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IS 14832 (2000): Animal Feeding Stuff - Determination of

Urea [FAD 5: Livestock Feeds, Equipment and Systems]

“!ान $ एक न' भारत का +नम-ण”

Satyanarayan Gangaram Pitroda
“Invent a New India Using Knowledge”

“!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता ह”

है”
ह
Bhartṛhari—Nītiśatakam
“Knowledge is such a treasure which cannot be stolen”
 lS 14832 :2000
1S06654 :1991

*w’m-
wpT1-m-’@”m=m T
Indian Standard
ANIMAL FEEDING STUFF — DETERMINATION
OF UREA

ICS 65.120

@ BIS 2000

BUREAU OF INDIAN STANDARDS

MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 110002

September 2000 Price Group 2

Livestock Feeds Sectional Committee, FAD 5

NATIONAL FOREWORD

This Indian Standard, which is identical with ISO 6654:1991 ‘Animal feeding stuffs — Determination
of urea content’ issued by the International Organization for Standardization (ISO) was adopted by
the Bureau of Indian Standards on the recommendation of the Livestock Feeds Sectional Committee
and approval of the Food and Agriculture Division Council.

Method of test for determination of moisture content was covered in 6 of IS 7874 (Part 1); after
publication of this Indian Standard the above referred clause of 1S 7874 (Part 1) will be deleted
through amendment.

In this adopted standard, certain terminology and conventions are not identical to those used in
Indian Standards. Attention is drawn specially to the following:

a) Wherever the words ‘International Standard’ appear referring to this standard, they should
be read as ‘Indian Standard’.

b) Comma (,) has been used as a decimal marker while in Indian Standards, the current
practice is to use a point (.) as the decimal marker.

In reporting the results of a test or analysis made in accordance with this standard, if the final value,
observed or calculated, is to be rounded off, it shall be done in accordance with IS 2:1960 ‘Rules
for rounding off numerical values (revised)’.
 IS 14832:2000
ISO 6654:1991

Indian Standard
ANIMAL FEEDING STUFF — DETERMINATION
OF UREA

-.

1 Scope 5 Reagents

This International Standard specifies a spectro- All reagents shall be of recognized analytical grade.
metric method for the determination of the urea The water used shall be distilled water or water of
content of animal feeding stuffs. at least equivalent purity.

5.1 Activated carbon, which does not adsorb urea.

2 Normative reference
5.2 4-dlmethyl-amino-benzaidehyde (4-DMAB), sol-
ution prepared as follows.
The following standard contains provisions which,
through reference in this text, constitute provisions Dissolve 1,6 g of 4-DMAB in 100 ml of 96 ?4. (V/Vj
of this International Standard. At the time of publi- ethanol, add 10 ml of concentrated hydrochloric acid
cation, the edition indicated was valid. All standards
(P20 = 1,49 glml) and mix.
are subject to revision, and parties to agreements
based on this International Standard are encour- This solution may be stored for a maximum of
aged to investigate the possibility of applying the 2 weeks.
most recent edition of the standard indicated below.
Members of IEC and ISO maintain registers of cur- 5,3 Carrez i soiution.
rently valid International Standards,
Dissolve In water 24 g of zinc acetate dihydrate
ISO 6498:1983, Animal feeding stuffs — Preparation [Zn(CH3COO)22H20] and 3 g of glacial acetic acid.
of (est samp/es. Make up to 100 ml with water and mix.

5.4 Carrez ii soiution.

3 Definition Dissolve in water 10,6 g of potassium hexa-

cyanoferrate(ll) trihydrate (potassium ferrocyanide
For the purposes of this International Standard, the trihydrate) {KJFe(CN)J3H20}. Make up to 100 ml
following definition applies. with water and mix.

urea content: The mass fraction of substances de-

5.5 Urea, standard solution corresponding to 1 g
termined using the procedure specified in this inter-
of urea per Iitre.
national Standard.

It is expressed as a percentage by mass.

6 Apparatus

Usual laboratory apparatus and, in particular, the

following.
4 Principle

Suspension in water of the test portion in the pres- 6,1 Rotary shaker, capable of being operated at a
ence of a decolorant, Stirring of the suspension, rotational frequency of about 30 rein-f to 40 rein-l.
then filtration. Addition to the filtrate of
4-dimethyl-amino-benzaldehyde (4-DMAB) and 6,2 Spectrometer, suitable for measuring
spectrometric measurement at 420 nm of the absorbance at a wavelength of 420 nm, and
absorbance of the solution thus obtained, equipped with cells of thickness 10 mm.

1
IS 14832:2000
ISO 6654:1991

6.3 Test tubes, 160 mm x 16 mm, fitted with ground 9.4 Blank test
glass stoppers.
Carry out a blank test in parallel with the determi-
6.4 Volumetric flasks, of 100 ml and 500 ml capac- nation using the same procedure and the same
ity. quantities of all reagents, but omitting the test por-
tion.

6.5 Water-bath, capable of being controlled at

20 ‘c. 9.5 Preparation of the calibration graph

9.5.1 Pipette into a series of five 100 ml volumetric

7 Sampling
flasks (6.4), 1 ml, 2 ml, 4 ml, 5 ml and 10 ml of the
urea standard solution (5.5). Make each flask up to
Sampling will be the subject of a future International
the mark with water, One millilitre of the standard
Standard.
solutions contains 10 Kg, 20 ~g, 40 pg, 50 pg and
100 ~g of urea respectively.
8 Preparation of the test sample
9.5.2 Pipette into a series of five test tubes (6.3)
Prepare the test sample in accordance with 5 ml of each of these solutions (9.5.1) (one dilution
1S0 6498. per test tube). Add to each test tube, by means of a
pipette, 5 ml of the 4-DMAB solution (5.2) and mix.
Transfer the solutions to spectrometer cells and
9 Procedure measure their absorbance at 420 nm, using the
spectrometer (6.2), against a compensation solution
9.1 Test portion containing 5 ml of 4-DMAB and 5 ml of water.

Weigh, to the nearest 1 mg, about 2 g of the test 9.5.3 Plot the calibration graph, with the
sample (clause 8). absorbance values on the ordinate and the corre-
sponding concentrations of urea, in micrograms per
For urea contents greater than 3 9!o (m/m), reduce millilitre, on the abscissa.
the test portion to 1 g or dilute the test solution
(see 9.2) so that it does not exceed a concentration
of 50 mg of urea per 500 ml. 9.6 Spectrometric measurement

For low urea contents, the test portion may be in- Transfer the soiution obtained in 9.3 to a
creased provided that the filtrate remains clear and spectrometer cell and measure its absorbance at
colorless, 420 nm, using the spectrometer (6.2), against the
blank test (9.4),
9.2 Preparation of the test solution
NOTE 1 If the sample contains simple nitrogenous
compounds, such as amino acids, carry out the measure-
9,2.1 Transfer the test portion (9,1) together with ment of absorbance at a wavelength of 435 nm.
1 g of the activated carbon (5.1) into a 500 ml
volumetric flask (6.4). Add 400 ml of water, 5 ml of
9.7 Number of determinations
the Carrez I solution (5.3) and 5 ml of the Carrez II
solution (5.4). Mix for 30 min using a rotary shaker
Carry out two determinations on test portions taken
(6,!). Make up to the mark with water, mix and filter
from the same test sample.
on a dense, slow, qualitative filter paper.

9.2,2 If the filtrate is coloured, prepare a fresh test 10 ExpressIon of results

solution in accordance with 9.2.1 but increase the
quantity of activated carbon added. The urea content, expressed as a percentage by
mass, of the sample is equal to
9.3 Colour development c
20xm
Transfer, by means of a pipette, 5 ml of the clear
colorless filtrate (9.2) into a test tube (6.3) and add, where
by means of a pipette, 5 ml of the 4-DMAB solution
(5.2). c is the urea content, in micrograms per
millilitre, of the filtrate of the test solution,
Mix and leave to stand for 15 min in a water-bath determined from the calibration graph
(6.5) controlled at 20 “C. (9.5.3);

2
 IS 14832:2000
ISO 6654:1991

m is the mass, in grams, of the test portion dard, or regarded as optional, together with details
(9.1). of any incidents which may have influenced the re-
sult.
11 Test report The test report shall ioclude all information necess-
ary for the complete identification of the sample.
The test report shall specify the method used ana
the result obtained. It shall also mention all operat-
ing details not specified in this International Stan-

3
Bureau of Indian Standards

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harmonious development of the activities of standardization, marking and quality certification of goods
and attending to connected matters in the country.

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Enquiries relating to copyright be addressed to the Director (Publications), BIS.

Review of Indian Standards

Amendments are issued to standards as the need arises on the basis of comments. Standards are also
rev iewed periodically; a standard along with amendments is reaffirmed when such review indicates -.
that no changes are needed; if the review indicates that changes are needed, it is taken up for revision.
Users of Indian Standards should ascertain that they are in possession of the latest amendments or
edit ion by referring to the latest issue of ‘BIS C@alogue’ and’ Standards: Monthly Additions’.

This Indian Standard has been developed from ,Doc : No. FAD 5 (979).

Amendments Issued Since Publication

Amend No. Date of Issue Text Affected

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