
[18. Concentrated Liquid Calcium 11%	(i) Calcium ( as Ca) per cent by weight minimum (ii) pH (iii) Specific gravity (kg/l)	11.0 9.5 +/- 1.00 1.3-1.4.]
[(h) NK Fertilisers]
1. Potassium Nitrate (13 - 0 - 45)	(i) Free flowing crystalline form
	(ii) Moisture per cent by weight, maximum	0.5
	(iii) Total nitrogen (all in Nitrate form), per cent by weight, minimum	13.0
	(iv) Water soluble Potash (as K₂o) per cent by weight, minimum	45.0
	(v) Sodium (as Na) (or dry basis) per cent by weight, maximum	1.0
	(vi) Total chloride (as Cl) (on dry basis) per cent by weight, maximum	1.5
	(vii) Matter soluble in water per cent by weight, maximum	0.05
2. Potassium Nitrate	(i) Free flowing crystalline form
	(ii) Moisture per cent by weight, maximum	0.5
	(iii) Total Nitrogen (all in Nitrate form), per cent by weight, minimum	13.0
	(iv) Water soluble Potash (as k2O) per cent by weight, minimum	45.0
	(v) Sodium (as Na) (on dry basis) per cent by weight, maximum	1.0
	(vi)Total Chloride (as Cl) (on dry basis) per cent by weight, maximum	1.5
	(vii)Matter insoluble in water per cent by weight, maximum	0.05
	(viii)Particle size—Not less than 95% material shall pass through 2.5 mm IS sieve and shall be retained on 1 mm IS sieve.
1. (i) P.K. Fertilisers
1. Mono - potassium Phosphate (0 - 52 - 34)	(i) Moisture per cent by weight, maximum	0.5
	(ii) Water soluble Phosphate (as P₂O₅) per cent by weight, minimum	52.0
	(iii)Water soluble Potash (as K₂O) per cent by weight, minimum	34.0
	(iv) Sodium (as NaCI) per cent by weight (on dry basis,) maximum	0.025
	(v) Particle size - 90% of the material shall pass through 0.5 mm IS sieve and he retained on 0.05 mm IS sieve."
[1(j) Potassium, Magnesium Fertilisers] :
Potassium Magnesium Sulphate	(i) Moisture, per cent by weight, maximum	0.5%
	(ii) Potash content (as K₂O) per cent by weight, minimum	22.0%
	(iii) Magnesium as MgO, per cent by weight, minimum	18.0%
	(iv) Total Chlorides (as Cl), per cent by weight (on dry basis), maximum	2.5%
	(v) Sodium (as NaCI), per cent by weight (on dry basis) maximum	2.0%
	(vi) Particle size : 90 per cent of the material shall pass through 3.35 mm IS sieve and shall be retained on 1 mm IS sieve.
1. (k) Nitrogen, Calcium Fertilizers
Calcium Nitrate	(i) Total Nitrogen per cent by weight, minimum	15.5%
	(ii) Ammoniacal Nitrogen per cent by weight, maximum	1.1%
	(iii) Nitrate Nitrogen as N per cent by weight, minimum	14.4%
	(iv) Water soluble Calcium as per cent by weight, minimum	18.8%
	(v) Water insolubles per cent by weight, maximum	1.5%
	(vi) Particle size : 90% of the material shall pass through 4.00 mm sieve and be retained on 1 mm IS sieve.
	Not more than 5% shall pass through 1 mm IS sieve.]

Part B

Tolerance limit in plant nutrient for various fertilizers

1. For fertilisers with definite compound like ammonium sulphate, urea, ammonium chloride, muriate of potash, sulphate of potash, super phosphate, decalcium phosphate, which contain more than 20 per cent plant nutrients	0.2
For those which contain less than 20 per cent plant nutrients	0.1
2. For calcium ammonium nitrate	0.3
3. [0.5 units each for N and P contents]		for individual nutrients subject to the maximum of 2.5 per cent. for all combined nutrients.
[4.] For nitrophosphates, ammonium sulphate, nitrates, urea, ammonium phosphate, ammonium phosphate sulphate, bonemeal, granulated mixture, compound/complex fertilisers [Physical mixtures of fertilisers (N.P.K. mixtures.] [mixtures of N.P.K. with micronutrients].		Tolerance varies with nutrient level in fertiliser, subject to the maximum of 2 per cent. for all combined nutrients :—
		Nutrient level (%)	Tolerance level (unit)
		5 or less	0.5
		16 to 20	0.6
		21 or more	1.7.
[5. For borax, chelated Zinc - EDTA and chelated iron - EDTA]	0.1
6. For solubor Copper Sulphate,Zinc Sulphate, Manganese Sulphate, and Ferrous sulphate.	0.2
7. For ammonium molybdate	0.5
[8. For Magnesium Sulphate]	0.1
[9. For mixture of micronutrien fertilisers]
		Tolerance varies with combined nutrient level in fertiliser.

		Nutrient level (%)	Tolerance level (unit)
		10 or less	0.1
		11 to 20	0.2
		21 or more	0.5]

Schedule II

[See Clause 28(1)(b) and 29]

Part A

Procedure for Drawal of Samples of Fertilizers

1. General requirements of sampling - In drawing samples, the following measures and precautions should be observed :

(a) samples shall not be taken at a place exposed to rain/sun;

(b) the sampling instruments shall be clean and dry when used ;

(c) the material being sampled, the sampling instrument and the bags of samples should be free from any adventitious contamination ;

(d) to draw a representative sample, the contents of each bag selected for sampling should be mixed as thoroughly as possible by suitable means ;

(e) the sample should be kept in suitable, clean dry and air tight glass or screwed hard polythene bottle of about 400 gms. capacity or in a thick gauged polythene bag. This should be put in a cloth bag which may be sealed with the inspector's seal after putting inside the detailed description as specified in Form 'J'. Identifiable details may also be put on the cloth bag like sample No/code No. or any other details which enables its identification ;

(f) each sample bag should be sealed air tight after filling and marked with details of sample type and brand of fertiliser, name of dealer/manufacturer and the name of Inspector who has collected sample.

2. Sampling from bagged material : -

(i) Scale of sampling. - (a) Lot (for manufacturers). - All bags, in single consignment of the material of the sample grade and type drawn from a single batch of the manufacture shall constitute a lot. If a consignment is declared to consist of different batches of manufacture, all the bags of each batch shall constitute a separate lot.

In the case of a consignment drawn from a continuous process, 2000 bags (or 100 tonnes) of the material shall constitute a lot.

(b) Lot (for dealers). - The lot is an identifiable quantity of same grade and type of fertiliser stored at an identifiable place subject to a maximum limit of 100 tonnes.

The lot shall be identified by the inspector based on visible appearance of bags, their packing and storage conditions. The stock of less than 100 tonnes with a dealer may also constitute one or more lots, if the material (fertilizer) of different sources and brand is available in such quantities.

(c) (i) Selection of bags for sampling. - The number of bags to be chosen from a lot shall depend upon the size of the lot as given in the table below :

Lot Size (no. of bags)

(N)

No. of bags to be selected for sampling

(N)

Less than 10

10 - 100

100 - 200

200 - 400

400 - 600

600 - 800

800 - 1000

1000 - 1300

1300 - 1600

1600 - 2000

All the bags of a lot should be arranged in a systematic manner. Start counting from any bag randomly, go on counting as 1,2,3 up to r and so on, r being equal to the integral of N/n. Thus, every rth bag counted shall be withdrawn and all bags shall constitute the sample bags from where the sample is to be drawn for preparing a composite sample.

(ii) Sampling from big godown/high stackings. - If the procedure given in Para 2(i)(c) is not possible to be adopted, the sample should be drawn from the randomly selected fertiliser bags from different layers, from top and from all open sides in a zig zag fashion.

(iii) Sampling from small godowns. - All the fertilizer bags of the same grade and type of each manufacturer though received on different dates shall be segregated and properly stacked. All bags of grade and type of fertiliser manufactured by particular manufacturing unit may be considered as one lot based on their physical conditions and the sample shall be drawn as per procedure laid down in Paras 2(i) (c) and 4.

(iv) Sampling from damaged stock. - (a) In case of torn or lumpy bags, damaged fertiliser bags or sweepings, the stock should be arranged according to identifiable lots. From each lot the number of bags shall be selected as per procedure (2) (i) (c) if the bags allow the use of Sampling probe conveniently, the samples should be drawn sampling probe.

(b) In case it is not possible to use the sampling probe, the bags may be opened and fertiliser material mixed together uniformally by hammering the big lumps or putting pressure if required, and then samples drawn by using suitable sample device.

3. Sampling probe -

(i) An appropriate sampling instrument to be used by the Inspectors for collection of a representative sample is called Sampling probe. The probe may comprise of a slotted single tube with solid conetip made of stainless steel or brass. The length of the probe may be approximately 60 to 65 cms. and the diameter of the tube may be approximately 1.5 cms. and the slot width may be 1.2 to 1.3 cms. The probe may be used if the physical condition of the fertilisers and the packing material permits its use.

(ii) In case of High Density Polyethylene Packings and also when the fertiliser material is not in free flowing condition, the use of sampling probe may not be possible, in such a case, selected bags for drawing samples may be opened and the fertilizers may be taken out of the bags and spread on a clean surface and samples drawn with the help of a suitable sampling device which may be made of stainless steel or brass cup.

4. Drawal of samples from bags -

(i) Drawal of sample and preparation of composites. Draw, with an appropriate sampling instrument, (sampling probe) small portions of the material from the selected bags as per procedure in Paras 2(i)(b), 2(11), 2(iii) and 2(iv) (a). The Sampling probe shall be inserted in the bag from one corner to another diagonally and when filled with fertiliser, the probe is withdrawn and fertiliser is emptied in a container/or on polythene sheet/or on a clean hard surface and made into one composite sample.

(ii) If the bags do not permit the use of sampling probe empty the contents of the bags on a level, clean and hard surface and drawn a composite sample by the process of quartering as described under Para 3(11) or 5.

[(iii) In case of chelated micronutrients and mixtures of micronutrients, the three identical containers of the batch, grade, type and manufacturer, shall be selected which shall constitute the composite samples, provided it is not possible to draw a composite sample of the size given under Paragraph 4 - A (iii)];

[4A. Weight of one sample. - One sample of fertiliser shall have the approximate weight, as specified below : -

(i) For straight micronutrient fertiliser	100 gms
(ii) For chelated micronutrient, fertiliser and mixture of or the maximum packing size of similar quantity.	50 gms
(iii) For other fertilisers and mixtures of fertilisers	400 gms]

5. Preparation of composite sample. - If the composite sample collected from the different selected bags is larger than [required weight] its size shall be reduced by method of quartering as detailed below : -

Spread the composite sample on a level, clean, hard surface, flatten it out and divide it into four equal parts. Remove any diagonally opposite parts. Mix the two remaining parts together to form a cone, flatten out the cone and repeat the operation of quartering till a composite sample of [required weight] is obtained.

6. Preparation of test sample and reference sample :

(i) The composite sample obtained above shall be spread out on a clean, hard surface and divided into three approximately equal portions [each of the weight as specified in paragraph 8]. Each of these samples shall constitute the test sample.

(ii) Each test sample shall be immediately transferred to a suitable container as defined under Para 1(e). The slip with detailed description may be put inside the sample bag. Each bag shall also be properly labelled as mentioned in Para 1(f).

(iii) Each test sample container shall then be sealed with the seals of the inspector. If possible, seal of the manufacturer/dealer or purchaser as the case may also be affixed.

[****]

7. Sampling from the bulk fertilisers in ships, bulk carriers and bulk containers :

(i) Sampling equipments : - (a) Sampling cup - The sampling cup can be fabricated from non - corrosive metal. Inside dimensions of cup mouth may be 3/4" x 10" as per the diagram of the sampling cup given in Figure - A below : -

Fig. A - Sampling Cup

(b) Sampling probe for bulk fertilisers. - The sampling probe should be made of non - corrodable material such as stainless steel or brass. It may be a slotted double tube with solid cone tip having a length of about 41/2" - 5" and diameter of about 11 1/4" - 5" to 11 1/2".

(c) Scope. - Samples from the hatch can also be collected by a suitable scoop made of stainless steel or brass.

(ii) Sampling Procedure: - (a) Drawal of samples from the ship of bulk fertilisers -

A. Sampling from conveyor belt. - When the material is unloaded from the ships and transported to temporary/storage godowns through conveyor belts. the sample can be drawn by passing the sampling cup through the entire stream of material as the material drops from a transfer belt or spout. The long slot in the top of sampling cup should be prependicular to the falling stream. Pass the cup through the complete stream at an uniform speed, so that the cup will collect approximately equal amounts in each pass but will never over.

A minimum of 10 equally timed and speed stream must be taken during the transfer operation. However, the stream samples are not applicable unless uniform continuous flow of fertiliser is maintained for more than 3 minutes while lot is being sampled.

B. Sampling from hatch. - In case of bulk fertilisers, from each hatch, at least 5 - 6 composite samples should be drawn from different depths and different points during unloading operation. To make one composite sample, minimum 5 samples should be drawn from different points at a specific depth. All these samples should be mixed to make one composite sample. The depths may be 0 - 5 m., 10 - 15 m. and 20 - 25 m. depending upon the depth of hatch. The samples at a particular depth should be drawn after removal of material up to the required depth. The samples may be drawn with the help of sampling probe or scoop.

(b) Drawal of samples from bulk carriers - trucks etc. - The Samples can be drawn as per vertical probing procedure of Association of official Analytical Chemists. The sampling probe should be about 41/2" to 5" length. Draw 10 vertical cuts from the following locations relative of the entire top of the conveyance. The 10 vertical cores are combined into a composite samples.

1		4		3
3	1		2		5
10		6		9

(b) Drawal of samples from bulk material in storage -

A. The bulk storage Piles (level or falt) up to 100 tonnes could be sampled as per figure B. Take 10 cores to the maximum. possible depth of the probe from the position indicated in the figure B and all cores are composited.

B. A one sided or sloped pile may be sampled at the points illustrated in Fig. C Withdraw one vertical core of material from locations 1 and 6 and two cores at locations 2,3,4 and 5. Composite all the probe samples and prepare the composite samples for analysis as per procedure laid down in Paras 1, 5 & 6.

Fig. B - Sample points for Coned or Ridged pile.

Fig.C - Sample points for 1/2 on one side Pile

8. Method of sampling of anhydrous ammonia : -

(i) Scoop. - This method is for use in obtaining samples of anhydrous ammonia. The method is based on the assumption that the material to be sampled is as claimed and contains only a small amount of impurity primarily water. It is recommended that duplicate samples be taken from each tank or vessel sampled.

(ii) Apparatus - (a) Tubes for sampling tubes, heat resistant glass, conical centrifuge type 200 ml with lower 100 ml. graduated from 0.2 ml in 0.05 ml division, 2 - 4 ml. in 0.1 division 4 - 10 ml. in 0.5 ml. divisions and 10 - 100 ml. 1.0 nil divisions. Tube type shown in figure E.

(b) Stoppers for tubes, rubber, with bent tube vent as in figure E.

(d) Sampling line and connection assembly constructed as in figure D, with flexible steel sampling nose 48" long, 1/4" NPT coupling at each end and 1/8" insulated steel delivery tip at one end.

(e) Sample tube adapter, constructed from rubber stopper and 6 mm. OD, glass or steel tubing as in figure E.

(f) Protective equipment : Rubber or other non - prorous gloves, offering complete protection to the hands and lower arms, full coverage goggles or approved gas mask.

Sampling Line & Connection

Fig. E

(iii) Reagents. - Charcoal, regent, 14 - 20 mesh.

Note. - If the sample is expected to contain excessive amount of water (one per cent or more), one piece of the charcoal may be added to each before introduction of the sample.

(iv) Procedure. - (a) Place two dry, clean sampling tubes in the sample carrier.

(b) Connect the sampling line connection assembly to the unloading valve of the tank, vessel or line to be sampled.

(c) Open the valves slowly and purge the sampling line connection assembly thoroughly by venting 3 to 4 litres of ammonia. Close the samples line globe valve.

(d) Remove the vented stoppers from the tubes and insert the adapter end of the sampling line connection assembly.

(e) Open the sample line valve and slowly fill the sampling tube to the 100 ml. mark, close the sample line valve.

(f) Remove the sampling line adapter and insert the vented stopper in the sampling tube.

(g) Repeat steps, d and e and f and fill the second tube.

(h) Close tank discharge valve and remove the sampling line connection assembly.

(i) Either know or note the vessel, container, or line pressure of material sampled.

(j) Tag the collected samples for identification and submit to the laboratory for processing by following methods in this section.

(v) Precaution. - (a) Liquid anhydrous ammonia causes severe burns on contact. It evaporates readily releasing the gas which may cause carrying degrees of irritation of the skin and mucus membrances and may injure severly the respiratory mucosa with possible fatal outcome.

(b) Avoid contacting liquid ammonia. In case of contact, immediately flush the affected parts with plenty of water for at least 15 minutes. Get medical attention at once in case of burns, especially to the eyes, nose and throat, or if the victim is unconscious.

(c) Ammonia gas in concentrations of 6,000 to 10,000 ppm. (by volume) is lethal within a few minutes. Irritation of the eyes, respiratory tract and throat results from concentrations as low as 500 to 1,000 ppm., a concentration of 2,000 ppm produces conclusive coughing and may be fatal after a short exposure, i.e., less than half an hour. The maximum concentration tolerated by the skin for more than few seconds is 2 per cent (i.e. when suitable respiratory protection is worn). The maximum allowable concentration for 8 hour working exposure is 50 ppm. This is the least detectable order.

(d) Obtain medical attention if exposure to the gas produce distress of any type.

(e) Rubber or other non - porous gloves, offering complete protection to the hands and lower arms must be worn when sampling anhydrous ammonia. Full coverage goggles must also be worn to protect the eyes unless an approved gas mask is used. The gas mask need only to be used if sampling cannot be done without possible inhalation of the vapours.

Part B

Methods of Analysis of Fertilizers

1. Preparation of sample for analysis in the laboratory :

(i) Procedure. - (a) Reduce gross sample to quantity sufficient for analysis or grand not 250 gms. of reduced sample without previous sieving.

(b) For fertiliser materials and moist fertiliser mixtures, that form a paste on putting pressure, grind in porcelain pestle and mortar to pass sieve with 1 mm. circular opening or No. 20 standard sieve.

(d) Grind as rapidly as possible to avoid loss or gain of moisture during operation.

(e) Mix thoroughly and store in tightly stoppered bottles.

2. Determination of moisture : -

(Not applicable to samples that yield volatile substances other than water at drying temperature).

(i) Procedure. - (a) Weight to the nearest mg. about 2 gms. of the prepared sample in a weighted, clean, dry squat form weighing bottle.

(b) Heat in an oven for about 5 hours at 99 - 101°C to constant weight. Cool in a desicator and weight.

(d) Report percentage loss in weight as moisture at temperature used.

Calculations

Free moisture percent by weight =

100 (B – C)
___________
B - A

A = Weight of the bottle.

B = Weight of the bottle plus material before drying.

C = Weight of the bottle plus material after drying.

(Reference "Methods of Analysis", AOAC, 1965)

(ii) Moisture in ammonium Chloride. - (a) Weight accurately about 5 gms. of prepared sample in a weighed shallow porcelain dish and dry for 24 hours in a vacuum desicator over sulphuric acid and re - weight.

(b) Preserve the dried material for subsequent tests.

Calculations

Moisture per cent by weight = 100 x W1/W3

W= Loss in weight in gm. in drying, and

W3 = Weight in gm. of the papered sample taken for test.

(iii) Samples. - Like urea, diammonium phosphate and ammonium nitrate which yield volatile substances other than water at drying temperature, the Karl Fishcher method given below is used for the determination of moisture.

(iv) Reagents. - (a) Iodine solution. - Added 125 gm. of iodine to a mixture of 650 ml. of methanol and 200 ml. of pyridine contained in flask, and immediately close the flask tightyly.

(b) Sulphur dioxide solution. - Pass dry sulphur dioxide into 100 ml. of pyridine contained in a 250 ml. graduated cylinder and cooled in an ice bath, until the volume reaches 200 ml.

(c) Fishcher reagent. - Slowly add iodine solution to cooled sulphur dioxide solution, stopper immediately and shake well until the iodine is dissolved. Transfer the solution to an automatic pipette, protected from absorption of moisture by a drying agent and allow to stand for 24 hours before standardising. The reagent deteriorates continuously and it should be standardized within one hour before use.

(d) Standard water solutions. - Measure exactly 2 ml. of water into a thoroughly dry 1 litre volumetric flask, dilute to volume with methanol. Retain sufficient quantity of the same methanol for the blank determination. Keep the solution in tightly closed containers.

(v) Procedure. - (a) Determination of end point in karl fischer titration. - In many cases the end point can be detected by the change of colour from a light brownish yellow to amber. But when the end point is not clearly defined, the electrometric method for determining the end point should be adopted. Adjust the potentiometer so that when a small excess (0.02 ml) of the reagent is present, a current of 50 to 150 microampers is recorded : The solution should be continuously and vigorously stirred. At the beginning of the titration, a current of only a few microampers will flow. After each addition of reagent, the pointer of the micro¬meter deflected but rapidly returns to the original position. At the end point a deflection is obtained which endures for a longer period.

(b) Standardization of the fischer reagent. - Pipette exactly 10 ml. of methanol into a dry titration flask and titrate with the Fischer reagent to the end point (V₁). Pipette exactly 10 ml. of the standard water solution into the flask and titrate to the end point (V₂).

(c) Titration of the material. - Transfer 25 ml. of methanol to the titration flask and titrate to the end point (V₃) with the Fischer reagent. Do not record the volume consumed. Quickly transfer to the titrated liquid an accurately weighed quantity of the material containing 10 to 50 mg. of water, stir vigorously and titrate to the end point.

Calculation

Moisture per cent by weight =

0.1 x W(V₃ - 2.5V₁)
_____________________
(V₂ - V₁) A

where,

W = Weight in mg. of water contained in 10 ml. of standard water solution

V₃ = Total volume in ml. of the reagent used in titration in (c).

V₁ = Volume in ml. of reagent used in titration of methanol in (b), and

V₂ = Total volume in ml. of the reagent used in titration in (b).

A = Weight in gm. of the material taken for the test in (c).

(Reference IS. Specification for urea, technical and pure. IS. 1781 - 1961)

3. Determination of nitrogen :

Methods of determination of total nitrogen, ammoniacal nitrogen, nitrate nitrogen and urea nitrogen have been described separately in this section. These methods can be adopted both for straight and mixed fertilisers. Scope of each method with various combinations have also been described with each method.

The relevant method of analysis which have been described are as given below :

(a) Total Nitrogen - for nitrate free samples.

(b) Total Nitrogen - for nitrate containing samples.

(d) Determination of ammoniacal nitrogen.

(e) Determination of ammoniacal and nitrate nitrogen.

(f) Determination of nitrate nitrogen.

(g) Determination of water insoluble nitrogen.

(h) Determination of urea nitrogen.

(i) Detection of nitrate. - For adopting a specific method as described above, it is necessary to detect the presence of nitrates in the sample before a particular method is adopted. The procedure for detection of nitrates is given below :

(a) Mix. 5 gm. sample with 25 ml. hot water and filter.

(b) To one volume of this solution add 2 volumes of sulphuric acid, free from HNO₃ and oxides of N, and let it cool.

(d) If the nitrates are present junction shows at first purple, afterward brown, or if only minute quantity is present. reddish colour.

(e) To another portion of solution add 1 ml. 1 per cent NaNO₃ and test as before to determine whether enough H₂SO₄ was added in the first test.

(Reference "Methods of Analysis" AOAC, 1965)

(ii) Reagents for determination of total nitrogen

(a) Sulphuric acid - 93 - 98 per cent H₂SO₄, N free.

(b) Copper sulphate - CuSO₄5H₂O reagent grade, N free.

(d) Salicylic acid - reagent grade, N free.

(e) Sulphide or thiousulphate solution - Dissolve 40 gm. commercial K₂S in 1 litre distilled water.

(Solution - of 40 gm. Na₂S or 80 gm. Na₂S or 80 gm. Na₂S₂O₃.5H₂O in 1 litre may be used.)

(f) Sodium hydroxide - Pellets or solution, nitrate free. For solution dissolve approximately 450 gms. solid NaOH in distilled water and dilute to 1 litre (Sp. gr. of solution should be 136 or higher).

(g) Zinc granule - reagent grade.

(h) Zinc dust - Impalpable powder.

(i) Methyl red indicator - Dissolve 1 gm. methyl red in 200 ml. alcohol.

(j) Hydrochloric or sulphuric acid standard solution. - 0.5 N or 0.1N when amount of N is small.

(k) Sodium hydroxide standard solution - 0.1N (or other specified concentration)

(1) Standardize each standard solution with primary standard and check one against another.

(2) Test reagents before using, by blank determination with 2 gms. sugar which ensures partial reduction of any nitrates present.

Caution : Use freshly opened sulphuric acid or add dry P₂O₅ to avoid hydrolysis of nitrites and cyanates. Ratio of salt to acid (wt : Vol) should be approximately 1 : 1 : at end of digestion for proper temperature control. Digestion may be incomplete at lower ratio, N may be lost at higher ratio.

(iii) Apparatus :

(a) For digestion - Use Kjeldahl flask of hard moderately thick, well annealed glass with total capacity approximately 500 - 800 ml. Conduct digestion over heating device, adjusted to bring 250 ml. water at 25°C to cooling boil in approximately for 5 minutes. Add 3 - 4 rolling chips to prevent superheating.

(b) For distillation - Use Kjeldahl or other suitable flask of 500 - 800 ml. capacity filled with rubber stopper through which passes lower end of efficient scrubber bulb or trap to prevent mechanical carryover of NaOH during distillation. Connect upper end of bulb tube to condenser tube by rubber tubing. Trap outlet of condenser in such a way as to ensure complete absorption of ammonia distilled over into acid in receiver.

(iv) Total nitrogen (in nitrate free samples) :

(a) Procedure -

(1) Place weighed sample (0.7 - 2.2 gm.) in digestion flask.

(2) Add 0.7 gm. copper sulphate, 15 gm. powdered K₂SO₄ or anhydrous Na₂SO₄ and 25 ml. H₂SO₄.

(3) If sample more than 2.2 gm. is used, increase sulphuric acid by 10 ml. for each gm. sample.

(4) Place flask in inclined position and heat gently until frothing ceases (if necessary add small amount of paraffin to reduce frothing).

(5) Boil briskly until solution clears and then for at least 30 minutes longer (2 hours for sample containing organic material)

(6) Cool add approximately 200 ml. distilled water, cool below 25°C.

(7) Add to the flask a layer of sodium hydroxide (25 gm. solid reagent or enough solution to make contents strongly alkaline) without agitation.

(8) Immediately connect flask to distilling bulk or condenser and with tip of condenser immerse in standard acid in receiver.

(9) Rotate flask to mix contents thoroughly, then heat until all ammonia has distilled (at least 150 ml. distillate)

(10) Titrate excess standard acid in distillate with standard sodium hydroxide solution, using methyl red as an indicator.

(11) Correct for black determination on reagents.

Calculations :

Per cent Nitrogen =

(ANa - BNb) x 0.01401 x 100
_____________________________
W

A = ml. of standard acid used.

B = ml. of standard NaOH used.

Na = Normality of standard acid.

Nb = Normality of standard NaOH.

weight of the sample taken in grams.

(Reference "Methods of Analysis", A.O.A.C., 1965).

(v) Total Nitrogen (for nitrate containing samples) :

(Not applicable to samples containing high concentrations of nitrate nitrogen and chlorides)

(a) Procedure -

(1) Place weighed sample (0.7 - 2.2 gm.) in digestion flask.

(2) Add 40 ml. H₂SO₄ containing 2 grams salicylic acid. Shake until thoroughly mixed and let stand, with occasional shaking, 30 minutes or more.

(3) Then add (i) 5 grams Na₂S₂O₃.5H₂O or (ii) 2 grams zinc dust (as impalpable powder not granulated zinc or filings).

(4) Shake the flask and let it stand for five minutes, then heat over low flame until forthing ceases.

(5) Turn off heat, add 0.7 grams copper sulphate, 15 gm. powdered K₂SO₄ (or anhydrous Na₂SO₄), and boil briskly until solution clears, then at least 30 minutes longer (2 hours for samples containing organic material).

Proceed further as in 6 - 11 of 3(iv).

Calculations :

Per cent Nitrogen =

(ANa - BNb) x 0.01401 x 100
_____________________________
W

A = ml of standard acid used.

B = ml. of standard NaOH used.

Na = Normality of standard acid.

Nb = Normality of standard NaOH.

weight of the sample taken in grams.

(Reference "Methods of Analysis", A.O.A.C., 1965).

(vi) Total nitrogen (for materials with high CI : NO₃ ratio and to materials containing only water soluble nitrogen).

(a) Reagents :

Reduced iron powder, electrically reduced, N.F. (National Formularly). For other reagents see 3(ii).

(b) Procedure -

Mixed Fertilisers :

(1) Place 0.5 - 2.0 gm. sample in Kjeldahl flask and add 2.5 grams reduced Fe (5 gms. is enough for 0.185 grams. NO₃).

(2) Add approximately 25 ml. distilled water, rotating flask at angle to wash down the sample.

(3) Let it stand for 15 minutes with occasional agitation, to ensure complete solution of all soluble salts.

(4) While rotating flask add 25 ml. cold H₂SO₄ (1 + 1) and let it stand until visible reaction ceases (use hood or vented digestion unit).

(5) Add boiling chip and boil 15 - 20 minutes, but do not take to dryness. (For samples containing organic matter, use 50 ml. cold H₂SO₄ (1 + 1), boil for 15¬20 minutes, add 0.7 grm. HgO add heat again for 30 minutes). Cool and proceed further as in (6) - (11) of 3(iv).

Calculations :

Per cent Nitrogen =

(ANa - BNb) x 0.01401 x 100
_____________________________
W

A = ml. of standard acid used.

B = ml. of standard NaOH used.

Na = Normality of standard acid.

Nb = Normality of standard NaOh.

weight of the sample taken in grams.

(Reference "Methods of Analysis", A.O.A.C., 1965).

(vii) Determination of ammoniacal nitrogen (Distillation Method).

(This method is for the determination of nitrogen present or available in the samples as ammonium ion. This method assumes that Urea is absent from the sample. Not applicable to MgNH₄PO₄ and Fe NH₄PO₄).

(a) Procedure -

(1) Place 0.7 - 3.5 grams; according to NH3 content of the sample, in distillation flask with approximately 300 ml. water and 2 grams of freshly ignited carbonate free MgO or NaOH solution.

(2) Connect the flask to condenser by Kjeldahl connecting bulb.

(3) Distill 100 ml. liquid into measured quantity of standard acid and titrate with standard NaOH solution, using methyl red as an indicator.

Calculations :

Per cent Nitrogen =

(ANa - BNb) x 0.01401 x 100
_____________________________
W

A = ml. of standard acid used.

B = ml. of standard NaOH.

Na = Normality of acid.

Nb = Normality of NaOH.

W = weight of the sample taken in gms.

(Reference - Recommended analytical methods of the National Plant Food Institute, Washington, D.C., 1961).

(viii) Determination of ammoniacal and nitrate nitrogen (Devarda Method)

(This method is for the determination of total nitrogen when only nitrate or mixture of nitrate and ammoniacal is present. This method assumes the absence of Urea, Calcium Cyanamide and organic matter from the sample).

(a) Procedure -

(1) Place 0.35 - 0.5 gms. samples in 600 - 700 ml. flask and add 300 ml. water, 3 grams Devarda Alloy, and 5 ml. NaOH solution (42 per cent by weight pouring later down side of flask so that it does not mix at once with contents).

(2) Allow the flask to stand for 15 minutes.

(3) By means of Division (J. Ind. Eng. Chem. 11,465 (1919) or other suitable scrubbing bulb that will prevent passing over of any spray connect with condenser, tip of which always extends beneath surface of standard acid in receiving flask.

(4) Mix contents of distilling flask by rotating.

(5) Heat slowly at first, and then at a rate to yield 250m1. distillate in 1 hour.

(6) Collect distillate in measured quantity of standard acid and titrate with standard NaOH solution, using methyl red as an indicator.

(7) In analysis of nitrate salts dissolve 3.5 or 5.0 grams in water, to 250 ml. and use 25 ml. aliquot.

Calculations :

Per cent Nitrogen =

(ANa - BNb) x 0.01401 x 100
_____________________________
W

A = ml. of standard acid used.

B = ml. of standard NaOH used.

Na = Normality of standard acid.

Nb = Normality of standard NaOH.

weight of the sample taken in grams.

(Reference "Methods of Analysis", A.O.A.C., 1965).

(ix) Determination of nitrate nitrogen

(Applicable in presence of calcium cyanamide and urea in the mixture).

(a) Procedure -

(1) Determine total nitrogen as in 3(v).

(2) Determine water insoluble N as in 3(x) but use 2.5 grams sample. Dilute to 250 ml.

(3) Determine ammoniacal N in 50 ml. filtrate as in 3(vii).

(4) Place another 50 ml. portion filtrate in 500 ml. Kjeldahl flask and 2 grams. Fe SO₄.7H₂O and 20 ml. H₂SO₄. (If total N is 5 per cent use 5 gm. Fe SO₄.7H₂O). Digest over hot flame until all water is evaporated and white fumes appear and continue digestion at least ten minutes to drive off nitrate N. If severe bumping occurs, add 10 - 15 glass beads. Add 0.65 grams Hg or 0.7 grams HgO and digest until all organic matter is oxidized cool, dilute add the K₂S solution and complete determination as 3(iv). Before distillation add a pinch of mixture of Zinc dust and granular "20 meash", Zinc to each flask to prevent bumping.

Calculations :

Total N (a) - Water insoluble N(b) - Water soluble N. Water Sol N - N obtained in (d) Nitrate N.

(Reference Methods of Analysis A.O.A.C. 1965).

(x) Determination of water insoluble nitrogen

(a) Procedure -

(1) Place 1 or 1.4 grams sample in 50 ml. beaker, wet with alcohol.

(2) Add 20 ml. water and let it stand for 15 minutes stirring occasionally.

(3) Transfer supernatant liquid to 11 cm. Whatman No. 42 paper in 60° long stem funnel 2.5" diameter and wash residue 4 to 5 times by decanting with water at room temperature (20 - 25°C).

(4) Finally, transfer all residue to filter and complete washing until filtrate measures 250 ml.

(5) Determine N as in 3(iv).

(xi) Determination of Urea nitrogen

(This method is for the determination of urea content of any mixed fertilisers).

(a) Reagents -

(1) Neutral urease solution - shake 1 gm. jack bone meal with 100 ml, water for 5 minutes. Transfer 10 ml. solution to 250 ml. (Erlemneyer flask, dilute with 50 ml. water and add 4 drops methyl purple indicator. Titrate with 0.1 N HCL to reddish purple, then back titrate to green with 0.1 N NaOH. From difference in ml., calculate amount of 0.1 N HCL required to neutralize remainder of solution (usually approximately 2.5 ml. per 100 ml.) add this amount of acid and shake well.

(b) Procedure -

(1) Weight 10±0.01 gm. sample and transfer to 15 cm. Whatman No. 12 fluted filter paper.

(2) Leach with approximately 300 ml. water into 500 ml. volumetric flask.

(3) Add 75 - 100 ml. saturated barium hydroxide solution to precipitate phosphates.

(4) Let it settle and test for complete precipitation with few drops of saturated barium hydroxide solution.

(5) Add 20 ml. 10 per cent sodium carbonate solution to precipitate excess barium and any soluble calcium salts.

(6) Let it settle and test for complete precipation.

(7) Dilute to volume, mix and filter through 15 cm. Whatman No. 12 fluted paper.

(8) Transfer 50 ml. aliquot (equivalent to I gm. sample) to 200 or 250 ml. Frlemneyer flask and add 1 to 2 drops methyl 1 purple indicator.

(9) Acidify solution with 2NHC 1 and add 2 to 3 dropse excess.

(10) Neutralize solution with 0.1 N NaOH to first change in colour indicator.

(11) Add 20 ml. neutral urease solution, close flask with rubber stopper and let it stand for 1 hour at 20 - 25°C.

(12) Cool the flask in ice water slurry and titrate at once with 0.1 N HCI to full purple colour, then add approximately 5 ml. excess.

(13) Record total volume added, back titrate excess HCI with 0.1 N NaOH to neutral end point.

Calculations:

Percentage urea = (ml. 0.1 N HCI - ml. 0.1 N NaOH) x 0.3003 wt. of sample (Reference "Methods of Analysis" A.O.A.C., 1965)

(xii) Determination of biuret

(a) Reagents :

(1) Alkaline tartarate solution - Dissolve 40 gm. NaoH in 50 ml. water cool, add 50 gm. of NaKC₄H₄O₆.H₂O and dilute to 1 litre. Let it stand for one day before use.

(2) Copper sulphate solution - Dissolve 15 gm. CuSO₄.5H₂O in CO₂ free water and dilute to 1 litre.

(3) Biuret standard solution - 1 mg./ml. Dissolve 100 mg. reagent grade biuret in CO₂ free water and dilute to 100 ml.

(4) Ion exchange resin. - Fill 50 ml. burette with 30 cm. column of Amberlite IR 120 (H) resin on glass wool plug. Regenerate column after each use by passing 100 ml. H₂SO₄ (1 + 9) or HCI (1+4) through the column approximately 5 ml. per minute and then washing with water until pH of effluent is 6:0.

(b) Preparation of standard curve :

(1) Transfer series of a liquots, 2 - 50 ml. of standard biuret solution to 100 ml. volumetric flask.

(2) Adjust volume to approximately 50 ml. with CO₂ free water. Add one drop of methyl red and neutralize with 0.1 N H₂SO₄ to pink colour.

(3) Add with swirling 20 ml. alkaline tartarate solution and then 20 ml. CuSO₄ solution.

(4) Dilute to volume shake for 10 seconds and place in water bath for 15 minutes at 30±5°C.

(5) Also prepare reagent blank.

(6) Determine absorbance of each solution against blank at 555 mw (instrument with 500 - 750 mw filter is also satisfactory) with 2 - 4 cm. cell, and plot standard curve.

A. In urea

(1) Stir continuously 2 - 5 gm. sample in 100 ml. approximately 50°C water for 30 minutes.

(2) Filter and wash into 250 ml. volumetric flask and dilute to volume.

(3) Transfer 25 ml. aliquot to 100 ml. volumetric flask and proceed as given under preparation of standard curve 3(xii).

B. In mixed fertilisers -

(1) Stir continuously 10 - 20 gm. sample in 150 ml. approximately (50°C) hot water for 30 minutes.

(2) Filter and wash into 250 ml. volumetric flask and dilute to volume.

(3) Transfer 25 ml. aliquot to column 3(xii) (a) (4) and adjust flow to 4 - 5 ml./ minute.

(4) Receive eluate in 100 ml. beaker.

(5) When liquid level falls to top of resin bed, wash with two 25 ml. portion water.

(6) To eluate and washings add two drops of methyl red and then 0.1 N NaOH to yellow colour.

(7) Add 0.1 NH₂SO₄ until solution just turns pink.

(8) Transfer to 100 ml. volumetric flask and dilute to volume with CO₂ free water.

(9) Transfer 50 ml. aliquot to 100 ml. volumetric flask and proceed as in preparation of standard curve given above.

Calculations :

From standard curve determine concentration of biuret in final dilution, then per cent of

biuret = Cx 100
________
W

Where C = Concentration in mg/ml. of biuret in final dilution obtained from standard curve.

W = Concentration of original sample in final dilution expressed as mg/ml. (Reference "methods of Analysis "A.O.A.C., 1965)

(xiii) Determination of free acidity in ammonium sulphate (as H₂SO₄)

(a) Reagents :

(1) Standard sodium hydroxide solution - 0.02 N.

(2) Methyl red indicator - Dissolve 0.15 gm. of water soluble methyl red in 500 ml. water :

(3) Methyl red - Methyl blue mixed indicator solution - prepared by mixing equal volumes of 0.2 per cent solution in rectified spirit of methyl red and 0.1 per cent solution in rectified spirit of methylene blue.

(b) Procedure -

(1) Dissolve about 20 gm. of prepared sample, accurately weighed in about 50 ml. cold natural water.

(2) Filter and make up the volume to about 200 ml.

(3) Titrate with standard sodium hydroxide solution, using one or two drops or methyl red as indicator.

(4) If satisfactory end point with methyl red is not obtained, methylene red - methylene blue mixed indicator may be used.

(5) Use preferably a micro biuret for this titration.

The filtering medium shall be neutral and shall not contain any alkaline material which would neutralize free acid.

Calculations :

Free acidity as H₂SO₄ per cent by weight =

4.904 AN
__________
W

A = Volume of ml. of standard NaOH solution.

N = Normality of standard NaOH solution.

W = Weight in gm. of prepared sample taken for the test.

(Reference - Specifications for ammonium sulphate, fertilizer grade IS : 826 - 1967)

(xiv) Determination of Arsenic in Ammonium Sulphate (A₂O₃)

(a) Reagents :

(1) Lead acetate solution - Prepare 10 per cent solution of lead acetate with sufficient acetic acid added to clear the solution.

(2) Dry lead acetate paper - Cut filter paper (Whatman No. 1 or equivalent) into strips 70 x 50 mm and keep them permanently suspended in lead acetate solution in a glass stoppered bottle. Before use, take out the strips and dry them in an atmosphere free from hydrogen sulphide.

(3) Mercuric Bromide solution - Dissolve 5 gm. of mercuric bromide in 100 ml. rectified spirit.

(4) Sensitized Mercuric Bromide paper strips - cut filter paper (Whatman No. 1 or equivalent) into strips 120 x 2.5 mm. Keep the strips permanently suspended in dark in a glass stoppered cylinder or amber bottle having mercuric bromide solution. Before use take out a strip, press it between sheets of filter paper and dry it in an atmosphere free from hydrogen sulphide.

(5) Dilute sulphuric acid - approximately 5 N.

(6) Concentrated hydrochloric acid.

(7) Potassium iodide solution - 15 per cent.

(8) Stannous chloride solution - Dissolve 80 gm. of stannous chloride in 100 ml. water containing 5 ml. of concentrated hydrochloric acid. If the solution is turbid add a few more ml. of hydrochloric acid and boil until clear solution is obtained. Add some metallic tin to the solution to prevent oxidation.

(9) Zinc - It is recommended that zink rods prepared as described below should be used for routine work, however, pellets described may be used.

(b) Preparation of zinc rods -

(1) Take a clean and dry hard glass test tube of 10 mm. internal diameter and 20 cm. length.

(2) Heat the test tube over a flame of Bunsen or blow pipe burner and add slowly arsenic free granulated zinc in small portions (1 to 2 gm. at a time) the next portion being added after the first one has completely melted.

(3) Continue heating and adding zinc until the melt is about 10 cm. high.

(4) Heat the clean melt for half an hour and then cool to room temperature.

(5) Break the tube to obtain the rod of zinc.

(6) Cut rod into pieces 20 mm. long.

(7) Coat the plane ends of the pieces with a paste of magnesium carbonate and gum arabic solution and dry.

(8) Coat the pieces all over with 1.5 mm. thick layer of paraffin wax.

(9) When required for use, scrap off the wax from the plane ends with a knife. protecting wax colour round the rods.

(10) Remove the past from the plane ends by soaking in water and activate the exposed surface by dipping in a solution containing one part of stannous chloride solution and seven parts of concentrated hydrochloric acid.

Treat zinc shots passing through IS sieve 570 (aperture 5600 microns) and retained on IS sieve 280 (aperture 2818 microns) with concentrated hydrochloric acid until the surface of zinc becomes clean and dull. Weight and keep under water, preventing contamination with dust.

(d) Standard sodium hydroxide solution - approximately 20 per cent.

(e) Standard arsenic trioxide solution.

(1) Dissolve 1.0 gm. of resublimed arsenic trioxide (AS₂O₃) in 25 ml. sodium hydroxide solution and neutralize with dilute sulphuric acid.

(2) Dilute with freshly distilled water containing 10 ml. of concentrated sulphuric acid per litre and make up the volume to 1 litre.

(3) Again dilute 10 ml. of this solution to 1 litre with water containing sulphuric acid and finally dilute 100 ml. of this solution to I litre with water containing sulphuric acid. 'One ml. of this solution contains 0.001 mg. of arsenic trioxide (AS₂O₃). The dilute solution shall be prepared freshly when required.

(f) Procedure -

(1) Dissolve 1.0 gm. of the prepared sample in 20 ml. water.

(2) Place dry lead acetate paper in the lower portion of the tube B (in figure 6) and glass wool moistened with lead acetate solution in its proper portion.

(3) Place the sensitized strips of mercuric bromide paper in tube A and connect the tubes together with a rubber stopper.

(4) Introduce the solution of the material into the bottle C (120 ml.) and then add 10 ml. of dilute sulphuric acid. Add 0.5 ml. of stannous chloride solution, 5 ml. potassium iodide solution and make up the volume with water to about 50 ml.

(5) Mix the contents and drop about 10 gm. of zinc. Immediately fit in position the rubber stopper carrying the tube B.

(6) Place the bottle in a warm place at about 40°C.

(7) At the end of two hours remove the test strip by means of tweezers.

(8) Carrying out the test prescribed above using a volume of standard arsenic trioxide solution containing 0.1 mg. of arsenic trioxide, in place of the solution of the material and compare the strain produced with the material with that produced with arsenic trioxide solution.

(9) The limit prescribed in the material specification shall be taken as not having been exceeded if the length of the strain as well as the intensity of its colour produced in the test with the material is not greater than those produced with arsenic trioxide solution.

(Reference "Modified Gutzet method of test for arsenic" IS - 2088 - 1962)

(xv) Determination of calcium nitrate (for calcium ammonium nitrate and nitrophosphates)

(a) Reagents :

(1) N = Amyl Alcohol

(2) Dilute hydrochloric acid - approximately 4 N :

(3) Standard calcium solution - weight 1.0 gm. of calcium carbonate dried at 120u±5()C and dissolve in the minimum quantity of dilute hydrochloric acid. Dilute the solution to I litre in a graduated flask.

(4) Ammonium chloride - hydroxide buffer solution. Dissolve 67.5 gm. ammonium chloride in a mixture of 570 ml. of ammonium hydroxide (sp. gr. 0.92) and 250 ml. water. Also dissolve separately a mixture of 0.931 gm. of disodium ethylene diamine tetra - acetate dihydrate and 0.616 gm. of magnesium sulphate (Mg SO₄.7H₂O) in about 50 ml. of water. Mix the two solutions and dilute to 1 litre.

(5) Standard disodium ethlylene diamine tetra - acetate (EDTA) solution - weigh 3.72 gm. of disodium ethylene diamine tetra - acetate dihydrate in water, and dilute in a graduated flask to 1 litre. The solution shall be standardized frequently against standard calcium solution following the procedure given below.

(6) Eriochrome Block T Judicator solution - Dissolve 01 gm in 20 ml. of rectified spirit. The solution shall be used for not more than a week.

(b) Procedure -

(1) Grind quickly about 5 gm of the material, accurately weighed, with about 50 ml. of amyl alchol in a pestle and mortar and transfer the contents to a conical flask.

(2) Wash the pestle and mortar with a few ml. of amyl alchol and add the washings to the flask.

(3) Shake the contents of the flask manually or in a mechanical; shaker for about half an hour and then filter.

(4) Transfer the filtrate to a separating funnel and extract calcium nitrate completely with water in five to six installments.

(5) A few drops of dilute hydrochloric acid may be added during the extraction with water to avoid formation of an emulsion of amyl alchol with water.

(6) Concentrate the water extract at low temperature to nearly half its volume.

(7) Transfer the concentrated solution to a conical flask, add 5 ml. of ammonium chloride - ammonium hydroxide buffer solution, 5 drops of eriochrome black T indicator solution and titrate against standard EDTA solution to a pure blue end point.

Calculations :

Calcium nitrate per cent by weight =

8.2 NV
________
W

where

N = Normality of standard EDTA solution.

V = Volume in ml. of standard EDTA solution used in the titration, and

W = Weight in gm. of the material taken for test.

(Reference : IS specification of CAN IS : 2409 - 1963)

(xvi) Determination of Chlorides other than ammonium chloride

(a) Reagents -

(1) Standard silver nitrate solution - 01. N.

(2) Concentrated nitric acid - conforming to IS : 264 - 1950.

(3) Ferrous ammonium sulphate solution saturated in water and stabilized by addition of 50 ml. nitric acid.

(4) Standard Ammonium Thiocyanate solution - 0.1 N.

(b) Procedure -

(1) Dissolve about 0.2 gm. of the prepared sample, previously dried as in procedure for determination of moisture and accurately weighed, in about 40 ml. water.

(2) Add exactly 50 ml. of standard silver nitrate solution and 5 ml. of concentrated nitric acid.

(3) Add 0.5 ml. of nitrobenzene and make up the volume of the. mixture of 100 ml. with water.

(4) Take exactly 50 ml. of the solution and add 2 ml. of ferric ammonium sulphate solution.

(5) Titrate the excess of silver nitrate in this portion with standard ammonium thiocyanate solution.

(6) Carry out a blank test following the procedure given as above but without using the material.

Calculations :

Total Chlorides (as Cl) per cent by weight (on dry basic) =

7.07 (V₁ - V₂)N
________________
W

….X

where

V₁ = Volume in ml. of standard ammonium thiocyanate used in the blank determination.

V₂ = Volume in ml. of standard ammonium thiocyanate solution used in the test with the material.

N = Normality of standard ammonium thiocyanate solution.

W = Weight in gm. of the derived prepared sample taken for the test.

Express the ammoniacal nitric content per cent by weight of the material determined earlier in terms of Cl as follows :

Chloride equivalent of the ammoniacal nitric content, per cent by weight = 2.531 x A Y where A is the ammoniacal nitric content determined earlier.

Balanced chloride equivalent to sodium chloride (NaCI) per cent by weight = 1.648 x (X - Y) = Balanced chloride other than ammonium chloride.

(Reference IS specification for ammonium chloride fertiliser grade (Revised) IS : 1114 - 1964 calculations notified).

4. Determination of phosphates -

Methods of determination of total phosphates, water soluble phosphates, citrate soluble phosphates , citrate insoluble phosphates and citric acid soluble phosphates have been described separately in this section. These methods are applicable to straight as well as mixed phosphatic fertilisers.

(i) Preparation of solution of the samples

(Separate methods of preparing solutions of the samples have been described according to the nature of the sample).

(a) Reagent :

Magnesium nitrate solution - Dissolve 950 gm. P - free Mg (NO₃)₂. 6H₂O in water and dilute to 1 litre.

(b) Procedure -

(1) Treat 1 gm. sample by (A), (B), (C), (D) or (E), method depending on the nature of the sample.

(2) Cool solution transfer to 200 to 250 ml. volumetric flask, dilute to volume, mix and filter through dry filter.

A. (Suitable for materials containing small quantities of organic matter). Dissolve in 30 ml. HNO₃ and 3 - 5 ml. HCI and boil until organic matter is destroyed.

B. (Suitable for fertilisers containing much Fe or Al phosphate and basic slag). Dissolve in 15 - 30 ml. HCI and 3 - 10 ml. HNO₃.

C. (Suitable for organic material like cotton seed meal alone or in mixture). Evaporate with 5 ml. of the Mg(NO₃)₂ solution, ignite and dissolve in HCI.

D. (Generally applicable to materials or mixtures containing large quantities of organic matter). Boil with 20 - 30 ml. H₂SO₄ in 200 ml. flask adding 2 - 4 gm. of NaNO₃ or KNO₃ at beginning of digestion and small quantity after solution is nearly colourless, or adding the nitric acid in small portions from time to time. When solution is colourless, cool add 150 ml. water boil for few minutes. Before adding NaNO₃ or KNO₃ let mixture digest, at gentle heat if necessary, until violence of reaction is over.

E. (Suitable for all fertiliser). Boil gently for 30 - 45 minutes with 20 - 30 ml. HNO₃ in a suitable flask (Preferably a Kjeldahl for samples containing large quantities of organic matter to oxidize all easily oxidizable matter). Cool and and 10 - 20 ml. of 70¬72 per cent perchloric acid. Boil very gently until solution is colourless or nearly so and white dense fumes appear in flask. Do not boil to dryness at any time. (Danger). (With samples containing large quantities organic matter temperature should be reised to fuming point approximately 170°C, cover a period of 1 hour at least). Cool slightly, add 50 ml. water and boil for few minutes.

(ii) Gravimetric Quinoline, Molybdate method for determination of total phosphorus

(a) Reagents :

(1) Citric molybdic acid reagent - Dissolve 54 gms. 100 per cent molybdic anhydride (MgO₃) and 12 gms. NaOH with stirring in 400 ml. hot water and cool. Dissolve 60 gms. citric acid in mixture of 140 ml. HCI and 300 ml. water and cool. Gradually add molybdic solution to citric acid solution with stirring. Cool, filter and dilute to 1 litre. (Solution may be green or blue, colour depends on exposure to light). If necessary add 0.5 per cent KBrO₃ solution drop by drop until green colour becomes pale. Store in dark in polyethlene bottle.

(2) Quinoline solution - Dissolve 50 ml. synthetic quinoline with stirring in mixture of 60 ml. HCl and 300 ml. water. Cool dilute to 1 litre, and filtre. Store in polyethylene bottle.

(3) Quimociac reagent - Dissolve 70 gms. of sodium molybdate dihydrate in 150 ml. water. Dissolve 60 gms. citric acid in mixture of 85 ml. HNO₃ and 150 ml. water and cool. Gradually add molybdate solution to citric acid - nitric acid mixture with stirring. Dissolve 5 ml. synthetic quinoline in mixture of 35 ml. HNO₃ and 100 ml. water. Gradually add this solution to molybdate citric - nitric acid solution mix and let it stand for 24 hours. Filter, add, 28 ml. acetone, dilute to 1 litre with water and mix well. Store in polythylene bottle.

(b) Procedure -

(1) Treat 1 gm. sample as prescribed in 4(i) and dilute it to 200 ml.

(2) Pipette into 500 ml. Erlenmeyer flask, aliquot containing not more than 25 mg. P₂O₅ dilute to approximately 100 ml. with water. Proceed with one of the following methods.

A. Add 30 ml. citric molybdic acid reagent and boil gently for 3 minutes. (Solution must be precipitate free at this stage). Remove from heat and swirl carefully. Immediately add from burette 10 ml. quinoline solution with continuous swirling (Add first 3 - 4 ml. dropwise and remainder in steady stream) or

B. Add 50 ml. quimociac reagent, cover with watch glass place on hot plate in well ventilated hood, and boil for a minute.

After treatment with A or B cool to room temperature, swirl carefully 3 - 4 times during cooling, filter into gooch with glass fibre filter paper previously dried at 250°C and weighed. and wash five times with 25 ml. portion of water. Dry cruicible and contents for 30 minutes at 250°C. Cool in desicator to constant weight as (C₉H₇N)₃ H₃PO₄. 12MoO₃. Subtract weight reagent blank. Multiply by 0.03207 to obtain weight of P205. Report as per cent P₂O₅.

(iii) Determination of water soluble phosphorus (a) Procedure -

(1) Place 1 gm. sample on 9 cms. filter paper and wash with small portion of water until filtrate measure approximately 250 ml.

(2) Let each portion pass through filter before adding more and use suction if washing would not otherwise be complete within 1 hour.

(3) If the filtrate is trubid, add 1 - 2 mi. HNO₃ dilute to 250 ml. and mix.

(4) Pipette into 500 ml. Erlenmeyer flash aliquot more than 25 mg. P₂O₅.

(5) Dilute if necessary to 50 ml.

(6) Add 10 ml. HNO₃(1 plus 1) and boil gently for 10 minutes. Cool and dilute to 100 ml. and proceed as 4(ii) (b) (2) (B). (Reference "Methods of Analysis" A.O.A.C., 1965).

(iv) Citrate insoluble phosphorus

(a) Reagents :

(1) Ammonium citrate solution - should have specific gravity of 1.09 at 20°C and pH of 7.0 as determined electrometrically.

Dissolve 370 gms. crystalline citric acid in 1.5 litre disstilled water and nearly neutralized by adding 345 ml. NH₄OH (28 - 29% NH₃). If concentration of ammonia is less than 28 per cent add correspondingly large volume, and dissolve citric acid in correspondingly smaller volume of water, cool and check pH. Adjust with NH₄OH (1 plus 7) or citric acid solution to pH - 7. Dilute solution if necessary to specific gravity of 1.09 at 20'C. (Volume will be approximately 2 litres). Keep in tightly stopped bottles and check pH from time to time. If pH has changed from 7.0, readjust.

(2) Other reagents and solutions as in 4(i) and 4(ii).

(b) Procedure (Acidulated samples and mixed fertilisers) -

(1) After removing water soluble P₂O₅ in 4(iii) transfer filter and residue within 1 hour to 200 or 250 ml. flask containing 100 ml. ammonium acetate solution previously heated to 65°C.

(2) Close flask tightly with smooth rubber stopper.

(3) Shake vigorously until paper is reduced to pulp and relieve pressure by removing stopper momentarily.

(4) Continuously agitate contents of stopper flask in apparatus equipped to hold contents of flask at exactly 65°C. (Action of apparatus should be such that dispersion of sample in citrate solution is continually maintained and entire inner surface of flask and stopper is continually bathed with solution).

(5) Exactly 1 hour after adding filter and residue, remove flask from apparatus, and immediately filter contents by suction as rapidly as possible through whatman No. 5 paper or equivalent, using buchner or ordinary funnel with platinum or other cone.

(6) Wash with distilled water and at 65°C until volume of filtrate is approximately 350 ml. allowing time for through draining before adding more water.

(7) If material is one that will yield coludy filtrate, wash with 2 per cent NH₄NO₃ solution.

(8) Determine P₂O₅ in citrate insoluble residue by one of the following methods :

A. Dry paper and contents, transfer to crucible, ignite until all organic matter is destroyed and digest with 10 - 15 ml. HCI until all phosphate dissolve, or

B. Treat wet filter and contents as in method prescribed in 4(i)(b) (2) (A) (C) (D) or (E). - Dilute solution to 250 ml. or other suitable volume, mix well filter through dry paper and determine P₂O₅ as in 4(ii).

(Reference "Methods of Analysis", A.O.A.C., 1965).

(v) Citrate soluble phosphorus

Substract [* * *] and citrate insoluble P₂O₅ from total P₂O₅ to obtain citrate soluble P₂O₅ -

(Reference "Methods of Analysis" A.O.A.C., 1965).

(vi) Citric acid soluble P₂O₅ in fertilisers other than basic slage

(a) Reagents :

(1) Concentrated hydrochloric acid.

(2) Concentrated citric acid.

(3) Calcium carbonate finely ground.

(4) 5 N Sodium hydroxide solution.

(5) Dilute hydrochloric acid - Dilute 240 ml. of concentrated hydrochloric acid with water to 1 litre.

(6) Citric molybdic acid solution - Stir 54 gms. of molybdic anhydride (MoO₃) with 200 ml. of water, add 11 gms. of sodium hydroxide and stir the mixture whilst heating to boiling point until the molybdic anhydride dissolves. Dissolves 60 gms. of citric acid in about 250 to 300 ml. of water and add 140 ml. of concentrated hydrochloric acid. Pour the molybdate solution into the acid solution, which is stirred throughout the addition. Then cool and if necessary filter the solution through paper pulp pad. Dilute the solution to 1 litre. If the solution is slightly green or blue in colour add dropwise a dilute (0.5 to 1.0 per cent) solution of potassium bromate until the colour is discharged. This reagent should be kept in dark.

(7) Quinoline solution - Measure 60 ml. of concentrated hydrochloric acid and 300 to 400 ml. of water into 1 litre beaker and warm to 70" - 80°C. Pour 50 ml. of quinoline in a thin stream into the dilute acid while stirring. When quinoline has dissolved, cool the solution dilute to 1 litre, and if necessary filter through a paper pulp filter.

(8) Sodium hydroxide solution - 0.5 N carbonate free.

(9) Indicator solution - Mix three volumes of thymol blue solution and two volumes of phenolphthalein solution prepared as follows :

A. Thymol blue solution - Dissolve 250 mgs. thymoi blue in 5.5 ml. of 0.1 N sodium hydroxide solution and 125 ml. of industrial methylated spirit. Dilute with water to 250m1.

B. Phenolphthalein solution - Dissolve 250 mgs. phenolphthalein in 150 ml. of industrial methylated spirit and dilute with water to 250 ml.

(10) Hydrochloric acid - 0.5 N

(11) Sodium hydroxide solution - 0.5 N.

(12) Hydrochloric acid - 0.1 N.

(13) Surface active agent - 0.5 per cent. solution of sodium dedecyl benezene sulphonate suitable.

(b) Preparation of solution -

(1) Weigh to the nearest mg. about 5 mgs. of the sample and transfer to a stoppered bottle of about 1 litre capacity.

(2) Dissolve 10 gms. of pure crystallized citric acid (monohydrate) in water. dilute to 500 ml. and adjust the temperature to 20°C.

(3) Add the solution to the sample in the bottle, shaking so as to avoid the possibility of caking.

(4) Shake the bottle continuously for 30 minutes.

(5) Pour the whole of the liquid at once on a large medium - fine - filter and collect the filtrate. If the filtrate is not clear, pass it again through the same filter.

(1) Transfer a volume of the solution prepared according to procedure given above containing less than 70 mg. of phosphoric acid and preferably about 50 mgs. to a 500 ml. stoppered conical flask marked at 150 ml.

(2) Dilute the solution with water to 100 ml.

(3) If the sample does not contain calcium add 100 to 200 mg. of calcium carbonate.

(4) Then add 5 N sodium hydroxide solution dropwise until a faint permanent turbidity or precipitate is formed.

(5) Dissolve the precipitate by the dropwise addition of dilute hydrochloric acid, but avoid and excess.

(6) Dilute to 150 ml. and add 50 ml. of the citricmolybdic acid reagent, and heat the solution to inticipient abolition, maintain it at this temperature for 3 minutes and then bring it to the boiling point.

(7) From burette slowly add 25 ml of the qui noline solution with constant swirling throughout the first few ml. being added dropwise, the rest in slow stream.

(8) Keep the solution gently boiling during the addition.

(9) Immerse the flask in boiling water for 5 minutes, then cool it to 15°C, in running water.

(10) Filter with suction the contents of the flask on a paper pad, and wash the flask precipitate and filter such successive small washes of cold water until they are free from acid.

(11) Transfer the filter pad and precipitate to the original flask, rinse the funnel with water and collect the rinsing in the flask.

(12) If necessary, wipe the funnel with small piece of damp filter paper to ensure complete removal of the precipitate, and place and paper in the flask.

(13) Add water to a total of about but not exceeding 100 ml. Stopper the flask and shake it vigorously until the pulp and precipitate are completely dispersed.

(14) Remove the stopper and wash in with water, returning the washing of the flask.

(15) Add a measured volume of 0.5 N sodium hydroxide solution sufficient to dissolve the precipitate and leave a few ml. in excess.

(16) Shake the flask vigorously until all the precipitate dissolves. (To facilitate the dispersal of the precipitate, after the addition of 0.5 N sodium hydroxide solution, a few drops of the surface active agent may be added if necessary).

(17) Add 0.5 - 0.1 ml. of the indicator solution, and titrate the excess of sodium hydroxide with 0.5 N hydrochloric acid, until the indicator changes from violet to green - blue, and then very sharply to yellow at the end point.

(18) Deduct the number of ml. of 0.5 N hydrochloric acid used from the number of ml. 0.5 N sodium hydroxide, to ascertain the volume of 0.5 N sodium hydroxide equivalent to the phosphoric acid.

(19) Carry out a blank determination on all the regents, omitting only the sample, and using 0.1 N standard alkali and acid instead of 0.5 N for the titration. Calculate the blank in terms of 0.5 N alkali and subtract in from the original result.

(20) Calculate the amount of phosphoric acid in the portion taken for analysis from the factor 1.0 ml. of 0.5 N sodium hydroxide = 1.366 mgs. P₂O₅.

(Reference Statutory Instruments 1960. No. 1165, Agriculture, the Fertiliser and Feeding Stuffs Regulations, 1960).

(vii) Free phosphoric acid as P205

(a) Reagents :

(1) Acetone - conforming to IS : 170 - 1950 specification for acetone.

(2) Standard sodium hydroxide solution - 0.1 N

(3) Bromocresol green indicator solution - dissolve 0.1 gm. of bromocresol green in 100 ml. of rectified spirit conforming to IS : 323 - 1959 specification for rectified spirit (Revised).

(b) Procedure -

(1) Weigh accurately about 2.5 mgs. of the prepared sample and transfer to a soxhlet extract or

(2) Add about 100 ml. of acetone and extract for three hours.

(3) Cool and distil off the acetone as far as possible.

(4) Take up the residue with water and make up the volume to 250 ml.

(5) Pipette out exactly 100 ml. of this solution and titrate with standard sodium hydroxide solution, using bromocreosol green as indicator until colour just changes from yellow to blue.

Calculations :

Free phosphoric acid (as P₂O₅) per cent by weight =

17.75 x V x N
________________
W

Where

V = volume in ml. of standard sodium hydroxide solution used.

N = normality of standard sodium hydroxide solution, and

W = weight in gm. of sample taken for the test.

(Reference - IS Specification for superphosphate (Revised) (IS : 294 - 1962).

5. Determination of potassium :

Determination of potassium in all kinds of fertilisers is given in this section. Two alternative methods have been described. Any of these two methods may be used depending upon the availability of reagent, and suitability of the method.

(i) Perchloric acid method

(This method depends on the insolubility of potassium perchlorate and the solubility of sodium perchlorate in alcohol and is applicable in presence of alkali metals chlorides and nitrates. Sulphates and ammonium salts must be absent on account of the low solubility of sodium sulphate of ammonium perchlorate in alcohol. Phosphates must be removed. Method are given for the elimination of the effect of interfering substances).

(a) Reagents :

(1) Concentrated hydrocloric acid.

(2) Barium chloride solution. Dissolve 100 gms. of barium chloride in water ; filter, the solution and dilute to 1 litre.

(3) Dilute hydrochloric acid - Dilute 250 ml. of concentrated hydrochloric acid with water to one litre.

(4) Calcium oxide - finely ground.

(5) Ammonium hydroxide - - sp. gr. 0.88

(6) Ammonium carbonate solution - saturated aqueous solution.

(7) Ammonium oxalate solution - saturated aqueous solution.

(8) 20 per cent perchloric acid solution.

(9) Alcohol - industrial methylated spirit 95 - 96 per cent V/V.

(10) Wash solution - add potassium perchlorate to alcohol and shake until saturated solution is obtained. Keep the solution over solid potassium perchlorate and filter immediately before use.

(b) Potassium salts free from sulphates and other interfering substances. -

(1) Dissolve in water a portion of the sample weighed to the nearest mg. equivalent to potassium content to 1.5 to 2.0 gms. of potash.

(2) Cool the solution to 20°C, dilute to 500 ml. in a volumetric flask, mix well and filter through a dry filter.

(3) Determine the potash in 50 ml. of solution by precipitating with perchloric acid as described under procedure.

(The following method is given by eliminating the interference caused by presence of sulphate. If salts contain phosphates, iron, manganese or substances °other than sulphate that interfere with the determination of potash, the method described for mixed fertiliser should be used).

(1) Weigh to the nearest mg. a portion of the sample equivalent in potassium content to 1.5 to 2.0 gms. of potash, into a 50 ml. beaker. Add about 300 ml. of water and 20 ml. concentrated hydrochloric acid and heat the solution to boiling.

(2) To the boiling solution cautiously add, drop by drop barium chloride solution in an amount slightly in excess of that previously determined as necessary to ensure the complete precipitation of sulphate.

(3) Cool the liquid to 20(C, transfer to a 500 ml. volumetric flask, dilute to 500 ml. mix and filter through a dry filter.

(4) Take 50 ml. of the filtrate and evaporate to dryness in a basin, moisten the residue with concentrated hydrochloric acid.

(5) Again evaporate to dryness, dissolve the residue with 5 - 10 ml. dilute hydrochloric acid and filter, if necessary. Determine the potash in solution by the method described under procedure.

(d) Potassium in mixed fertilizers. -

(1) Weigh to nearest centigram about 10 gm. of the sample and if organic matter is present, gently incinerate at temperature not exceeding 500°C.

(2) Transfer the weighed portion of the sample or the incinerated residue to a 500 ml. beaker with a little water, and 10 ml. concentrated hydrochloric acid and then warm for ten minutes.

(3) Dilute with water to about 300 ml. and bring gradually to boiling point.

(4) Add 10 gms. of calcium oxide made into a paste with water.

(5) Bring contents again gently to the boiling point, and keep so heated for about half an hour with frequent stirring.

(6) Cool to 20°C, transfer to a 500 ml. volumetric flask, dilute to 500 ml. and after thoroughly shaking filter through a dry filter paper.

(7) Transfer 250 ml. of the filtrate to another 500 ml. volumetric flask, make just acid with hydrochloric acid and heat to boiling point.

(8) To the boiling solution cautiously add drop by drop, barium chloride solution until there is no further precipitation of barium sulphate.

(9) Render the contents of the flask alkaline with ammonium hydroxide solution and precipitate the calcium and any excess of barium by adding ammonium carbonate solution until no further visible precipitation occurs followed by the addition of about 1 ml. of ammonium oxalate solution.

(10) Cool to 20°C, dilute with water to 500 ml. and after thoroughly shaking, filter through a dry filter paper.

(11) Measure 100 ml. of the filtrate and evaporate to dryness in a basin. Expel the ammonium salts from the residue by gently heating the basic over a low flame, being careful to keep the temperature below that of faint redness.

(12) Cool the residue, moisten with concentrated hydrochloric acid and again evaporate to dryness.

(13) Take up the residue with water the filter if necessary.

(14) Determine the potash in the solution by precipitation with perchloric acids described under procedure.

(e) Procedure -

(1) Transfer the solution obtained as described about into a basin and add about 7 ml. of perchloric acid solution.

(2) Place the basin on a hot plate or sand bath and evaporate the contents until the white fumes are copiously evolved.

(3) Cool and dissolve the precipitate in a little hot water.

(4) Add about 1 ml. of perchloric acid solution and again concentrate to the fuming stage.

(5) Thoroughly cool the residue in the basic and stir in 20 ml. of alcohol.

(6) Allow the precipitate to cool and settle, then pour the clear liquid through a dry filter paper, draining the precipitate in the basin and stir in 20 ml. of alcohol.

(7) Allow the precipitate to cool and settle, then pour the clear liquid through a dry filter paper, drain the precipitate in the basin as completely as possible.

(8) Redissolve the precipitate on the paper and that remaining in the basin with hot water.

(9) Add 2 ml. of perchloric acid solution to the combined solution and evaporate the whole down to the fuming stage.

(10) Cool the residue in the basin and thoroughly stir the contents with 20 ml. of alcohol.

(11) Allow the precipitate to cool and settle and pour the clear liquid through a weighed gooch or sintered glass crucible, draining the precipatate as completely as possible from the liquid before adding 5 ml. of wash solution.

(12) Wash the precipitate by decantation with several similar small portions of the wash solution, pouring the washing through the crucible.

(13) Transfer the precipitate to the crucible and wash it well with the wash solution until free from acid.

(14) Dry the precipitate at 100°C and weigh.

(15) Regard the precipitate as potassium perchlorate (KC1O₄), and calculate its equivalent as potash (K₂O) by multiplying its weight by 0.34.

(Reference - Statutory Instruments 1960, No. 1165. Agriculture. The Fertiliser and Feeding Stuffs Regulation, 1960).

(ii) Sodium tetraphenyl boron method

This method is applicable to both mixed and straight potassium fertilisers.

(a) Reagents :

(1) Sodium hydroxide solution - 20 per cent. Dissolve 20 gms. NaOH in 100 distilled water.

(2) Formalchyde solution - 37 per cent.

(3) Sodium tetraphenyl boron (STPB) solution approximately 1.2 per cent. Dissolve 12 gms. sodium tetraphenyl boron in approximately 800 ml. water. Add 20 - 25 ml. Al (OH₃), stir for 5 minutes, and filter (Whatman No. 42 paper or equivalent) into 1 litre volumetric flask. Rinse breaker sparingly with water and add to filter. Collect entire filtrate, add 2 ml. 20 per cent NOH solution, dilute to volume with water, and mix. Let it stand for 48 hours and standardize. Adjust to that 1 ml. STPB - 1 per cent K₂O. Store at room temperature.

(4) Quaternary ammonium chloride solution - approximately 0.625 per cent Dilute 50 ml. of 12.8 per cent Zephiran chloride to 1 litre with water, mix and standardize. Cetyltrimethyl ammonium bromide may be substituted for Zephiran chloride. If other concentrated is used, adjust volume.

(5) Clayton yellow - 0.04 per cent. Dissolve 40 mgs. in 100 m1. water.

(b) Standardization of solution. -

(1) Zephiran chloride - To 1.0 ml. STPB solution in 125 ml. Erlenmeyer flask, add 20 - 25 ml. water, 1 ml. 20 per cent NaOH 2.5 ml. HCHO 1.5 ml. 4 per cent (NH₄)₂ C₂O₄ and 6 - 8 drops of indicator (5) above.

(2) Titrate to pink end point with Zephiran chloride solution, using 10 ml. semimicro burette. Adjust Zephiran chloride solution so that 2.0 ml. - 1.0 ml. STPB solution.

(3) Sodium tetraphenyl boron solution - Dissolve 2.5 gms. of KH₂PO₄ in water in 250 ml. volumetric flask, add 50 ml. 4 per cent (NH₄)₂ C₂O₄ solution, dilute to volume with water and mix. (It is not necessary to bring to boil). Transfer 15 ml. aliquote. (51.92 mgs. K₂O 3.10 mgs. K) to 100 ml. volumetric flask, add 2 ml. 20 per cent NaOH, 5 ml. HCHO and 43 ml. STPB reagent dilute volume with water, mix thoroughly, let stand 5 - 10 minute and pass through dry filter. Transfer 50 ml. aliquot of filtrate to 125 ml. Erelenmeyer flask, add 6 - 8 drops of indicator (5) above and titrate excess reagent with Zephiran solution. Calculate titration values as follows :

F = 34.61/(43 ml. = ml. Zephiran) = %K20/ml. STPB reagent. Factor applies to all fertilizers if 2.5 gms. sample is diluted to 250 ml. nd 15 ml. aliquot is taken for analysis. If results are to be expressed as K rather than K20. substitute 28.73 for 34.61 in calculating the value of F.

(1) Mixed fertilisers

A. Place 2.5 gms. sample or factor weight 2.430 gms. in 250 ml. volumetric flask.

B. Add 125 ml. water and 50 ml. saturated (NH4) C,04 solution. Add 1 ml. of diglycol stearate solution if needed to prevent foaming.

C. Boil for 30 minutes, add slight excess of NH4OH and after cooling dilute to 250 ml. Mix and pass through dry filter.

(2) Potassium salts (potassium chloride sulphate, potassium magnesium sulphate, and kainite).

A. Dissolve 2.5 gms. or factor weight 2.430 gms. and dilute to X 250 ml. without adding NH4OH and (NH4) C204.

B. When interfering substances such as NH3Ca, Al, etc., are present proceed as in (1) above.

(d) Procedure -

(1) Transfer 1.5 ml. aliquot of sample solution to 100 ml. volumetric flask and add 2 ml. 20 per cent NaOH and 5 ml. HCHO.

(2) Add 1 ml. standard STPB solution for each I per cent K20 expected in sample plus additional 8 ml. excess to ensure complete precipitation.

(3) Dilute to volume with water, mix thoroughly, let it stand for 5 - 10 minutes and pass it through dry filter (Whatrnan 12 or equivalent).

(4) Transfer 50 ml. filtrate to 125 ml. Erlenmeyer flask, add 6 - 8 drops of indicator

(5) and titrate excess reagent with standard Zephiran solution. per cent K2O in the sample = (ml. STPB added =ml. Zephiran XF where F = per cent - K2O/ ml. STPB = reagent.

(Reference "Methods of Analysis", AOAC, 1985).

6. Method of analysis of anhydrous ammonia -

(i) Estimation of Water and Ammonia

(a) Scoop : - This method is for the determination of water at 20"C (or room temperature), and usually ammonia by difference of any essentially anhydrous ammonia liquid product. Normally, the water will be low (less than 0.5 per cent) or sample of synthetic ammonia.

(b) Apparatus :

(1) Ammonia sampling tubes.

(2) Fume hood with exhaust dust.

(3) Sample tube holder rack in hood fabricated as convenient to hold number of tubes desired.

(4) Gloves protective, elbow length.

Charcoal, reagent, 14 - 20 mesh.

(d) Procedure, -

(1) Remove the sample tubes from the sample carrier and place in the provided sample tube holders in the fume hood.

(2) Allow the ammonia sample to boil and to evaporate (approximately 30 minutes) spontaneously in the fume hood. Because of the low temperature of the boiling ammonia, atmospheric moisture freezes on the sample container. Exercise care to see that one of this ice contaminates the sample.

(3) When the boiling of the sample ceases, remove the tube from the holder. Wipe the outer surface of the tube dry and read and record the volume of liquid residue in the tube.

(4) From the volume recorded, calculate the weight per cent water in the sample. Ignore the small piece of charcoal, if used.

Calculation :

per cent water =

A x F x 0.890 x 0.684
__________________
V x 0.682

x 100

where

A = volume of residue in tube (ml.)

V = Volume of sample taken (m1.)

F = Evaporation factor, taken from Table I corresponding to noted sample pressure when sampling was obtained.

0.890 = density of residue (gm./ml.)

0.684 = weight fraction of water in residue.

0.682 = density of sample (gm./ml.)

(e) Precaution. -

The evaporation in the fume hood should be conducted with the window lowered to protect personnel from possible spray. Whenever it is necessary to handle the sample tubes, protective gloves should be worn.

Note. - Determination of residue on evaporation will not usually be necessary for normal ammonia sample. If sample is clear and water residue is clear and colourless, the residue may be considered to be nil. However, should it become necessary to determine the dissolved solids and suspended solids in anhydrous ammonia, proceed as follows :

(f) Sampling. -

Thoroughly clean and dry the sample tubes, weigh (W₁) each selected tube to the nearest milligram with a similar tube as counterpoise. Obtain the samples as desired earlier.

(g) Apparatus. -

(1) Analytical balance.

(2) Ammonia sampling tubes.

(3) Rubber tubing, 1/4 OD length desired.

(h) Reagents :

Air dry, carbon dioxide - free.

(i) Procedure. -

(1) Start test following elimination of the water content of the sample as given earlier.

(2) Connect one end of a rubber hose to a source of dry, carbon dioxide - free air. Insert the other end of the hose into the sample tube and gently force air through the sample tube until all liquid has been evaporated and no ammonia gas remains in the tube.

(3) Wipe a counterpoise and the sample tube containing the residue to substantially the same extent to remove moisture which may have condensed on their outer surfaces.

(4) Weigh the sample tube and residue to nearest milligram.

(5) Calculate the weight per cent residue of the sample.

Calculation :

Per cent residue =

(W₂ - W₁) x F x 100
___________________
V x 0.682

Where

W₁ = weight (gm.) of tube (before sampling).

W₂ = weight (gm.) of tube and residue.

V = volume of sample taken (m1.)

F = Evaporation factor, taken from Table 1 corresponding to noted sample pressure when sample was obtained.

0.682 density of sample (gm./m1.)

(j) Precaution. -

A check of completeness of evaporation may be made by visual inspection and by carefully smelling the air in the tube making sure that no carbon dioxide or moisture from the breath enters the sample tube.

(k) Per cent ammonia content (by difference) -

In general, the ammonia content of the sample will : per cent ammonia = 100 - (Per cent water - per cent residue, if determined).

(l) Precaution. -

(ii) Determination of oil content

(a) Apparatus :

(1) 500 ml. conical flask calibrated to hold 300 ml. (about 200 gms.) of sample and with a bunk fitted with glass through which the exist gas can be led away to a safe place. Ensure that the glass tube and exist line are free from constructions.

(2) Platinum dish 75 mm. in diameter.

(b) Reagent :

The reagent used shall be of a recognised analytical reagent quality like petroleum spirit with a boiling range of 40°C to 60°C.

(1) Quickly running the sample (as per the sample procedure) up to the calibration mark, insert the bunk.

(2) Immerse the flask in a continuous stream of cold water and allow the ammonia to evaporate slowly.

(3) When the evaporation is complete, remove the bunk and gently blow out the last faces of ammonia with a small jet of filtered air free from carbon dioxide.

(4) Dry the outside of the flask.

(5) Place the flask in an oven at 105±5°C for 15 minutes to remove the moisture blowing out the last traces with a gentle stream of filtered air at the end of this period ; allow to cool.

(6) Add to the flask approximately 100 ml. of the petroleum spirit and swirl to dissolve the oil.

(7) Filter the solution through a small filter paper (a Whatman No. 31 paper is suitable) direct into the platinum dish previously traced to the nearest 0.1 mg.

(8) Repeat the operation with two more successive 10 ml. portions of petroleum spirit, filtering through the same paper as before into the same dish.

(9) Evaporate the combined extracts to dryness on a water both in a fume cup board, taking care to avoid naked lights and finally dry in an over at 105±°C for 30 minutes.

(10) Allow the dish to cool thoroughly in a descicator and weight against to the nearest 0.1 mg.

(11) At the same time, can), out a blank determination on the petroleum spirit and filter paper.

(12) Make an appropriate correction in the calculation.

Calculation :

Oil content, parts per million by weight =

(W₁ - W₂) - W₃
__________________
0.68 V₂

x 10⁶F

Where

W₁ = weight in grams of platinum dish and residue.

W₂ = weight in grams of dish alone.

W₃ = weight in grams of oil in blank determination.

V₂ = volume in ml. of sample taken.

F = evaporation factor taken from Table corresponding to noted sample pressure when sample was obtained.

Table

Evaporation factor for different vessels or line pressures.
Vessel or Line Pressure (Psig.)	Evaporation Factor (F)
1	2
0	1.000
10	0.963
20	0.940
30	0.920
40	0.900
50	0.885
60	0.870
70	0.860
80	0.850
90	0.840
100	0.830
110	0.821
120	0.813
130	0.805
140	0.797
150	0.789
160	0.782
170	0.776
180	0.770
190	0.764
200	0.758

Note. - When a sample of liquid ammonia is transferred to an open flask/tube from a container in which the pressure is higher than the atmospheric pressure that sample quickly reaches the thermodynamics equilibrium with its new environment. Some of the liquid ammonia will evaporate and since the ammonia so lost as vapour contains non - volatile constituents present in the original product, the concentration of these constituents will increase in the liquid sample taken. This may be significant factor in subsequent analysis and fortunately it is possible to calculate from the known thermodynamic properties of ammonia an appropriate correction. This correction is usually known in this context at the evaporation factor of flask factor and is simply that fraction by weight of the original liquid ammonia which remains as liquid in the sample Multiplication of the determined quantity of a non - volatile constituents (oil, water dissolved or suspended solids, etc.) by the evaporation factor gives a result which will be closer to the true figure. The Table above presents the evaporation factor for different vessel or line pressure.

When a sample is transferred (example by gravity) from a container of a cylinder or the sampler at the same pressure, there should be no loss by evaporation and in this case it is not necessary to use evaporation factor :

7. Method of analysis of zinc sulphate, [Both Heptahydrate and Monohydrate]

(i) Quality of reagents

Pure chemicals and distilled water shall be used in tests.

Note. - "Pure chemicals" shall mean chemicals that do not contain impurities which effect the results of analysis.

(ii) Determination of matter insoluble in water

(a) Reagents :

Dilute Sulphuric Acid - 10 percent.

(b) Procedure. -

Dissolve 25.0 gms. of the material in 125 ml. of water and add 1 ml. of dilute sulphuric acid. Heat the solution to boiling, filter through a weighed and prepared Gooch crucible or sintered glass crucible (G.No. 4) and wash the residue thoroughly with hot water. Dry the crucible at 110" ±5" to constant mass.

Calculation:

Matter insoluble in water,

per cent. by mass weight = 4A

Where,

A = weight in gm. of the residue

[(iii) Determination of Zinc in Zinc Sulphate by modified EDTA titration method] -

(a) Reagents :

(1) EDTA Solution. - Dissolve 3.72 gms. of Di - sodium Ethylene Diamine Tetraacetate dihydrate in distilled water and make up the volume to 1 litre.

(2) Standard Zinc metal solution. - Weight about 1.0 gm. of zinc metal and record the weight accurately. Express this as W. Add HCl (1:1) @20 ml.per gm. of zinc metal. Keep it for few hours and allow it to dissolve completely. Make up the volume of the solution to exactly 100 ml.

(3) Ammonium Hydroxide (20 per cent) (m/m/).

(4) Ammonium Chloride - AR grade salt.

(5) [Sodium Cyanide] - AR/GR grade salt.

(6) Eriochrome Black (T) indicator mixture.

Mix thoroughly 1 gm. of Eriochrome Black (T) indicator with 100 gm. of AR Grade sodium chloride.

(7) Formaldehyde - Acetic Acid solution (4 per cent.). - Dissolve 100 ml. of Formaldehyde (37 - 40 per cent) in about 100 ml. of distilled water. Add 40 ml. glacial Acetic acid and make volume to 1 litre with distilled water.

(8) Hydroxylamine Hydrocloride . - AR Grade solid salt.

Note. - Distilled water means glass distilled water.

(b) Procedure. -

(1) Standardization of EDTA solution :

(A) Take 10 ml. of Zinc metal solution (standard).

(B) Add about 0.1 gm. of ammonium chloride and 30 ml. of ammonium hydroxide solution (20 per cent).

(D) Add a pinch of eriochrome black (T) indicator mixture. It will give red colour.

(E) Titrate it with EDTA solution to obtain clear blue end point. Note the volume of EDTA used as V₁ ml.

(2) Estimated of Zinc in sample :

(A) Weigh accurately 1.0 gm. of given zinc sulphate sample and dissolve it in 100 ml. of distilled water in a volumentric flask.

(B) Take 10 ml. of alquote in beaker. Add 0.1 gm. of hydroxylamine hydrocholoride and 0.1 gm of ammonium chloride.

(C) Cautiously add small quantity of sodium cyanide. White precipitate will appear. Continue adding sodium cyandic till white precipitate disappears while swirling the beaker with hand. Add about 0.5 gm. excess of sodium cyanide.

(D) Dilute it by adding about 30 ml. of ammonium hydroxide (20 per cent.) and add about 30 ml. of distilled water.

(E) Add a pinch of eriochrome black (T) indicator mixture. It will give red colour.

(F) Titrate with EDTA solution till there is a sharp change to violet colour. Note the volume of EDTA used as V₂ (ml.).

(G) Add 20 ml. of Formaldehyde - acetic acid solution into above titrate solution and mix well. Red colour will reappear.

(H) Titrate it with EDTA solution to get blue end point without red tinge. Note the volume of EDTA used in second titration as V₃ ml.

Calculation :

Zinc % = 10/V₁= X V₃ X x W

W = Weight in gm. of piece of Zinc metal taken for preparation of standard zinc solution.

V₁= Volume of EDTA solution (in ml.) used for 10 ml. of Zinc metal solution.

V₃= Volume of EDTA solution (in ml.) used for second titration.]

(iv) Calorimetric Method for Determining of Copper

(a) Outline of the method - Sodium diethydithio carbonate reacts with slightly acidic or ammoniacal solution of copper in low concentration to produce a brown colloidal suspension of the cupric diethyl - dithiocarbonate. The suspension can to be used with extreme care. be extracted with an organic solvent and the colour extracted determined spectrophotometrically.

(b) Apparatus :

Photometer - Any suitable photoelectric calorimeter.

(1) Ammonium Citrate Solution - 40 per cent (m/v)

(2) Sodium Diethydithiocarbamate Solution - 0.1 per cent (m/v).

(3) Standard Copper Solution - Clean the surface of a copper wire with sand paper. Dissolve 100 gms of pure copper in dilute nitric acid. Heat the solution nearly to dryness to drive off the acid. Add about 10 ml. of water and heat again nearly to dryness. Take up the residue in water , add 25 ml. of IN sulphuric acid and dilute to 1000 ml. One millilitre of this solution contains 0.1 mg. of copper (as Cu).

Alternatively, dissolve 3.928 gms. of cupric sulphate (CuSO₄.5H₂O) in sufficient water containing 1 or 2 ml. of concentrated Sulphuric acid and dilute to 1000 ml. One millilitre of the solution contains 1.0 mg. of copper (as Cu).

(4) Ammonium Hydroxide - 18 N.

(5) Carbon Tetrachloride.

(d) Preparation of sample. -

(1) Weigh 5 gms. of sample accurately, dissolve in distilled water and add 1 ml. of sulphuric acid. Filter the solution and make up to 250 ml. with water in a volumetric flask.

Take 50 ml. of the above solution in a beaker, heat, pass hydrogen sulphide gas or add sodium sulphide solution and ensure complete precipitation. Filter hot and keep the filtrate for subsequent analysis.

(2) Boil the reside with dilute nitric acid and filter, if necessary. To the filtered solution add sulphuric acid, evaporate, dilute and filter. Keep the residue for determination of lead.

(e) Procedure of determination of copper -

Determine copper in the filterate form (d) (2) above by diethyldithiocarbonate method as detailed below. -

Transfer an aliquot containing not more than 0.65 mg. of copper to a 100 ml. separating funnel. Add 2 ml. of ammonium citrate solution and adjust the pH to 8.5 with ammonium hydroxide. The presence of ammonium ions helps to dionize iron, if present and to prevent rapid fading of developed colour. Add 10 ml. of sodium diethyldithiocarbamete solution and develop the colour. Extract the coloured complex with 5 - 10 ml. of carbon tetrachloride. Separate the carbon tetrachloride layer from aqueous layer and centrifuge for 5 minutes to separate the water droplets. Transfer the carbon tetrachloride solution to a photometric cell and measure its absorbance at the wavelength of maximum absorption 440 mu relative to reagent blank. Calculate the corrected absorbance by subtracting the reading obtained for the solution containing - no copper.

Transfer to a series of 100 ml. separating funnels aliquots of standard copper solution corresponding to 0, 0.1, 0.2, 0.5, 1.0, 2.0, 2.5 gms. of copper and proceed exactly as prescribed above. Plot a graph of corrected, absorbance of solutions against their copper contents.

Note. - If Bismuth is suspected it has to be separated or otherwise accounted for.

Calculation. - Calculate the corrected absorbance of substracting the value obtained for the black from that obtained for the test solution and read from the calibration curve the corresponding mass of copper.

Copper (as Cu). per cent by mass =

M₁
_____
M₂

x 100

Where

M₁= mass in gm. of copper as determined in the given aliquot of the test solution,and

M₂ = mass in gm. of the material present in the aliquot of the test solution.

(v) Determination of lead by colorometric method using dithizone.

(a) Apparatus :

Nessler Cylinders - 50 ml. capacity.

(b) Reagents :

(1) Standard Lead Solution. - Dissolve 0.40 gm. of lead nitrate [Pb (NO₃)₂] in water containing 2 or 3 ml. of concentrated nitric acid and make up the volume to 1000 ml. with water. Transfer 10 ml. of this solution to a volumetric flask, add 2 or 3 ml. of concentrated nitric acid and dilute with water to 1000 ml. one millilitre of this solution contains 2.5 mgs. of lead (as Pb). The diluted solution shall be freshly prepared.

(2) Dilute Ammonium Hydroxide. - IN, approximately.

(3) Reagent A - Dissolve 25 gms. of triammonium citrate or 22 gms of citric acid and 4 gms. of hydroxylamine hydrocholoride in about 200 ml. of water. Add dilute ammonium hydroxide to bring the pH to 8.5 Dilute the solution to 500 ml. Purify this solution by extracting with 15 ml. portions of 0.01 per cent dithizone solution until the final colour of the dithizone extract is green. Wash the aqueous reminder portion three times with 25 ml. portions of chloroform and finally with 25 ml. portion of carbon tetrachloride.

(4) Dithizone (Dephenyl thiocarbazone or phenylazothiomoformic Acid) Solution - Dissolve 0.01 gm. of dithizone in 100 ml. of carbon tetrachloride, shaking intermittently for 1 hour. Allow to stand overnight and shake once again before using. This shall be kept in a cool and dark place. This gives a 0.01 per cent solution. Filter, if necessary. Dilute 10 ml. of this solution to 100 ml. with carbon tetrachloride in a 100 ml. volumetric flask. This shall be prepared fresh before determination. This gives a solution of 0.001 per cent.

Note 1. - Carbon tetrachloride used should be further purified. One litre of carbon tetrachloride is extracted with two portions of 25 ml. dilute ammonium hydroxide and then kept over 100 gms. to activated carbon. Before use, it is decanted and distilled at about 80°C over a little fresh lime.

Note 2 - Sometimes dithzone solid and its 0.01 per cent solution deteriorate on storage. The 0.01 per cent solution should, therefore, be tested before further dilution, by shaking 2 ml. of the solution with 5 ml. of 1 per cent ammonium hydroxide. If the organic layer is only faintly yellow under these conditions, the solution may be used. If it is deeply coloured it shall be discarded and fresh solution be prepared. The solution as well as the reagent should be stored in a refrigerator and exposure to sunlight should be avoided during analytical work. To increase the stability of 0.01 per cent solution, it should be covered with a thin aqueous layer saturated with sulpur dioxide.

(5) Thymol Blue Indicator Solution. - 1 per cent (m/v) solution in rectified spirit.

(1) Take several aliquots of standards lead solution into a series of separating funnels and 5 ml. of the water and 10 ml. of the Reagent A. Then add 2 drops thymol blue indicator and bring the pH of the solution to 8.5 by addition of dilute ammonium hydroxide. Add 5 ml. of the dithizone solution and shake well for about 10 seconds. Drain the organic layes into stoppered Nessler cylinders.

(2) Dissolve the residue obtained in (iv) (d) (2) above, in dilute nitric acid and dilute with water. Take a suitable aliquot of the solution and transfer to a separating funnel. Develop the colour as prescribed above. Drain the organic layer into a stoppered Nessler cylinder, add 10 ml. of dilute ammonium hydroxide, and shake for about 10 seconds. Compare the colour developed with those of the standard solutions. Note the volume of the standard with which the colour of test solution matches.

Note. - If the colour of the test solution is intermediate between two standard solutions, then the experiment is repeated by taking more number of standard solutions in that range and exact colour matching is arrived at.

Calculation :

Heavy metals as (PB), per cent by mass =

100 x V x f
_____________
M

where

V= Volume in ml. of standard lead solution matching with the test solution.

f= mass in gm. of heavy metals (as Pb) equivalent to 1ml of standard lead solution, and

M = mass in gm. of the material in the aliquot taken for the test.

(vi) Determination of magnesium in the sample material

(a) Reagents :

(1) Eriochrome Black T Indicators. - Dissolve 0.1 gm. of eriochrome black T in 25 ml. or methyl alcohol.

(2) Ammonium Hydroxide Ammonium Chloride Buffer Solution - Mix. 350 ml. of ammonium hydroxide (20 per cent w/w) with 54 gms. of ammonium chloride. Dilute with water and make up the volume to 1000 ml. (The PH of the solution should not be more than 10).

(3) Standard Magnesium Solution. - 0.01 M Weigh 2.4640 gm. of magnesium sulphate (MgSO₄.7H₂O) and dissolve it in water. Make up the volume to one litre.

(4) Ethylenediamine Tetra Acetate (EDTA) Solution. - Dissolve 3.72 gms. of disodium ethylenediamine tetra acetate dihydrate in water and make up the volume to one litre.

(b) Procedure. -

Standardization of EDTA Solution. - Take 10 ml. of standard magnesium solution in a conical flask. Add 20 ml. of water, one millitre of eriochrome black T indicator and 25 mi. of ammonium hydroxide ammonium chloride buffer solution. Heat at 40°C to 50°C and then titrate with EDTA solution, maintaining the temperature between 40°C and 50°C until the colour changes from wine red to distinct blue. IOM

Molarity of EDTA solution =

10M₁
______
V₁

where

M₁ = molarity of standard magnesium solution, and

V₁ =Volume in ml. to EDTA solution used for titration.

(c) Determination of Magnesium in the sample material - Take the filterate from (iv)(d) (1) (after removal of copper) add a few drops of concentrated nitric acid, boil and cool and then add solid ammonium chloride (about 2 grams). boil and cool add ammonium - hydroxide till the strong smell of ammonia comes and filter the precipitate through sintered crucible, take filterate and add dilute sulphuric acid till the solution is acidic (text with methyl red), heat the solution to boil and add excess of di - ammonium - hydrogen phosphate with continuous stirring. Add 10 percent ammonia solution with continuous stirring till the solution is just alkaline (test with methyl red) while precipitate of zinc ammonium phospate will be formed the optimum pH for precipitation is 6 - 7, allow it to stand for 3 - 4 hours. Filter through Whatman filter paper No. 40. Collect the filterate in a volumetric flask. Make up the volume (say 100 ml.) Take a suitable alquote (say 10 ml.) for the determination of magnesium. Add 20 ml. of water, one ml. of aricohrome black - T indicator and 20 ml of ammonia hydroxide ammonia chloride buffer solution. Heat to 40° to 50°C and titrate with standard EDTA solution maintaining the temperature between 40°C and 50°C until the colour changes from wine red to distinct blue.

1 ml. of 0.01 M - EDTA = 0.2432 mg. of "Mg" Mg =

X x 0.2432
___________
5

where

(vii) Determination of pH

Dissolve 5 gm. of the material in freshly boiled and cooled water. Dilute to [100 ml.]

and mix. Determine the pH value of the solution with a pH meter.

[(viii) Determination of Iron in the sample material (Spectro - Photometric Method)].

(This method is applicable for Iron contents up to 1mg. in the final aliquot)

(a) Outlines of the method - In test solution, iron is determined absorptions metrically as Ferrous Iron form red complex with Ortho - phenanthroline.

(b) Apparatus. - Photo - electric colorimeter.

(1) Bromophenol - blue indicator solution - 0.4 per cent solution in 95 per cent ethanol.

(2) Sodium Citrate solution - 25 per cent.

(3) Hydroquinone solution - 1 per cent in a Acetic acid buffer of pH 4.5 obtained by mixing 65 ml. of 0.1 M Acetic acid with 35 ml. of 0.1 M Sodium Acetate solution.

(4) Ortho - phenantroline solution - 0.25per cent in 25 per cent ethanol.

(5) Dilute Hydrochloric acid - 1:3 (VN).

(6) Standard Iron solution - Dissolve 0.7002 gm. of Ammonium Ferrous Sulphate (AR) in distilled water containing 2 ml. of perchloric acid and dilute to 100 ml. with water. One ml. of this solution contains 1 mg. of Iron

(d) Preparation of sample solution. - Boil 1 gm. sample in 20 ml. of Hydrochloric Acid (1:1) for 30 minutes and filter through Whatman No. 40 filter paper in 100 ml. volumetric flask. Follow with repeated extracted with boiling dilute Hydrochloric acid (1:5) decanting the solution each time through the same filter paper. Dilute the filtrate to 100 ml.

(e) Procedure -

(1) Take 5 ml. of the sample solution, add bromophenol blue indicator solution and titrate with Sodium Citrate solution until yellow colour changes to blue.

(2) Take another 5 ml. of sample solution in a 25 ml. volumetric flask. Add 1 ml. of Hydroquinone solution, 3 ml. of Othrophenanthroline solution and an amount of Sodium Citrate solution equivalent to the above titration in step (1), Dilute with distilled water to 25 ml. Allow the solution to stand for one hour.

(3) Measure the absorbance of test solution at 510 nm using 1 cm. cell and also blank with water.

(4) Place 4, 8, 12, 16 and 20 ml. of standard Iron solution in a series of 100 ml. volumetric flasks. To each add 40 ml. of diluted Hydrochloric acid and dilute to 100 ml. with water.

(5) Using ml. aliquote proceed as for sample solution.

(6) Measure the absorbance of the solution and plot a graph relating to absorbance to number of mgs. of Iron.

Calculation :

Per cent Fe = 2x/W

where,

X= Concentration (in mg.) of Iron in final sample solution aliquot as determined from the graph.

W= Weight in gm. material taken for the preparation of sample solution.]

8. Alternate method of analysis of zinc - sulphate, [Both Heptahydrate and Monohydrate]

(i) Quantity of reagents

(1) Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests.

Note. - "Pure chemicals" means chemicals that do not contain impurities which affect the results of analysis.

"Demineralised water" means the water obtained after passing distilled water through a cation and a anion exchange resins or a combined cationanion exchange resin.

(ii) Determination of Zinc

(a) Reagents :

(1) Standard Zinc solution. - Weigh 0.4398 gm. of zinc sulphate (ZnSO₄.7H₂O - A.R. grade) on a clear watch glass and transfer it to one litre flask through the funnel giving several washings to watch glass and funnel with glass distilled or demineralised water. Add one ml. of 10 per cent sulphuric acid (A.R. Grade) and make the volume upto the mark. Stopper the flask and shake the solution well. This is 100 ppm Zinc solution hereinafter called Standard A. This solution should be stored in a clean bottle for further use. Dilute 10 ml. of 100 ppm solution of zinc (Standard A) to 100 ml. to get 10 ppm standard zinc solution designated as Standard B.

(2) Glass distilled or mineralised water of pH 2.5 ±0.5. - Dilute 1 ml. of 10 per cent sulphuric acid to one litre with glass distilled or mineralised water and adjust the pH to 2.5. with a pH meter using H₂O₄ or NaOH. This solution is called acidified water and and 5 to 10 litre of this solution should be prepared at a time.

(3) Preparation of working standards. - Pipette the following volume of Standard B in 50 ml. numbered volumetric flask and make the volume with acidified water.

Flask No.	Volume of Standard B taken (ml.)	Concentration of zinc after making volume to 50 ml. (ppm)
1	0.0	0.0
2	1.0	0.2
3	2.0	0.4
4	3.0	0.6
5	4.0	0.8
6	5.0	1.0
7	6.0	1.4
8	9.03	1.8
9	10.0	2.0

Stopper the flasks and shake them well. Prepare the standard in duplicate. The same acidified water should be used for preparing the solution of unknown fertilizer samples. Fresh standards should be prepared every time when a fresh lot of acidified water is prepared.

(b) Procedure. -

(1) Preparation of Zinc Sulphate fertiliser samples. - Weigh 0.25 gm. of the material on a clean watch glass and transfer it to one litre volumetric flask through the funnel giving repeated washings with glass distilled water and dissolve the material by shaking well. Then make the volume up to mark with glass distilled water and shake well.

(2) Take 5ml. of the prepared solution in 250 ml. volumetric flask and make the volume with acidified water. Shake the solution well and filter through Whatman No. 42 filter paper in dry clean flasks. The flasks should be rinsed with a 10 to 15 ml. of the filterate and then continue filtration.

(3) Flaming the solutions. - Flame the standards and the filtered samples on atomic absorption spectrophotometer as a wavelength of 213.8 mu (Zn line of the instrument).

Calculations : Prepare a standard curve of known concentration of zinc solution by plotting the absorbance values on Y - axis against their respective zinc concentration on X - axis. Calculate the percentage zinc in zinc fertilizer by multiplying zinc concentration value calculated from standard curve by 20.

Example:

Weight of fertiliser sample	= 0.25 gm.
Volume made	= 1000sml.
Further dilution	= 5.0 times
Reading of the samples from Atomic Absorption	= Y
Corresponding, concentration value of zinc from standard	= X ppm
curve against Y absorbance
Percentage Zinc in the fertilizer	= 20(X)

(d) Precautions

(1) Weighing must be done on an electric balance.

(2) All the glass apparatus to be used should be of corning make and washed with dilute hydrochloric acid (1:4) and washed thoroughly with distilled and then with demineralised water.

(3) The pipette should be rinsed with the same solution to be measured.

(4) The outside of the pipette should be wiped with filter paper after taking out from the solution to be measured.

(5) After using the pipette, place than on a clean dry filter paper in order to prevent contamination.

(6) To start filtration, only a few drops should be added first in order to the one filter paper and then continue further filtration.

(iii) Determination of magnesium

(a) Reagents :

(1) Strontium chloride. - Dissolve 7.5 gms. of strontium chloride (SrCl₂.6H₂O) in one litre of glass distilled water.

(2) Standard Magnesium solution. - Weigh 0.507 gm. of magnesium sulphate (MgSO₄.7H₂O) one a clean watch glass and transfer it to one litre, flask through the funnel giving several washings to watch glass and the funnel with glass distilled or demineralised water. This is 50 ppm Mg. solution Dilute 10 ml. of 50 ppm solution of Mg to 10 ml. to get 5 ppm standard Mg. solution.

(3) Preparation of working standards. - Pipette the following volume of 5 ppm standard Mg. solution in 50 ml. numbered volumetric flasks. Add 10 ml. of strontium chloride solution to each flask and make up the volume to 50 ml.

Flask No.	Volume of 5 ppm Mg. solution taken (m1.)	Volume of strontium chloride added (ml.)	Concentration of magnesium after making the volume to 50 ml. (ppm)
1	0.0	10.0	0.0
2	2.0	10.0	0.2
3	4.0	10.0	0.4
4	6.0	10.0	0.6
5	8.0	10.0	0.8
6	10.0	10.0	1.0

Stopper the flask and shake them well. Prepare fresh standards every fortnight.

(b) Procedure. -

(1) Pipette 20 ml. of the solution which was prepared for the determination of zinc by dissolving 0.25 gm. of the fertilizer sample in one litre flask (Step A - 2.2.1). Add 10 ml. of strontium chloride. Make up the volume to 50 ml.

(2) Flame the standards and the samples on atomic absorption spectrophotometer at a wavelength of 285.5 mu (Mg. line of the instrument).

Calculations :

Prepare a standard curve of known concentrations of Mg. solutions by plotting the absorbance value on Y - axis against their respective concentration values on X - axis. Percentage magnesium in the zinc fertilizer will correspond to the concentration values calculated from the standard curve.

Example :

Weight of the fertiliser	=0.25gm.
Volume make	=1000 ml.
Further dilution	= 2.5 time
Reading of the sample from atomic absorption spectrophotometer	=Y
Corresponding, concentration of Mg. from standard curve against Y absorbance	= X ppm
Percentage magnesium in the fertiliser	=X

(iv) Determination of copper

(a) Reagents :

(1) Standard Copper Solution. - Weigh 0.1965 gm. of copper sulphate (CuSO₄.5H₂O) on a clean watch glass and transfer it to one litre flask through the funnel giving several washings to watch glass and the funnel with glass distilled water. Add one ml. of 10 per cent sulphuric acid and made up the volume up to the mark. Stopper the flask and shake the solution well. This is 50 ppm Cu solution and should be stored in a clean bottle for further use. Dilute 10 ml. of 50 ppm solution of copper to 100 ml.to get 5 ppm standard copper solution.

(2) Glass distilled or mineralised acidified water of pH 2.5± 0.5 (same as given in 8(ii) (a)(2)).

(3) Preparation of working standards. - Pipette the following volume of 5 ppm standard copper solution in 50 ml. numbered volumetric flask and make the volume with acidified water.

Flask No.	Value of 5 ppm standard Cu solution taken (ml.)	Concentration of copper after making volume to 50 ml. (ppm)
1	0.0	0.0
2	2.0	0.2
3	4.0	0.4
4	6.0	0.6
5	8.0	0.8
6	10.0	1.0

Stopper the flasks and shake them well. Prepare fresh standards every fortnight.

(b) Procedure. -

(1) The solution with was prepared for the determination of zinc by dissolving 0.25 gm. of the fertilizer sample in one litre flask Step 8(ii)(b)(1) should be used for the determination of copper.

(2) Flame the standards and the samples on an atomic absorption spectro - photometer at the wavelength of 324.8 mu (Cu line of the instrument).

Calculation:

Prepare a standard curve of known concentrations of copper solutions by plotting the absorbance values on Y - axis against their respective concentration values on X - axis. Calculate the percentage copper in the zinc fertilizer by multiplying the copper concentration value calculated from the standard curve by 0.4.

Example :

Weight of the fertilizer sample	=0.25gm.
Volume made	=1000 ml
Reading of the samples from atomic absorption spectrophotometer	=Y
Corresponding, concentration of copper from standard curve against Y	= X ppm
absorbance percentage copper in the fertilizer	=0.4X

(v) Determination of lead

(a) Reagents :

(1) Standard Lead Solutions. - Weight 0.1965 gm. of lead nitrate [Pb(NO₃)₂] on a clean watch glass and transfer it to one litre flask through the funnel giving several washings to watch glass and funnel with glass distilled or demineralised water. Add 10 ml. of concentrated distilled nitric water. Add 10 ml. of concetrated distilled nitric acid and make the volume up to the mark. Stopper the flask and shake the solution well. This is 100 ppm lead solution and should be stored in a clean bottle for the further use. Dilute 10 ml. of 100 ppm solution of lead to 100 ml. with 1 per cent nitric acid solution to get 10 ppm standard lead solution.

(2) per cent nitric acid solution. - Dilute 10 ml. of concentrated distilled nitric acid to one litre with glass distilled water.

(3) 20 per cent zinc sulphate solution. - Weigh 20 gms. of zinc sulphate (ZnSO₄.7H₂O) and dilute to 100 ml. with 1 per cent nitric acid solution.

(4) Preparation of working standards. - Pipette the following volume of 10 ppm standard lead solution in ml. numbered volumetric flask. Add 5 ml. of 20 per cent zinc sulphate solution to each flask and make the volume with 1 per cent nitric acid solution.

Flask No.	Volume of 10 ppm lead solution taken (ml.)	Volume of 20% zinc sulphate solution added (ml.)	Concentration of lead after making the volume to 50 ml. (ppm)
1	0.0	5.0	0.0
2	2.0	5.0	0.4
3	4.0	5.0	0.8
4	6.0	5.0	12
5	8.0	5.0	1.6
6	10.0	5.0	2.0

Stopper the flasks and shake them well.

(b) Procedure. -

(1) Preparation of zinc sulphate fertiliser samples. - Weigh 1 gm. of the material on a clean watch glass and transfer to 50 ml. volumetric flask through the funnel giving washings with 1 per cent nitric acid solution. Dissolve the material and make the volume with 1 per cent nitric acid solution. Samples should be prepared in duplicate.

(2) Flaming the solutions. - Flame the standards and samples on atomic absorption spectrophotometer at a wavelength of 217 mu (Lead line of the instrument.)

Calculations : Prepare a standard curve of known concentrations of lead solution by plotting the absorbance values on Y - axis against their respective lead concentration on X - axis. calculate the percentage lead in zinc fertilizer by multiplying lead concentration value calculated from standard curve by 0.005.

(vi) Determination of pH

(a) Procedure :

Dissolve 5 gm. of the material in freshly boiled water. Dilute to [100 ml] and mix. Determine the pH value of the solution with pH meter.

(vii) Determination of matter insoluble in water

(a) Procedure :

Dissolve 25.0 gms. of the material in 125 ml. of water. Filter through a weighed and prepared Gooch crucible or sintered glass crucible (G. NO. 4) and wash the residue thoroughly with water. Dry the crucible at 110°±8° to constant mass.

Calculations :

Matter insoluble in water per cent by weight = 4 - A.

Where,

A = Weight in gm. of the residue.

[(viii) Determination of Iron (Atomic Absorption Spectrophotometric method)]

(a) Reagents. -

(1) Standard Iron Solution (1000 ppm.) - - Weigh accurately 1 gm. pure Iron wire and put it in approximately 30 ml. of 6N HC1 in a beaker and boil. Transfer it to one litre volumentric flask through the funnel giving several washings to the beaker and funnel with glass distilled water. Make the volume up to the mark. Stopper the flask and shake the solution well. This is 1000 ppm Iron solution.

(2) Glass distilled or demineralised water of pH 2.5±5.0.

(3) Preparation of working standards. - Pipette 10 ml. Iron stock solution in 100 ml. volumentric flask and dilute to volume. This is 100 ppm Iron solution. Pipette the following volumes of 100 ppm Iron solution in 50 ml. volumetric flask and make the volume with acidified water.

Flask No.	Volume of 100 ppm standard Iron solution	Constriction of Iron atter making volume 50 ml. (ppm)
1	0.0	0.0
2	1.0	2.0
3	2.0	4.0
4	3.0	6.0
5	4.0	8.0
6	5.0	10.0
7	6.0	12.0
8	7.0	14.0
9	8.0	16.0
10	9.0	18.0
11	10.0	20.0

Stopper the flask and shake the solution well.

(b) Procedure. -

(1) Preparation of fertiliser sample solution. - Weigh exactly I gm. of the material on a clean watch glass and transfer it to a one volumetric flask through the funnel giving repeated washing with acidified water and dissolve the material by shaking well. Make the volume up to the mark with acidified water and shake well and filter through. Whatman No. 42 filter paper in dry clean flask. The flask should be rinsed with a 10 - 15 ml. of filtrate and then continue filtrarion.

(2) Flaming the solution. - Flame the standards and the filtered sample on Atomic Absorption Spectrophotometer at a wavelength of 248.3 mm. using clean in air acetylene flame.

Calculations : Prepare a standard curve of known concentrations of Iron solution by plotting the absorbance value of Y - axis against the respective iron concentration on X - axis: Determine the concentration of Iron in the sample solution from the graph.

Total Iron (as Fe) per cent =X/10

Where X= Concentration of Iron (in ppm) obtained from the standard curve.

Note. - In case a sample has been analyzed by both the methods. viz. indicated under the heading '7' and '8' the result obtained by the method indicated under the heading `8'shall prevail.

9. Method of analysis of manganese sulphate, agricultural grade -

(i) Quality of Reagents

Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests.

Note. - "Pure chemicals" means chemicals that do not contain impurities which affect the results of analysis. "Demineralised water" means the water obtained after passing distilled water through a cation and a anion exchange resins or a combined cation anion exchange resin.

(ii) Determination of Manganese

(a) Reagents :

(1) Standard Manganese Solution. - Weigh 3.0763 gms. of manganese sulphate (MnSO₄.H₂O) - A.R. grade) on a clean whatch glass and transfer it to one litre flask through the funnel giving several washings to watch glass and funnel with acidified water and make the volume up to the mark. This solution will be 100 ppm Mn. A secondary dilution of 5 ml. to 100 ml. with acidified water gives 1 50 ppm working standard.

(2) Glass distilled or demineralised water of pH 2.5±0.2 - Dilute 1 ml. of 10 per cent sulphuric acid to one litre with glass distilled or demineralised water and adjust the pH to 2.5 with a pH meter using 10% H₂SO₄ of NaOH. This solution is called acidified water.

(3) Preparation of working standard. - Pipettee the following volume of working standard solution in 50 ml. numbered volumetric flasks and make the volume with acidified water.

Flask No.	Volume of working standard taken (ml.)	Concentration of maganese after making volume to 50m1. (ppm)
1	0.0	0.0
2	0.5	5.0
3	1.0	1.0
4	1.5	1.5
5	2.0	2.0
6	25	2.5
7	3.0	3.0
8	3.5	35
9	4.0	4.0

Stopper the flasks and shake them well. Prepare the standard in duplicate. The same acidified water should be used for preparing the solution of unknown fertiliser samples. Fresh standards should be prepared every time when a fresh lot of acidified water is prepared.

(b) Procedure. -

(1) Preparation of manganese sulphate fertiliser samples. - Weight exactly 0.4 gm. of the material on the clean watch glass and transfer it to a one litre volumetric flask through the funnel giving repeated washings with acidified water and dissolve the material by shaking well. Make the volume up to the mark with acidified water and shake well.

(2) Take 5 ml. of the prepared solution in 250 ml. volumetric flask and make the volume with acidified water. Shake the solution well and filter through What man No. 42 filter paper in dry clean flasks. The flasks should be rinsed with a 10 to 15 ml. of the filtered and then continue filtration.

(3) Flaming the solution. - Flame the standards and the filtered samples on atomic absorption spectrophometer at a wavelength of 279.5 mu(Mn line of the instrument).

Calculation :

Prepare a standard curve to known concentration of manganese solution by plotting the absorbance value on Y - axis against their respective manganese concentrations on X - axis. Calculate the percentage manganese in manganese fertiliser by multiplying manganese concertraion value calculated from the standard cover by 12.5.

(b) Precaution :

(1) Weighing must be done accurately with a precision balance.

(2) All the glass appratus to be used should be conrning/pyrex make and washed with dilute hydrocholoride acid (1:4) and washed thoroughly with distilled and then with demineralised water.

(3) The Pipette should be rinsed with the same solution to be measured.

(4) The outside of the pipette should be wiped with filter paper after pipetting the solution to be measured.

(5) After using the pipette place than on a clear dry filter paper, in order to prevent contamination.

(6) To start filtration, only a few drops should be added first in order to wet the filter paper and then continue further filteration.

10. Alternative method of analysis of managenese sulphate agricultural grade. -

(i) Quality of Reagents

Unless specified otherwise, pure chemicals and glass distilled or demineralized water shall be used in tests.

Note. - "Pure Chemicals" means chemicals that to not contain impurities which affect the results of analysis.

"Demineralised water" means the water obtained after passing distilled water through a cation and anion exchange resins or a combined cation - anion exchange resin.

(ii) Determination of manganese

(a) Reagents :

(1) Hydrogen peroxide (H₂O₂) - 30 per cent.

(2) Nitric acid (HNO₃) - Concentrated.

(3) Orthophsperic acid (H₂PO₄ - From steel industry) - 85 per cent.

(4) Potassium periodate (KIO₄)

(5) Glass distilled or mineralised water of pH 2.5±0.5 Dilute 1 ml. of 10 per cent H₂SO₄ to one litre with glass distilled or demineralised water and adjust the pH to 2.5 with a pH meter using 10% H₂SO₄ or NaOH.

(6) Standard Manganese solution. - Weigh 3.0763 gms. of manganese sulphate (MnSO₄.H₂O - A.R. grade) on a clear watch glass and transfer it to one litre flask through the funnel giving several washings to the watch glass and the funnel with acidified water and make the volume up to the mark. This solution will be 1000 ppm Mn. A secondary dilution of 5 ml. to 100 ml. with acidified water gives a 50 ppm working standard.

(7) Preparation of working standards. - Pipettee 0.0., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.5 and 5.0 ml. of standard solution of manganese in a series of 11 different 100 ml. beakers. Evaporate the solutions to dryness at low temperature on a hot plate or steam bath. Add 5 ml. of HNO₃ and 2 ml. of 30% H₂O₂ Cover the beakers with watch glass and digest the contents on a steam bath or hot plate for 30 minutes. Remove the glass cover and evaporate the contents to dryness. Cool the beakers, add 2 ml. of HNoO₃ 5m1. of phosphoric acid and heat till boiling. Again cool and mix the contents with 10 ml. of demineralised water. Add 0.3 of KlO₄ and heat the solutions on water bath until pink colour appears. Add about 20 ml. water and heat for about 40 minutes or until there is no more increase in colour intensity. Cool the beakers to room temperature, transfer the solutions to 50 ml. volumetric flasks. Make the volume with deiognised water and shake thoroughly. The concentration of manganese ion will be as follows :

Flask No.	Volume of working standard taken (ml.)	Concentration of manganese after making volume to 50 ml. (ppm)
1	0.0	0.0
2	0.5	0.5
3	1.0	1.0
4	1.5	1.5
5	2.0	2.0
6	2.5	2.5
7	3.0	3.0
8	3.5	3.5
9	4.0	4.0
10	4.5	4.5
11	5.0	5.0

Measure the light transmittance at 540 mu (green filter). Make the standard curve by plotting per cent transmittance against the concentration of manganese.

(b) Procedure. -

(1) Analysis of manganese sulphate fertiliser sample. - Weight exactly 0.500 gm. of the material on a clean watch glass and transfer it to a one liter volumetric flask through the funnel giving repeated washings with acidified water and dissolve the material by shanking well. Make the volume up to the mark with acidified water and shake well. Pipette one ml. solution in a 100 ml. pyrex/ corning beaker and proceed further as described above for preparation of standard curve and make the volume to 50 ml. The standard and the test samples should be prepared and processed for estimation at the same time.

Calculation :

The percentage of manganese in the manganese sulphate fertiliser sample is calculated by multiplying manganese concentration value calculated from standard curve by 10.

[11. Method of analysis of borax (sodium tetraborate decahydrate fertiliser)]

(i) Quality of reagents

Unless specified otherwise, pure chemicals & glass distilled water shall be used in tests.

Note : Pure chemicals means chemicals that do not contain impurities which affects the results of analysis.

(ii) Determination of boron :

(A) Atomic Absorption spectrophotometer method :

(a) Reagents :

(1) Standard Boron Solution. - Weigh 44.095 gms. of sodium tetraborate decahydrate (Na₂B₄O₇.10H₂O) - A.R. grade on clean watch glass and transfer it to one litre volumetric flask through the funnel giving several washings to watch glass and funnel with glass distilled water. Make the volume up to the mark. Stopper the flask and shake the solution well. This is 5000 ppm boron solution hereinafter called standard A. This solution should be stored in clean bottle for further use.

(2) Preparation of working standard. - Pipette the following volume of standard A in 50 ml. numbered volumetric flasks and make the volume with water.

Flask No.	Volume of standard A taken (ml.)	Concentration of boron after making volume to 50 ml. (ppm)
1.	0.0	0.0
2.	4.0	400
3.	6.0	600
4.	8.0	800
5.	10.0	1000
6.	12.0	1200
7.	14.0	1400
8.	16.0	1600

(b) Procedure. -

(1) Preparation of sample solution. - Weight 2.500 gms. of the material on a clean watch glass and transfer it quantitatively in a 250 ml. volumetric flask through the funnel giving repeated washing with water. Make up the volume stopper the flask and shake well.

(2) Flaming the solution. - Flame the standards and the sample solution on atomic absorption spectrophotometer at a wavelength of 249.8 nm using nitrous oxydeacetylene flame,.

Calculations :

Prepare a standard curve of known concentrations of boron solutions by plotting the absorbance value on Y - axis against their respective boron concentration on X - axis. Calculate the percentage boron in the given sample by multiplying boron concentration value obtained from standard curve by 0.01.

Example :

Weight of the fertiliser sample =2.50 gm.

Volume made = 250 ml.

Dilution factor 2 50/2.5 =100

Reading of sample from atomic absorption = Y Corresponding value of boron from standard curve against Y absorbance = X ppm percentage of boron in the sample = 0.01X.

(B) Titrametric method :

(a) Outline of the method :

Borax is determined by first converting it to boric acid with hydrocholoride acid and then titrating against Sodium hydroxide solution after complexing boric acid with mannitol or sorbitol.

(b) Reagents :

(1) Standard hydrochloric acid - 0.5 N.

(2) Standard sodium hydroxide - 1.0 N.

(3) Methyl red indicator - Dissolve 0.1 gm. of the material in 60 ml. of rectified spirit and dilute with water to 100 ml.

(4) Phenolphthlein indicator - Dissolve 1 gm. of the material in 100 ml. of rectified spirit.

(5) Mannitol or sorbitol.

(1) Dissolve 3.0 gm. of the material accurately weighed in 60 ml. of water and titrate with hydrochloric acid, using methyl red solution as indicator.

(2) Boil and cool the solution.

(3) Add 20 gm. of mannitol or sorbitol, and titrate with sodium hydroxide, using phenophthalein solution as indicator.

Calculation :

Boron content per cent by Weight = 1.0819 V. N/W

V= Volume in ml. of standard sodium hydroxide used.

N = Normality of sodium hydroxide, and

W= Weight in gm. of the material taken for the test.

(Ref - ISI - 110 - 1980)

(iii) Determination of matter insoluble in water :

Same as method No. 8(viii) (a);

(iv) Determination of pH

Dissolve 3.8 gm. of material in water and make it to 100 ml. Measure the pH value of the solution with the help of suitable pH meter, using glass electrodes.

(v) Determination of lead:

Same as method No. (v) Substitute Zinc sulphate by borax.

12. Method of analysis of copper sulphate fertiliser (CuSO₄.5H₂O)

(i) Quality of reagents :

Unless specified otherwise pure chemicals and glass distilled or demineralised water shall be used in tests.

Note. - "Pure chemicals" means chemicals which do not contain impurities which affect the result of analysis. Demineralised water, means the water obtained after passing distilled water through a cation and anion exchange resin or a combined cation anion exchange resin.

(ii) Determination of copper :

(A) Atomic absorption Spectrophotometer method

(a) Reagents :

As specified in 8(iv)(a), (1), (2) and (3).

(b) Procedure. -

(1) Weigh 0.25 gm. of the material on a clean watch glass and transfer it to one litre volumetric flask through the funnel giving repeated washing with glass distilled water. Add one ml. of 10 per cent sulphuric acid and make up the volume.

(2) Take 5 ml. of the prepared solution in 500 ml. volumetric flask and make up the volume. Shake the solution well and filter through Whatman No. 42 filter paper in dry clean flask. The flask should be rinsed with a 10 to 15 ml. of the filterate and then continue filteration.

(3) Flaming the solution : Flame the standards and the filtered samples on an atomic absorption spectrophotometer at a wavelength of 324.8 mm. using air - acetylene flame.

Calculation:

Prepared a standard curve of known concentrations of Copper solution by plotting the absorbance value on Y - axis against their respective concentration values on X - axis :

Per cent, copper in simple = 10 x/w

where

W= Weight in gm. of the material taken for test.

X = Cone of copper (in ppm) as determined from the graph.

(B) By Iodometric Titration method :

(a) Outline of the method :

Copper is determined with the addition of postassium iodide and titrating the liberated iodine against standard sodium thio - sulphate solution.

(b) Reagents :

(1) Sodium Carbonate - A.R. Grade.

(2) Potassium Idodide - A.R. - Crystals.

(3) Acetic Acid - A R Grade.

(4) Standard Sodium Thiosulphate solution. - 0.1 N.

(5) Starch indicator solution. - Triturate 5 gm. of starch and 0.01 gm. of mercuric iodide with 30 ml. of cold water and slowly pour it with stirring into one litre of boiling water. Boil for there minutes. Allow to cool and decant off the supernatant clear liquid.

(6) Potassium thiocyanate crystals.

(c) Procedure. -

(1) Dissolve about I gm. of the test sample (accurately weighed), in 50 ml. of water.

(2) Add a pinch of sodium carbonate till a slight trubidity appears. Then add 5 ml. of acetic acid , 3 gms. of potassium iodide and titrate the liberated iodine with sodium thiosulphate solution, using starch as an indicator, until only a faint blue colour remains.

(3) Add about 2 gms. of potassium thiocyanate, shake and continue the titration until the blue colour disappears.

Calculation:

Copper per cent mass =

6.35 V.N.
_________
M

where

V= Volume in ml. of standard sodium thiosulphate solution.

N = Normality of standard thiosulphate solution, and

M = Mass in gm. of the sample taken for the test.

[Ref. IS - 261(1982)].

(iii) Determination of lead (Pb) - Same as method No. 8(v). Substitute zinc sulphate by copper sulphate.

(iv) Determination of soluble Iron and Aluminium compounds (expressed as Fe) :

(a) Outline of the method - iron and aluminum are detemihed gravimetrically by precipitation ammonium hydroxide.

(b) Reagents :

1. Concentrated nitric acid - A R grade.

2. Ammonium Chloride - A R grade.

3. Dilute Ammonium Hydroxide - approximately 15 per cent NH₃(m/v).

4. Dilute hydrochloric acid - 33 per cent (m/v).

(1) Take 10 gms of the test sample and add 25 ml. of water, 2 ml. of nitric acid and 5 gms. of ammonium chloride.

(2) Make the solution alkaline by adding ammonium hydroxide solution.

(3) Keep it on a water bath until the precipitate has floculated, keeping the solution alkaline by the addition of more ammonium hydroxide, if necessary.

(4) Filter and wash the resided with dilute ammonium hydroxide.

(5) Dissolve the residue in hot dilute hydrochloric acid.

(6) Make the solution again alkaline by adding ammonium hdroxide and allow the precipitate to settle.

(7) Filter and wash residue with water.

(8) Dry the residue, ignite and weigh till a constant mass is obtained.

Calculation :

Soluble iron and aluminum compounds (as Fe) per cent by mass = 70

M₁
___
M₂

where

M₁ = Mass in gm. of the residue obtained, and

M₂ = Mass in gm,. of the sample taken for the test. (Ref. : IS : 261 =1982)

(v) Determination of matter insoluble in water :

(a) Reagents :

(1) Concentrated sulphuric acid. - A R Grade

(b) Procedure. -

Weigh accurately about 10 gms. of the test sample and dissolve in 100 ml. of water, Add 3 ml. of sulphuric acid and stir thoroughly at room temperature. Filter through a tared filter paper or tared Gooch or Sintered glass crucible (G. No. 4). Wash the residue with water till it is free from acid. Dry the filter paper or crucible in an oven maintained at a temperature of 105 to 110C till constant mass is obtained.

Calculation :

Insoluble matter, per cent by mass =

M₁
____
M₂

x 100

where

M₁ = Mass in gm. of the residue obtained for the test.

M₂ = Mass in gm. of the material taken for the test.

(vi) Determination of pH:

(a) Procedure :

Dissolve 5 gm. of the test sample in water and make up the volume to 100 ml. Determine

the pH with glass electrodes using a suitable pH meter.

13. Method of analysis of ferrous sulphate (heptahydrate) FeSO₄.7H₂0

(i) Quality of reagents:

Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests.

Note. - "Pure Chemicals" means chemicals tht do not contain impurities which affect the results of analysis. Demineralised water means the water obtained after passing distilled water through a cation and an anion exchange resin or a combined cation - anion exchange resin.

(ii) Determination of ferrous iron :

(a) Outline of the method - Ferrous iron content is determined by titration with standard potassium permanganate solution.

(b) Reagents :

1. Dilute Sulphuric acid - 4 N approximately.

2. Standard Potassium permanganate solution. - 0.1 N.

3. Orthophospheric acid.

1. Weigh accurately about 5 gm. of the prepared sample and dissolve it in water.

2. Add 1 to 2 ml. of dilute sulphuric acid and make up the solution to 250 ml. in a volumetric flask.

3. Pipette out exactly 50 ml.of the solution in a flask.

4. Add 10 ml. of dilute sulphuric acid and 2 ml. of Orthophospheric acid and titrate with standard potassium permanganate solution.

Calculation :

Ferrous iron, per cent by weight =

27.92 V. N.
__________
W

Where

V = Volume of standard potassium permanganate used.

N = Normality of standard potassium permanganate solution.

W= Weight in gm. of the material taken for the test.

(Ref. : IS : 262 - "1982)

(iii) Determination of total iron

(A) Titrametric method using potassium dichromate :

(a) Outline of the method :

Total iron is determined by reducing ferric iron with stannous chloride and then titrating the total ferrous iron with standard potassium dichromate solution.

(b) Reagents :

(1) Concentrated hydrochloric acid.

(2) Stannous chloride solution. - Dissolve 6.0 gms. of stannous chloride crystals (SnCl₂.2H₂O) in 60 ml. of concentrated hydrochloric acid and dilute with water to 100 ml. Keep this solution in stoppered bottle.

(3) Mercuric chloride solution. - saturated.

(4) Concentrated sulphuric acid.

(5) Diphenylamine indicator - Dissolve 0.1 gms. of Diphenylamine in 100 ml. of concentrated sulphuric acid.

(6) Standard potassium dichromate solution. - 1.0 N.

(1) Weigh accurately about 5 gm. of the prepared sample and dissolve it in water.

(2) Add 1 to 2 ml. of the dilute sulphuric acid and make up the volume to 250 ml. in volumetric flask.

(3) Take 50 ml. aliquot of this solution and add 100 ml. of concentrated hydrochloric acid and heat to boiling.

(4) Reduce the iron by adding stannous chloride solution until the yellow colour of the ferric chloride disappears. Do not add more than 2 to 3 drops of stannous chloride in excess.

(5) Cool the solution, add rapidly mercuric chloride solution and stir so that a white precipitate of mercurous chloride forms slowly, incidating that a slight excess of stannous chloride is present.

(6) Allow to stand for 5 to 10 minutes, dilute to 150 to 200 ml. and add 5 ml. of phosphoric acid and 5 drops of diphenylamine indicator. Ti trate with standard potassium dichromate solution until the colour of the solution changes to a deep blue which does not fade on stirring.

Calculation :

Total iron, per cent by weight =

27.92 V. N.
__________
W

Where,

V = Volume in ml. of potassium dichromate used in titration.

N = Normality of standard potassium dichromate solution, and

W = Weight in gm. of the material present in the aliquot.

(Ref. : IS 262 - 1967)

(B) Atomic Abrosption Spectrophotometer Method:

(a) Reagents :

(1) Standard Iron Solution. - (1000 ppm) - Weigh accurately 1 gm. pure iron wire and put it in approximately 30 ml. 6N HO., in a beaker and boil. Transfer it to one litre volumetric flask through the funnel giving several washings to the beaker and funnel with glass distilled water. Make the volume up to the mark. Stopper the flask and shake the solution well. This is 100 ppm iron solution.

(2) Glass distilled or mineralised water of pH 2.5±0.5.

(3) Preparation of working standards. - Pipette 10 ml. iron stock solution in 100 ml. volumetric flask and dilute to volume. This is 100 ppm iron solution. Pipette the following volumes of 100 ppm iron solution in 50 ml. numbered volumetric flask and make the volume with acidified water.

Flask No.	Volume of 100 ppm standard iron solution taken (ml.)	Concentration of iron after making volume to 50 ml. (ppm)
1	2	3
1.	0.0	0.0
2.	1.0	2.0
3.	2.0	4.0
4.	3.0	6.0
5.	4.0	8.0
6.	5.0	10.0
7.	6.0	12.0
8.	7.0	14.0
9.	8.0	16.0
10.	9.0	18.0
11.	10.0	20.0

Stopper the flask and shake the solution well.

(b) Procedure.

(1) Preparation of ferrous sulphate fertiliser sample: Weigh exactly 1.000 gm. of the material on a clean watch glass and transfer it to a one litre volumetric flask through the funnel giving repeated washing with acidified water and dissolve the material by shaking well. Make the volume up to the mark with acidified water and shake well.

(2) Take 5 ml. of the prepared solution in 100 ml. volumetric flask and make up the volume with acidified water. Shake the solution well and filter through Whatman No. 42 filter paper in dry clean flasks. The flasks should be rinsed with a 10 to 15 ml. of the filterate and then continue filteration.

(3) Flaming the solution. - Flame the standards and the filtered sample on atomic absorption spectrophotometer at a wavelength of 248.3 nm using clean air acetlylene flame.

Calculation :

Prepare a standard curve of known concentrations of iron solutions by plotting the absorbance value on Y - axis against their respective iron concentrations on X - axis. Determine the concentration of iron in the sample solution from the graph.

Total iron = 2X/W

Where,

X= Concentration of Fe (in ppm) obtained from the standard curve. W = Weight in gm. of the material taken for the test.

(iv) Determination of ferric iron :

Substract the value of ferrous iron per cent from total iron per cent to obtain the Ferric iron per cent in the sample.

(Ref. IS : 262 - 1967)

(v) Determination of pH :

Dissolve 5.0 gms. of the sample in 100 mi. of freshly boiled and cooled water and determine the pH by means of a pH meter using glass electrode.

(vi) Determination of matter insoluble in water:

(a) Procedure :

Dissolve 25.0 gms. of the material in 125 ml. of water. Filter through a weighed and prepared Gooch crucible or sintered glass crucible (G.No. 4) and wash the residue thoroughly with water. Dry the crucible at 100°±81' to constant mass.

Calculation:

Matter insoluble in water per cent by weight = 4A. Where A = Weight in gm. of the residue.

Note. - In case a sample has been analysed by both the methods, viz. indicated under the hearing '7' and '8' the result obtained by the method indicated under the heading '8' shall prevail.

(vii) Determination of of lead :

(a) Reagents :

(1) Standard Lead Solutions. - Weight 0.1599 gm. of lead nitrate [Pb(NO₃)₂] on a clean watch glass and transfer it to one litre flask through the funnel giving several washings to watch glass and funnel with glass distilled or demineralised water. Add 10 ml. of concentrated distilled nitric water Add 10 ml. of concetrated distilled nitric acid and make the volume up to the mark. Stopper the flask and shake the solution well. This is 10 ppm lead solution and should be stored in a clean bottle for further use. Dilute 10 ml. of 100 ppm solution of lead to 100 ml. with 1 per cent nitric acid solution to get 100 ppm standard lead solution.

(2) 1 per cent nitric acid solution. - Dilute 10 mi. of concentrated distilled nitric acid to one litre with glass distilled water.

(3) 20 per cent zinc sulphate solution. - Weigh 20 gms of zinc sulphate (ZnSO₄.7H₂O) and dilute to 100 ml. with 1 per cent nitric acid solution.

(4) Preparation of working standard. - Pipette the following volume of 10 ppm standard lead solution in ml. numbered volumetric flask. Add 5 ml. of 20 per cent zinc sulphate solution to each flask and make the volume with 1 per cent nitric acid solution.

Flask No.	Volume of 10 ppm lead solution taken (ml.)	Volume of 20% zinc sulphate solution added (ml.)	Concentration of lead after making the valume to 50ml. (ppm)1
1	0.0	5.0	0.0
2	2.0	5.0	0.4
3	4.0	5.0	0.8
4	6.0	5.0	1.2
5	8.0	5.0	1.6
6	10.0	5.0	2.0

Stopper the flask and shake them well.

(b) Procedure. -

(2) Flaming the solution. - Flame the standards and the samples on atomic absorption spectrophotometer at a wavelength of 217 mu (Lead line of the instrument).

Calculations: Prepare a standard curve of known concentrations of lead solution by plotting the absorbance values on Y - axis against their respective lead concentration on X - axis. Calculate the percentage lead in zinc fertilizer by multiplying lead concentration value calculated from standard curve by 0.005.

Substitute zinc Sulphate by ferrous sulphate.

14. Method of analysis of ammonium molybdate (NH₄)₆Mo₇O₂₄4H₂O

(i) Quality of reagents :

Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests :

Note. - "Pure Chemicals" means chemicals that do not contain impurities which affect the result of analysis. "Demi neralised Water" means the water obtained after passing distilled water through a cation and anion exchanges resins or a combined cationtanion exchange resin.

(ii) Determination of molybdenum:

(A) Atomic Absorption spectrophotometer method :

(a) Reagents :

(1) Potassium Sulphate. - Dissolve 25 gm. of AR Potassium sulphate in 1 litre of distilled water.

(2) Standard molybdenum solution. - Weigh 1.5 gm. of molybdenum trioxide (MoO₃) and transfer it to one litre volumentric flask through the funnel giving several washings to beaker and funnel with glass distilled. Make up the volume up to the mark. This is 1000 ppm standard molybdenum solution.

(3) Preparation of working standards. - Pipette the following volume of 1000 ppm standard Mo solution in 100 ml. numbered volumentric flasks. Add 20 ml. of potassium sulphate solution to each flask and make up the volume to 100 ml. Stopper the flask and shake them well.

Flask No.	Volume of 1000 ppm Mo Solution taken (ml.)	Volume of Potassium sulphate added (ml.)	Concentration of molybdenum after making the volume to 100 ml. (ppm)
1.	0.0	20.0	0
2.	2.0	20.0	20
3.	3.0	20.0	30
4.	4.0	20.0	40
5.	5.0	20.0	50
6.	6.0	20.0	60

(b) Procedure. -

(1) Preparation of Ammonium Molybdate fertiliser sample : Weigh exactly 0.1 gm. of the material on the clean watch glass and transfer it to a 100 ml. volumetric flask through funnel giving repeated washing and dissolve the material by shaking well. Make the volume upto the mark.

(2) Take 10 ml. of the prepared solution in a 100 ml. volumetric flask, add 20 ml. of potassium sulphate solution and make the volume upto the mark.

(3) Flaming the solution : Flame the standards and the filtered samples on an atomic absorption spectrophotometer at a wavelength of 313.3 nm using nitrousoxideacetylene flame.

Calculation:

Prepare a standard curve of known concentration of molybdenum solution by plotting the absorbance value on Y - axis against their respective molybdenum concentration on X - axis.

per cent of Mo = X/10W

Where

X =Concentration of molybdenum (ppm) as obtained from the caliberation curve. W= Weigh in gm. of the material taken for the test.

(B) Calorimetric Method for Determination of Molybdenum :

(a) Outline of Method :

Molybdenum (vi) in acid solution when treated with stannous chloride (best in the presence of a little ferrous ion) is converted largely into molybdenum (v). This forms a complex with thiocyanate ion, probably largely Mo (SCN)₅, which is red in colour. The latter may be extracted with solvents possessing donor oxygen atoms (3 - Methylbutanol is preferred). The colour depends upon the acid concentration (optimum concentration 1M) and the concentration of thiocyanate ion (K 1.0 per cent but colour intensity is constant in the range 2 - 10 per cent); it is; little influenced, by excess of stannous chloride. The molybdenum complex has maximum absorption at 465 nm.

(b) Apparatus :

1. Photometer. - Any suitable photoelectric colorimeter

1. Standard molybdenum solution. - (0.001 per cent molybdenum) - Weigh 0.184 of AR Ammonium molybdate (NH₄)₆Mo₇O₂₄.4H₂O on a clean watch glass and transfer it to one litre volumetric flask through the funnel giving several washings to watch glass and funnel with glass distilled water. Make the volume upto the mark. Pipette out 10 ml. of this solution into 100 ml. volumetric flask and make the volume upto the mark, This gives 0.001 per cent Mo solution.

2. Ferrous Ammonium Sulphate. - Dissolve 10 gm. of AR ferrous ammonium sulphate salt in 100 ml. of every dilute H₂SO₄.

3. Stannous chloride solution. - Dissolve 10 gm. of AR stannous chloride dihydrate in 100 ml. of 1 M - hydrochloric acid.

4. Potassium thiocyanate solution - Dissolve 10 gm. of AR potassium thiocyanate salt in 100 ml. of distilled water.

5. Iso - amly alcohol.

(d) Preparation of Sample :

Weigh 0.1 gm. of sample on a clean watch glass and transfer it to one litre volumetric flask, through the funnel giving several 'washings to watch glass and funnel with glass distilled water. Make the volume up to the mark. Pipette out 5 ml. of this solution into a 100 ml. of volumetric flask and make up the volume up to the mark.

(e) Procedure. -

(1) Place 1.0, 2.0, 3.0, 4.0 and 5.0 ml. of the 0.001 per cent molybdenum solution (containing 0.01 mg., 0.02 mg., 0.03 mg., 0.04 mg. and 0.05 mg. molybdenum) severally in 50 ml. capacity separating funnels and diluting each with an equal volume of water.

(2) Add to each funne 12.0 ml. of conc. Ha. 1.0 ml. of ammonium ferrous sulphate and 3.0 ml. of the potassium thiocyanate solution.

(3) Shake gently and then introduce 3.0 ml. of the stannous chloride solution.

(4) Add water to bring the total volume in each separating funnel to 25 ml. and mix.

(5) Pipette 10 ml. of redistilled 3 - methylbutanol (iso - amyl alcohol) into each funnel and shake individually for 30 seconds.

(6) Allow the phases to separate and carefully run out the lower aqueous layer.

(7) Remove the glass stopper and pour the alcoholic extract through small plug of purified glass wool in a small funnel and collect the organic extract in a 1.0 cm. absorption cell.

(8) Measure the absorbance at 465 nm in spectrophottimeter against a 3 - methylbutanol blank.

(9) Plot absorbance against concentration of standard molybdenum solutions and draw and draw the calibration curve.

(10) Take 10 ml. of the sample solution and determine to absorbance of it by subjecting it to the same treatment as the standard solutions using calibration curve, determine the corresponding concentration of molybdenum in the sample solution.

Calculation :

% Mo = 2000X / Y.W.

Where

Y= Volume of sample solution taken for the test.

W= Weight in gm. of the material taken for the preparation of the sample solution. (Ref. : Vogal's Text Book of Quantitative Analysis).

(iii) Determination of matter insoluble in water :

(a) Procedure :

Dissolve 25.0 gm. of the material in 125 ml. of water. Filter through a weighed and prepared Gooch crucible of sintered glass crucible (G.No. 4) and was the residue thoroughly with water. Dry the crucible at 1108" to constant mass.

Calculation :

Matter insoluble in water percent by weight= 4A Where A =Weight in gm. of the residue.

(iv) Determination of lead :

(a) Reagents :

(1) Standard Lead Solutions. - Weight 0.1599 gm. of lead nitrate (Pb(NO₃)₂ on a clean watch glass transfer it to one litre flask through the funnel giving several washings to watch glass and funnel with glass distilled or demineralised water. Add 10 ml. of concentrated distilled nitric water. Add I 0 ml. of concentrated distilled nitric acid and make the volume up to the mark. Stopper the flask and shake the solution well. This is 100 ppm lead solution and should be stored in a clean bottle for further use. Dilute 10 ml. of 100 ppm solution of lead to 100 ml. 1 per cent nitric acid solution to get 10 ppm standard lead solution.

(2) 1 per cent nitric acid solution. - Dilute 10 ml. of concentrated distilled nitric acid to one litre with glass distilled water.

(3) 20 per cent zinc sulphate solution. - Weigh 20 gms. of zinc sulphate (ZnSO₄.7H₂O) and dilute to 100 ml. with 1 per cent nitric acid solution.

(4) Preparation of working standards. - Pipette the following volume of 10 ppm standard lead solution in ml. numbered volumetric flask. Add 5 ml. of 20 per cent zinc solution to each flask and make the volume with 1 per cent nitric acid solution.

Flask No.	Volume of 10 ppm Lead solution taking (ml.)	Volume of 20% zinc sulphate solution added (ml.)	Concentration of lead after making the volume to 50 ml. (ppm).
1	0.0	5.0	0.0
2	2.0	5.0	0.4
3	4.0	5.0	0.8
4	6.0	5.0	1.2
5	8.0	5.0	1.6
6	100	5.0	2.0

Stopper the flask and shake them well;.

(b) Procedure. -

(1) Preparation of zinc sulphate fertiliser samples. - Weigh I gm. of the material on a clean watch glass and transfer to 50m1. volumetric flask through the funnel giving washings with 1 per cent nitric acid solution. Dissolve the material and make the volume with 1 per cent nitric acid solution. Samples should be prepared in duplicate.

(2) Flaming the solution. - Flame the standards and the samples on atomic absorption spectrophotometer at a wavelength of 217 mu (Lead line of the instrument.)

(3) Calculations. - Prepared and standard curve of known concentrations of lead solution by plotting the absorbance values on Y - axis against their respective lead concentration on X - axis. Calculate the percentage lead in zinc fertilizer by multiplying lead concentration value calculated from standard curve by 0.005.

Substitute zinc sulphate by ammonium molybdate.

15. Method of analysis of chelated zinc (As Zn - EDTA)

(i) Reagents :

(a) EDTA Solution. - (0.05M) - Dissolve 18.612 gm. of disodium ethylene diamine tetraacctate dehydrate (EDTA) in distilled water and make up the volume to 1 litre.

(b) Standard Zinc solution (1000 ppm) : Weight accurately 1.0 gm. of zinc metal in a beaker. Add 20 ml. HCL (1:1). Keep it for few hours and allow it to dissolve completely. Transfer the solution to 1 litre volumentric flask. Make the volume up the mark.

(d) Ammonium Nitrate. - AR grade salt.

(e) Buffer solution (pH - 10) - Dissolve 8.0 gm. AR grade ammonium nitrate in 65 ml. of water and add 35 ml. of concentrated ammonia solution (sp. gr. - 0.88)

(f) Eriochrome black (T) indicator mixture - Mix thoroughly 1 gm. of eriochrome back (T) indicator with 100 gms. of AR grade potassium nitrate.

(g) Hydroxylamine Hydrocloride - AR grade.

(h) Potassium cyanide - AR grade (to be used with extreme care) 15 per cent. aq.solution.

(i) Manganese sulphate solution.Dissolve 11.15 gm. of AR grade manganese sulphate in 1 litre of distilled water.

(j) Sodium Fluoride AR grade.

(ii) Preparation of sample solution :

Weigh accurately 1.0 gm. of the sample and transfer it to 100 ml. volumetric flask. Make up the volume with distilled water. Keep it overnight.

(iii) Procedure :

(a) Standardization of EDTA solution.

(1) Take 10 ml. of zinc solution (standard).

(2) Dilute it by adding 30 ml. distilled water.

(3) Add 10m1. of buffer solution and 30 - 40 mg. of indicator mixture.

(4) Titrate with EDTA solution till clear blue end point is obtained. Note the volume of EDTA used as V₁ ml.

(b) Standardisation of manganese sulphate solution.

(1) Take 25 ml. of manganese sulphate solution.

(2) Dilute it by adding 100 ml. distilled water.

(3) Add 0.25 gm. of hydroxylamine hydrocloride and 10 ml. of buffer solution.

(4) Add 30 - 40 mg. of indicator mixture.

(5) Titrate with EDTA solution till clear blue end point is obtained. Note the volume of EDTA used as V₂ ml.

(1) Take 10 ml. of sample solution.

(2) Dilute it by adding 100 ml. of distilled water.

(3) Add 0.25 gm. of hydroxylamine hydrocloride.

(4) Add 10 ml. of buffer solution and 30 - 40 mg. of indicator mixture.

(5) Warm to 40° and titrate with standard EDTA solution (Preferably stirring magnetically) to clear blue end point. Note the volume of EDTA used as V₃ ml.

(6) After the end point add 2.5 gms. of sodium flouride and stir for one minute.

(7) Titrate the solution with standard manganese sulphate solution, slowly, till a permanent red colour is obtained. Note the volume of manganese sulphate added as V₄ ml.

(8) Stir for 1 minute.

(9) Titrate the excess of manganese ions with EDTA solution until the colour changes to pure blue. Note the volume of EDTA used as V₅ ml.

(10) After the second end point (step 9) add 4 - 5 ml. of 15 per cent aqueous potassium cyanide solution.

(11) Titrate it with manganese sulphate solution till colour changes sharply from blue to led. Note the volume of manganese sulphate solution added as V₆ ml

Calculation:

Molarity of standard zinc. Molarity of EDTA solution

(M1) = solution x volume of standard zinc solution taken.

Volume of EDTA used (V₁)

Molarity of standard manganese sulphate solution

M₁V₂

Volume of standard manganese sulphate taken.

Nos. of millimoles of EDTA used in titrating Zn + other metals (A) = M₁V₃

No. of millimoles of EDTA liberated by Sodium fluoride (B) = M₂V₄ - M₁V₅

Hence Nos. of millimoles of EDTA used for titrating zinc (C) = A - B

But Nos. of millimoles of EDTA liberated by KCN (D) = M₂V₆

Hence No. of millimoles of EDTA contained by Zn - EDTA sample =D - C EDTA

percent = 372.24 (D - C)

Per cent of magnesium in the sample = 24.31 x B

Per cent of free zinc = 65.38 x c C

Per cent Zinc chelated with EDTA = 65.38 (D - C)

(Ref. Vogal's Text Book of Quantitative Inorganic Analysis).

16. Method of analysis of chelated iron (as Fe - EDTA) :

(i) Quality of reagents :

Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests.

Note. - "Pure chemicals" means chemicals that do not contain impurities which affect the results of analysis. "Demineralised Water" means the water obtained after passing distilled water through a cation and an anion exchange resins or a combined cation - anion exchange resin.

(ii) Determination of chelated iron:

(a) Reagents :

(1) Sodium hydroxide solution. - 0.5N. Dissolve 20 gm. NaOH in water & dilute to 1 litre.

(2) Disodium EDTA solution.

(3) Iron standard solution

(A) Stock solution. - (1000 ppm). Dissolve. 1.000 gm. pure Fe wire in approximately 30 ml. 6N HCL with boiling. Dilute to 1 liter in a volumetric flask with distilled water.

(B) Intermediate solution. - (100 ppm) Pipette 10 ml. iron stock solution and 10 ml. Na₂H₂ EDTA solution in 100 ml. volumetric flask and dilute to volume.

(C) Working solution. - Pipette the following volumes of 100 ppm. intermediate solution in 50 ml. numbered volumetric flask and make the volume with 0.5N HCL.

Flask No.	Volume of 100 ppm. standard iron solution taken (ml.)	Concentration of iron after making volume to 50 ml. (ppm)
1.	1.0	2.0
2.	2.0	4.0
3.	3.0	6.0
4.	4.0	8.0
5.	5.0	10.0
6.	6.0	12.0
7.	7.0	14.0
8.	8.0	16.0
9.	9.0	18.0
10.	10.0	20.0

(b) Apparatus - Atomic Absorption Spectrophotometer - with air/aceilylene flame.

(1) Preparation of sample solution :

(A) Weigh sample containing approximately 40 mg. Fe into 200 ml. tall form beaker.

(B) Wet with 2 - 3 drops of alcohol and dissolve in 100 ml. of water.

(C) Add 4 drops of 30 per cent H₂O₂ mix and adjusts pH of solution to 8.5 with 0.5N NaoH. If pH drifts above 8.8 discard solution and repeat analysis.

(D) Transfer solution to 200 ml. volumetric flask, dilute to volume with water and mix

(E) Filter solution through quantitative paper.

(F) Pipette 10 ml. filtrate into 200 ml. volumetric flask and dilute to volume with 0.5 N HCI.

(2) Flaming the solution :

Flame the standards and the sample solution on atomic absorption spectrophotometer and a wavelength of 248.3nm using air acetlylene flame. In same manner determine Fe blank on all reagents used.

Calculation :

Prepare a standard curve of known concentration of Fe solution by plotting the absorbance value on Y axis against their respective Fe concentration on X axis.

Chelated Iron % =

(ppm Fe in sample ppm - Fe in blank x 0.4)
__________________________________
g sample

(Ref.: Method of analysis AOAC, 1984)

(iii) Determination of pH:

(a) Procedure :

Dissolve 5 gm. of the material in freshly boiled water. Dilute to 25 ml. and mix. Determine the pH value of the solution with pH meter.

[17. Determination of Sodium in Potassium Chloride and Potassium Sulphate by Atomic Absorption Spectrophotometric Method] -

(i) Quantity of Reagents. - Unless specified otherwise, pure chemicals and glass distilled or demineralised water shall be used in tests.

(ii) Reagents :

(1) Ammonium Oxalate Solution. - Dissolve 40g of Ammonium Oxalate in 1 litre of distilled water.

(2) Standard Sodium Solution. - (1000 ppm) - Weigh accurately 2.5421g of dried NaCl on a clean watch glass and transfer it to one litre volumetric flask. Make up the volume up to the mark. Stopper the flask and shake the solution well. This is 1000 pm. sodium solution.

(3) Preparation of working standard. - Pipette the following volumes of 1000 ppm. standard sodium solution in 100 ml. volumetric flask and make up the volume with glass distilled water.

Flask No.	Volume of 1000 ppm standard sodium solution taken (ml.)	Concentration of sodium after making volume to 100 ml. (ppm).
1	0.0	0.0
2.	2.0	20.0
3.	4.0	40.0
4.	6.0	6(10
5.	10.0	100.0
6.	12.0	120.0
7.	14.0	140.0
8.	16.0	160.0
9.	18.0	180.0
10.	20.0	200.0

Stopper the flask shake the solution well.

(iii) Procedure :

(1) Preparation of sample solution. - Weigh exactly 2.5 grams sample on a clean watch glass and transfer it to 250 ml. volumetric flask through the funnel giving repeat washings with glass distilled water (about 125 ml. of water should be used). Add 50 ml. of Ammonium Oxalate solution. Boil for 30 minutes, cool dilute to volume mix and pass through dry filter paper (Whatman No. 1 or equivalent).

(2) Flaming the solution. - Flame the standards and the filtered sample on Atomic Absorption Spectro - photometer at a wavelength of 330.3 mm using clean air acetylene flame.

Calculation :

Prepare a standard of known concentrations of Sodium solution by plotting the absorbance value on Y - axis against their respective sodium concentration on X - axis. Determine the concentration of sodium in the sample solution from the graph.

Sodium (Na) as Sodium Chloride % = 0.0254 X.

Where X is the concentration of Sodium in ppm obtained from the standard curve.

(Ref.: AOAC , 1984)]

18. Determination of Total Chlorides in Potassium Sulphate by Silver Nitrate Volumetric Method :

(i) Reagents :

(1) Standard Silver Nitrate (AgNO₃) solution - 0.1N.

(2) Potassium Chromate (K₂Cro₄) Indicator solution. - 5 per cent.

(ii) Procedure :

(1) Weigh accurately about 10 grams of the prepared sample and transfer into 500 ml. beaker.

(2) Add about 250 ml. of water and warm gently for about 20 minutes.

(3) Cool and dilute the solution to 500 ml. in a volumetric flask.

(4) Allow to stand to let any insoluble matter settle.

(5) Transfer with pipette 100 ml. of the clear solution into a conical flask.

(6) Titrate with standard AgNO₃ solution (0.1N) using 1 ml. of Potassium Chromate Indicator solution.

Calculation :

Total Chlorides (as Cl), per cent by Weight (on dry basis). =

17.73 N.V.
__________
W

Where

N = Normality of standard AgNO3 solution.

V= Volume of standard AgNO3 used.

W= Weight in gram of the material taken for test.

[19. Method of analysis of magnesium sulphate]:

(i) Quality of Reagents :

Unless specified otherwise pure chemicals and glass distilled or demi neralised water shall be used in tests.

(ii) Determination of Magnesium :

(A) Atomic Absorption Spectrophotometeric method.

(a) Reagents :

(1) Hydrochloric Acid 0.5 N

(2) Magnesium Standard Solution : - Dissolve 1.013 gm Magnesium Sulphate (MgSO₄.7H₂O) in 0.5N Hydrochloric acid solution and dilute to 100 ml. with this acid in a volumetric flask. This is 1000 ppm magnesium stock solution.

(3) Strontium Chloride solution : - Dissolve 15 gm Strontium Chloride (SrCl₂.6H₂O) in 0.5 N Hydrochloric acid and dilute to 100 ml. with the same solvent.

(b) Preparation of working standards :

(1) Pipette 1 ml. of 1000 ppm Magnesium stock solution to 100 ml. capacity volumetric flask and make up the volume with 0.5 N Hydrochloric acid. This is 10 ppm Magnesium Solution.

(2) Pipette the following volume of 10 ppm Magnesium solution in 100 ml. numbered volumetric flask. Add 10 ml. of Strontium Chloride solution to each flask and make up the volume to 100 ml. with 0.5 N Hydrochloric acid.

Flask No.	Volume of 10 ppm. Mg. solution taken (ml.)	Volume of strength chloride added (ml.)	Concentration of Mg (ppm) after making the volume to 100 ml.
1.	0.0	10	0.0
2.	2.0	10	02
3.	4.0	10	0.4
4.	6.0	10	0.6
5.	8.0	10	0.8
6.	10.0	10	1.0

(1) Preparation of sample solution. -

(A) Weigh I gm of the sample aid place in a 500 ml. volumentric flask. Add about 300 ml. water and boil for 30 minutes. Allow to cook, dilute to the mark with water, mix and filter.

(B) Pipette 5 ml. of the filtrate (A) into a 100 ml. volumetric flask, make up to the the mark with water and mix.

(C) Transfer by pipette 5 ml. of the diluted filtrate (B) into 1(X) ml. volumetric flask and make up to the mark with 0.5 N.H. 1.

(2) Blank solution. - Prepare a blank solution from which only the sample has been omitted.

(3) Flaming of solution. - Flame the standard and sample solutions on Atomic Absorption Spectrophotometer at wavelenght of 285.2 nm using Air Acetylene flame.

Calculation:

Plot the calibration curve using the mean absorbances on Y - axis and the corresponding concentrations of Magnesium (ppm) on X - axis. Determine the concentration of Magnesium in the sample by reference to the calibration curve.

Magnesium per cent - 20X

Where X - concentration of Magnesium (in ppm) obtained from the standard curve.

(B) Titrametric method (EDTA) Titrations

(Applicable to the samples which do not contain phosphate as impurity).

(a) Reagents :

(1) Buffer solution (pH - 10.0) - Dissolve 67.5 gm ammonium chloride in 200 ml distilled water add 570 ml ammonia solution and dilute to 1 litre.

(2) Potassium hydroxide - Potassium cyanide solution - Dissolve 280 gm pottasium hydroxide and 66 gm potassium cyanide in 1 litre of distilled water.

(3) Potassium cyanide solution (2%) - Dissolve 2 gm potassium cyanide in 100 ml. of distilled water.

(4) Eriochrome Black T indicator solution - Dissolve 0.2 gm. of indicator in 50 ml of methyl alcohol containing 2 gm of hydroxylamine hydrocloride.

(5) Calcium standard solution (1 mg/m1) - Dissolve 2.4973 gm calcium carbonate, primary standard grade, previously dried for 2 hours at 285" in HCl (1+10). Dilute to 1 litre with distilled water.

(6) Calcine indicator mixture - Grind together 1 gm calcein indicator 2', 7' - bis [bis (Corboxnymethyl) amino] methyl - (fluorescein, sodium derivate sodium salt.) 10 gm charcoal and 100 gm potassium chloride.

(7) Disodium dihydrogen ethylene diamine tetra acetic acid standard solution (0.4%) - Dissolve 4 gm NH₂H₂ - EDTA in 1 litre of distilled water.

(8) Triethanolamine (1 +1)

(9) Potassium ferrocyanide solution (4%) - Dissolve 4 gm potassium ferrocyanide in 100 ml of distilled water.

(b) Standardization of Calcium solution :

(1) Pipette 10 ml. calcium standard solution into 250 ml erlenamayer flask.

(2) Add 100 ml. of distilled water, 10 ml. KOH - KCN solution, 2 drops of triethanolamine solution, 5 drops of potassium ferrocyanide solution and 15 ±1 mg of calcein indicator.

(3) Immediately place the flask on a magnetic stirrer in front of daylight fluorescent light with white background.

(4) While stirring, titrate with EDTA solution to disappearance of all fluorescent green and until solution remains pink, titrate more than 3 aliquots. From average, calculate calcium titer value.

Calcium Titer - Volume of calcium standard (mg/ml) Volume EDTA solution used (ml.)

From calcium titer, calculate magnesium titer value as follows :

Magnesium titer - Calcium titer x 0.6064

(1) Weigh 1 gm magnesium sulphate fertiliser sample into 250 ml volumetric flask.

(2) Add 200 ml of distilled water and boil for 30 minutes.

(3) Cool dilute to volume with water and mix.

(d) Procedure :

(1) Titration for Ca + Mg

(A) Pipette 25 ml of aliquot in 250 ml erlenmayer flask.

(B) Dilute with 100 ml. of distilled water.

(C) Add 5 ml of buffer solution (pH 10), 2m1potassium cyanide solution, 2 drops of triethanolamine solution, 5 drops of potassium ferrocyanide solution, and 8 drops of eriochrome black T indicator solution.

(D) Titrate immediately with EDTA solution, stirring and lighting as in standardisation. Colour changes are wine, red, purple, dark blue, to clear blue end point, becoming green if over titrated. Note the volume of EDTA used as V₁ - ml.

(2) Titration of Calcium :

(A) Pipette 25 ml of aliquot in 250 ml erlenmayer flask.

(B) Dilute with 100 ml. of water.

(C) Add 10 ml COH - KCN solution, 2 drops of triethanolamine solution, 5 drops of potassium ferrocyanide solution and 15 ±1 mg of calcein indicator.

(D) Titrate immediately with EDTA solution as in standardization. Note the volume the EDTA used as V₂ - ml.

Calculation:

Magnesium per cent = (VI - V2) x Mg Titer EDTA (Reference : AOAC, 14th edition, 1984).

Ministry of Human Resource Development (Department of Women and Child)

[20. Determination of particle size in different fertilizers]:

(i) Apparatus :

(A) Indian Standard sieves of 20 cms diameter and 5 cms in width with lid and bottom pan of required sizes.

(B) Sieve shaker or vibrator with automatic timer and variable cycle control

(D) Weighing pan, round suitable for holding sample and approximately 23 cms. in diameter.

(E) Brush either soft, fine wire or stiff bristle.

(ii) Procedure :

(A) Take the sieve of requires sizes and stack them in progressive order. The biggest sieve sizes should be on the top and the smallest in the bottom. Place the empty pan at the bottom.

(B) Put 200 gms. of ungrined sample on the top sieve and place lid on the top of stack.

(D) Weighing pan, round suitable for holding matic timer is used, set timer for 5 minutes. If vibrator is used, also set vibrator control at 3000 cycle per minutes.

Note. - If mechanical sieve shaker is not available, use hand sieving Conduct sieving by appropriate lateral and vertical motions accompanied by jarring action. Continue until no appreciable change is noted in sieve fractions.

(E) After completion of shaking time, transfer material from each sieve to weighing pan with brush and weight to ±0.1 gm.

(F) Record weight from each sieve (sum of weights from each sieve should agree closely with original sample weight.)

Calculations :

Per cent weight on sieve =

Weight on sieve (gm.)
___________________________
Total weight of sample (gm.)

[Reference. - Method IV - A TH - 1982).]

[[[21A.] Method of Analysis of Calcium Nitrate]

(i) Determination of total nitrogen

By the method as specified in 3(viii)

(ii) Determination of Ammoniacal Nitrogen

By the method as specified in 3 (vii)

(iii) Determination of nitrate

By the method as specified in 3(viii) & (vii)

(iv) Determination of water insoluble matter

By the method as specified in 8(vii)

(v) Determination of water soluble calcium

A. Titrametric Method (EDTA) Titrations

(a) Reagents

(i) Buffer solution (pH - 12.0) - Dissolve 225 gm. KOH (AR) in 250 ml water. Add 15 ml Triethanolamine and make volume to one litre after cooling the solution.

(ii) Pattan and Reeder's indicator (HHSNNA) - Dissolve 0.2 gm indicator in 100 ml. methanol.

(iii) Calcium Standard Solution (mg/m1) - - Dissolve 2.4973g calcium carbonate primary standard grade, previously dried for 2 hours at 285°C, in HCl (1+10). Dilute to 1 litre with distilled water.

(iv) Disodium dihydrogent ethylene diamine tetra acetic standard solution (0.01M) - Dissolve 3.7224 gm Na₂H₂ - EDTA (AR) previously dried at 105 to 110°C for 1 hr. in distilled water and make volume to one litre.

(b) Standardisation of Calcium Solution

(i) Pipette 10 ml. calcium standard solution into 250 ml. Erlenmayer Flask.

(ii) Add 10 - 15 ml. distilled water and 20 ml. Buffer solution.

(iii) Add few drops of indicator's solution. A red rose colour will appear.

(iv) Titrate with EDTA till a blue colour is obtained. Note the volume of EDTA used as V₁ ml.

Calcium Titer (mg/m1) =

Volume of Calcium standard solution (ml)
______________________________
Volume of EDTA solution used (ml) (V₁)

(i) Weigh 2.5g calcium nitrate fertiliser sample into 250 Volumetric Flask.

(ii) Make up the volume with distilled water Shake well.

[(iii) In case of Potassium Magnesium Calcium Sulphate weighed 2.5 gm of fresh sample in 250 ml volumetric flask and add 5 ml concentrated HCI, make up the volume and shake well.]

(d) Procedure

(i) Pipette 5 ml. of aliquot in 250 ml. Erlenmayer Flask.

(ii) Add 10 - 15 ml. distilled water & 20 ml Buffer solution.

(iii) Add few drops of indicator's solution. A red rose colour will appear.

(iv) Titrate with EDTA till a blue colour is obtained. Note the volume of EDTA used as V₂ ml.

Calculation

Calcium per cent = V₂ x Calcium Titer x 2

B. Atomic Absorption Spectrophotometric Method

(1) Reagent. - Unless specified otherwise pure chemicals and glass distilled or demineralised water shall be used in test.

(a) Hydrochloric Acid = 3N

(b) Preparation of stock solution : -

(i) Calcium stock solution. - Dissolve 2.498 gram CaCO, in a minimum amount of 3 N Hcl. Dilute to 1liter (1000 ppm Ca). Dilute 10 ml to 100 ml. This is 100 ppm stock solution.

(ii) Lanthanum stock solution. - 50g La/L. Dissolve 58.65 gram La₂O₃ in 250 ml. HCI, adding acid slowly. Dilute to 1 liter.

[or, Strontium Chloride Solution: Dissolve 15g, Strontium chloride (SrCI₂6H₂O) in 0.5NHCI and dilute to 100 ml.]

[(C) Preparation of working standard: - Pipette the following volume of 100 ppm Calcium stock solution in 100 ml numbered volumetric flasks. Add 10 ml of Strontium Chloride solution or 20 ml of Lanthanum solution to each flask and make the volume to 100 ml with distilled water.

Flask No.	Volume of 100 ppm, Ca solution Taken (ml.)	Concentration of Calcium (ppm) after making the colume 100 ml.
1	0	0
2	1	1
3	2	2
4	3	3
5	4	4
6	5	5]

[(1) Weigh exactly 1.0g. of sample on a clean watch glass and transfer it to 1000 ml. volumetric flask through the funnel giving repeated washings with glass distilled water and make up the volume.

(2) Take 1 ml. of prepared solution in 100 ml. volumetric flask, add 20 ml. of Lanthanum solution or 10 ml. of strongflum chloride solution and make up the volume. Filter, if required.]

(3) Flaming of the solution. - Flame the standard and sample solution on AAS at wave length of 422.7 nm using Air - Acetylene flame.

(4) Calculation. - Plot the calibration curve using the maen absorbance on Y axis and the corresponding concentration of Ca (ug/ml) at X axis. Determined the concentration of calcium in the sample by reference to the calibration curve.

% Ca= Concentration (ug/ml) x 10^-2

[[22A.] Analysis of Micronutrients Fertilisers Mixtures by Atomic Absorption Spectrophotometric Method.

(a) Reagents:

(1) Acidified water : Dissolve 10 ml of 10% Sulphuric Acid in 10 litre of double distilled water and adjust the pH to 2.5 (10.5) with help of a pH meter using H2So4 or NaOH.

(2) Strontium Chloride Solution : Dissolve 25 gm of AR Potassium Sulphate in one litre of distilled water.

(b) Procedure:

(i) Preparation of standard stock solution:

Dissolve the specific quantity of standard material for specific element with the help of suitable solvent as indicated below and make up the volume 1000 ml with distilled water. Further dilute it to suitable flaming range with acidified water in case of Fe, Cu, Mn and Zn. In case of Mg. add 20 ml of Strontium Chloride and in case of Mo, add 20 ml of Potassium Sulphate before make up the volume up to the mark, in 100 ml volumetric flask with acidified water.

Element	Material	Solvent	Concentration in ppm	Flaming range of standard ppm.	Wave length in mm	Flame
zinc	1.0g Metal	30 ml 1.1 HCI	1000	0.2-2.0	213-9	Air/C₂H₂
Iron	1.0g Iron wire	30 ml 1.1 HNO₂	1000	2.20	248.3	Air/C₂H₂)
Manganese	1.0g Metal	50ml Conc. HCI	1000	0.5-5	279.5	Air/C₂H₂
Copper	1.0g Cooper Turning	30 ml (1:1) HNO₃	1000	0.5-1	324.8	Air/C₂H₂
Boron	28.5720g Boric Acid	Warm distilled water	5000	400-1600	248.8	N₂O/C₂Hs
Molybdenum	1.5 Molybdenum Trioxide	10ml HCI	1000	20-60	313.3	Air/C₂H₂ (Rich) or N₂O/C₂N2
Magnesium	1.0 Metal Powder	30ml 1:1 HCI	1000	0.2-1	285.2	Air/C₂H₂

[(ii) Preparation of sample solution:

Weigh 10 grams or sample in 500 ml volumetric flask, Add 10 ml of 1:1 HCI and 100ml distilled water, warm gently. Cool and make up the volume up to the mark with distilled water. Shake well and keep it stand for atlas 1 hour. Dilute it to further suitable working standard range with acidified water in case of Fe, Cu, Mn and Zn. In case of Mg. add 20 ml of strontium chloride and in case of Mo, add 20 ml of potassium sulphate before making up the volume upto mark, in 100 ml flask with acidified water.]

(iii) Flaming of Solutions:

Flame the standard solution of specific element at concentration mode. Then flame the sample solution to observe the concentration (in ppm) of specified element.

Calculation:

23. Determination of Beron in Micronutrient Fertiliser Mixture-Volumetric Method

(a) Reagents

(1) Mannitol

(2) Blemuth Nitrate Solution: Dissolve 22 gm of Bismuth Nitrate (BiNO)s 5H2O in 8 ml of conc. Nitric Acid Warm slightly and dilute to 100 ml with distilled water.

(3) Nitric Acid:- Concentrated AR Grade.

(4) Dilute Nitric Acid:- (1:20)

(5) Sodium Hydroxide Solution:- 10%M/V

(6) Sodium Hydroxide Solution=0.02M

(7) Bromothymol Blue Indicator Solutions.

(b) Procedure:

(1) Weigh accurately 2.5 gms of prepared sample into 250 ml. beaker.

(2) Add 2 ml of cone. Nitric Acid and 50 ml of water and warm gently and dilute to 100 ml with water.

(3) warm the solution (but do not boil).

(4) Slowly add 5ml of Bismuth Nitrate Solution for each 1% of P2O5 percent in the sample, from a burette with continuous stirring and maintaining the liquid hot during addition.

(5) Cool and transfer the contents in to 250 ml volumetric flask and dilute upto the mark with distilled water.

(6) Allow precipitate to settle and dilute upto the mark with distilled water.

(7) Filter through a dry filter paper (No.40) rejecting the first few drops into a breaker.

(8) Pipette 100 ml of the filtrate in 250 ml beaker, add a few drops of indicator solution.

(9) Add few drops of 10% NaOH solution with thorough stirring until the indicator turns blue.

(10) Filter through a dry filter paper (No. 40) into 250 ml beaker and carefully was the residue several times with cold water. The total volume should be 150 to 200 ml.

(11) Adjust the pH to about 5 by adding diluted HNO3. Warm to about 90C (don not boil), and stir vigorously to avoid Co2 Cool the solution.

(12) Place in the solution, the electrodes of pH meter and adjust the pH to exactly 6.3 using NaOH solution.

(13) Add 10 gms. of mennitol and again adjust pH to exactly 6.3 with 0.02 NNaOH solution.

(14) Continue adding mannitol in 10 gms portion till pH remains constant at 6.3 Note the total volume of 0.02 NNaOH used for the 1st addition of Mannitol (Step 13).

(15) Carry out a blank determination on fertilizers mixture to which Borate in not added.

%B=(A-B)x0.05405/W

Where :A=Volume of 0.02 N NaOH solution used in the sample.

B= Volume of 0.02 N NaOH solution used in the blank.

Note: In case phosphate is not present in the sample, proceed as follows:-

(i) Take 2.5 gms of the sample in 250 ml volumetric flask and make up the volume with distilled water.

(ii) Proceed as given from step (B) onwards.]

[24. Determination of total Sulphur

(The total sulphur includes sum of sulphur present in Sulphate and elemental form).

(A) Determination of Sulphate Sulphur : Method of determination of sulphur present in the sulphate form in various fertilizers for nitrate free and nitrate containing samples is given below:-

Classification of procedures

Procedure (a) : For nitrate free samples such as ammonium sulphate, potassium sulphate, zinc sulphate, copper sulphate, ferrous sulphate, manganese sulphate, N.P. and NPK complexes and mixtures.

Procedure (b) : Super phosphate

Procedure (c) : For nitrate containing fertilizers such as ammonium phosphate sulphate nitrate 20 : 20: 0

(i) Quality of Reagents

Unless specified otherwise, pure chemicals, glass, distilled or demineralised water shall be used in tests.

Note: (1) "pure chemicals" means chemicals which do not contain impurities which affect the results of analysis.

(2) "demineralised water" means water obtained by water passing through a cation and anion exchange resins or a combined cation anion exchange resins.

(ii) Reagents

(1) Hydrochloric acid-concentrated

(2) Dilute hydrochloric acid-Dilute 250 ml. of concentrated hydrochloric acid to 1000ml. of water.

(3) Barium chloride solution-2 percent Dissolve 20g of barium chloride in 1000 ml of water.

(4) Silver nitrate solution-5 percent. Dissolve 5g silver nitrate in 100 ml water.

(iii) Procedure (a)

(1) Weight about 2.5g of sample and transfer to 250ml capacity volumetric flask with the help of dilute hydrochloric acid.

(2) Make up the volume with dilute hydrochloric acid.

(3) Apply stopper, shake well and filter through Whatman Filter paper No. 40 or equivalent part in a dry beaker if the solution is not clear and transparent.

(4) Take 25ml of the filtered aliquot in a beaker of 250 ml capacity. Add 100ml of water and heat to boil. While stirring add in a slow stream 1ml of hot barium Chloride solution for each one percent. Sulphur expected in sample plus additional 10 ml in excess to ensure complete precipitation of sulphur as barium sulphate. Boil for a minute.

(5) Digest the precipitate on a hot plate or water bath for 2 hours at low temperature such that the solution does not boil. Ensure the supernatent liquid to be clear and transparent. Cool to room temperature. Filter into a 30ml capacity G4 grade sintered Gooch crucible previously dried at 250C cooled and weighed. Wash the precipitate 10-12 times with hot water to ensure the precipitate to make it free from barium chloride. The filtrate may be tested with silver nitrate solution to confirm that the precipitate is free from chloride.

(6) Dry the crucible and its content to 250C for two hours in a furnace by raising the temperature of the furnace slowly from room temperature. After drying, cool to room temperature in a desiccator to a constant weight taken.

Calculations
Sulphate Sulphur (as S) per cent by weight=	137.4 x M W
Sulphate Sulphur (as S) per cent by weight=
Where
M = weight of precipitate
W = weight of the sample taken

Procedure (b) : Weigh about 2.5g of sample and transfer to a beaker of 250 ml capacity. Add 25ml of concentrated hydrochloric acid and 25 ml of water. Heat to boil the solution gently. Boil for 5 minutes and cool. Add 75 ml of water and transfer quantitatively into a volumetric flask of capacity 250ml with dilute hydrochloric acid solution and make up the volume. Apply stopper, shake will and filter about 50ml or prepared solution through Whatman filter paper No. 40 or equivalent and proceed as at step (2) of Procedure(a) onward.

Procedure (c) : (1) Weigh about 2.5g of sample and transfer to a beaker of 250ml capacity. Add 10ml of concentrated hydrochloric acid and grind the sample with the help of a glass rod. Evaporate to dryness on a hot plate. Soak the mass again with few drops of concentrated hydrochloric acid and evaporate to dryness.

(2) Add 100ml of dilute hydrochloric acid, heat to boil and cool to room temperature. Transfer the content to a 250ml volumetric flask. Filter a portion of the solution through a whatman No.40 filter paper or equivalent and proceed at step (2) of Procedure (4) onwards.

Note: Based on AOAC 1995.

(B) Determination of elemental Sulphur

(a) Reagents

(1) Hydrochloric Acid-concentrated (4N)

(2) Sulphur powder (AR Grade)

(3) Acetone saturated with sulphur -Add 2-3g of sulphur powder in 250ml of acetone in stoppered bottle. Shake well and allow it to stand for 1 hour. Filter through whatman filter paper No.42 & collect filtrate in a stoppered glass bottle.

(4) Carbon disulphide (AR Grade)

(b) Procedure

Take 1g of prepared sample in a beaker. Add 50ml of 4 N HCI boil for 5 minutes. Cool and filter through a sintered glass crucible (G4). Wash thoroughly with distilled water. Wash the residue 5-6 times with 10ml portion of Acetone saturated with sulphur. Dry at 100C for 1hour. Cool in a desiccator and take weight of the crucible. Let the weight be W1.

After taking he weight of the crucible (W1) wash the crucible 8-10 times with 10ml portion of carbon disulphide thoroughly under slow suction, Dry the crucible at 100C for 1 hour. Cool in a desiccator and weight Let this weight be W2.

Calculations
Elemental sulphur as (S) % per cent by weight=	(W1-W2) x 100
Elemental sulphur as (S) % per cent by weight=	W
Where "W" is the weight of the sample taken.

(C) Total sulphur (as S) per cent by weight = Sulphate sulphur (as per procedure 24)+elemental Sulphur (as per procedure 24A).]

25. Method of Analysis of Zincated Phosphate (Suspension).

Determination of total phosphate

By the method as specified in 4(ii)

Determination of Citrate soluble phosphates

By the method as specified in 4(v)

Determination of total Zinc

Procedure:

(ii) Cool, filter and make up the volume suitably and proceed as per the procedure specified at Method No. 7(iii)(b)(2) step (b) onwards or Method No. 8(ii)(b) step (2) onwards.

[26. Method of analysis of Zincated Oxide (suspension)

[(i) Determination of total zinc

Scope: Total extraction of Zinc (Zn), Arsenic (As), Lead (Pb) and Cadmium (Cd) from Zinc Oxide Suspension Concentrate.

Principle: Zinc and heavy metal impurities are extracted from the sample with boiling Aqua Regia.

(a) Sample preparation. - The details of the procedure for drawl of samples of fertilizers have been provided in Schedule II Part A, Serial No. 9 (Method for sampling of liquid fertilizers (other than anhydrous ammonia), however importance of proper shaking and mixing of contents before withdrawal of sample and before drawing aliquot for analysis is particularly emphasized. Since in suspension formulation insoluble materials might settle down over time, thorough mixing of contents would ensure representative composition along the entire depth of the container.

(b) Reagents. - All the reagents should be of analytical grade to ensure negligible concentration of the elements to be determined.

(1) Glass double distilled water (free from micronutrients)

(2) Hydrochloric acid 37% HCI (HCI) = 12 mol/1, p = 1.18 g/ml)

(3) Nitric acid 65% HNO (c(HNO₃) = 14.3 mol/1, p = 1.4 g/ml)

Apparatus for thermal heating digestion - with reaction vessel and reflux condenser.

The vessel should be at least 5 times the volume of the aqua regia used. In case reaction vessel with reflux condenser is not available, Erlenmeyer flask or high beakers covered with watch glass can be used for the purpose. Ash free filter paper is required if filtration is necessary.

(d) Procedure:

(1) Weight one gram (1 ± 0.001g) of the sample and transfer quantitatively to the reaction vessel

(2) Moisten the sample with about 0.5 to 1.0ml distilled water.

(3) Mix the contents well and 21 ml of HCL) followed by 7 ml of HNO₃ (reagent both drop wise to reduce foaming.

(4) Connect the condenser to the reaction vessel and let the mixture stand at laboratory room temperature until effervescence ceases.

(5) Turn on the heating device and slowly raise the temperature of the reaction mixture to reflux condition. Maintain for 2 hours.

(6) Ensure that the condensation zone is lower than half of the height of the condenser.

(7) After 2 hours of reflux, allow to cool and rinse the condenser with 10ml of distilled water.

(8) Transfer the contents quantitatively into a 500 ml volumetric flask and dilute to the mark with water. The test solution corresponds to a 500 times dilution of the sample.

(9) Test solution can be filtered, if necessary (should not be required for Zinc Oxide Suspension concentrate). If filtered, discard the first 20ml (approx) portion for analysis.

(10) Prepare a blank test solution following the same procedure as the sample. This is to be used for background correction of analysis, in case of any possible contamination through reagents.

(11) Measurement can be carried out immediately, or can be stored in tightly closed plastic vessels for up to 15 day.

Note. - Addition of one drop of octanol to the reaction vessel can be used as an antifoaming agent.

(e) Determination of Zinc in the extracted material:

Process for analysis of Zinc shall be as per the procedure specified at Method no. 7 (iii) (b) (2) step (B) onwards or method No. 8 (ii) (b) step (2) onward.

Note. - Calculation need to be adjusted accordingly, in view of extent of dilution of original sample.]

(ii) Determination of total Zinc

By the method as [specified in 8(v) after extraction through aqua regia as specified in sub-item (d) of item (i)]

(iii) Determination of specific gravity

By the method as specified in 21

[(iv) For determination of Arsenic

By the method specified as 8 (ix) after extraction through aqua regia as specified in sub-item (d) of item (i).

(v) For determination through Cadmium

By the method specified as 8 (x) after extraction through aqua regia as specified in sub-item (d) of item (i).]

27. Determination of Boron in Colemanite

(a) Reagents

(1) Mannitol

(2) Bismuth nitrate solution; Dissolve 22g of bismuth nitrate [(BiNO3)3 5H2O] in 8ml of cone. Nitric acid. Warm slightly and dilute to 100 ml with distilled water.

(3) Nitric acid-Concentrated AR grade.

(4) Dilute nitric acid-(1:20).

(5) Sodium hydroxide solution-10% M/V.

(6) Sodium hydroxide solution-0.5M.

(7) Bromothymol blue indicator solutions.

(b) Procedure-

(i) Weigh accurately 2.5g of sample into 250 ml of beaker.

(ii) Add 5ml of concentrated Nitric acid and 50ml of water, warm gently and dilute to 100ml cool and transfer the contents into 250 ml volumetric flask ad dilute to the mark with distilled water.

(iii) Filter through a dry filter paper Whatman No. 40 rejecting the first few drops into a beaker.

(iv) Pipette 100 ml of the filtrate into 250 ml beaker, add a few drops of indicator solution.

(v) Add few drops of 10% NaOH solution with thorough stirring until the indicator turns blue.

(vi) Filter through a dry filter paper into 250ml beaker and carefully wash the residue several times with cold water. The total volume should be 150-200ml.

(vii) Adjust the pH to about 5 by adding dilute nitric acid. Warm to about 90C (do not boil) and stir vigorously to avoid carbon dioxide. Cool the solution.

(viii) Adjust the pH to exactly 6.3 using NaOH solution.

(ix) Add 10g of mannitol and again adjust pH to exactly 6.3 with 0.5 NaOH solution.

(x) Continue adding mannitol in 10g portion till pH remains constant at 6.3. Note the total volume 0.5 NaOH use for the first addition of mannitol.

(xi) Carry out a blank determination to which borate is not added.

Percentage boron =

(A-B) x 1.3512

Where A is the volume of 0.5 N NaOH solution used in the sample

B is the volume of 0.5 N NaOH solution used in the blank.

28. Method of Analysis of Customized and Fortified Fertilizers

(i) Determination of nitrogen by method 3(iv) or 3(v).

(ii) Determination of ammoniacal nitrogen by method 3(vii).

(iii) Determination of neutral ammonium citrate soluble phosphate by method 4(v).

(iv) Determination of water soluble [Phosphorus (Available Phosphorus)] by method 4(iii).

(v) Determination of water soluble potash by method 5(i).

(vi) Determination of sulphur by method 24.

(vii) Determination of micronutrients in Customized or Fortified Fertilisers.

(a) Sample Preparation:-

1. Weigh accurately 2.5g sample into a 250ml beaker. Add 50ml of 1+1 HCI and cover with a glass.

2. Heat to boiling and continue to boil until volume is reduced to about 25ml.

3. Dilute to about 100ml with water and bring to boil.

4. Cool, transfer to a 500 ml volumetric flask and dilute to volume with water.

5. Mix thoroughly and allow to stand until a clear solution is obtained or filter a portion through a dry Whatman No. 40 filter paper.

6. Transfer 25ml aliquot to 250ml volumetric flask dilute to volume and mix thoroughly.

7. Dilute it further to suitable standard working range with acidified water.

8. Prepare a blank solution in the same manner, omitting the sample.

(b) Preparation of Standard Stock Solution, working standards and Determination.

As given in Method 22A

[29. Determination of Plant Available Silicon (S1(OH)4)

CaCl2 extraction of Silicon fertilizer amendment

Analysis with UV

(a) Reagents : Do not store any reagent in glass containers, and do not expose to glassware any longer than absolutely necessary while making up reagents, e.g., dissolve reagent/prepare solution in plastic beaker with slightly less than the final volume of solvent, and use volumetric glassware only for a short interval at the very end to make up to volume.

1. Extractant 0.01 M CaCl2. Dissolve 7.35g CaCl2 2H2O in de-ionised water, and make up to 5L.

2. Tartaric Acid: Dissolve 100g of tartaric acid and make up to 500ml with deionized water. Store in polyethylene bottle. Make up fresh solution when appreciable sediment forms. Store n refrigerator.

3. ANSA reducing agent: Dissolve 25g of sodium bisulphite (NaHSO3) in 200 ml water, and combine with a solution containing 2g of anhydrous sodium sulfite (Na2SO3) and 0.4g of 1-amino-2-napthol-4-sulfonic acid in 25ml of water. Dilute the combined solution to 250ml and store in a polyethylene bottle in fridge. Discard if solution darkens in colour.

4. Ammonium para molybdate tetrahydrate: Dissolve 54g ammonium molybdate (see note 2) in about 800 ml deionised water in a plastic beaker. Adjust pH to 7 with 5N NaOH or 0.5M H2SO4 depending on whether the pH needs to be raised or lowered. Make up to 1L. and store in polyethylene bottle in refrigerator.

5. 0.5M Sulfuric Acid: 56ml of cone. sulfuric acid (from plastic bottle) in 21 deionised water.

6. 5NNaOH: 20g of NaOH pellets in 100 ml of deionised water (Make and store in plastic container.

(b) Standards:

ml secondary standard in 50 ml flask	mg/L.Si in solution	Equivalent Si fertiliser mg/kg Si
0	0	0
0.5	0.5	5
1	1	10
2	2	20
3	3	30
4	4	40
5	5	50
6	6	60

Method:

1. Dry approximately 2-3 grams of Silicon fertiliser in a 105C oven overnight, Gind the sample if not already in powder form.

2. Weigh 0.4 of dried Silicon fertiliser into a 70ml container, and add 40 ml 0.01M CaCl2, Lid, and shake in an end over end shaker overnight (16hours). Centrifuge sample extract at 2000 rpm for 10 minutes. The extract should be clear.

3. Transfer 1ml of filtrate into a 20ml capacity test tube (See note 4)

4. Add 2.5ml of 0.5M sulfuric acid

5. Add 2.5ml ammonium molybdate solution

6. Shake well on a vortex stirrer and wait 5 minutes.

7. Add 1.25ml tartaric acid solution.

8. Add 0.25ml ANSA reducing solution Mix well.

9. Prepare a sample blank for each sample in a similar way, but using water instead of the ammonium molybdate solution. Also prepare a reagent blank, using 0.01M CaCl2 instead of sample and water instead of the ammonium molybdate solution. The reagent blank is used to zero the spectrophotometer prior to calibration.

10. Prepare a standard curve from the working standards by using the same aliquot for the standards as is used for the samples and treat the same way. (See note 5).

11. The spectrophotometer should be set to 820 mm wavelength. After 30 minutes, transfer reagent blank to spectrophotometer cell, place in machine and zero absorbance. Remove, transfer sample to cell, place in machine and record absorbance reading (ie. colour intensity due to amount of soluble silica Si in sample). Wash cell before next sample with a small amount of the next sample blank.

12. Read the concentration of silica Si in the soil sample, mg/kg Si, directly from a regression equation prepared from the standard curve.]

[Note: - 1. Discard ANSA solution if it does not completely dissolve, or yields a dark solution immediately. Try again after changing source of de-ionised water.
2. Molybdate compounds are considered as environmental hazard. Waste containing those elements has to be collected and disposed of according to regional requirements.
3. Avoid use of glass whenever possible. All standards and reagents should be kept in polyethylene or other plastic bottles, particularly NaOH to avoid any possibility of it being stored in glass. (NaOH is able to actively dissolve silica in glass)
4. Don not start filling glass test tubes in advances with any of the reagents or sample, at the glass can leach silicon.
5. As the same amount of each reagent is used for both standard solutions and samples, a small amount of Si contamination can be tolerated. The calibration line will allow for minor background contamination. However, if the blue colour in the zero stand becomes too intense, the working range may be pushed beyond the linear range of the calibration line, and all contain solutions must be prepared again. Si Contamination may occur in one or more of the following solutions; de-ionised water (used to make up one or all of the solutions), 0.5 sulfiuric acid, ammonium para molybdate, tartaric acid or ANSA. Another deionised water supply should be used, if available. If Si is still present, reagents and extractant may need to be prepared in distilled, no de-ionised water.]

[30. Method of analysis of Sodium Silicate

(i) Determination of Silicon

(a) Chemicals and Glasswares:

(1) Cone. HCI

(2) Cone. HNO₃

(3) Hot Plate

(4) Platinum Crucible

(5) Muffle Furnace (Temp, capacity by 10000C)

(6) Desiccators and other routine laboratory glasswares

(b) Procedure:

(1) Take 1 gm of prepared sample in Teflon or coming beaker, add 2-3 ml HCI solution (1:1) and 2-3 ml HNO₃ solution (1:1).

(2) Digest the solution on a hot plate till it becomes semi-solid, cool it and again add 3-4 ml HNO₃ solution. Dry it completely on hot plate.

(3) Add 2-3 ml HCI solution (1:1) and boil it till yellow fumes cease. Filter it with Filter Paper No. 1, wash with 10 ml HCI solution (1:1) one time and 2-3 washing with hot water of 10 ml portion each till yellow colour disappear.

(4) Transfer the filter paper alongwith residue in pre-weighed platinum crucible, dry for one and half hour in muffle furnace at 250°C temperature and finally ignite the residue at 950°C temperature for 30 minutes.

(5) Cool the crucible in desiccator and re-weigh and calculate Calculation:

% Si as SiO₂ = Final weight of crucible- empty weight of crucible x 100

Weight of sample

(i) Determination of sodium By the method as specified in Serial Number 17

(ii) Determination of Specific gravity By the method as specified in Serial Number 21

31. Method of analysis of nano Nitogen.

(i) Determination of Nitrogen - Total Kjeldahl Nitrogen

(a) Procedure:

(1) Liquid Nano Fertilizer sample : Take aliquot quantity (10 ml) of sample in the Kjeldahl flask.

(2) In case of solid Nano Fertilizer Sample : Take one gram of powdered nano-fertilizer sample is transferred to the Kjeldahl flask

(3) Add 15g K₂SO₄ or 12g anhydrous Na₂SO₄, 0.4g anhydrous N₂SO₄, or 0.6g CuSO₄.5H₂O, and approximately 0.8g alundum granules.

(4) Add 37ml diluted Sulfuric acid with water H₂SO₄+H₂O(1 + 1,v/v) or 20 ml concentrated Sulfuric acid, if adequate ventilation is available.

(5) Add sufficient test portion mass, precisely 0.1000 to 2.800 g for fertilizers with 30 to 5% nitrogen, respectively. Rinse the inner wall with about 10ml water.

(6) Transfer the flask to a preheated (400°C) Kjeldahl block digestor and digest test portions for 75 minutes.

(7) Remove the flask from the heating block and upon cooling (the reaction mixed must be near room temperature). Wash the inner wall with 20-30 ml water and mix.

(8) Prepare the distillate receiving flask (300ml Erlenmeyer flask) by adding 30ml of 0.25 N standardized Sulfuric acid to trap the expected total Nitrogen in the test portion.

(9) Add 2-3 drops of Methyl purple indicator and install the receiver on the outlet tube of the distillation unit, being sure that the distillate outlet tube end is totally immersed in the standardized acid solution.

(10) Install the digestion tube on the distillation unit. Initiate steam generation and slowly dispense about 80ml (30 -35%) Sodium Hydroxide into the flask.

(11) Continue steam distillation until about 250 ml or more of steam condensate has been collected in the receiving flask. This usually requires about 6-8 minutes.

(12) If color changes to green, add more 0.25 NH₂SO₄ to bring the color back to purple and record the amount of acid added.

(13) Titrate to a grey end point (pH5.7) with 0.25 N Standard NaOH. The color of the distillate depends upon the amount of total nitrogen in the test portion, which is a function of the amount of ammonia trapped in the receiver flask.

(14) A green color indicates that the acid in the trap was neutralized by the Ammonia. At this point, add an additional known amount of standardized H₂SO₄ to get to the grey end point.

(15) The net volume (in ml) of standardized acid would be equal to the total amount of acid initially added to the receiving flask plus the amount of the acid added, after distillation, to reach the grey end point. A blue or Purple colour indicates that there is still acid in the receiving flask, and back titration with NaOH is required.

(16) The net volume standardized acid would be equal to the amount of acid in the receiving flask minus the amount of base added, after distillation, to reach to the grey end point.

(b) Calculations:

Weight percent total nitrogen is calculated as follows:
Total N %= (net mL std acid x N of std acid)- (net mL std base x
N of std base)x 1.4008
Sample weight, g

(ii) Physical Particle Size (as per Transmission Electron Microscope (TEM) Analysis)

(a) Equipment and Apparatus:

(1) Transmission Electron Microscope

(2) Sample grid

(3) Tweezers, Petri dish. Ethanol and Deionized water

(b) Procedure:

(1) TEM sample grids carbon coated film that is electron transparent supported by copper mesh or equivalent are suitable.

(2) Glassware and apparatus used for sample preparation should be cleaned with filtered, demineralized water and stored dry.

(3) The apparatus used for dispersion and deposition of particles consists of a small glass vial with a screw-on cap, a teflon pillar about 10 mm high that may be inserted into the vial, a petri dish, and a teflon block about 40 mm by 40 mm square.

(4) Hold the grid with tweezers, dip rinse the grid thoroughly with ethanol. Wick the excess liquid off the grid using filter paper. Place the dried grid onto the clean Teflon block.

(5) Place a 10pL drop of the nanoparticle solution onto the grid.

(6) Cover the grid with a petri dish lid and let stand at room temperature for getting the grid dry, typically from 5 to 30 minutes, depending on solvent type.

(7) Transfer the grid on TEM sample holder. Record enough micrographs to image a minimum of 200 nanoparticles per sample per grid square from a minimum of 2 widely separated regions of the grid using a well-aligned and stable TEM, operated at a fixed magnification that allows a large number of nanoparticles to be visible within the micrograph field of view, while ensuring that each individual nanoparticle is recorded with a large number of image pixels.

(1) Transmission Electron Microscope grids (especially thin film membranes) are very fragile and must be held by their edges with fine tweezers so as not to damage or crack the membrane.

(2) Exposure of Transmission Electron Microscope grids to the ambient environment should be minimized to reduce the likelihood of dust contamination. Grids should be stored in suitable boxes in dust-free or desiccating cabinets.

(3) A well-aligned Transmission Electron Microscope is essential to obtain accurate particle size results.

(4) A minimum of 200 discrete particles should be measured from each of at least two widely separated regions of the sample (that is, different grid squares or membrane regions). Foreign debris in a given image (e.g., dust particles or residues from the rinsing and drying process) should be avoided.

(5) Particle size results obtained from Transmission Electron Microscope measurements may not coincide with those obtained from other techniques (e.g., dynamic light scattering). This is due in part to differences in the weighted averages determined in each case (e.g., number for Transmission Electron Microscope versus intensity for dynamic light scattering), as well as differences in the physical property that is actually measured (e.g., projected area versus hydrodynamic diffusion area).

(iii) Hydrodynamic particle size (as per Dynamic Light Scattering (DLS) Analysis

(a) Equipment and Apparatus:

(1) Dynamic Light Scattering Equipment

(2) Sample Cuvettes of size

(3) Deionized water

(b) Procedure:

(1) For liquid nano-fartilizer sample, 10 ml is sonicated for one minute and used for the analysis. In case for solid nano-fertilizer, fertilizer: distilled water (1:10 ratio) suspension is prepared, sonicated for one minute.

(2) Known standard samples (either one of the nano particles such as Ag, Au or TiO₂) are used to set the machine.

(3) Load sample into the Cuvette.

(4) Pre-rinse filter with solvent (at least 1ml, depending on filter size and dead volume of filter holder or cartridge).

(5) After loading syringe with sample and inserting syringe filter, allow the first 4 drops to go to waste. Use the next 4 drops to pre-rinse the cuvette, and discard. The remainder can be used for the sample measurement.

(6) Load sample into cuvette using minimum amount necessary to ensure liquid level is at least 2mm above the entrance height of the laser beam for your particular instrument configuration.

(7) Take care not to touch the cuvette windows with your bare hands while loading. Wipe outside of quartz or glass cuvette with lens paper if needed. Cap the cuvette to prevent dust contamination and solvent evaporation.

(8) Inspect the cuvette to ensure that air bubbles are not clinging to the optical window area.

(9) Load the sample into Dynamic Light Scattering machine.

(10) Perform 3 to 10 independent measurements per sample.

(1) To measure the size of solid phase nano nitrogen particles, 1 gram of nanoparticles sample should be suspended in 20ml water followed by 1 minute ultra-sonication before the size measurement carried out by the Dynamic Light Scattering.

(2) Measurement cuvettes should be cleaned with filtered demineralized water and stored dry.

(3) The choice of pore size depends on the maximum dimension of the test particles and their tendency to adhere to the filter membrane, µ? Suspended medium (such as solvent, dispersant, solution) should be filtered prior to sample preparation using a 0.1 or 0.2 .

(4) A typical starting sample concentration is 1 mg/ml.

(5) Use cuvette with quartz or equivalent optical-quality windows.

(6) Pre-rinse cuvette with filtered solvent at least 3 times.

(iv) Zeta Potential Analysis

(a) Equipment and Apparatus:

(1) Dynamic Light Scattering (DLS).

(2) Sample Cuvettes of zeta.

(3) Deionized water.

(b) Procedure:

(1) For liquid nano-fertilizer sample, 10 ml is sonicated for one minute and used for the analysis and in case for solid nano-fertilizer, fertilizer : distilled water (1:10 ratio) suspension is prepared, sonicated for one minute.

(2) Known standard samples (either one of the nano particles such as Ag, Au or TiO₂ are used to set the machine.

(3) Loading Sample into the Cuvette.

(4) Pre-rinse filter with solvent (at least 1ml, depending on filter size and dead volume of filter holder or cartridge).

(5) After loading syringe with sample and inserting syringe filter, allow the first 4 drops to go waste. Use the next 4 drops to pre-rinse the cuvette, and discard. The remainder can be used for the sample measurement.

(6) Load sample into cuvette using minimum amount necessary to ensure liquid level is at least 2mm above the entrance height of the laser beam for your particular instrument configuration.

(7) Take care not to touch the cuvette windows with your bare hands while loading. Wipe outside of quartz or glass cuvette with lens paper if needed.

(8) Cap the cuvette to prevent dust contamination and solvent evaporation.

(9) Inspect the cuvette to ensure that air bubbles are not clinging to the optical window area.

(10) Load the sample into Dynamic Light Scattering machine.

(11) Perform 3 to 10 independent measurements per sample.

(1) Measurement cuvettes should be cleaned with filtered demineralized water and stored dry. The choice of pore size depends on the maximum dimension of the test particles and their tendency to adhere to the filter membrane. µ suspending medium (such as solvent, dispersant, solution) should be filtered prior to sample preparation using a 0.1 or 0.2

(2) A typical starting sample concentration is 1 mg/ml.

(3) Use cuvette with quartz or equivalent optical-quality windows.

(4) Pre-rinse cuvette with filtered solvent at least 3 times.

(v) Viscosity Measurement:

(a) Equipment and Apparatus:

(1) Demineralised water.

(2) Viscometer.

(3) Measuring cylinder.

(4) Bottle Adapter.

(b) Procedure:

(1) Calibrate the viscometer with distilled water to set the machine as 1 cps.

(2) Viscometer Assembly.

(3) Attach vertical pole to the base using the wrench provided in the case.

(4) Attach the viscometer to the vertical pole.

(5) Connect power cable to the viscometer.

(6) Turn on the viscometer and calibrate.

(7) Starting up - Select and attach required spindle.

(8) Raise the viscometer to the highest level using a screw on the vertical pole.

(9) Place beaker with liquid under the spindle.

(10) Lower the viscometer until the spindle is submerged to the spindle's mark.

(11) Viscometer Operation-This depend on the software of the instrument make. The analyst should follow the manufacturer's operating instructions for a particular instrument. Set the viscometer using distilled water to measure lcps at room temperature.

(1) Wait for 30-60 sec before reading.

(2) Good results are in range 60%-80% of torque.

(3) Results depend on beaker and volume of liquid so use the same beakers for comparison measurements.Viscosity of a Liquid nano-fertilizer N is measured directly.

(4) For solid nano-fertilizer, fertilizer: water suspension should be in 1:10 ratio with distilled water.

(vi) Measurement of pH

(a) Apparatus: pH meter, vacuum pump, beaker, pipette, glass rod, china dish, spatula etc.

(b) Reagents: Buffer solutions of pH 4.0, 7.0 and 9.2: One buffer tablet of the respective pH is dissolved in water and the volume is made to 100 ml.

(1) Take 10 ml of liquid sample, homogenise it and take the pH measurement. For solid/powder samples (1g dry sample/10 ml water) homogenisation or ultrasonic agitation of the sample for 1 minute followed by pH measurement should be taken after settling of the samples

(2) pH meter is set at room temperature and calibrated by immersing the electrodes in different buffer solutions of pH 4.0, 7.0 and 9.2.

(3) Take the beaker of homogenised samples and dip the electrodes into it and note the pH reading.

(4) After each determination the electrodes must be washed with distilled water and wiped out by ordinary filter paper.

(d) Precautions:

(1) Proper homogenisation/sonication must be done.

(2) The glass and reference electrode of pH meter should always remain dipped in water.

(3) Buffer solutions should be prepared accurately and stored well in glass container.

(4) It is desirable to prepare fresh buffer solutions after few days. Connect the pH meter to the stabilizer to avoid the fluctuations in pH readings. Adjust the temperature knob of pH meter at room temperature for correct pH determination]

32. [-]

[33. Method of analysis of potassium Thiosulphate and Calcium Thiosulphate (Liquid Fertilizers)

(i) Determination of potassium by the method as specified in 5 (ii);

(ii) Determination of Specific Gravity by the method as specified in 21 (ii);

(iii) Determination of Calcium by the method as specified in 21 A (v);

(iv) Determination of Arsenic by the method as specified in 8 (ix);

(v) Determination of Lead by the method as specified in 8(v);

(vi) Determination of Sulphur (as S) in potassium thiosulphate and calcium thiosulphate (Liquid Fertiliser)-

(a) Apparatus

(i) Electric hot plate, water bath, muffle furnace, Oven;

(ii) Electronic balance;

(iii) Desiccators;

(iv) Gooch crucible;

(v) Vacuum pump for filtration;

(vi) Electronic balance (0.001 g capacity);

(vii) Whatman No 40 and 50 filter papers;

(viii) Routine laboratory glassware, instruments;

(b) Reagents

(i) Cone. Hydrochloric acid (37% HCL);

(ii) Dilute hydrochloric acid - Take 250 ml of concentrated hydrochloric acid in one litre volumetric flask. Make the volume upto 1000 ml water. Shake and mix it well';

(iii) Barium chloride solution: Dissolve 120 gm barium chloride in 1000 ml of water;

(iv) silver nitrate (5 percent solution): Dissolve 5 g of silver nitrate in 100 ml of distilled water;

(v) Hydrogen per oxide 30 percent (fresh) solution: Dilute liquid hydrogen peroxide to bring final concentration 30 percent;

(vi) Sodium hydroxide solution: Dissolve 40 g sodium hydroxide salt in 100 ml distilled water;

(vii) Methyl orange indicator: Dissolve lg methyl orange indicator in 100 ml alcohol. Shake it and store in cool dry Place;

(viii) Sodium Thiosulphate : AR Grade.

1. Oxidation of liquid fertilizer test sample;

(a) Weight 5.0 g sample of potassium thiosulphate (or calcium thiosulphate) liquid fertilisers (up to 0.001 g) accurately in a dry, clean 500 ml graduated flask on an electronic balance. Denote "W" to this as weight of sample. Fill the flask to the mark of 500 ml by using double distilled water'.

(b) Add exactly 50 ml (of the diluted sample from (a) to a 500 ml beaker and add approximately 100 ml of water. Add 2 ml of 40% NaOH solution to it. Add now slowly 2 ml of 30% hydrogen per oxide (H202) solution. Cover the beaker with watch glass carefully. Heat the solution on low heat on hot plate slowly for about 30 minutes. Cool it and then add slowly 2 ml of hydrogen peroxide 30% solution again. Heat the solution on hot plate slowly. If required add more 1 to 2 ml of hydrogen per oxide solution. Total hydrogen peroxide solution addition should not be more than 6 ml for complete oxidation. Cool the beaker; add 100 ml of water and then mix.

(c) Add a few drops of methyl orange indicator to the solution. Add 2-5 ml of dilute hydrochloric acid and mix it until colour changes to red uniformly. Add a few more drops of acid and mix well with glass rod. Then, keep the beaker on hot plate (below boiling temperature, (800C) to bring the solution to colorless within 10 minutes. Note: If necessary, add a few drops of 30% H202 to the beakers to bring the solution to colourless.

2. Precipitation of sulphur for gravimetric determination of total sulphur,-

(a) Add slowly 15 ml barium chloride to the oxidized sample to ensure complete precipitation of sulphur as barium sulphate;

(b) Digest the precipitation on a hot plate or water bath for minimum 5 hours at such a temperature that the solution does not boil (around 800C). Put a watch glass above the beaker to prevent complete evaporation of the solution. Ensure the supernatant liquid to be clear and transparent. Cool it to room temperature and keep it over night to settle precipitates;

(c) Take 30 ml capacity G4 grade Sintered Gooch crucible. Dry it in furnace at 2500C; cool it and record weight of empty crucible(Gl);

(d) Pour the sample with precipitate into 30 ml capacity G4 grade Sintered Gooch Crucible previously weighed. Filter it. Wash the precipitate 10-12 times with hot water to ensure the precipitate to make it free from excess barium chloride. The filtrate may be tested with silver nitrate solution to confirm that the precipitate is free from chloride.

(e) Dry the crucible having barium sulphate precipitate at 2500 C for two hours in a furnace by raising the temperature of the furnace slowly from room temperature to constant weight taken and note the weight (G2).

3. Calculations -

Total sulphur (as S) percent = (G2-.G1) X 13.74 X DF

Whereas

Gl= weight of Gooch crucible (+ filter paper if used)

G2 = Weight of Gooch crucible + barium sulphate precipitate (+ filter paper if used) W= Weight of sample

Factor 13.74= Per cent total sulphur (w/w) in barium sulphate = 32 x 100/233.39

34. Method of analysis of Phospho Gypsum. - 1. Determination of calcium as calcium Sulphate Dihydrate in Phospho Gypsum

(i) Standarisation of calcium solution-as per method specified in 21A (b)

(ii) Procedure -

(a) Take 1 gram of sample in 250 ml dry beaker. Add 50 ml dilute Hydrochloric acid (1:1)

(b) Boil for 5 minutes on slow heater and cool to room temperature. Add 50 ml distilled water.

(d) Take 25 ml aliquot in 250 ml washed conical flask and 20 ml distilled watter and 20 ml buffer solution. Add few drops of indicator and tighted with EDTA solution till blue color is obtained. Note the volume of EDTA used as V2.

Note - if the solution is not clear after digestion filter it and then take aliquot.

2. Calculation - Calcium (as Calcium Sulphate Dihydrate) per cent.= V2 X Calcium titer X 3. 4 X 1]

[Schedule III]

[Refer clauses 2(h) and (q)]

[Part-A]

Specifications of Bio Fertilisers

1.	Rhizobium
	(i) Base	=	Carrier based' in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	(ii) Viable cell count	=	CFC minimum-5x10 cell/g of powder granules or carrier material or 1x10 cell/ml of liquid.
	(iii) Contamination level	=	No contamination at 10 dilution
	(iv) pH	=	[5.0-7.0]
	(v) Particle size in case of carrier based material	=	All material shall pass through 0.15-0.212 mm IS sieve
	(vi) Moisture percent by weight, maximum in case of carrier bases	=	30-40%
	(vii) Efficiency Character	=	Should show effective modulation on all the species listed on the packet.
Type of carrier:
The carrier material such as peal, lignite, peat soil, humus, wood charcoal or similar material favoring growth of the organism.
2.	Azotobacter
	(i) Base	=	Carrier based' in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	(ii)Viable cell count	=	CFU minimum 5x10 cell/g of carrier material or 1x10 cell/ml of liquid
	(iii) Contamination level	=	No contamination at 10 dilution
	(iv) pH	=	[5.0-7.0]
	(v) Particle size in case of carrier based material	=	all material shall pass through 0.15-0.212mm IS sieve
	(vi) Moisture percent by weight maximum	=	30-40%
	(vii) Efficiency Character	=	The strain should be capable of fixing at least 10mg of nitrogen per g of sucrose consumed.
Type of carrier:
The carrier material such as peal, lignite, peal soil, humus, wood charcoal or similar material favoring growth of the organism.
3.	Azospirillum
	(i) Base	=	Carrier based' in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	(ii) Viable cell count	=	CFU minimum 5x10 cell/g of powder/granules or carrier material or 1x10 cell/ml of liquid material
	(iii) Contamination level	=	No contamination at 10 dilution
	(iv) pH	=	[5.0-7.0]
	(v) Particle size in case of carrier based material	=	All material shall pass through 0.15-0.212 mm IS Sieve
	(vi) Moisture percent by weight, maximum in case of carrier based	=	30-40%
	(vii) Efficiency Character	=	Formation of white pellicle in semisolid Nitrogen free bromothymol blue media.
Type of Carrier
The carrier material such as peat, lignite, peat soil, humus, wood charcoal or similar material favoring growth of the organism.
4.	Phosphate Solubilising Bacteria
	(i) Base	=	Carrier based' in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	(ii) Viable cell count	=	CFU minimum 3x10 cell/g of carrier material or 1x10 cell/ml of liquid material
	(iii) Contamination level	=	No contamination at 10 dilution
	(iv) pH	=	[5.0-7.0] for moist/dry powder granulated carrier based and 5.0-7.5 for [liquid based or capsule in gelatin base]
	(v) Particle size in case of carrier based material	=	All material shall pass through 0.15-0.212mm IS Sieve
	(vi) Moisture percent by weight, maximum in case of carrier based	=	30-40%
	(vii) Efficiency Character	=	The strain should have phosphate solubilizing capacity in the range of minimum 30% when rested spectro photometrically. In terms of zone formation, minimum 5 mm solubilization zone in prescribed media having at least 3 mm thickness
[5.	Mycorrhizal Biofertilisers
	(i) Form/base	=	Fire powder/tablets/granules/mot blomass mixed with growing substrate
	(ii) Particle size for carrier based	=	90% should pass through 250 micron IS sieve powder formulations (60 BBS)[In case of granules, 90 percent of the material shall pass through 6mm IS Sieve.]
	(iii) Moisture content percent maximum	=	[12]
	(iv) pH	=	[5.0-7.0]
	[(v) Total viable propagules/gram of product	=	100gm of finished product with minimum 60 spores per gram
	(vi) Infectivity potential	=	Inoculum Potential : 1200 IP/g determined by MPN method with 10 fold dilution)]]
Type of carrier-
The carrier material such as peat, lignite, peat soil, humus, wood charcual or similar material favouring growth of organism.
[6.	Potassium Mobilizing Biofertilizers (KMB)
	1. Base		Carrier based in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	2. Viable cell count		CFU minimum 5x10 cells/g of powder, granules or carrier material on dry weight basis or 1x10 cell/ml of liquid
	3. Contamination		No contamination at 10 dilution
	4. pH		[5.0-7.0] for carrier based in form of powder or granules and 5.0-7.5 for [or capsule in gelatin base]
	5. Particle size in case of carrier based moist powder		Powder material shall pass through 0.15 to 0.212 mm IS sieve.
	6. Moisture percent by weight, maximum in case of powder based		30-40
	7. Efficiency character		Minimum 10mm solubilization zone in prescribed media having at least 3mm thickness.
Type of carrier-The carrier material such as peat, lignite, peat soil, humus, tale or similar material favouring growth of micro-organisms.
7.	Zinc Solubilizing Biofertilizers (ZSB)
	1. Base		Carrier based in form of moist/dry powder or granules, or [liquid based or capsule in gelatin base]
	2. Viable cell count		CFU minimum 5x10 cells/g of powder granules or carrier material on dry weight basis or 1x10 cell/ml of liquid
	3. Contamination		No contamination at 10 dilution
	4. pH		[5.0-7.0] for carrier based in form of powder or granules and 5.0-7.5 for [liquid based or capsule in gelatin base]
	5. Particle size in case of carrier based moist powder		Powder material shall pass through 0.15 to 0.212 mm IS sieve
	6. Moisture content percent by weight, maximum in case of carrier based		30-40
	7. Efficiency character		Minimum 10 mm solubilization zone in prescribed media having at least 3 mm thickness.]
[8.	Azotobacter
	(i) Base		Carrier based' in form of moist/dry powder or granules of [liquid based or capsule in gelatin base]
	(ii) Viable cell count		CFU minimum 5x10 cells/g of power/granules or carrier material of 1x10 cells/ml of liquid
	(iii) Contamination level		No contamination at 10 dilution
	(iv) pH		5.5-6.0 for moist/dry power, granulated or carrier based and 3.5-6.0 for liquid.
	(v) Particle size in case of carrier based material		All material shall pass through 0.15-0.212 mm IS sieve
	(vi) Moisture per cent by weight, maximum in case of carrier based		30-40%
	(vii) Efficiency character		Formulation of yellowish pellicle in remisolid medium N free medium]
[9.	Carrier Based Consortia
	(i) Base		Carrier based in form of moist powder or granules
	(ii) Viable count		CFU minimum in a mixture of any 2 or maximum three of following micro organisms;
			CFU minimum
			Rhioblum or Azotobacter or Azospirillum 1x10 per g
			CFU minimum PSB 1x10 per g
			CFU minimum KSB 1x10 per g
	(iii) Particle size in case of carrier based moist powder		All material shall pass through 0.15 to 0.212 mm IS sieve
	(iv) Total viable count of all the biofertilizer organisms in the product		CFU minimum 5x10 cells per gm of carrier/powder
	(v) Moisture percent by weight, maximum in case of carrier based		30-40%
	(vi) Contamination		No contamination at 10 dilution for carrier based/granule based inoculants
	(vii) Efficiency character Azotobacter		The strain should be capable of fixing at least 10mg of Nitrogen fixation/g of C-source
	Azospirillum		The strain should be capable of fixing at least 10mg of N-fixation/g of malate applied.
	PSB		Minimum 5mm zone of solubilization zone on PSB media having at least 3 mm thickness
	KMB		Minimum 5mm zone of solubilization of KSB media having at least 3mm thickness
	Rhizobium		Nodulation test positive
10.	Liquid Consortia
	(i) Individual viable count in Liquid based		CFU minimum in a mixture of any 2 or more of following micro-organisms
			CFU minimum
			Rhizobium or Azotobacter or
			Azospirillum 1x10 per ml
			CFU minimum PSB 1x10 per ml
			CFU minimum KSB 1x10 per ml
	(ii) Total viable count of all the biofertilizer organisms in the product		CFU minium 5x10 cells per ml of [liquid based or capsule in gelatin base]
	(iii) Contamination		No contamination at any dilution
	(iv) pH		5.0-7.0
	(v) Efficiency character Azotobacter		The strain should be capable of fixing at least 10 mg N-fixation/g of C-source
	Azospirillum		The strain should be capable of fixing at least 10 mg of N-fixation/g of malate applied
	PSB		Minimum 5mm zone of solubilization zone on PSB media having at least 3 mm thickness
	KMB		Minimum 5 mm zone of solubilization of KSB media having at least 3mm thickness
	Rhizobium		Nodulation test positive.]
[11.	Phosphate solubilizing fungal biofertilizer
I.	Base		Carrier based in the form of moist/dry powder or granules or [liquid based or capsule in gelatin base]
II.	Moisture percentage by weight, maximum in case of carrier based		10
III.	Spore Count (per ml or gram)		Minimum 1x10 spores/g 1x10 viable fungal spores/ml of the liquid
IV.	Contamination		Nil for liquid inoculums 1x10 cells/gm for carrier based preparation
V.	pH		Liquid : 3.5 to 5.5
			Carrier : 6.0 to 7.0
VI.	Efficiency character		The strain should have phosphate solubilization capacity in the range of 30% when tested spectrometrically. In terms of zone formation, minimum 10 mm solubilization zone in prescribed media having least 3mm thickness.]

Part-B

Tolerance Limit of Biofertilisers

[1. In case of Rhizobium, Azotobacter, Azospirillum and Phosphate Solubilising. Bacteria, the total viable count shall not be less than 1x10 CFU/gm of carrier material in the form of powder or guanules or 5x10 CPU/ml in case of liquid formulations during the entire period of shelf life.

2. In case of Mycorrhizal Biofertilisers, the viable propagules shall not be less than No.]

[Part-C]

Procedure for drawal of sample of bio-fertilisers procedure for sampling of bio-fertilisers

1. General Requirements of Sampling

1.0. In drawing, preparing and handling the samples, the following precautions and directions shall be observed.

1.1 Sampling shall be carried out by a trained and experienced person as it is essential that the sample should be representative of the lot to be examined.

1.2 Samples in their original unopened packets should be drawn and sent to the laboratory to prevent possible contamination of sample during handling and to help in revealing the true conditions of the material.

1.3 Intact packets shall be drawn from a protected place not exposed to dampness, air, light, dust or soot.

2. Scale of Sampling

2.1 Lot

All units (containers in a single consignment of type of material belonging to the same batch of manufacture) shall constitute a lot. If a consignment consists of different batches of the manufacture the containers of the same batch shall be separated and shall constitute a separate lot.

2.2 Batch

All inoculant prepared from a batch fermentor or a group of flasks (containers) constitute a batch.

2.3 For ascertaining conformity of the material to the requirements of the specification, samples shall be tested from each for separately.

2.4 The number of packets to be selected from a lot shall depend on the size of the lot and these packets shall be selected at random and in order to ensure the randomness of selection procedure given in IS-4905 may be followed.

3. Drawal of Samples

3.1 The Inspector shall take three packets as sample from the same batch. Each sample constitutes a test sample.

[3.2 Of the sample of biofertilisers drawn by the Inspector, the label affixed therein shall be defaced and a label containing generic name of bio-fertilisers or organic fertilisers, expiry date, code number and date of sampling shall be pasted on the upper margin of sample packet/container. The sample shall be kept in cloth bag and be sealed with Inspector seal. The sealed cloth bag containing sample along with form 'P' shall be kept in another cloth bag and be sealed with inspector seal. The identifiable detail such as code number, date of sampling name of bio fertiliser shall be put on cloth bag containing biofertiliser with form 'P'.

3.3 Out of three samples collected one sample so sealed shall be sent to Incharge of the laboratory notified by the State Government under clause 29 or the Director National Centre of Organic Farming at Ghaziabad. In case where the sample is sent to Director National Centre of Organic Farming at Ghaziabad, or his authorized officer shall recode the sample and forward to any of the Regional Centre of Organic Farming at Bengaluru, Bhubaneshwar, Gaziabad, Imphal, Jabalpur, Nagpur and Panchkula for analysis. Another sample shall be give to manufacturer or dealer, as the case may be. The third sample shall be sent by the inspector to the next higher authority for safe custody. The appellate authority shall be sent any of the latter two samples for referee analysis under sub-clause (2) of clause 29B.]

3.4 The number of samples to be drawn from the lot

Let/Batch	Number of Samples
Upto 5,000 packets	03
5,001-10,000 packets	04
More than 10,000 packets	05

Part-D

Methods of Analysis of Biofertilisers

1A. Method of analysis of rhizobium bio-fertilisers

1. Apparatus

1.1 Pipettes Graduated 1ml and 10ml

1.2 Dilution Bottles or Flasks

1.3 Petri Dishes Clear, Uniform, flat-bottomed.

1.4 Hot-Air Oven

Capable of giving uniform and adequate temperature, equipped with a thermometer, calibrated to read up to 250C and with vents suitably located to assure prompt and uniform heating.

1.5 Autoclave

1.6 Incubator

1.7 Hand Tally of Mechanical counting Device

1.8 pH meter

2. Reagents

2.1 Congo Red-one percent aqueous solution

2.2 Medium

Use a planting medium of the following composition:

Agar	20g
Yeast Extract	1g
Mannitol	10g
Potassium hydrogen phosphate (K2HPO4)	0.5g
Magnesium sulphate (MgSO47H2O)	0.2g
Sodium Chloride (NaCI)	0.1g
Congo red	2.5ml
Distilled water	1000ml
pH	7.0

2.3 Sterilizing and preparation procedure for plates:

2.3.1 Sterilize the sampling and plating equipment with dry heat in a hot air oven at not less than 160C for not less than 2 hours.

2.3.2 Sterilize the media by autoclaving at 120C for 20min. To permit passage of steam into and from closed containers when autoclaved, keep stopper slightly loosened or plugged with cotton. Air from within the chamber of the sterilizer should be ejected allowing steam pressure to rise.

Preparation of planting Medium and Pouring

2.3.3 Prepare growth medium in accordance with the composition of the specific biofertiliser

2.3.4 Melt the required amount of medium in boiling water or by exposure to flowing steam in partially closed container but avoid prolonged exposure to unnecessarily high temperature during and after melting. Melt enough medium which will be used within 3h. Re-sterilisation of the medium may cause partial precipitation of ingredients.

2.3.5 When holding time is less than 30 min, promptly cool the molten medium to about 45C, and store until used, in a water bath or incubator at 43 to 45C. Introduce 12 to 15 ml of liquefied medium or appropriate quantity depending on size of the petridish at 2 to 44C into each plate. Gently lift the cover of the dish just enough to pour in the medium, Sterilise the lips of the medium containers by exposure to flame.

(a) immediately before pouring.

(b) Periodically during pouring, and

(c) When pouring is complete for each batch of plates, if portions of molten medium remain in containers and are to be used without subsequent sterilization of pouring additional plates. As each plate is poured thoroughly mix the medium with test portions in the Petri dish.

2.3.6 By rotating and tilting the dish and without splashing the medium over edge, spread the medium evenly over the bottom of the plate. Provide conditions so that the medium solidifies with reasonable promptness (5-10 min) before removing the plates from level surface.

3. Preparation of serial dilutions for plate counts

3.1 Dispense 30g of Inoculant to 270ml of sterile distilled demineralized water and shake for 10 min on a reciprocal shaker or homogeniser. Make serial dilutions up to 10 Take 0.1 ml or suitable alliquots of 10 to 10 dilutions using sterile pipettes and deliver to Petri dishes containing set medium as given in 2.1 and spread it uniformly with a spreader. Invert the plates and promptly place them in the incubator.

4. Incubation of plates

4.1 Label the plates and incubate at 28+/2C for 3 to 5 days for fast growing Rhizobia and 5 to 10 days for slow-growing ones.

4.2 Colony Counting aids.

Count the colonies with the aid of magnifying lens under uniform and properly controlled, artificial illumination. Use a colony counter, equipped with a guide plate and rules in centimeter square. Record the total number of colonies with the hand tally. Avoid mistaking particles of undissolved medium or precipitated matter in plates for pin-point colonies. To distinguish colonies from dirt, specks and other foreign matter, examine doubtful objects carefully.

4.3 Count all plates but consider for the purpose of calculation plates showing more than 30 and less than 300 colonies per plate. Disregard colonies which absorb congo red and standout as reddish colonies. Rhizobium stands out as white, translucent, glistening and elevated colonies. Court such colony numbers and calculate figures in terms of per litre, of carrier. Also check for freedom contamination at 10 dilution.

5. Test for Nodulation

5.1 Pot Culture Test

Plant Nutrient Solution

Composition	Concentration	g/l
(a) Potassium chloride	0.001M	0.0745
(b) Potassium hydrogen phosphate (K₂HPO₄)	0.001M	0.175
(c) Calcium sulphate (CaSO₄₂H₂O)	0.002M	0.344
(d) Magnesium sulphate (MgSO₄₇H₂O)	0.001M	0.246
(e) Trace elements solution:
(1) Copper sulphate (CuSO₄SH₂O)	0.01mg/kg	0.78
(2) Zinc sulphate (ZnSo₄₇H₂O)	0.25mg/kg	2.22
(3) Manganese sulphate (MnSO₄4H₂O)	0.25mg/kg	2.03
(4) Ammonium molybdate [(NH4)+MO₇O₂₄H₂O]	0.0025mg/kg	0.01
(5) Boric acid (H₂BO₄)	0.125mg/kg	1.43
Prepare the solution No. (e) consisting of trace elements in one litre of stock solution and add final nutrient solution at the rate of 0.5 ml per litre.
(f) Iron solution	g/100ml
(1) ferrous-sulphate	5
(2) Citric acid	5
Prepare the solution No. (f) as 100 ml of stock solution and add final nutrient solution at the rate of 0.5 ml per litre.

Preparation

Notes: 1. The nutrient solution may be prepared in the tap water provided the water is soft.
2. The nutrient solution should be shaken well to disperse calcium sulphate before dispensing.
3. If the solution is made up with distilled water, the pH is about 7.2 before autoclaving and falls to 5.5 on autoclaving and rises slowly on standing to about 5.8. However, there is no need to adjust pH. For most tropical legumes, pH of about 6.0 is adequate.

5.3 Procedure

5.3.1 Immerse the seeds in 95 percent alcohol and follow by surface-sterilization in freshly prepared chlorine water (for 15 to 20 min) or 0.1 percent mercuric chloride solution 3 min in a suitable container such as a screw-capped bottle or a test-tube with a rubber hung. In case of seeds with tough seed coat, concentrated sulphuric acid may be used as a surface sterilant for 20 to 30 min. It is recommended that the seeds should be placed overnight in a desiccator containing calcium chloride before surface sterilization with sulphuric acid. Pour out the sterilant and wash the seeds in several changes of sterile water and wash the seeds in several changes of sterile water (at least ten times) to get rid of the sterilant. Fill earthenware or glazed pots with soil (2 parts soil and 1 part washed coarse sand) (pH 6 to 7) and autoclaving for 2h at 120 C. After two days incubation at room temperature, repeat autoclaving to ensure complete sterility of soil. Inoculate surface-sterilized seeds with a water slurry of the inoculant taken from a culture packet (15to 100g seeds per gram of inoculant depending on the size of the seed) and sow the seeds. Keep a set of pots with uninoculated seeds as control and also a set of pots with ammonium nitrate at the rate of 100kg N/he as control aid incubate them in a pot-culture house during appropriate seasons for appropriate plants, taking care to separate the inoculated pots from the control pots. If growth rooms or cabinets having facilities to adjust temperature and light are available, the post may be incubated in such controlled environmental conditions. Sterilize the nutrient solution at 120C for 20 min and irrigate each pot once to the mosisture holding capacity of soil. Subsequently, water the seedling periodically with sterilized water preferably through a plastic tube, taking care to prevent splashing of water from inoculated pots to uninoculated ones. Maintain required number of replicated pots (4 to 6) for each botanical species for statistical analysis.

5.3.2. After two to three weeks of growth, thin down the number of plants in each pot to four uniform plants. At the end of 6 to 8 weeks, take one set of pots from both the control and inoculated series and, separate the plants carefully from the soil under slow-running water. Obtain date on the number, colour (effective nodules are pink or red) and mass of nodules. At the end of 6 to 8 weeks, harvest the shoot system, dry at 60C for 48 H and determine dry mass. For the above purpose, maintain adequate replications of pots (4 to 16).

5.3.3 Record the Nodulation data regarding formation of pink colour of nodules as revealed visually when nodules are cut open by a razor blade. After computing the data, based on the dry mass of plants and nodulation data decide the effectiveness of cultures. If good effective pink modulation is obtained in inoculated plants together with local absence or sometimes presence of stray nodules in controls and if there is a 50 percent increase in the dry mass of plants over the uninoculated control without nitrate, if may be concluded that the culture is of the required quality.

1B. Method of analysis of azotobacter bio-fertiliser

1. Apparatus-same as of Rhizobium

2. Reagents

2.1 Medium

use a planting medium of the following composition

Agar	20g
Sucrose (C₁₂H₂₂O₁₁)	20.0g
Ferric sulphate Fez (SO₄)₃	0.1g
Dibasic potassium phosphate (K₂HPO₄)	1.0g
Magnesium sulphate (MgSo₄,₇H₂O)	0.5g
Sodium Chloride (NaCl)	2.0g
Sodium Molybdate (Na₂MoO₄)	0.005gms
Distilled water	1000ml
pH	6.8 to 7.2

2.2 Sterilization & preparation procedure for plates : same as for Rhizobium

Preparation of Planting Medium and pouring

3. Preparation of serial dilutions for plate counts

4. Incubation of plates: Same as Rhizobium

4.1 Label the plates and incubate at 28+/-3^°C for 4 to 6 days.

4.2 Colony counting aide Same as Rhizobium

Azolobactor chrococcum colonies are gummy, raised with or without striations, viscous and often sticky. The pigmentation varies from very light brown to black Count the colony number and observe the cyst formation as given below and calculate number per gram of the carrier material.

Grow the vegetative cells at 30^°C on Burks agar medium comprising sucrose 20g, dipotassium hydrogen phosphate 0.64g, dihydrogen potassium phosphate 0.20g, sodium chloride 0.20g; calcium sulphate 0.05g, sodium molybdate 0.001g; ferric sulphate 0.0003g, agar 20g and distilled water 1.000ml. Look for vegetative cells after 18 to 24h either by simple staining method or through a phase contrast microscope.

Grow the cyst cells on Burks agar medium as given above with 0.3 percent n-butanol in place of the carbon source. Look for cyst formation after 4 to 5 days in cubation.

5. Test for nitrogen fixation in pure culture

5.1 Pure culture medium

5.1.1 Prepare medium as given for Azotobacter (2.1 under 1B), excluding agar.

5.2 Procedure

Select from each Azotobacter colony, of the type that has been counted as Azotobacter chroococcum in. One colony and plate on the medium given in. Use this pure culture for inoculating the broth for nitrogen fixation. For this purpose, take 50-ml aliquots of broth in 250-ml conical flasks for inoculation. After 12 days growth at 28^°C, test the contents on the flasks for purity by streaking on fresh medium and concentrating over a water-bath (50 to 60^°C) to dryness. Wash the dried culture and take it as a sample. The contents of the flasks in inoculated control series should be processed in a similar manner.

5.3 Determination by Kjeldahl Method

(i) Reagents
(ii) Sulphuric acid- 93-98 percent, N-free
(iii) Digestion mixture-Mix copper sulphate and potassium sulphate in the ratio-1:10 and grind them to a fine powder.
(iv) Sodium hydroxide pellets or solution, N-free-For solution, dissolve about 450g of sodium hydroxide in water, cool, and dilute to 1 litre (sp gr of the solution should be at least 1.36)
(v) Zinc granules-reagent grade
(vi) Indicators:-
(a) Methyl red indicator	-	Dissolve 1g of methyl red in 200 ml of ethanol.
(b) Mixed indicator	-	Prepare mixed indicator by dissolving 0.8 of methyl red and 0.2g of methyl blue in 500ml of ethanol.
(vii) Hydrochloric or sulphuric acid	-	standard solution 0.5 or 0.1N when amount of nitrogen is small.
(viii) Sodium hydroxide standard solution	-	0.1N (or other specified concentration).

Note. - Ratio of salt to acid (m/v) should be about 1:1 at the end of the digestion for proper temperature control. Digestion may be incomplete at a lower ration, and nitrogen may be lost at higher ratio. Each gram of fat consumers 10ml of sulphuric acid and each gram of carbohydrate 4.0 ml of sulphuric acid during digestion.

5.4 Apparatus

(i) For digestion- Use Kjeldahl's flasks of had, moderately thick, well-annealed glass with total capacity approximately 500 to 800 ml. conduct digestion over heating device adjusted to bring 250 ml of water at 25^°C to rolling boll in about 5 minutes. To test the heaters, preheat for 10 minutes in the case of gas burners and for 30 minutes in the case of electric heaters. Add 3 to 4 boiling chips to prevent superheating.

(ii) For distillation-Use 500 to 800 ml Kjeldahl's flask fitted with rubber stopper through which passes the lower end of an efficient scrubber bulb or trap to prevent mechanical carry-over of sodium hydroxide during distillation. Connect the upper end of the bulb tube to a condenser by a rubber tubing. Trap the outlet of the condenser in such a way as to ensure absorption of ammonia distilled over with the receiver.

(a) Procedure: Place 0.25g of the sample in the digestion flasks. Add 0.7gm mercuric oxide, 15gm potassium sulphate followed by 25ml of sulphuric acid. Share, let stand for about 30 minutes and heat carefully until frothing ceases. Boil briskly until the solution clears and continue boiling further for 90 minutes. Cool, add about 200ml of water, cool to room temperature and add a few zinc granules.

(b) Tilt the flask and carefully add 50ml of sodium hydroxide solution without agitation. Immediately connect the flask to the distillation bulb on the condenser whose tip is immersed in 50ml of standard 0.1N acid in the receiving flasks. Rotate the digestion flask carefully to mix the contents. Heat until 150ml of the distillate collects and titrate excess acid with 0.1N base using methyl red or mixed indicator. Carry out blank determination on reagents.

Note. - Check the ammonia recording periodically, using inorganic nitrogen control, for example, ammonium sulphate.

(i) Nitrogen content, percent by mass:-

(Milliters of 0.1N acid for sample-mililiters of 0.1N acid for blank)x 0.14 mass of sample taken

(ii) Total nitrogen in culture-Total dry mass of sample x percent nitrogen.

(d) Take a 1.0g of accurately weighed sample each from the inoculated series and from the controls. Put them separately in 250ml volumetric flask, add 150ml water, mix the content and make up the volume to 250ml water. Shake for 5 minutes and centrifuge for 15 minutes at 40000 rev/min. Estimate glucose in the supernatant in triplicate. The difference between the two provides the data of actual amount of glucose consumed. Calculate the amount of nitrogen foxed per gram of sucrose consumed.

(i) Reagents

(ii) Soxplet modification of Fehling solution:-Prepare by mixing equal volumes of Solution A and Solution B immediately before using.

(iii) Copper sulphate solution (Solution A)- Dissolve 34.639g of copper sulphate crystals (CuSO₄5H₂O) in a water, dilute to 500 ml ad filter through glass wool of filter paper.

Standardization of copper sulphate solution: - Using separate pipettes, pipette accurately 5 ml of Solution A and 5 ml of Solution B into a conical flask of 250ml capacity. Heat this mixture to boiling on an asbestos gauze and add standard invert sugar solution from a burette, about 1 ml less than the expected volume which will reduce the Fehling solution completely (about 48ml). Add 1 ml of methylene blue indicator while keeping the solution boiling. Complete the titration within 3 minutes, the end point being indicated by change of colour from the blue to red. From the volume of invert sugar solution used, calculate the strength(s) of the copper sulphate solution by multiplying the titre value by 0.001 (mg/ml of the standard invert sugar solution). This would give the quantity of invert sugar required to reduce the copper in 5ml of copper sulphate solution.

(iv) Potassium sodium tartrate (Rochelle salt) Solution (Solution B):- Dissolve 173g of potassium sodium tartrate and 50g of sodium hydroxide in water, and dilute to 500ml. Let the solution stand for a day, and filter.

(v) Hydrochloric acid-sp gr 1.18 at 20^°C (approximately 12N)

(vi) Standard invert sugar solution-Weigh accurately 0.95g of sucrose and dissolve it in 500ml of water. Add 32 ml of concentrated hydrochloric acid, boil gently for 30 minutes and keep aside for 24 hours. Neutralize with sodium carbonate and make the final volume to 1000ml: 50ml of this solution contains 0.05g of invert sugar.

(vii) Methylene blue indicate-0.2 percent in water.

(viii) Procedure: - Place about 1g (M), accurately weighed, of the prepared sample of A1 into a 250ml volumetric flask and dilute with about 150ml or water. Mix thoroughly the contents of the flask and make the volume of 250 ml with water. Using separate pipettes, take accurately 5 ml each of Solution A and Solution B in a porcelain dish. Add about 12ml of A1 solution from a burette and heat to boiling over an asbestos gauze. Add 1 ml of methylene blue indicator and while keeping the solution boiling complete the titration within 3 minutes, the end point being indicated by change of colour from blue to red. Note the volume (H) in ml of A1 solution required for the titration.

(ix) Calculation

Total reducing sugars, percent by mass =

250 x 100 x S
___________
H x M

Where

S= strength of copper sulphate solution.

H= volume in ml of A1 solution required for titration, and

M- mass in g of A1 taken for the test.

(i) Procedure: To 100 ml of the stock A1 solution, add 1 ml of concentrated hydrochloric acid and heat the solution to near boiling. Keep aside overnight. Neutralize this solution with sodium carbonate and determine the total reducing sugars as described in.

(ii) Calculation

(a) Sucrose, percent by mass=(reducing sugars after inversion, percent by mass)-(reducing sugars before inversion, percent by mass) x 0.95

(b) Nitrogen, mg per gram of sucrose consumed =2(a-b)-C

a= initial quantity of sucrose taken for the test,

b= mass of sucrose as calculated in (a), and

c= amount of nitrogen fixed per gram of glucose.

[1C. Method of analysis of azospirillum bio-fertilisers

1. Apparatus: same as Rhizobium

2. Reagents

3. Medium

Use N-free semisolid medium (Nib) of the following composition for preparation of MPN tubes

DL-Malic acid	5.0
K2HPO4	0.5
MgSo47H2O	0.2
NaCl	0.1
CaCl₂	0.02
Trace element Soln	2.0ml
Fe EDTA (1.64% Soln.)	4.0ml
Vitamin soln.	1.0ml
KOH	4.0ml
Bromothymol blue (0.5% aq.)	2.0ml
Adjust pH to 6.8-7.0 with KOH For seml solid add agar	1.75g
For solid medium add agar	15.0g
21.1 Trace element solution (g/litre)
Na2M0O4H2O	0.2
MnSO4H2O	.0235
H₃BO₃	0.28
CuSo₄5H₂O	0.008
ZnSO₄7H₂O	0.024
Distilled water	1000ml
Use 2 ml of this solution in one litre of Nib media Vitamin solution (g/litre)
Biotin	0.01
Pyridoxin	0.02
Distilled water	1000ml
Use one ml of this sol. In one litre of Nfb media

2.2 Setrilization and preparation of MPN tubes

2.2.1 Prepare Nitrogen free Bromothymol Blue malate medium as mentioned at paragraph 2.1. Boil to dissolve agar. Quickly dispense 10ml molten media in 15x150 ml test tubes or screw capped culture tubes and close either with cotton plugs or screw caps. Minimum of 25 tubes shall be needed for each sample.

2.2.2. Sterlize the tubes by autoclaving at 121C for 20 minutes, as in Rhizobium at paragraph 2.3.2

3. Preparation of serial dilution of MPN count

Dispense 30g of Azospirillum biofertilizers in 270 ml of sterile water and shake for 10 minutes on a reciprocal shaker. Maker serial dilutions up to 10 dilution. Pipette out 1 ml aliquots of 104 ti 10 dilution and deliver it to screw cap tubes or test tubes containing N-free semi solid Nfb media.

4. Incubation of tubes

Label the tubes and incubate t 36+1C for 3-4 days in vertical position in a test tubes stand. Do not disturb the medium during the entire period of incubation.

5. Counting

5.1 Count the tubes which have turned blue and have developed typical white sub-surface pellicle.

5.2 Count the tubes as +ve or -ve for the presence of sub-surface pellicle and consider for the purpose of calculation.

5.3 Method for Estimating MPN Count

5.3.1 To calculate the most probable number of organisms in the original sample, select as P1 the number of positive tabes in the least concentrated dilution in which the tubes are positive or in which the greatest number of tubes in +ve, and let P2 and P3 represent the numbers of positive tubes in the next two higher dilutions.

5.3.2 Then find the row of number in Table 1 in which P1 and P2 correspond to the values observed experimentally. Follow that row of numbers across the table to the column headed by the observed value of P.

5.3.3 The figures at the point of intersection is the most probable number of organisms in the quantity or original sample represented in the inoculum added in the second dilution, Multiply this figure by the appropriate dilution factor to obtain the MPN value.

5.3.4 Azospirillum count/g carrier =

Value from MPN table x Dilution level
____________________________

Dry mass of product

Table 1

Most Probable Numbers for use with 10 fold dilution and 5 tubes per dilution

(Cochran, 1950)

P1	P2		Most probable umber of indicated value of P3
P1	P2	0	1	2	3	4	5
0	0	-	0.018	0.036	.0.54	0.072	0.090
0	1	0.018	0.036	0.055	0.073	0.091	0.11
0	2	0.037	0.055	0.074	0.092	0.11	0.13
0	3	0.056	0.074	0.093	0.11	0.13	0.15
0	4	0.075	0.094	0.11	0.13	0.15	0.17
0	5	0.094	0.11	0.13	0.15	0.17	0.19
1	0	0.020	0.040	0.060	0.080	0.10	0.12
1	1	0.040	0.06	0.081	0.10	0.12	0.14
1	2	0.061	0.082	0.10	0.12	0.16	0.17
1	3	0.089	0.10	0.13	0.16	0.17	0.19
1	4	0.11	0.13	0.15	0.17	0.19	0.22
1	5	0.13	0.15	0.17	0.19	0.22	0.24
2	0	0.046	0.068	0.091	0.12	0.14	0.16
2	1	0.068	0.068	0.091	0.12	0.14	0.16
2	2	0.093	0.12	0.14	0.17	0.19	0.22
2	3	0.12	0.14	0.17	0.20	0.22	0.25
2	4	0.15	0.17	0.20	0.23	0.25	0.28
2	5	0.17	0.20	0.23	0.26	0.29	0.32
3	0	0.078	0.11	0.13	0.16	0.20	0.23
3	1	0.11	0.14	0.17	0.20	0.23	0.27
3	2	0.14	0.17	0.20	0.24	0.27	0.31
3	3	0.17	0.21	0.24	0.28	0.31	0.35
3	4	0.21	0.24	0.28	0.32	0.36	0.40
3	5	0.25	0.29	0.32	0.37	0.41	0.45
4	0	0.13	0.17	0.21	0.25	0.30	0.36
4	1	0.17	0.21	0.26	0.31	0.36	0.42
4	2	0.22	0.26	0.32	0.38	0.44	0.50
4	3	0.27	0.33	0.39	0.45	0.52	0.59
4	4	0.34	0.40	0.47	0.54	0.62	0.69
4	5	0.41	0.48	0.56	0.64	0.72	0.81
5	0	0.23	0.31	0.43	0.58	0.76	0.95
5	1	0.33	0.46	0.64	0.84	1.1	1.3
5	2	0.49	0.70	0.95	1.2	1.5	1.8
5	3	0.79	1.1	1.4	1.8	2.1	2.5
5	4	1.3	1.7	2.2	2.8	3.5	4.3
5	5	2.4	3.5	5.4	9.2	16.0	-]

1D. Method of analysis of phosphate solubilising bacteria bio-fertilisers

1. Apparatus: Same as Rhizobium

2. Reagents

2.1 Medium

Use a planting medium of the following composition

Glucose	10.0g
Tri-calcium phosphate	5.0g
Ammonium sulphate	0.5g
Magnesium sulphate	0.1g
Sodium chloride	0.2g
Yeast extract	0.5g
Manganese sulphate	Trace
Ferrous sulphate	Trace
Distilled water	1000ml
Agar	15.0g
pH adjusted to 7+/-0.2

2.2 Sterilising and preparation procedure for plates : Same as Rhizobium

Preparation of plating medium and pouring

Same as Rhizobium

3. Preparation of serial dilutions for plate counts

Same as Rhizobium

4. Incubation of plates

4.1 Label the plates and incubate at 28+1C for 4 to 6 days.

4.2 Colony counting aids : Same as Rhizobium

Counting

Count the total number of colonies on the plates including colonies with solubilisation zone with the help of colony counter.

Methods for counting solubilisation zones

(a) Take 10g of PSBI (BF) in 90 ml in water

(b) Make a ten fold dilution series up to 10

(c) Take 0.2 ml aliquote of 10 to 10 dilution using sterile pipettes and deliver to Petri dishes containing pikowskeyi media.

(d) Spread it uniformly, Invert the plates and incubate them up to 2 weeks at 28+/2C

(e) Count the colonies showing hallow cones and measure their diameter. Minimum acceptable zone is 10 mm in diameter.

5. Determination of Soluble Phosphorous using ascorbic acid

5.1 Apparatus

Spectrophotometer capable of transmission measurements at 840 to 880 mm.

Extractant: It is olsen extract.

5.2 Reagents

Ammonium Molybdate [NH₄)6MO₇O₂44H₂O]

L-Ascorbic Acid

p-Nitrophenol

4NH₂SO₄

5.3 Preparation of reagents

5.3.1 Sulphomolybdic Acid

- Take 20g of ammonium molybdate and dissolve in 300ml of distilled water.

- Add slowly 450ml of 10N H₂SO₄

- Cool the above mixture and add 100 ml of 0.5 percent solution of antimony potassium tartrate.

- Cool and and make the volume to one litre. Store in glass bottle away from direct sunlight.

5.3.2 Preparation of Mixed Reagent

Add 1.5g of L-ascorbic acid in 100 ml of the above stock solution and mix. Add 5ml of this solution to develop colour. Mixed reagent is to be prepared fresh as it does not keep for more than 24h.

5.3.3 Procedure

[(i) Preparation of Sample

Pure culture medium same as at 2.1 above excluding agar.

Prepare broth medium in 100ml aliquots in 6 no, 250 ml conical flasks and sterilize in autoclave at 121C for 20 min.

(ii) Inoculation of Medium

Select one PSB colony of the type that has been counted as PSB (showing sufficient zone of solubilization) and streak on set medium as described at 2.1 in a Petri dish. Use this pure culture for inoculating the broth. Inoculate 3 flasks and keep 3 flasks as uninoculated control. Incubate the flasks over rotary shaker for 12 days at 28+1C.

After 12 days, filter the contents of each flask separately through Whatman No. 42 filter paper or centrifuge at 10,000 rpm for 15 min.

(iii) Add 10 ml of filtrate/centrifugate to 50 ml of olsen extractant and shake for 30 min. over rotary shaker.]

(iv) Filter the suspension through Whatman filter paper No. 40. If the filtrate is coloured then add a tea spoon of Dacro-60 (activated phosphorous free carbon), reshake and filter.

(v) Take a known aliquot (5 to 25 ml) of the extract in a 50 ml volumetric flask.

(vi) Add 5 drops of p-nirophenol indicator (1.5 per cent solution in water) and adjust the pH of the extract between 2 and 3 with the help of 4 NH₂SO₄. The yellow colour will disappear when the pH of the solution becomes 3. Swirl gently to avoid loss of the solution along with the evolution of CO₂.

(vii) When the CO₂ evolution has subsided, wash down the neck of the flack and dilute the solution to about 40ml.

(viii) Add 5 ml of the sulphomolybdic acid mixed reagent containing ascorbic acid, swirl the content and make up the volume.

(ix) Measure the transmission after 30 min at 880 mm using red filter. The blue colour developed remains stable upto 60 minutes.

(x) Record the concentration of phosphorous (P) in the extract form from the standard curve and calculate the concentration of soluble phosphorous as follows.

5.3.4 Calculations

(a) Weight of the substance taken=xg

(b) Volume of the extractant added=50ml

(d) Volume made after colour developed=50ml

(e) Reading from the standard curve against percent transmission recorded =x ppm

(f) Soluble Phosphorus, percent P =

z x 50 x 10^-6 x 50
_____________________ x 100
yx

5.3.5 Preparation of Standard Curve

Prepare standard curve using 0.1 to 0.6 ppm P in 50 ml volumetric flask. Plot the standard curve by taking concentration of soluble P on x-axis and percent T on y-axis using a semi-log graph paper. It is a straight line relationship between the soluble P and percent T when plotted on a semi-log graph paper.

[5-A. Qualification of P solubilization activity.

- The P solubilization is quantified by inoculating the test culture in Pikovskaya's broth for 5 day at 30°C. Culture suspension is centrifuged at 8000 rpm for 15 min and the supernatant was used (Murphy and Riley, 1962).

(a) Reagents:

(i) Sulfuric acid (5 N): Dilute 70 ml of concentrated sulfuric acid to 500 ml.

(ii) Ammonium molybdate: Dissolve 20 gm of ammonium molybdate in water and dilute to 500 ml. Store the solution in a Pyrex glass bottle.

(iii) Ascorbic acid (0.1 M): Dissolve 1.32 g of ascorbic acid in 75 ml of water. This solution should, if possible, be prepared on the day it is required as the ascorbic acid easily becomes oxidized. If it is necessary to keep the solution it can probably be stabilized by addition of 25 mg of ethylenediaminetetraacetic acid (disodium salt) and 0.5 ml of formic acid per 75 ml of solution.

(iv) Potassium antimonyl tartrate: Dissolve 0.2743 g of potassium antimonyl tartrate in distilled water and dilute to 100 ml.

(v) Mixed reagent: Mix thoroughly 125 ml of 5 N sulfuric acid and 37.5 ml of ammonium molybdate. Add 75 ml of ascorbic acid solution and 12.5 ml of potassium antimonyl tartrate solution. This reagent should be prepared as required as it does not keep for more than 24 h.

(b) Standard phosphate solution: Stock phosphate solution: Prepare a solution containing 0.1757 g of potassium dihydrogen phosphate per L. This solution contains 40 mg P (as phosphate)/L. Prepare solutions of different concentrations as given below-

Volume from stock (40 mg/L)	Distilled water (mL)	Final Volume (mL)	Final Concentration (mg/L)
5.0	45.0	50.00	4.0
4.0	46.0	50.00	3.2
3.0	47.0	50.00	2.4
2.0	48.0	50.00	1.6
1.0	49.0	50.00	0.8
0.0	50.0	50.00	0.0

1. Pipette 40 ml of the culture supernatant into a 50-ml calibrated flask, add 8 ml of the mixed reagent from a tilt measure and make the volume to 50 ml with distilled water, and mix well.

2. Dilute the reaction mixture if required.

3. After not less than 10 min measure the optical density of the solution at 880 nm.

4. Determine the reagent blank in the same manner using uninoculated Pikovskaya medium.

5. The amount of soluble P in the culture supernatant is calculated (z) by plotting the OD values against standard curve prepared using different concentrations of P as phosphate as detailed in the table. The calibration curve only needs occasional checking as it remains constant and appears to be independent of changes in the batches of reagents.

6. Calculate the solubilized P as:-

mg P043--P/L = z X (Dilution factor used at step 2)].

6. Maintenance and preparation of culture and quality control at broth stage

Rhizobium

1. Maintenance of pure cultures

1.1 Maintain pure cultures of rihizobia on yeast extract mannitol agar (YEMA) slants of the following composition:

Mannitol	10.0g
Potassium hydrogen phosphate (K2HPO4)	0.5g
Magnesium sulphate (MgSO₄7H₂O)	0.2g
Sodium chloride (NaCl)	0.1g
Calcium carbonate (CaCo₃)	1.0g
Yeast extract	1.0g
Agar	18.0g
Distilled water	1 litre
pH	6-8-7.0

1.2 Transfer a loopful of the pure culture to each of the agar slants aspectically in an inoculation room and incubate at 28+/-2C for 3 to 10 days depending upon the species of Rhizobium. Always keep pure cultures at 4C.

2. Preparation of Inoculum Cultures

2.1 Prepare yeast mannitol broth of the composition as given in 1.1 minus the agar.

2.2 Transfer a loopful of the culture.

3. Quality Control Tests Recommended at Broth Stage

3.1 Qualitative Tests

3.1.1 Check for freedom from visible contaminants

3.1.2 The pH of the bacterial broth shall normally be between 6.5 and 7.5

3.1.3 Smear and Gram stain

3.1.3.1 Reagents

(a) Ammonium oxalate crystal violet stain weigh 0.2g of crystal violet and dissolve in 20 ml of 95 percent ethyl alcohol. Dissolve separately 0.8g of ammonium oxalate in 80 ml of distilled water. Mix the two solutions and filter through a filter paper.
(b) Iodine solution
Iodine	1.00g
Potassium Iodine	2.00g
Distilled water	300m;
Weigh the ingredients and dissolve in water. Filter through a filer paper.
(c) Erythrosine	1.00g
Phenol	5.00g
Distilled water	100ml
Weigh the ingredients, dissolve in distilled water and filter through a filter paper.

3.1.3.2 Procedure

Prepare a smear on a clean microscope slide, fix over a flame by gentle and intermittent hearing, air cool and flood with ammonium oxalate crystal violet stain for 1 min. After removing the excess of ammonium oxalate crystal violet, wash the slide under a gentle stream of running tap water. Flood the slide with iodine solution for half a minute remove excess stain wash with 95 percent ethyl alcohol ad finally wash under a gently stream of running tap water. Flood the slide with erythrosine stain for about 3 min, wash under a gentle stream of running tap water and dry between the folds of a filter paper. Examine the slide under a compound microscope using an oil immersion objective.

Note: A smear prepared from undiluted broth should be free from Gram positive cells. The presence of a few gram positive sells in occasional fields which may be due to dead cells in the medium may be disregarded.

3.1.4 Absence of Growth on Glucose- Peptone Agar

The composition of the glucose-Peptone agar is as follows:

Glucose	10.0g
Peptone	20.0g
Sodium chloride (NaCI)	5.0
Agar (IS 6850)	15.0
Distilled water	1000ml
Bromocresol purple	10 ml of 1.6 persons ethly alcohol solution
pH	7.2

Note: When a loopful of the broth is streaked into this medium and incubated at 28+/-2^°C for 24 h, the purple-violet colour of the medium (due to the indicator bromocresol purple) shall not change. If the colour changes to yellow (acidic reaction) or blue (alkaline reaction) the broth is grossly contaminated.Hence, the broth should be rejected.

3.1.5 Streak on yeast Extract mannitol Agar with Congo Red.

When a loopful of broth culture is streaked to a plate of this medium ad incubated at 28+/-2^°C for 3 to 10 days, it shall show colonies of bacteria with growth characteristics same as that of the pure culture used in preparation of the broth, otherwise, the broth should be rejected.

3.2 Quantitative Test

3.2.1 Viable or Plate Counts

Serially dilute one mililiter of the broth to obtain dilutions of the order of 10 to 10. Plate 0.2 ml aliquots of the dilutions on YEMA plates and incubate at 28+/-2C for 2 to 6 days, depending on the species of Rhizobium. The counts of viable Rhizobium in the final broth from shake culture of termentors shall be not less than 10 to 10 cells/ml. otherwise, the broth should be rejected.

Azospirillum

1. Maintenance of pure cultures

2. Preparation of Inoculum culture and Mass culture

3. Quality Control Test Recommended at Broth stage

Azotobactor

1. Maintenance of pure cultures

Agar	20gm
Sucrose	20gm
Ferrous Sulphate	0.1
Dibasic Potassium Phosphate	1.0gm
Magnesium Sulphate	0.5gm
Calcium carbonate	2.0gm
Sodium molybdate	0.005gm

2. Preparation of inoculum culture.

3. Quality control Tests recommended by Broth stage.

4. Packing, Marking, Storage and use.

[Biofertiliser shall be packed in suitable plastic bags/packets, thickness of which shall not be less than 75-100 micron or in suitable plastic bottles.]

(a) Name of the product;

(b) Name and address of the manufacturer;

(d) Type of the carrier used;

(e) Batch number;

(f) Date of manufacture;

(g) [Expiry date which shall not be less than 6 months from the date of manufacture in case of carrier based powdered/granulated formulation of Rhizobium, Azotobacter, Azospirillum and PSB biofertilisers and liquid based Rhizobium biofertliser, while it shall not be less than twelve months from the date of manufacture in case of liquid based Azotobacter, Azospirillum and PSB biofertilisers.]

(h) Net mass in kg/gram and area meant for;

(i) Storage instruction worded as under:

Store in cool place away from direct sun light and heat

(j) Any other information required under the Standards of Weights and Measures (Packaged Commodities) Rules, 1977.

"The contents of the packet are sufficient enough for seed treatment on to the given area to be broadcasted or given seedlings for root dipping depending on the specified crops as denoted on the packet. Mix the inoculants with seed gently with the minimum amount of water taking care to avoid damage to seek coat. Dry the inoculated seed under shade over clean surface gunny bag and sow them immediately.

Use only for the crops mentioned. Use before the expiry date and do not expose to direct sun light or heat.

Biofertiliser is not a chemical fertiliser hence do not mix inoculated seeds or inoculant with agro-chemicals."

^°

[1E. Method of analysis for mycorrhizal bio-fertilisers

1. Estimation of pH- As specified in Schedule IV Part D at serial number 1.

2. Estimation of moisture contents- As specified in Schedule IV Part D at serial number 2 of Fertiliser (Control) Order, 1985.

3. Estimation of total viable propagules

3.1 Harvesting of spores from finished product

A. By sieving

(a) Equipment and Reagent:- Stalking sieves with nylon or stainless steel mesh and a large range of pore sizes of isolating spores from the carrier or soil sample.

(i) 40-50 micron (0.04 mm) sieve for small sized spores.

(ii) 100 micron (0.14 mm) sieve for medium sized spores.

(iii) 250 micron (0.25 mm) sieve for very large sized spores and sporocarps.

(iv) Wash bottles containing water.

(v) Jars for collecting the sieving.

(vi) Stereo zoom (stereomicroscope).

(vii) Patri dishes (11cm) for observing the sieving under stereomicroscope.

(viii) Micropipettes for spore picking.

(ix) Centrifuge.

(b) Procedure- Mix the soil in a substantial volume of water and decant through a series of sieves arranged in descending order of mesh size. Roots and coarse debris collected on a coarse (60-ISS) sieve, while spores are captured on one or more finer sieves. Vigorous washing with water is necessary to free spores from aggregates of clay or organic materials. Collect the sieving in jars. Transfer the sieving onto the grided petri dishes/plate and observe under stercomicroscope. Count the number of spores in plate/dish and express it as spores/g of the sample.

(B) By sucrose gradient

(i) Collect the sieving by the method described above. Transfer the sieving into centrifuge tubes and centrifuge for 5 minutes at 1,750 rpm in a horizontal rotor.

(ii) Decant the supernatant liquid carefully and resuspend pellet in 60% sucrose solution. Again centrifuge for 2-5 minutes.

(iii) Pour the supernatant (with spores) onto a 300 BSS sieve size and rinse with water to remove the sugar. Transfer the sieving onto the grided petri dishes/plate and observe under steromicroscope. Count the number of spores in plate/dish and express it as spores/g of the sample.

3.2 Spore staining

(a) Equipment and Reagent

(1) Equipments and reagents for spore extraction as described previously.

(2) 2.5-diphenyl-2N-tetrazolium bromide (MTT)

(3) Distilled water

(4) Eppendort

(5) Stereomicroscope

(6) Petri-dishes

(b) Procedure

(i) Prepare 0.25% solution of MTT (2,5-diphenyl-2N-tetrazolium bromide).

(ii) Avoid exposure of MTT solution to light, as the stain is light sensitive.

(iii) Add freshly collected AMF spores (approximately 100 in number) collected by any of the two methods described above, to the staining solution and incubate at 27C in sterile eppendoif in dark.

(iv) Observe the spores for different colour reactions using stereomicroscope under dark field after 24 hours, 48 hours and 72 hours of incubation.

(v) Spores, which stained red or pink, are treated as viable as per the following formulae:

% Spore viability=

No. of spores which stained red or pink x 100
____________________________

Total number of spores

4. Assessment of Infectivity Potential - The bloassay is used to determine the number of infective propagules present in the product. Once the infective propagules (spores, myoelia and vesicles in the root fragments) come in contact with the host roots they give out a turgid mycelial structure- the appressoria, which is the initial step in the penetration event. This appressoria enters the root through an entry point. This entry point can be visualized by staining and enumerated as a measure of the infectivity of the inoculum. Host plants are grown from pre germinated seeds and a known weight of the inoculum is applied to experimental host plant in post. These pots are maintained for 14 days after which they are harvested, the root length measured and then stained. The resulting entry points are counted to ascertain the infectivity potential.

(A) Equipments and Reagents

(a) Pots (5 x 7) cm in size)

(b) Sorghum seeds (Serghum vulgare)

(d) Petri dish (grided)

(e) Water bath

(f) Glass slides and cover slips

(g) Compound microscope

(h) Coarse sieve to prevent root loss during washing/changing solutions

(i) Plastic vials with fight-sealing lids for storage of stained samples in 50% glycerol

(j) Potassium hydroxide solution (5-10%)

(k) Alkaline H2O2 (25% Ammonia solution : 3ml + 10% H2O2 : 30 ml + Distilled water 67 ml)

(l) 1% HCI

(m) 50% glycerol-water (v/v) solution for de-staining and storage of stained roots.

(n) Lactoglycerol (Lactic acid : 876 ml + Glycerine : 64 ml + Distilled water : 60 ml)

(B) Staining solutions

(a) 0.01% acid fuschin: 0.01g acid fuschin in 100 ml acetoglycerol

(b) 0.05% trypan blue : 0.05g trypan blue in 100 ml acetoglycerol

(d) Dissolve CBE in water before adding equal volumes of lactic acid and glycerol.

(C) Procedure

(i) Place 100g test sample in a pot

(ii) Dilute the inoculum with sterilized sand if the inoculum is very rich

(iii) Plant 10-12 pre germinated seeds of Sorghum and grow for 14 days

(iv) Harvest the pots and recover roots (fine roots can be rescued using sieve) completely.

(v) Chop the roots equally 1cm 1 length

(vi) Measure and record he root length (using grid line intersect method described below) from each sample/dilution.

(vii) Clear the roots in KOH solution and stain the root pieces (described below)

(viii) Count the number of infection points/ entry points formed on randomly picked 100 segments.

(ix) Calculate the average number of entry points formed in 1 cm segment

(x) Calculate the total number of infection points/infective propagules (IP) by multiplying the average number of entry points formed in 1 cm segment by the total root length

(xi) Extrapolate the IP present as numbers per gram of substrate/inoculum

(D) Estimation of root length

(1) Equipment

(a) Scissors

(b) Petri dish (9cm in size consisting 1.33 cm x 1.33 cm grids

(d) Stereo zoom microscope]

[1F. Method of analysis for [Potash Mobilising Biofertilisers] (KSB)

1. Estimation of total viable count and contamination

1. Apparatus -

1.1 Pippettes graduated 1 ml and 10 ml

1.2 Dilution bottles or flasks

1.3 Petri dishes clear, uniform, flat-bottomed

1.4 Hot-air oven

Capable of giving uniform and adequate temperature, equipped with a thermometer, caliberated to read upto 250C and with venus suitably located to assure prompt and uniform heating.

1.5 Autoclave

1.6 Incubator

1.7 Had tally or mechanical counting device

1.8 pH meter

2. Regeant

2.1 Medium

Use plating medium of the following composition for total viable count and contamination

Medium for analysis of total viable count and contamination (Ingredients g/lit)

Marital	15.0
Yeast extract	3.0
Peptone	2.0
Agar	18.5
Trace element solution	1ml
Distilled Water	1000ml
Trace element solution (Ingredients g/lit)
Sodium molybdate	0.20
Boric acid	0.28
Manganese sulphate	0.23
Copper sulphate	0.01
Zinc sulphate	0.03
Distilled Water	1000ml
Medium for studying zone of solubilization in KSB (Ingredients g/lit)
Glucose	5.0
Magnesium sulphate	0.005
Ferric chloride	0.1
Calcium carbonate	2.0
Potassium mineral (mica powder)	2.0
Calcium phosphate	2.0
Distilled water	1000ml.

(a) Immediately before pouring.

(b) Periodically during pouring, and

(c) When pouring is complete for each batch of plates, if portions of molten medium remain in containers and are to be sued without subsequent sterilization for pouring additional plates. As each plate is poured thoroughly mix the medium with test portions in the petri dish.

3. Preparation of Serial Dilution for Plate Counts.

4. Incubation of Plates.

Count the colonies with the aid of magnifying lens under uniform and properly controlled, artificial illumination. Use a colony counter, equipped with a guide plate and rules in centimeter square. Record the total number of colonies with the hand tally. To distinguish colonies from dirt, specks and other foreign matter, examine doubtful objects carefully.

5. Counting.

Count the total number of colonies on the plates including colonies with solubilisation zone with the help of a colony counter.

6. Method for estimation of K solubilization zones.

[7. Method of Analysis for estimation of Potassium solubilisation:

Determination of soluble potash using Flame Photometer

1. Preparation of Sample Prepare Aleksandrov broth media comprising of 5.0 g glucose; 0.5 g MgS04.7H20; 0.1 g CaC03; 0.006 g FeC13; 2.0 g Ca3(P04)2; 3.0 g potassium aluminium silicate in 1: 1 of distilled water. The pH of this media is adjusted to 7.2+0.2byusingl N NaOH. Prepare broth media in 100 ml aliquots in 6 no., 250 ml conical flasks and sterilize in autoclave at 121°C for 20 min.

2. Inoculation of Media Select one Potash Mobilising Biofertilisers colony of the type that has been counted as KSB (showing sufficient zone of solubilization) and streak on Aleksandrov agar medium as described above in a Petri dish. Use this pure culture for inoculating the broth. Inoculate 3 flasks and keep 3 flasks as uninoculated control. Incubate the flasks over rotary shaker for 7 days at 30+l°C.

3. Determination of Soluble K

(1) After 7 days, centrifuge the broth at 10,000 rpm for 15 min. Collect the supernatant for estimation of potassium by Flame Photometer.

(2) Add 1 ml of filtrate/supernatant to 50 ml volumetric flask and make up the volume to 50 ml. Estimate K content by feeding the solution to Flame Photometer. The amount of potassium solubilized is calculated from the standard curve by multiplying the observed value with dilution factor.

4. Preparation of Standard curve

(1) Make a stock solution of 1000 ppm K by dissolving 1.909 g. of AR grade potassium chloride (dried at 60°C for 1 h) in distilled water in a 1000 ml volumetric flask; and make up to 1 litre with distilled water. Prepare 100 ppm standard by diluting 100 ml of 1000 ppm stock solution to 1 litre with distilled water.

(2) Pipette 0.5,1.0,1.5 and 2.0 ml of 100 ppm solution into 100 ml volumetric flasks and make up the volume up to the mark. These solutions contain 0.5, 1.0, 1.5 & 2.0 ppm K (mg/Kg) respectively. Plot standard curve using concentrations against flame photometer values. Express K solubilized as mg/lit of broth].

1G. Method of Analysis for zinc solubilizing biofertilizers

2. Estimation of total viable count and contamination.
1. Apparatus
1.1 Pippettes graduated 1 ml and 10 ml
1.2 Dilution bottles or flasks
1.3 Petri dishes clear, uniform, flat-bottomed
1.4 Hot-air oven
Capable of giving uniform and adequate temperature, equipped with a thermometer, calibrated to read upto 250C and with venus suitably located to assure prompt and uniform heating.
1.5 Autoclave
1.6 Incubator
1.7 Hand tally or mechanical counting device
1.8 pH meter
2. Reagents
2.1 Medium
Use plating medium of the following composition for total viable count and contamination.
Medium for analysis of Total Viable Count, Contamination and zone of solubilization for Zn solubilizing biofertilser
(Ingredients g/lit)
Glucose	10.0
Zinc oxide	1.0
Amm sulphate	0.5
Potassium chloride	0.2
Yeast extract	0.5
Ferrous sulphate	0.01
Manganese sulphate	0.01
Di Pot Hyd. phosphate	0.5
Distilled water	1000 ml.

(a) Immediately before pouring

(b) Periodically during pouring, and

(c) When pouring is complete for each batch of plates, if portions of molten medium remain in containers and are to be used without subsequent sterilization for pouring additional plates. As each plate is poured thoroughly mix the medium with test portions in the petri dish.

3. Preparation of Serial Dilution for Plate Counts.

4. Incubation of Plates.

5. Counting.

Count the total number of colonies on the plates including colonies with solubilization zone with the help of a colony counter.

6. Method for estimation of Zinc solubilisation zones.

(a) Randomly disperse root in dish with grid lines

(b) Count the intersects on roots across the horizontal and vertical lines

Total number of intersects =HI+VI=8+7 (example) = 15

Thus, the root length = 11/14 x 15 x 1 (as the grid size is 1cm) (example)

(3) Clearing and staining root specimens - Clearing and staining procedures requires root samples that should be washed free of soil. It is important that KOH and staining solution volumes are sufficient for the amount of roots being processed and that, roots are not tightly clumped together for uniform contact with solutions. To ensure uniform staining, the roots should be chopped into smaller (1-2 cm) segments.

(a) Wash root specimens under running tap water thoroughly. Place them in beaker containing 5-10% KOH solution for about 15-30 minutes. The concentration of KOH and time of incubation of roots depend upon the age and tendernesa of the roots.

(b) Four off the KOH solution and rinse the roots well in a beaker using at least three complete changes of tap water or until no brown colour appears in the rinse water.

(d) Rinse the roots thoroughly using at least three complete changes of tap water to remove the H₂O₂

(e) Cover the roots with 1% HCI and soak for 3-4 minutes and then pour off the solution Do not rinse after this step because the specimens must be acidified for proper staining.

(f) Incubate the roots with staining solution (0.01% acid funchsine in lactoglycerol or 0.05% trypan blue in lacto phenol) and keep them ovemight for staining.

(g) Place the root specimens in glass petriplate/multi well plate for destaining. The destaining solution (50% glycerol) is the standard used in step 4, but of course, without the stain

(4) Sample storage and slide preparation: -

(1) If clearing and staining is not possible immediately then fresh roots can be kept moist and stored at 5C (for several days), or may be preserved in 50% ethanol for months together in tightly sealed vials.

(2) Staining quality is subsequently improved by destaining roots in 50% glycerol for several months prior to observation to allow excess stain to leach from roots. Semi-rmanent slides of stained roots can be made with PVLG mountant. For temporary slide the stained roots can be observed in plain lactoglycerol.]

[7. Determination of soluble Zinc using Atomic Absorption Spectrophotometer

(1) Preparation of Sample

Prepare Zinc solubilizing broth media comprising of glucose-lO.Og; ammonium sulphate-l.Og; potassium chloride- 0.2g, dipotassium hydrogen phosphate-0.lg, magnesium sulphate-0.2g and zinc carbonate/zinc oxide lgm in 1: 1 of distilled water. The pH of this media is adjusted to 7.0 by using 1 N NaOH. Prepare broth media in 100 ml aliquots in 6 no., 250 ml conical flasks and sterilize in autoclave at 121°C for 20 min.

(2) Inoculation of Media

Select one ZnSB colony of the type that has been counted as ZnSB (showing sufficient zone of solubilization) and streak on ZnSB agar media as described above in a Petri dish. Use this pure culture for inoculating the broth. Inoculate 3 flasks and keep 3 flasks as uninoculated control. Incubate the flasks over rotary shaker for 7 days at 30+1 °C.

(3) Determination of Soluble Zinc

(a) After 7 days, centrifuge the broth at 10,000 rpm for 15 min. Collect the supernatant for estimation of solubilized Zinc.

(b) Add 1 ml of filtrate/supematantto 50 ml volumetric flask and make up the volume to 50 ml. Estimate Zn content by feeding the solution to Atomic Absorption Spectrophotometer. The amount of zinc solubilized was estimated by subtracting the soluble zinc of the inoculated sample from the uninoculated control.

(c) Standard curve should be made by diluting 0.5, 1.0, 1.5, 2.0 and 3.0 ml of lOOppm standard stock solution (commercially available) of Zinc Sulphate with doubled distilled water in volumetric flasks and make up the volume to 100ml to obtain standards having concentrations of 0.5, 1.0, 1.5, 2.0, 3.0 ppm.

(d) The amount of zinc solubilized is calculated from the standard curve by multiplying the observed value with dilution factor (50). Express Zn solubilized as mg/lit of broth].

[1H. Methods of analysis for acetobacter (SPP)

1. Apparatus - As specified in the Method of analysis of Rhizobium at serial number 1A.

2. Reagents

2.1 Medium

Use plating medium of the following composition for total viable count and contamination: -

Medium for analysis of total viable count and contamination (ingredients gram/litre)

Sucrose	100g
K₂HPO₄ (Dl-Potassium Hydrogen Phosphate	0.4g
KH₂PO₄ (Potassium di-Hydrogen Phosphate)	0.6g
MgSO₄ (Magnesium Sulphate)	0.2g
Calcium Chloride	0.02g
Sodium Molybdate	0.02g
Ferric Chloride	0.01g
Bromothymol blue solution (0.5% in 0.2m KOH)	5.0ml
Distilled water	1000ml
pH	5.5g
agar agar	18.5g

3. Sterilising and preparation procedure for plates

(Same as specified in the Method of Analysis of Rhizobium)

3.1 Preparation of plating medium and pouring

(Same as specified in the Method of Analysis of Rhizobium)

4. Preparation of serial dilutions for plate counts

(Same as specified in the Method of Analysis of Rhizobium)

5. Incubation of plates

(Same as specified in the Method of Analysis of Rhizobium)

5.1 Colony counting Aids

Count the colonies with the aid of magnifying lens under uniform and properly controlled, artificial illumination. Use a colony counter, equipped with guide plate and rules in centimetre square. Record the total number of colonies with had tally. Avoid mistaking particles of undissolved medium or precipitated matter in plates for pin point colones. To distinguish colonies from dirt, specks and foreign matter, examine doubtful objects carefully.

5.2 Court all plates but consider for the purpose of calculation only those plates showing more than 30 and less than 300 colonies per plate. Acetobacter a nitrogen fixing bacteria stand out as irregular 2-3 mm diameter, smooth flat with bright yellow or yellow with orange centre colour. Court such colony numbers and calculate figures in terms of per litre, of carrier. Also check freedom from contamination at 10.

6. Test for confirmation.

1. Apparatus (Same as specified in the Method of Analysis of Azospirillum at serial number 1C)

2. Reagent

2.1 Medium (semi solid for pellicle formation) (ingredients gm per litre)

Sucrose	100g
K₂HPO₄ (Dl-Potassium Hydrogen Phosphate	0.4g
KH₂PO₄ (Potassium di-Hydrogen Phosphate)	0.6g
MgSO₄ (Magnesium Sulphate)	0.2g
Calcium Chloride	0.02g
Sodium Molybdate	0.02g
Ferric Chloride	0.01g
Bromothymol blue solution (0.5% in 0.2m KOH)	5.0ml
Distilled water	1000ml
pH	5.5g
agar agar	1.75g

3. Sterilization and preparation of MPN tubes

(Same as specified in the Method of Analysis of Azospirillum at serial number 1C)

4. Preparation of serial dilution for MPN count

(Same as specified in the Method of Analysis of Azospirillum at serial number 1C)

5. Incubation of tubes

(Same as specified in the Method of Analysis of Azospirillum at serial number 1C)

6. Counting-Yellowish pellicle formation below 1 mm of upper surface of nitrogen free semi solid media. Counting the tubes or plates which have turned yellowish in colour after inoculation and ascertained the presence of pellicle in undistributed medium. To determine usual contamination on the same examine doubtful objects carefully.

7. Method for Estimating MPN count

Count all tubes which have turned yellowish and consider them for the purpose of calculation. Count such type of tubes and tally this count with MPN table (as specified in the Method of Analysis of Azospirillum at serial number 1C in Table 1) to get the number of cells per gram of carrier or number of cells per ml of liquid.]

[1-I. Method of analysis of carrier based consortia of bio-fertiliser and liquid consortia of bio-fertiliser

(I) Methods of Analysis of Rhizobium biofertiliser-Same as specified for Rhizobium at Serial number 1A.

(II) Methods of Analysis of Azotobacter-Same as specified for Azotobacter at serial number 1B.

(III) Methods of Analysis of Azospirillum - Same as specified for Azospirillum at serial number 1C.

(IV) Methods of Analysis of Potash Mobilising Bacteria (KMB)-Same as specified for Phosphate Solubilising Bacteria at serial number 1F.]

[(1-L) Method of analysis of phosphate solubilizing fungal bio-fertiliser

1. Apparatus required:

1.1 Pipettes, graduates, 1 ml and 10 ml

1.2 Conical Flasks, 150ml and 250ml

1.3 Screw-Capped Tubes, 10ml

1.4 Incubator 1.5 Petri Dishes

1.5 Hot Air Oven

1.6 pH Meter

1.7 Autoclave

1.8 Haemocyshonete

1.9 Compound Microscope

1.10 Glass Slides and Cover Slips

1.11 Forceps

Note. - Entry 1-1 already exists as inserted vide SO 1117(F), date 30.4.2014. Hence it should be read as 1.1]

1.12 Needles

1.13 Glass Rods

2. Preparation of serial dilution for plate count (spore/chi)

2.1 Dispense 30g of PSFI to 270 ml of sterile distilled/demineralized water and shake for 10 min on a reciprocal shaker or homogenizer. Make serial dilutions up to 10 by suspending 10 ml aliquot of previous dilution to 90ml of water. Take 0.1 ml or suitable aluquotes upto 10 dilutions using sterile pipettes and deliver to Petri dishes containing set medium as given in section 1.2 and spread it uniformly with a spreader. Invert the plates and promptly place them in the incubator.

2.2 Incubation of plates.

2.2.1 Label the plates and incubate at 28+1C for 2 to 4 days.

2.2.2 Colony counting aids

Count the colonies with the aid of magnifying lens under uniform and properly controlled, artificial illumination. Use a colony counter, equipped with a guide plate and rules in Centimeter Square. Record the total number of colonies with the hand tally. Avoid mistaking particles of undissolved medium or precipitated matter, in plates for pinpoint colonies. To distinguish colonies form dirt, specks and other foreign matter, examine doubtful objects carefully.

2.2.3 Count all plates but consider for the purpose of calculation plates showing more than 30 and less than 300 colonies per plate. Count such colony numbers and calculate figures in terms of per gram/litre of carrier. Also check for freedom from contamination at 10 dilution

2.2.4 Counting

2.2.4.1 Count the colonies showing hallow Zones and measure their diameter.

3. Determination of soluble phosphorus using ascorbic acid method:

3.1 Apparatus

Spectrometer capable of transmission measurement at 840 to 880 mm.

Extractant: It is Olsen extract.

3.2 Reagents

Ammonium Molybdate [(NH₄)₆ Mo₇O₂₅4H₂O]

L-Ascorbic Acid

p-Nitro phenol

4NH₂SO₄

3.3 Preparation of reagents

3.3.1 Sulphomolybdic Acid:-

3.3.1.1. Take 20g of ammonium molyhdate and dissolve in 300 ml of distilled water.

3.3.1.2. Add slowly 450 ml of 10N-H₂SO₄

3.3.1.3 Cool the above mixture and add 100 ml of 0.5 percent solution of antimony Potassium tartrate.

3.3.1.4 Cool and make the volume to one liter. Store in glass bottle away from direct sunlight.

3.3.2 Preparation of Mixed Reagent Add 1.5g of L-ascorbic acid in 100ml of the above stock solution and mix. Add 5ml of this solution to develop colour. Mixed reagent is to be prepared fresh as it does not keep for more than 24h.

3.3 Procedure

(i) Preparation of Sample

Prepare broth medium in 100ml aliquots in 6no., 250 ml conical flasks and sterilize in autoclave at 121 C for 20min.

(ii) Inoculation of Medium

Select one PSFI colony of the type that has been counted as PSFI (showing sufficient zone of solubilization) and streak on set medium in Petri dish. Use this pure culture for inoculating the broth Inoculate 3 flasks and keep 3 flacks as uninoculated control. Incubate the flasks over rotary shaker for 12 days at 28+1C

After 12 days, filter the contents of each flask separately through Whatman No. 42 filter paper or centrifuge at 10,000 rpm for 15 min.

(iii) Add 10ml of filtrate/centrifugate to 50ml of olsen extractant and shake for 30min over rotary shaker.

(iv) Filter the suspension through Whatman filter paper No. 40. If the filtrate is coloured then add a tea spoon of Dacro-60 (activated phosphorous free carbon), reshake and filter.

(v) Take a known aliquot (5 to 25 ml) of the extract in a 50 ml volumetric flask.

(vi) Add 5 drops of p-nitrophenol indicator (1.5 per cent solution in water) and adjust the pH of the extract between 2 and 3 with the help of 4NH₂SO₄. The yellow colour will disappear when the pH of the solution becomes 3. Swirl gently to avoid loss of the solution along with the evolution of CO₂.

(vii) When the CO₂ evolution has subsided, was down the neck of the flask and dilute the solution to about 40ml.

(viii) Add 5 ml of the sulphomolybdic acid mixed reagent containing ascorbic acid, swirl the content and make up the volume.

(ix) Measure the transmission after 30 min at 880 mm using red filter. The blue colour developed remains stable upt 60 minutes.

(x) Record the concentration of phosphorus (P) in the extract form the standard curve and calculate the concentration of soluble phosphorous as follows:

3.4 Calculations

(a) Weight of the substance taken = x g

(b) Volume of the extract added = 50ml

(d) Volume made after colour developed = 50 ml

(e) Reading from the standard curve against percent transmission recorded = z ppm

(f) Soluble Phosphorous percent P =

Z x 50 x 10 x 50 x 100
____________________________

Y.N

3.5 Preparation of standard Curve

Prepare standard curve using 0.1 to 0.6 ppm P in 50 ml volumetric flask. Plot the standard curve by taking concentration of soluble P on x-axis and Percent T on Y-axis using a semi log graph paper. It is a straight line relationship between the soluble P and percent T when plotted on a semi-log graph paper.]

Schedule IV

[Refer clauses 29(h) and (q)]

[Part A]

Specifications of Organic Fertilisers

1.	City compost:
	[(i) Moisture, per cent by weight,	maximum 25]
	[*	*]
	[*	*]
	(iv) Particle size	Minimum 90% material should pass through 4.0mm IS sieve
	(v) Bulk density (g/cm³)	<1.0
	(vi) Total organic carbon, per cent by weight, minium	12.0
	[(vii) Total of Nitrogen (as N), Phosphate as P₂O₃ and Potash as K₂O per cent by weight, minimum	1.2]
	[***]
	[****]
	(x) CN ration	<20