DIN EN ISO 17180-2013 Animal feeding stuffs - Determination of lysine methionine and threonine in commercial amino acid products and premixtures (ISO 17180 2013) German version EN .pdf

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1、August 2013 Translation by DIN-Sprachendienst.English price group 10No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS

2、 65.120!%(2“2052915www.din.deDDIN EN ISO 17180Animal feeding stuffs Determination of lysine, methionine and threonine in commercial aminoacid products and premixtures (ISO 17180:2013);English version EN ISO 17180:2013,English translation of DIN EN ISO 17180:2013-08Futtermittel Bestimmung von Lysin,

3、Methionin und Threonin in handelsblichen aminosurehaltigenProdukten und Vormischungen (ISO 17180:2013);Englische Fassung EN ISO 17180:2013,Englische bersetzung von DIN EN ISO 17180:2013-08Aliments des animaux Dtermination de la teneur en lysine, mthionine et thronine dans les acides aminsindustriels

4、 et les pr-mlanges (ISO 17180:2013);Version anglaise EN ISO 17180:2013,Traduction anglaise de DIN EN ISO 17180:2013-08www.beuth.deDocument comprises pagesIn case of doubt, the German-language original shall be considered authoritative.1608.13 DIN EN ISO 17180:2013-08 2 A comma is used as the decimal

5、 marker. National foreword This document (EN ISO 17180:2013) has been prepared by Technical Committee ISO/TC 34 “Food products” (Secretariat: AFNOR, France), Subcommittee SC 10 “Animal Feeding Stuffs” (Secretariat: ISIRI, Iran). Based on the results of parallel voting, Technical Committee CEN/TC 327

6、 “Animal feeding stuffs Methods of sampling and analysis” (Secretariat: NEN, Netherlands) decided to adopt ISO 17180:2013 as a European Standard. The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Produ

7、cts Standards Committee), Working Committee NA 057-03-03 AA Futtermittel. EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 17180 April 2013 ICS 65.120 English Version Animal feeding stuffs - Determination of lysine, methionine and threonine in commercial amino acid products and premixtures (

8、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) Futtermittel - Bestimmung von Lysin, Methionin und Threonin in handelsblichen aminosurehaltigen Produkten und Vormischungen (ISO 1718

9、0:2013) 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 bibliographic

10、al 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

11、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 M

12、acedonia, 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 COMIT EUROPEN DE NORMALISATION

13、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 national Members. Ref. No. EN ISO 17180:2013: EContents Page DIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 2 Foreword 3 1 S

14、cope 4 2 Principle 4 3 Reagents and materials 4 4 Apparatus .6 5 Procedure .7 5.1 Preparation of samples .7 5.2 Number of prepared extracts 7 5.3 Extraction using dilution by weighing .7 5.4 Extraction using volumetric dilution .8 5.5 Chromatography 8 6 Calculation of results 8 6.1 Principle of the

15、method and control of the calibration 8 6.2 Calculation if dilution by weighing is applied .9 6.3 Calculation if volumetric dilution is applied .9 7 Precision .10 7.1 Interlaboratory test 10 7.2 Repeatability .10 7.3 Reproducibility .11 8 Test report 11 Annex A (informative) Data gathered from colla

16、borative study .12 Bibliography 14 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. ” Th

17、is 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

18、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

19、 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, Slo

20、vakia, 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. “DIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 3 WARNING This document can involve the use of hazardous m

21、aterials, 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 pri

22、or to use.1 ScopeThis 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

23、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 PrincipleThe samples are treated in dilute hydrochloric acid and then diluted with sodium citrate buffer. Norleucine internal sta

24、ndard 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

25、or by fluorescence detection after post-column reaction with ortho-phthaldialdehyde (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 chromato

26、graphy. Check the analyser and repeat all affected measurements.7.3 ReproducibilityThe 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

27、will be, in 95 % of cases, less than the reproducibility limit R given 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, wi

28、th reference to this International Standard (ISO 17180:2013);d) all operating 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

29、been checked, the final quoted result obtained.DIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 11 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 co

30、ncentrated amino acid premixes. Analysed in duplicate were 13 samples, four pure amino acids and six premixes including three Youden matched pairs.See Table A.1.DIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 12 Table A.1Trade product or premixAmino acidaExpected value %No. of labsMean %Standard devi

31、a-tion of repeata- bility srCV of 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 1,8

32、5 1,16 1,37 0,76Thr 15,00 16 14,85 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 1

33、4,00 17 14,21 0,13 0,93 0,27 1,92 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

34、0,26 2,13 0,58 0,73 0,78Met 30,72 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 premixLy

35、s 10,24 17 10,12 0,12 1,21 0,15 1,5 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,

36、99 0,42 0,84 0,75N10 pre-mixLys 23,28 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,

37、42 1,52 2,22 3,98 0,75N13 l-ThreonineThr 95,95 17 95,46 1,11 1,17 2,07 2,16 3,12 5,78 1,07aLys lysine; Met methionine; Thr threonineDIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 13 Bibliography1 AOAC Official Method 999.13, Lysine, methionine, and threonine in feed grade amino acids and premixes2 Fontaine J., Eudaimon 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. 771783DIN EN ISO 17180:2013-08 EN ISO 17180:2013 (E) 14