1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN ISO 17180:2013Animal feeding stuffs Determination of lysine,methionine and threoninein commercial amino acidproducts and premixtures (ISO17180:2013)BS EN ISO 17180:2013 BRI
2、TISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO17180:2013.The UK participation in its preparation was entrusted to TechnicalCommittee AW/10, Animal feeding stuffs.A list of organizations represented on this committee can beobtained on request to its secretary.T
3、his publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2013. Published by BSI StandardsLimited 2013ISBN 978 0 580 76077 8ICS 65.120Compliance with a British Standard cannot confer immu
4、nity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 30 April 2013.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 17180 April 2013 ICS 65.120 English Version
5、Animal feeding stuffs - Determination of lysine, methionine and threonine in commercial amino acid products and premixtures (ISO 17180:2013) Aliments des animaux - Dtermination de la teneur en lysine, mthionine et thronine dans les acides amins industriels et les pr-mlanges (ISO 17180:2013) Futtermi
6、ttel - Bestimmung von Lysin, Methionin und Threonin in handelsblichen aminosurehaltigen Produkten 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 f
7、or 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 thr
8、ee 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 bo
9、dies 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,
10、Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN
11、 national Members. Ref. No. EN ISO 17180:2013: EBS EN ISO 17180:2013EN ISO 17180:2013 (E) 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
12、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
13、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 organi
14、zations 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, M
15、alta, 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. BS EN ISO 17180:2013ISO 17180:2013(E) ISO 2013 Al
16、l rights reserved iiiContents PageForeword iv1 Scope . 12 Principle 13 Reagents and materials . 14 Apparatus . 35 Procedure. 45.1 Preparation of samples 45.2 Number of prepared extracts . 45.3 Extraction using dilution by weighing 45.4 Extraction using volumetric dilution . 55.5 Chromatography 56 Ca
17、lculation of results . 56.1 Principle of the method and control of the calibration 56.2 Calculation if dilution by weighing is applied 66.3 Calculation if volumetric dilution is applied . 67 Precision . 77.1 Interlaboratory test. 77.2 Repeatability . 77.3 Reproducibility . 88 Test report . 8Annex A
18、(informative) Data gathered from collaborative study 9Bibliography .11BS EN ISO 17180:2013ISO 17180:2013(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is nor
19、mally 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
20、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
21、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 o
22、f 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 reservedBS EN
23、ISO 17180:2013INTERNATIONAL STANDARD ISO 17180:2013(E)Animal feeding stuffs Determination of lysine, methionine and threonine in commercial amino acid products and premixturesWARNING This document can involve the use of hazardous materials, operations, and equipment. This document does not purport t
24、o 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 ScopeThis International Standard specifies a method for t
25、he 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 St
26、andard, the term “amino acids” used in Clause 2 onwards refers to lysine, methionine, and threonine.2 PrincipleThe 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 ana
27、lyser 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-ph
28、thaldialdehyde (OPA).3 Reagents and materialsUse only reagents of recognized analytical grade, unless otherwise specified.3.1 Water, double distilled water or equivalent purity (conductivity 1,5 %), the error is due to the chromatography. Check the analyser and repeat all affected measurements.7.3 R
29、eproducibilityThe absolute difference between two test results (averages of two determinations) obtained using the same method on identical test material in different laboratories with different operators using different equipment will be, in 95 % of cases, less than the reproducibility limit R give
30、n in Annex A.8 Test reportThe test report shall contain at least the following information:a) all information necessary for the complete identification of the sample;b) the sampling method used, if known;c) the test method used, with reference to this International Standard (ISO 17180:2013);d) all o
31、perating details not specified in this International Standard, or regarded as optional, together with details of any incidents which may have influenced the test result(s);e) the test result(s) obtained;f) if the repeatability has been checked, the final quoted result obtained.8 ISO 2013 All rights
32、reservedBS EN ISO 17180:2013ISO 17180:2013(E)Annex A (informative) Data gathered from collaborative studyA total of 17 laboratories participated in a collaborative study for determination of lysine, methionine, and threonine in trade products or concentrated amino acid premixes. Analysed in duplicat
33、e were 13 samples, four pure amino acids and six premixes including three Youden matched pairs.See Table A.1. ISO 2013 All rights reserved 9BS EN ISO 17180:2013ISO 17180:2013(E)Table A.1Trade product or premixAmino acidaExpected value %No. of labsMean %Standard devia-tion of repeata- bility srCV of
34、repeata- bility CV,r%Standard deviation of repro-duci- bility sRCV of reproduci- bility CV,R%Repeata- bility limit r = 2,8SrReproduci- bility limit R = 2,8SRHorratN1 Biolys Lys 17 45,89 0,39 0,84 1,06 2,31 1,08 2,96 1,03N2 premixMet 27,00 16 26,55 0,41 1,56 0,49 1,85 1,16 1,37 0,76Thr 15,00 16 14,85
35、 0,19 1,29 0,35 2,33 0,54 0,97 0,88N3 premixLys 20,80 17 20,56 0,27 1,33 0,51 2,49 0,77 1,44 0,98Met 11,00 17 11,26 0,14 1,24 0,21 1,84 0,39 0,58 0,66Thr 8,00 17 8,22 0,05 0,65 0,16 1,9 0,15 0,44 0,65N4 premixLys 16,80 17 16,63 0,22 1,29 0,39 2,32 0,6 1,08 0,89Met 14,00 17 14,21 0,13 0,93 0,27 1,92
36、0,37 0,76 0,72Thr 11,00 17 11,19 0,09 0,82 0,25 2,22 0,26 0,7 0,8N5 premixLys 12,80 17 12,58 0,09 0,74 0,23 1,86 0,26 0,65 0,68Met 32,00 17 32,04 0,16 0,5 0,83 2,59 0,45 2,32 1,09Thr 22,00 17 22,1 0,18 0,81 0,43 1,94 0,5 1,2 0,77N6 premixLys 12,29 17 12,3 0,21 1,68 0,26 2,13 0,58 0,73 0,78Met 30,72
37、17 30,59 0,32 1,06 0,76 2,5 0,9 2,14 1,05Thr 21,12 17 21,25 0,25 1,18 0,4 1,87 0,7 1,11 0,74N7 premixLys 10,40 17 10,27 0,12 1,21 0,19 1,81 0,35 0,52 0,64Met 9,00 17 9,1 0,08 0,92 0,13 1,48 0,23 0,38 0,52Thr 14,00 17 14,03 0,1 0,7 0,29 2,07 0,27 0,81 0,77N8 premixLys 10,24 17 10,12 0,12 1,21 0,15 1,
38、5 0,34 0,42 0,53Met 8,86 17 8,91 0,08 1,17 0,17 1,89 0,29 0,47 0,66Thr 13,79 17 13,83 0,19 1,35 0,31 2,25 0,52 0,87 0,84N9 premixLys 24,00 17 23,48 0,26 1,13 0,5 2,15 0,74 1,41 0,86Met 19,00 17 19,09 0,19 0,99 0,33 1,73 0,53 0,93 0,67Thr 15,00 17 15,09 0,15 1 0,3 1,99 0,42 0,84 0,75N10 pre-mixLys 23
39、,28 17 22,85 0,19 0,84 0,41 1,78 0,54 1,14 0,71Met 18,43 17 18,65 0,13 0,68 0,29 1,53 0,36 0,8 0,59Thr 14,55 17 14,65 0,14 0,96 0,31 2,15 0,4 0,88 0,81N11 l-LysineHClLys 76,04 17 74,11 0,69 0,93 1,32 1,78 1,93 3,7 0,85N12 dl-MethionineMet 93,55 17 93,27 0,79 0,85 1,42 1,52 2,22 3,98 0,75N13 l-Threon
40、ineThr 95,95 17 95,46 1,11 1,17 2,07 2,16 3,12 5,78 1,07aLys lysine; Met methionine; Thr threonine10 ISO 2013 All rights reservedBS EN ISO 17180:2013ISO 17180:2013(E)Bibliography1 AOAC Official Method 999.13, Lysine, methionine, and threonine in feed grade amino acids and premixes2 Fontaine J., Euda
41、imon M., Liquid chromatographic determination of lysine, methionine, and threonine in pure amino acids (feed grade) and premixes: Collaborative study. J. AOAC Int. 2000, 83 pp. 771783 ISO 2013 All rights reserved 11This page deliberately left blankThis page deliberately left blankBSI is the independ
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