Integrated Plant Nutrient Management909 204 5811hrj@nihausa.com

PAKISTAN STANDARD FOR BIO ORGANO

PS: 5295-2014
ICS: 65.080

PAKISTAN STANDARD SPECIFICATION
FOR
BIO ORGANO PHOSPHATE

PAK

PAKISTAN STANDARDS AND QUALITY CONTROL AUTHORITY,
STANDARDS DEVELOPMENT CENTRE,
Plot No. ST-7A, Block-3, Scheme 36, Gulistan-e- Johar
Karachi.

PAKISTAN STANDARDS SPECIFICATION
FOR
BIO ORGANO PHOSPHATE (BOP) FERTILZER

0.0 FOREWORD
0.1 This Pakistan Standard was adopted by Pakistan Standards & Quality Control Authority on
26-11-2014 after the draft finalized by the Fertilizers and Allied Product Technical Committee had
been approved by the National Standards Committee for Chemical Division. 0.2 Bio Organo phosphate (BOP) fertilizer product provides 20% (±2) organic P2O5 25% (±5) organic
matter. The product provides multiple benefits to the farmer by improving soil and crop health. The
organic P component is more efficient with low P fixationation tendency in soil and prolonged
availability to crop than chemical fertilizer, under soil and climatic condition. The product can
positively contribute in promoting balanced fertilizer use and breaking current stagnancy in crop
yields thus improving agricultural productivity.
Bio Organo Phosphate (BOP) produced by an innovation and has better use efficacy due to less
probability of fixation, precipitation or insolubilization than current commercial soluble
chemical/inorganic fertilizers. This will help increase P use efficiency and optimize ever increasing
per hectare use of phosphorous fertilizers.
0.3 The fertilizer committee responsible for the preparation of this draft felt that it is necessary to lay
down the specifications on BOP fertilizer to safeguard the interests of the farmer community and to
protect them from using wrong types or grades of BOP fertilizer as well safe guard the interest of
the manufactures. 0.4 While preparing this standard the views of all stake holders including manufacturers, importers,
testing authorities, technologist/experts and consumers have been taken into consideration and also
the existing trade practice in this field in the country, by the Technical Committee. Furthermore, due weightage had given to the need for international co-ordination among standards prevailing in
different countries of the world. 0.5 For the purpose of deciding whether a particular requirement of this standard is complied with, the
final value, observed or calculated, expressing the result of a test or analysis shall be rounded off in
accordance with PS: 103 – 1991 “Rules for Rounding Off Numerical Values”. The number of
places retained in the rounded off value should be the same as those of the specified value in the
standard.
0.6 In order to keep abreast with the progress of trade and Industry Pakistan Standards are revised
periodically. Suggestions from the members are welcomed and will be placed before the
Technical Committees for consideration at the time of revision.
0.7 This standard is intended mainly to cover the technical provisions relating to the supply of the
material, and it does not cover all the necessary provision of a contract.

1.0 SCOPE
1.1 This standard provides requirement and methods for Bio Organo Phosphate for use as fertilizer. The material is known as BOP.

2.0 REQUIREMENT:
2.1 The material shall consist essentially of BOP and shall be in the form of free flowing granules. It
shall be free from visible foreign matters and shall be according to the requirements specified in
Table 1, when tested according to the methods prescribes in column 4 of same table.

Table-1:
REQUIREMENTS FOR BIO ORGANO PHOSPHATE (BOP) FERTILZER

3.0 PACKING AND MARKING:
3.1 Packing: The material shall be packed and supplied in sound, strong, moisture proof packages or
container as agreed to between producer/marketer/distributor and purchaser/vendor, such as natural
or synthetic fiber bags of multi wallpaper with bitumen or polyethylene moisture proofing layer. Mono film bags of heavy polyethylene (0.15-0.2 mm thickness) or in such other suitable containers.
Jute or woven polypropylene bags with mono-film polyethylene liners.
3.2. Marking: The container/sack/bag shall be securely closed and marked with the following
information:
a. Name of the material, namely “Bio Organo Phosphate (BOP)”
b. Minimum organic P content (as %P2O5)
c. Name and Address of the manufacturer/importer/distributor; trade mark if any
d. Net Weight in kg of the material in the container
e. Any information required by law enforcement agencies or by the buyer.
3.3 Storage Requirements:
a. Store in cool, clean, dry and well-ventilated area at room temperature. b. Avoid contact with moisture, as it may cause product-handling problems
c. Store away from oxidizers, acids, and food and drinkable items.

4.0 SAMPLING:
4.1 Representative sample of the material shall be drawn as prescribed in Appendix – A.

APPENDIX – A
SAMPLING OF BIO ORGANO PHOSPHATE (BOP) FERTILIZER

A-1 GENERAL REQUIREMENTS OF SAMPLING
A-1.0 Following precautions and directions shall be observed in drawing, preparing, storing and handling
test samples.
A-1.1 Samples shall be taken at a place protected from damp air, dust and soot.
A-1.2 The sampling instruments shall be clean and dry when used.
A-1.3 Precautions shall be taken to protect the samples, the material being sampled, the sampling
instruments and the containers for samples from adventitious contamination.
A-1.4 To draw a representative sample, the contents of each container/bag selected for sampling shall be
mixed as thoroughly as possible by suitable means.
A-1.5 The samples shall be placed in clean, dry and airtight glass or other suitable containers on which the
material has no action.
A-1.6 The sample containers shall be of such a size that they are almost completely filled by the sample.
A-1.7 Each sample container shall be sealed airtight after filling and marked with full details of sampling,
date of sampling and other important particulars of the consignment.
A-1.8 Samples shall be stored at dry place.

A-2 SCALE OF SAMPLING:
A-2.1 Lot – All the containers in a single consignment of the material drawn from a single batch of
manufacture shall constitute a lot. If a consignment is declared to consist of different batches of
manufacture, the batches shall be marked separately and the groups of containers in such batch
shall constitute separate lots.
A-2.2 The number of sampling containers/bags to be chosen from a lot shall depend on the size of the lot
and shall be in accordance with column 1 and 2 of Table II.

TABLE – II
NUMBER OF CONTAINERS TO BE SELECTED FOR SAMPLING

niha

A-2.3 These containers/bags shall be chosen at random from the lot, and in order to ensure randomness of
selection the following procedure may be adopted.
A-2.4 Arrange all the containers in the lot in a systematic manner and starting from any container, count
them 1,2,3, ……… etc up to r and so on, r being equal to the integral part of N/n. Every rth
containers thus counted shall be withdrawn and all such containers shall constitute the sample.

A-3 TEST SAMPLE AND REFEREE SAMPLE:
A-3.1 Draw with an appropriate sampling instrument small portions of the material from different parts of
the containers selected, the total quantity taken out from each container being sufficient to conduct
the tests for all characteristics given in table-1.
A-3.2 Mix thoroughly all portions of the material drawn from the same container to form an individual
test sample. Equal quantities from all individual test samples so formed shall be mixed together to
form a composite test sample.
A-3.3 All the individual test samples and the composite test sample shall be divided into three equal parts,
thus forming three sets of test samples. These parts shall be immediately transferred to thoroughly
dried bottles, which shall then be sealed air tight with stopper. One of these sets of test sample shall
be sent to the purchaser and another to the vendor.
A-3.4 Referee Sample – The third set of test samples bearing the seals of the purchaser and the vendor,
shall constitute the referee sample and shall be used in case of dispute between the purchaser and
the vendor. It shall be kept at a place agreed to between the purchaser and the vendor.

A-4 NUMBER OF TEST:
A-4.1 Test for the determination of organic phosphate as P2O5% content shall be conducted on each of the
individual test samples.
A-4.2 Test for the determination of bioavailability of P after soil incubation over a period of time shall be
conducted on each of the individual samples only to differentiate BOP from other chemical
phosphatic fertilizers. This test should be conducted only if required by any of the stakeholders as
defined in 0.4 of foreword. A-4.3 Test for the determination of moisture % shall be conducted on each of the individual test samples.
A-4.4 Test for the remaining characteristics given in table-1 shall be conducted on the composite test
sample.

A-5 CRITERION FOR CONFORMITY:
A-5.1 The test results for organic phosphate as P2O5% shall be recorded as shown in Table III. The mean
and the range of the test result shall be calculated as follows:

Mean (X) = The sum of the test results
                      Number of test results

Range (R) = the difference between the maximum and the minimum values of the test results.
A-5.1.1 The appropriate expression as shown in col. 6 of Table III shall be calculated for the
characteristic. If the condition given in col. 6 of Table III is satisfied, the lot shall be declared to
have satisfied the requirement for this characteristic.
A-5.2 For the remaining characteristics, the test results on the individual and composite test sample shall
satisfy the requirements specified in table-1.
A-5.3 A lot shall be declared as conforming to the specification only when it has satisfied each of the
requirements specified in table-1 except bioavailability of P after soil incubation, as this test is to
differentiate BOP from other chemical phosphatic fertilizers and shall be conducted on special
request from any of the stake holders as defined in 0.4 of foreword.

TABLE – III
CRITERION FOR CONFORMITY

APPENDIX - B
DETERMINATION OF MOISTURE IN BOP (OVEN METHOD)

B-1 APPARATUS:
a. Weighing bottle- size 50 mm x 30 mm, fitted with ground glass stopper with a hole.
b. Air oven; heated electrically with temperature control system.
c. Pestle and Mortar – made of porcelain or glass, size 102 mm (4 inch internal diameter).
d. Balance

B-2 PROCEDURE
B-2.1 Preparation of Sample – Place 1 gram of sample in a mortar and grind quickly particle size required
is less than 1 mm, weigh 5 g of the ground sample into a weighing bottle using an analytical
balance.
B-2.2 Determination – Place the weighing bottle containing the sample in the air oven maintained at
100 + 20C. After 4 hours take the sample bottle out, and cool in a desiccator for 15 – 20 minutes.
Silica gel is desirable as desiccating agent. Reweigh the sample using an analytical balance.

B-3 CALCULATION:

APPENDIX – C
ANALYSIS OF BIO ORGANO PHOSPHATE (BOP) FERTILIZER

QUALITY OF REAGENTS:
C-1.1 Unless specified otherwise, Analytical grade chemicals and distilled water (see PS: 593-1996*)
shall be used in tests.
NOTE: – ‘Analytical grades Chemicals’ shall mean chemicals that do not contain impurities that
affect the results of analysis.

C-2 PREPARATION OF SAMPLE:
C-2.1 Crush 50 g of BOP to pass through 500 microns PS Sieve. Dry to constant weight to obtain the
prepared sample and keep in clean glass-stoppered weighing bottle in a desiccator for subsequent
tests.
C-3 ANALYSIS OF ORGANIC P CONTENTS (P2O5) IN BOP FERTLIZER

C-3.1 PRINCIPLE : Determination of organic P contents (P2O5) in the BOP thorough Spectro
photometric method
C-3.2 APPARATUS

. Analytical Balance
. Glassware : volumetric flasks, pipettes, pipette fillers, beakers, etc.
. Spectro Photometer
. Digestion Block
. Fume Hood
C-3.3 REAGENTS (According to ICARDA manual)
C-3.3.1 Ammonium Heptamolybdate – Ammonium Vanadate in Nitric Acid
a. Dissolve 22.5 grams of ammonium heptamolybdate [(NH4)6 Mo7 O24. 4H2O] in 400 mL
distilled water.
b. Dissolve 1.25 grams of ammonium metavenadate [NH4VO3] in 300 mL hot distilled water. c. Add (b) to (a) in a one-liter volumetric flask, and let the mixture cool to room temperature.
d. Slowly add 250 mL concentrated nitric acid (HNO3) to the mixture, cool the solution to
room temperature, and bring to 1-L volume with Distilled water.
C-3.3.2 Preparation of stock standard solution
a. Dry about 2.5 g potassium di-hydrogen phosphate [KH2PO4] in an oven at 105°C for
1hour, cool it in desiccator and store in a tightly stopper bottle.
b. Dissolve 0.2197 g dried potassium di-hydrogen phosphate in Distil water, and bring to 1-
liter volume with distilled water. c. This solution contains 50 ppm P (Stock Solution).
C-3.4 Method
C-3.4.1 Digest 0.1 g dried material with sulfuric acid and hydrogen peroxide according to the method of
McGill and Figueiredo (1993).
a. Place 0.1 g of BOP fertilizer in digestion tubes, add 2mL of concentrated H2SO4 and incubate
digestion tubes overnight at room temperature.
b. After incubation, pour 1 mL of H2O2 (35% A. R. grade extra pure) through the sides of digestion
tubes and rotate.
c. Place tubes in digestion block and heat up to 350°C until fumes are produced and continued to
heat for another 30 minutes. After that remove tubes from digestion block and cool.
d. Add 1 mL of H2O2 slowly. Place tubes back into digestion block until fumes are produced for 20
minutes.
e. Repeat above process until colorless solution is obtained.
f. Make up extract volume up to 50 mL by adding distilled water.
g. Filter the made up volume and use for determination of total phosphorus.

C-3.4.2 Measurement of organic P contents (P2O5) by spectrophotometer (According to ICARDA manual)
a. Pipette 10 mL of digested filtrate into a 100 mL volumetric flask.
b. Add 10 mL ammonium vanadomolybdate reagent, dilute solution to volume with distilled water.
c. Prepare a standard curve:
 Pipette 1, 2, 3, 4, and 5 mL standard stock solution into 100 mL volumetric flasks to
prepare 0.5, 1.0, 1.5, 2.0, and 2.5 ppm working standards of P. Add 10 mL
ammonium- vanadomolybdate reagent in each flask and make volume up to mark
with distilled water.
 Make a blank with 10 mL ammonium-vanadomolybdate reagent and proceed as for
the samples and working standards.
 Read absorbance of blank, standards, and samples after 30 minutes at 410 nm
wavelengths.
 Prepare a calibration curve for standards, plotting absorbance against the respective
P concentrations.
d. Read organic P concentration in the unknown samples from the calibration curve.

C-3.5 Calculation of organic P contents (P2O5%)

niha

C-4 ANALYSIS OF ORGANIC MATTER IN BOP FERTILIZER
C-4.1 PRINCIPLE : Loss of weight on ignition.
C-4.2 APPARATUS
a. Crucible
b. Balance
c. Muffle Furnace
d. Desiccator

C-4.3 Method
C-4.3.1 Weigh clean dry crucible (W1).
C-4.3.2 Add 5 g of sample in a tarred crucible and oven dry at 105 oC for 4 hours.
C-4.3.3 Weigh sample and crucible. This yields the oven dry weight (W2).
C-4.3.4 Place crucible with same oven dried sample in furnace at 550 to 600 oC for 6 hours to burn sample
and form ash. Cool the crucible and ash in desiccator and weigh. This yields the ash weight (W3).
C-4.4 Calculation

niha

C-5 DETERMINATION OF BIOAVAILABLE P IN SOIL SPIKED WITH BOP FERTILIZER
C-5.1 PRINCIPLE: Bioavailability of P after soil spiked with BOP fertilizer over a period ranging from 2
to 13 weeks through sodium bicarbonate extractions and colorimetric analysis. C-5.2 APPARATUS
a. Analytical balance
b. UV visible Spectrophotometer
c. Orbital shaker
d. Centrifuge
e. Extraction bottle, 250 mL with stopper
f. Glassware of standardized laboratory Beakers, volumetric flasks, pipettes, funnels etc.
g. Incubator and plastic beakers for incubation

C-5.3 REAGENTS
C-5.3.1 Sodium Bicarbonate Solution (NaHCO3), 0.5 M. 42 grams of sodium bicarbonate is dissolved in
900 mL of distilled water. pH is adjusted to 8.5 by using 5 N NaOH solution and make the
volume 1L with distilled water. Store the solution in glass or plastic container.
C-5.3.2 Sulfuric Acid Solution (H2SO4), 5 N. 148 mL of concentrated sulfuric acid is diluted with distilled
water in the fume hood. Mix and cool it, then make the volume 1L by using distilled water.
C-5.3.3 Standard Stock Solution of Phosphorus. a. Dry potassium dihydrogen phosphate (KH2PO4) in oven at 105 0C for an hour and is cooled in
desiccator and store it in airtight bottles.
b. Weigh 2.197 g (KH2PO4) and dissolve in to a 1L volumetric flask by using distilled water and
the concentration of this stock solution is 500 ppm
c. Take 50 mL from stock solution into a 250 mL volumetric flask and make the volume by
using distilled water, the concentration of this sub stock is 100 ppm
d. Now prepare working standards of 2, 4, 6, 8, 10, 12, 14, 16 and 18 ppm by dissolving 2, 4, 6,
8, 10, 12, 14, 16 and 18 mL in 100 mL volumetric flask from sub stock solution (100 ppm).
C-5.3.4 Color Developing Reagent (CDR)

a. Reagent A
 Dissolve 12 g of ammonium heptamolybdate (NH4)6 Mo7O24.4H2O in 250 mL of distil water.
 Dissolve 0.2908 g of antimony potassium tartrate (KSbO.C4H4O6) in 100 mL of distil water.
 Add and dissolve both reagents in a 2 L volumetric flask. Also add 5 N Sulfuric acid solution, mix it well and make the volume with distilled water and keep away from light and
heat.

b. Reagent B
 Dissolve 1.056 g of L. ascorbic acid (C6H8O6) in 200 mL of reagent A and mix thoroughly. Reagent B is prepared according to the need as it become ineffective after 24 hours.

C-5.4 PROCEDURE
a. Take six samples of 500 g sieved soil in separate plastic beakers. Label three beakers as
“Fertilizer”, three beakers as “Soil” and make three replications. Spike the soil of beakers
labeled as “Fertilizer” with 100 g of BOP, any other P fertilizer or any other material that
needs to be checked. The beakers labeled as “Soil” should be maintained without addition or
spiking with any fertilizer material. Maintain soil water holding capacity at 50%. Incubation
should be carried out for 13 weeks at 28 ±1°C.
b. Take 5 g well mixed soil sample from each beaker periodically at 7 days interval.
c. Transfer air-dried soil samples in separate flasks of 250 mL each and add 100 mL of sodium bicarbonate (NaHCO3) solution (0.5 M) with pH 8.5 in each flask.
d. Shake each flask at 200 – 300 rpm for 30 minutes.
e. Filter each solution, separately, by using filter paper Whatman No. 42.
f. Transfer 10 mL of each filtrate in separate 100 mL volumetric flasks and add 10 mL of reagent B and make the volume 100 mL with distilled water in each flask.
g. Shake each flask well and take readings against absorbance by feeding; first blank, then
Standards, followed by 6 samples of “Fertilizer” and “Soil”.
h. Make calibration curve for Standards by plotting absorbance against P concentrations.
i. Measure P concentration of “Fertilizer” and “Soil” samples from the calibration curve.

j. Calculate average extracted P from “Fertilizer” and “Soil” (6 samples, 3 replications); and
record average of “Fertilizer” under column A and of “Soil” under column B in table IV.
k. Calculate Bioavailable P (ppm) as per following formula. = Average extracted P from “Fertilizer” – Average extracted P from “Soil” samples
l. Compare Bioavailable P (ppm) value of column C in table IV with corresponding standard
value given in column D. If values of column C and D are in close proximity then the product
will be BOP, if not then it is either chemical fertilizer or some other material.
m. This test may be continued for minimum of 2 weeks to maximum of 13 weeks until the type of
product is clearly identified by the testing authority. If values of column C follows the trend of
values given in column D then the product is BOP if not then it is either chemical fertilizers or
some other material.

niha

APPENDIX-D
DETERMINATION OF SIZE DISTRIBUTION OF BOP

D-1 APPARATUS / EQUIPMENT
D-1.1 Stainless steel sieves of the required mesh size with lid & bottom pan.
D-1.2 Sieves Shaker
D-1.3 Balance
D-1.4 Brush

D-2 METHOD:
D-2.1 Arrange the individually tare sieves in descending order of mesh size from top to bottom.
D-2.2 Place receiving pan on the bottom of stack
D-2.3 Weigh about 200 to 300g of sample taken through sample divider.
D-2.4 Transfer sample onto the top sieve place lid on top of stack.
D-2.5 Place the sieve stack on shaker and tighten the belts evenly on both sides.
D-2.6 Set timer of vibrator to 5 minutes amplitude at 3.0 mm and start the vibrator.
D-2.7 After shaking stops switch-off the vibrator and remove the sieves one by one.
D-2.8 Weigh sieve + sample on top loading balance.
D-2.9 Calculate the weight of samples retained on each sieve.

D-3 CALCULATION
D-3.1 Calculate weight percent on each sieve by following formula