EN ISO 17180:2013

SLOVENSKI STANDARD
01-junij-2013
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Animal feeding stuffs - Determination of lysine, methionine and threonine in commercial
amino acid products and premixtures (ISO 17180:2013)
Futtermittel - Bestimmung von Lysin, Methionin und Threonin in handelsüblichen
aminosäurehaltigen Produkten und Vormischungen (ISO 17180:2013)
Aliments des animaux - Détermination de la teneur en lysine, méthionine et thréonine
dans les acides aminés industriels et les prémix (ISO 17180:2013)
Ta slovenski standard je istoveten z: EN ISO 17180:2013
ICS:
65.120 Krmila Animal feeding stuffs
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 17180
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2013
ICS 65.120
English Version
Animal feeding stuffs - Determination of lysine, methionine and
threonine in commercial amino acid products and premixtures
(ISO 17180:2013)
Aliments des animaux - Détermination de la teneur en Futtermittel - Bestimmung von Lysin, Methionin und
lysine, méthionine et thréonine dans les acides aminés Threonin in handelsüblichen aminosäurehaltigen Produkten
industriels et les pré-mélanges (ISO 17180:2013) und Vormischungen (ISO 17180:2013)
This European Standard was approved by CEN on 21 March 2013.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 17180:2013: E
worldwide for CEN national Members.

Contents Page
Foreword .3
Foreword
This document (EN ISO 17180:2013) has been prepared by Technical Committee ISO/TC 34 "Food products"
in collaboration with Technical Committee CEN/TC 327 “Animal feeding stuffs - Methods of sampling and
analysis” the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by October 2013, and conflicting national standards shall be withdrawn at
the latest by October 2013.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 17180:2013 has been approved by CEN as EN ISO 17180:2013 without any modification.

INTERNATIONAL ISO
STANDARD 17180
First edition
2013-04-15
Animal feeding stuffs —
Determination of lysine, methionine
and threonine in commercial amino
acid products and premixtures
Aliments des animaux — Détermination de la teneur en lysine,
méthionine et thréonine dans les acides aminés industriels et les pré-
mélanges
Reference number
ISO 17180:2013(E)
©
ISO 2013
ISO 17180:2013(E)
© ISO 2013
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
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Published in Switzerland
ii © ISO 2013 – All rights reserved

ISO 17180:2013(E)
Contents Page
Foreword .iv
1 Scope . 1
2 Principle . 1
3 Reagents and materials . 1
4 Apparatus . 3
5 Procedure. 4
5.1 Preparation of samples . 4
5.2 Number of prepared extracts . 4
5.3 Extraction using dilution by weighing . 4
5.4 Extraction using volumetric dilution . 5
5.5 Chromatography . 5
6 Calculation of results . 5
6.1 Principle of the method and control of the calibration . 5
6.2 Calculation if dilution by weighing is applied . 6
6.3 Calculation if volumetric dilution is applied . 6
7 Precision . 7
7.1 Interlaboratory test. 7
7.2 Repeatability . 7
7.3 Reproducibility . 8
8 Test report . 8
Annex A (informative) Data gathered from collaborative study . 9
Bibliography .11
ISO 17180:2013(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 17180 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 10, Animal
feeding stuffs.
iv © ISO 2013 – All rights reserved

INTERNATIONAL STANDARD ISO 17180:2013(E)
Animal feeding stuffs — Determination of lysine,
methionine and threonine in commercial amino acid
products and premixtures
WARNING — This document can involve the use of hazardous materials, operations, and
equipment. This document does not purport to address all the safety risks associated with its use.
It is the responsibility of the user of this document to establish appropriate safety and healthy
practices and determine the applicability of local regulatory limitations prior to use.
1 Scope
This International Standard specifies a method for the quantitative determination of free (non-protein-
bound) lysine, methionine, and threonine in commercial products and premixtures containing more than
about 10 % mass fraction of the respective amino acid. It does not distinguish between d- and l-forms.
NOTE For the purposes of this International Standard, the term “amino acids” used in Clause 2 onwards
refers to lysine, methionine, and threonine.
2 Principle
The samples are treated in dilute hydrochloric acid and then diluted with sodium citrate buffer. Norleucine
internal standard is added and the amino acids are separated by an amino acid analyser or high performance
liquid chromatography (HPLC), using a cation exchange resin and sodium citrate buffer eluent solutions.
The amino acids are measured colourimetrically following post-column reaction with ninhydrin reagent
or by fluorescence detection after post-column reaction with ortho-phthaldialdehyde (OPA).
3 Reagents and materials
Use only reagents of recognized analytical grade, unless otherwise specified.
3.1 Water, double distilled water or equivalent purity (conductivity <10 µS/cm).
3.2 Standard substances.
3.2.1 Lysine·HCl crystals, purity superior to 99 % mass fraction dried under vacuum in a desiccator
for 2 days over P O prior to use.
2 5
3.2.2 Threonine crystals, purity superior to 99 % mass fraction dried under vacuum in a desiccator
for 2 days over P O prior to use.
2 5
3.2.3 Methionine crystals, purity superior to 99 % mass fraction dried under vacuum in a desiccator
for 2 days over P O prior to use.
2 5
3.3 Norleucine crystals, for use as internal standard, purity superior to 99 % dried under vacuum in a
desiccator for 2 days over P O prior to use.
2 5
3.4 Sodium hydroxide solution, c(NaOH) = 7,5 mol/l, for pH adjustment of sodium citrate buffer.
Carefully dissolve 300 g sodium hydroxide in water (3.1) and make up to 1 l.
ISO 17180:2013(E)
3.5 Hydrochloric acid, γ(HCl) = 370 g/kg.
3.6 Sodi
...