CD/K/725:2010

ICS 67.120.10

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EAST AFRICAN STANDARD

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Meat and meat products — Determination of nitrite content
(Reference method)

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EAST AFRICAN COMMUNITY

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© EAC 2010 First Edition 2010

 CD/K/725:2010

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Foreword

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Development of the East African Standards has been necessitated by the need for harmonizing
requirements governing quality of products and services in East Africa. It is envisaged that through

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harmonized standardization, trade barriers which are encountered when goods and services are
exchanged within the Community will be removed.

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In order to meet the above objectives, the EAC Partner States have enacted an East African
Standardization, Quality Assurance, Metrology and Test Act, 2006 (EAC SQMT Act, 2006) to make

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provisions for ensuring standardization, quality assurance, metrology and testing of products
produced or originating in a third country and traded in the Community in order to facilitate industrial

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development and trade as well as helping to protect the health and safety of society and the
environment in the Community.

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East African Standards are formulated in accordance with the procedures established by the East

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African Standards Committee. The East African Standards Committee is established under the
provisions of Article 4 of the EAC SQMT Act, 2006. The Committee is composed of representatives of
the National Standards Bodies in Partner States, together with the representatives from the private

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sectors and consumer organizations. Draft East African Standards are circulated to stakeholders
through the National Standards Bodies in the Partner States. The comments received are discussed
and incorporated before finalization of standards, in accordance with the procedures of the

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Community.

Article 15(1) of the EAC SQMT Act, 2006 provides that “Within six months of the declaration of an
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East African Standard, the Partner States shall adopt, without deviation from the approved text of the
standard, the East African Standard as a national standard and withdraw any existing national
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standard with similar scope and purpose”.

East African Standards are subject to review, to keep pace with technological advances. Users of the
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East African Standards are therefore expected to ensure that they always have the latest versions of
the standards they are implementing.
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© East African Community 2010 — All rights reserved*

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East African Community

P O Box 1096
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Arusha
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Tanzania
Tel: 255 27 2504253/8
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Fax: 255-27-2504481/2504255
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E-Mail: eac@eachq.org
Web: www.each.int
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© 2010 EAC — All rights of exploitation in any form and by any means reserved worldwide for EAC Partner States’ NSBs.
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ii © EAC 2010 — All rights reserved

 CD/K/725:2010

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Introduction

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In the preparation of this East African Standard, the following sources were consulted extensively:

ISO 2918:1975, Meat and meat products — Determination of nitrite content (Reference method)

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Codex Alimentarius website: http://www.codexalimentarius.net/mrls/pestdes/jsp/pest_q-e.jsp

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USDA Foreign Agricultural Service website: http://www.mrldatabase.com

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USDA Agricultural Marketing Service website: http://www.ams.usda.gov/AMSv1.0/Standards

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USDA Plant Inspectorate Service website: http://www.aphis.usda.gov/import_export/plants

European Union: http://ec.europa.eu/sanco_pesticides/public

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Assistance derived from these sources is hereby acknowledged.

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© EAC 2010 — All rights reserved iii

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INTERNATIOSTANDARD
N AL 2918.

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INTERNATIONAL ORGANIZATION F O R STANDARDIZATION #MEXfiYHAPOAHAII OPTAHM3AUMR I-IO CTAHAAPTM3ALWi~ORGANISATION INTERNATIONALE DE NORMALISATION

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Meat and meat products - Determination of nitrite content

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(Reference method)
Viandes et produits ;i base de v iande - De’termination
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de la teneur en nitrites (M&ode de re’fkrence)
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F irst edition - 1975-09-01

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UDC 637.5 : 546.17 Ref. No. IS0 2918-1975 (E)

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Descriptors : meat, meat products, chemical analysis, determination of content, nitrites.

Price based on 3 pages

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FOREWORD

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IS0 (the International Organization for Standardization) is a worldwide federation
of national standards institutes (IS0 Member Bodies). The work of developing

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International Standards is carried out through IS0 Technical Committees. Every

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Member Body interested in a subject for which a Technical Committee has been set
up 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.

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Draft International Standards adopted by the Technical Committees are circulated
to the Member Bodies for approval before their acceptance as International ed
Standards by the IS0 Council.
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International Standard IS0 2918 was drawn up by Technical Committee
ISO/TC 34, Agricultural food products, and circulated to the Member Bodies in
April 1974.
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It has been approved by the Member Bodies of the following countries :

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Australia France Romania

Austria Germany South Africa, Rep. of
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Belgium Hungary Spain

Bulgaria India Turkey
Chile Israel United Kingdom
Czechoslovakia Netherlands Yugoslavia
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Denmark New Zealand

Egypt, Arab Rep. of Poland
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The Member Body of the following country expressed disapproval of the document
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on technical grounds :
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Canada
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@ International Organization for Standardization, 1975 l

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Printed in Switzerland
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 INTERNATIONAL STANDARD IS0 29184975 (E)

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Meat and meat products - Determination of nitrite content

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(Reference method)

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1 SCOPE AND FIELD OF APPLICATION 5 1.3 Borax solution, saturated

This International Standard specifies a reference method Dissolve 50 g of disodium tetraborate decahydrate

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for the determination of the nitrite content of meat and (Na2B~07~10H20) in 1 000 ml of tepid water and cool to
room temperature.

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meat products.

5.2 Sodium nitrite standard solutions

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2 REFERENCE
Dissolve 1,000 g of sodium nitrite (NaN02) in water and
IS0 3100, Meat and meat products - Sampling. dilute to 100 ml in a one-mark volumetric
ed flask. Pipette
5 ml of the solution into a 1 000 ml one-mark volumetric
flask. Dilute to the mark.
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3 DEFINITION Prepare a series of standard solutions by pipetting 5 ml,

10 ml and 20 ml of this solution into 100 ml one-mark
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nitrite content of meat and meat products : The nitrite volumetric flasks and diluting to the mark with water.
content determined according to the procedure described in These standard solutions contain respectively 2,5 rug, 5,0 ,ug
this International Standard and expressed as milligrams of and 10,Opg of sodium nitrite per millilitre.
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sodium nitrite per kilogram (parts per million).

The standard solutions and the dilute (0,05 g/l) sodium
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nitrite solution from which they are prepared shall be made

up on the day of use.
4 PRINCIPLE
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Extraction of a test portion with hot water, precipitation of

the proteins and filtration. In the presence of nitrite, 5.3 Solutions necessary for colour development
development of a red colour by the addition of
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sulphanilamide and N-I-naphthylethylenediamine dihydro-

chloride to the filtrate and photometric measurement at
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5.3.1 Solution I
a wavelength of 538 nm.
Dissolve, by heating on a water bath, 2 g of sulphanilamide
(NH2CeH4S02NH2) in 800 ml of water. Cool, filter, if
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necessary, and add 100 ml of concentrated hydrochloric

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5 REAGENTS acid solution (p2c I,19 g/ml), while stirring. Dilute to

1 000 ml with water.
All reagents shall be of analytical quality. The water used
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shall be distilled water or water of at least equivalent

purity. 5.3.2 Solution II
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Dissolve 0,25 g of N-I-naphthylethylenediamine dihydro-

5.1 Solutions for precipitation of proteins
chloride (C,oH7 NHCH2CH2NH2m2HCI) in water. Dilute to
250 ml with water.
or

5.1.1 Reagent I

Dissolve 106 g of potassium ferrocyanide trihydrate Store the solution in a well-stoppered brown bottle. It shall
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[ K~Fe(CN)e~3H201 in water and dilute to 1 000 ml. be kept in a refrigerator, for not longer than one week.
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5.1.2 Reagent II
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5.3.3 Solution Ill

Dissolve 220 g of zinc acetate dihydrate
[Zn(CH3C00)2m2H 2 01_ and 30 ml of glacial acetic acid in Dilute 445 ml of concentrated hydrochloric acid solution
water and dilute to 1 000 ml. (p2c I,19 g/ml) to 1 000 ml with water.

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lSO2918-1975(E)

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6 APPARATUS 8.3 Deproteination

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Usual laboratory equipment and the following items :
8.3.1 Transfer the test portion quantitatively into the

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conical flask (6.8) and add successively 5 ml of saturated
6.1 Mechanical meat mincer, laboratory size, fitted with a borax solution (5.1.3) and 100 ml of water at a

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perforated plate with holes not greater than 4 mm in temperature not below 70 “C.
diameter.

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8.3.2 Heat the flask for 15 min on the boiling water bath
6.2 Analytical balance. (6.5) and shake repeatedly.

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6.3 One-mark volumetric flasks of 100 ml, 200 ml and 8.3.3 Allow the flask and its contents to cool to room

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1 000 ml, complying with ISO/R 1042, Class B. temperature and add successively 2 ml of reagent I (5.1 .I )
and 2 ml of reagent II (5.1.2). Mix thoroughly after each
addition.

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6.4 One-mark pipettes of 10 ml and, if necessary, with
another capacity according to the aliquot of filtrate (8.4.1),

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8.3.4 Transfer the contents to a 200 ml one-mark
complying with ISO/R 648, Class A.
volumetric flask (6.3). Dilute to the mark with water and
mix. Allow the flask to stand for 30 min at room

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6.5 Boiling water bath. temperature.

6.6 Photoelectric calorimeter or spectrophotometer with ed

8.3.5 Carefully decant the supernatant liquid and filter it
cells of 1 cm optical path length. through the fluted filter paper (6.7) so as to obtain a clear
solution.
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6.7 Fluted filter paper, diameter about 15 cm, free of

nitrite. 8.4 Colour measurement
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8.4.1 Pipette an aliquot portion of the filtrate (V ml), but

6.8 Conical flask, 300 ml. not more than 25 ml, into a 100 ml one-mark volumetric
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flask (6.3) and add water to obtain a volume of about

60 ml.
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7 SAlMPLE 8.4.2 Add 10 ml of solution I (5.3.1), followed by 6 ml of

solution I I I (5.3.3), mix and leave the solution for 5 min at
7.1 Proceed from a representative sample of at least 200 g. room temperature, in the dark.
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See IS0 3100.

8.4.3 Add 2 ml of solution I I (5.3.2), mix and leave the
7.2 Prepare the test sample (8.1) immediately or, if this
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solution for 3 to 10 min at room temperature in the dark.

cannot be done, store the sample at a temperature of 0 to Dilute to the mark with water.
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5 ‘C, for not longer than 4 days.

8.4.4 Measure the absorbance of the solution in a 1 cm
cell using a photoelectric calorimeter or a spectro-
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photometer (6.6), at a wavelength of about 538 nm.

8 PROCEDURE
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NOTE - If the absorbance of the coloured solution obtained from

8.1 Preparation of test sample the test portion exceeds that obtained for the standard solution
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with the highest concentration, repeat the operations described

Make the sample homogeneous by passing it at least twice in 8.4, reducing the quantity of filtrate pipetted in 8.4.1.

through the meat mincer (6.1) and mixing. Keep it in a

completely filled, air-tight, closed container under 8.5 Number of determinations
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refrigeration.
Carry ou t two ind ependen t determinations, beginning with
Analyse the test sample as soon as possible, but always different test porti ons take n from the same test sample.
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within 24 h.
8.6 Calibration curve
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NOTE - In the case of uncooked products, analyse immediately

after homogenization.
8.6.1 Pipette respectively into four 100 ml one-mark
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volumetric flasks (6.3) 10 ml of water and 10 ml

8.2 Test portion
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of each of the three sodium nitrite standard solutions (5.2),

Weigh, to the nearest 0,001 g, about 10 g of the test containing 2,5 rug, 5,0 pg and IO,0 ,ctg of nitrite per
sample. millilitre.

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 ISO2918-1975 (E)

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8.6.2 To each flask add water to obtain a volume of about c is the concentration of sodium nitrite, in micrograms

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60 ml and proceed as described in 8.4.2 to 8.4.4. per millilitre, read from the calibration curve, that
corresponds with the absorbance of the solution

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8.6.3 Draw the calibration curve by plotting the measured prepared from the test portion (see 8.4.4).
absorbances against the concentrations, in micrograms per Take as the result the arithmetic mean of the two

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millilitre, of the standard solutions. determinations, provided that the requirement for
repeatability (see 9.2) is satisfied. Express the result to the

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nearest 1 mg per kilogram of product.

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9 EXPRESSION OF RESULTS 9.2 Repeatability

The difference between the results of two determinations

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9.1 Method of calculation and formula carried out simultaneously or in rapid succession, by the
Calculate the nitrite content of the sample, expressed as same analyst, shall not be greater than IO % of the mean

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milligrams of sodium nitrite per kilogram, using the value.
formula :

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2 000 10 TEST REPORT
NaNO* = c- x-
mxV The test report shall show the method used and the result

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obtained; it shall also mention all operating conditions not
where
specified in this International Standard, or regarded as
m is the mass, in grams, of the test portion; ed
optional, as well as any circumstances
influenced the result.
that may have

V is the volume, in mil ilitres, of the aliquot portion of

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the filtrate (see 8.4.1 taken for the photometric The report shall include all details necessa ry for camp
determination; identification of the sample.
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CD/K/725:2010

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