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INTERNATIONAL ISO
STANDARD 5983-1

First edition
2005-07-01

Animal feeding stuffs — Determination of

nitrogen content and calculation of crude
protein content —
Part 1:
Kjeldahl method
Aliments des animaux — Détermination de la teneur en azote et calcul
de la teneur en protéines brutes —
Partie 1: Méthode Kjeldahl

Reference number
ISO 5983-1:2005(E)

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ISO 5983-1:2005(E)

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ISO 5983-1:2005(E)

Contents Page

Foreword............................................................................................................................................................ iv
1 Scope ..................................................................................................................................................... 1
2 Normative references ........................................................................................................................... 1
3 Principle ................................................................................................................................................. 1
4 Reagents and materials ....................................................................................................................... 1
5 Apparatus .............................................................................................................................................. 2
6 Sampling ................................................................................................................................................ 2
7 Preparation of test sample................................................................................................................... 3
8 Procedure .............................................................................................................................................. 3
9 Calculation and expression of results................................................................................................ 5
10 Precision ................................................................................................................................................ 6
11 Test report ............................................................................................................................................. 7
Annex A (informative) Results of interlaboratory test.................................................................................... 8
Bibliography ..................................................................................................................................................... 10

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ISO 5983-1:2005(E)

Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.

International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.

The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.

Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.

ISO 5983-1 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 10, Animal
feeding stuffs.

This first edition of ISO 5983-1, together with ISO 5983-2:2005, cancels and replaces ISO 5983:1997, which
has been technically revised.

ISO 5983 consists of the following parts, under the general title Animal feeding stuffs — Determination of
nitrogen content and calculation of crude protein content:

 Part 1: Kjeldahl method

 Part 2: Block digestion/steam distillation method

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INTERNATIONAL STANDARD ISO 5983-1:2005(E)

Animal feeding stuffs — Determination of nitrogen content

and calculation of crude protein content —
Part 1:
Kjeldahl method

1 Scope

This part of ISO 5983 specifies a method for the determination of the nitrogen content of animal feeding stuffs
by the Kjeldahl process, and a method for the calculation of the crude protein content.

The method does not measure oxidized forms of nitrogen or heterocyclic nitrogen compounds.

This method does not distinguish between protein nitrogen and non-protein nitrogen. If it is important to
determine the content of non-protein nitrogen, an appropriate method should be used.

2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.

ISO 6498, Animal feeding stuffs — Preparation of test samples

3 Principle
The organic matter is digested by sulfuric acid in the presence of a catalyst. The reaction product is rendered
alkaline, then the liberated ammonia is distilled and titrated. The nitrogen content is calculated and the result
is multiplied by the conventional factor to obtain the crude protein content.

4 Reagents and materials

Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or deionized water
or water of equivalent purity.

The reagents [except the standard materials (4.6)] shall be practically free from nitrogenous compounds.

4.1 Potassium sulfate.

4.2 Catalyst, either 4.2.1 or 4.2.2.

4.2.1 Copper(II) oxide (CuO).

4.2.2 Copper(II) sulfate pentahydrate (CuSO4·5H2O).

4.3 Sulfuric acid, c(H2SO4) = 18 mol/l, ρ20(H2SO4) = 1,84 g/ml.

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ISO 5983-1:2005(E)

4.4 Paraffin wax.

4.5 Saccharose.

4.6 Standard materials, either 4.6.1 or 4.6.2.

4.6.1 Acetanilide, with melting point 114 °C; nitrogen (N) content 103,6 g/kg.

4.6.2 Tryptophan, with melting point 282 °C; nitrogen (N) content 137,2 g/kg.

Dry before use.

4.7 Sodium hydroxide solution, w(NaOH) = 33 % (mass fraction).

4.8 Collecting liquid, either 4.8.1 or 4.8.2.

4.8.1 Sulfuric acid, standard volumetric solution, c(H2SO4) = 0,05 mol/l or c(H2SO4) = 0,125 mol/l.

4.8.2 Boric acid, ρ (H3BO3) = 40 g/l.

4.9 Solutions for titration.

4.9.1 Sodium hydroxide, standard volumetric solution, c(NaOH) 0,1 mol/l or c(NaOH) = 0,25 mol/l.

4.9.2 Sulfuric acid, standard volumetric solution, c(H2SO4) = 0,05 mol/l or c(H2SO4) = 0,125 mol/l.

The molarity of standard volumetric solutions should be known to the fourth decimal point.

4.10 Mixed indicator, neutral point at pH 4,4 to 5,8.

Dissolve 2 g of methyl red and 1 g of methylene blue in 1 000 ml of ethanol [ϕ(C2H5OH) = 95 % (volume
fraction)].

4.11 pH indicator paper.

4.12 Boiling aids, such as granulated pumice stone, or glass beads of diameter 5 mm to 7 mm, or
carborundum chips, washed in hydrochloric acid and in distilled water, and ashed.

5 Apparatus
Usual laboratory apparatus and, in particular, the following.

5.1 Analytical balance.

5.2 Digestion, distillation and titration apparatus.

6 Sampling
A representative sample should have been sent to the laboratory. It should not have been damaged or
changed during transport or storage.

Sampling is not part of the method specified in this part of ISO 5893. A recommended sampling method is
given in ISO 6497.

Store the sample in such a way that deterioration and change in its composition are prevented.

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