1、BS EN ISO 12228-1:2014Incorporating corrigenda January 2015 and March 2017BS EN ISO 12228-1:2014Determination of individualand total sterols contents Gas chromatographic methodPart 1: Animal and vegetable fats and oilsIncorporating corrigendum January 2015(ISO 12228-1:2014)BSI Standards PublicationW
2、B11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06BS EN ISO 12228-1:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO 12228-1:2014. Together with BS EN ISO 12228-2:2014 it supersedes BS EN ISO 12228:1999, which is withdrawn.The UK participation in it
3、s preparation was entrusted to Technical Committee AW/307, Oilseeds, animal and vegetable fats and oils and their by-products.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of
4、 a contract. Users are responsible for its correct application. The British Standards Institution 2017. Published by BSI Standards Limited 2017ISBN 978 0 580 97675 9ICS 67.200.10Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under
5、 the authority of the Standards Policy and Strategy Committee on 31 July 2014.Amendments/corrigenda issued since publicationDate Text affected31 January 2017 Supersession statement in National foreword updated31 March 2017 Implementation of ISO corrected text 15 May 2015: See ISO foreword for detail
6、sEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 12228-1 July 2014 ICS 67.200.10 Supersedes EN ISO 12228:1999English Version Determination of individual and total sterols contents - Gas chromatographic method - Part 1: Animal and vegetable fats and oils (ISO 12228-1:2014) Dtermination de la
7、 teneur en strols individuels et totaux -Mthode par chromatographie en phase gazeuse - Partie 1: Corps gras dorigines animale et vgtale (ISO 12228-1:2014) Bestimmung der individuellen und der Gesamtsterine - Gaschromatographisches Verfahren - Teil 1: Tierische und pflanzliche Fette und le (ISO 12228
8、-1:2014) This European Standard was approved by CEN on 14 June 2014. 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
9、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
10、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
11、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
12、EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 12228-1:2014 EBS EN ISO 12228-1:2014EN ISO 12228-1:2014 (E) 3 Foreword This do
13、cument (EN ISO 12228-1:2014) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNOR. This
14、 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 January 2015, and conflicting national standards shall be withdrawn at the latest by January 2015. Attention is drawn to the possibility that some of th
15、e 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. This document supersedes EN ISO 12228:1999. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
16、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, Netherlan
17、ds, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12228-1:2014 has been approved by CEN as EN ISO 12228-1:2014 without any modification. BS EN ISO 12228-1:2014EN ISO 12228-1:2014 (E) 3 Foreword Thi
18、s document (EN ISO 12228-1:2014) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal f s and oils and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNOR. T
19、his 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 January 2015, and conflicting national standards shall be withdrawn at the latest by January 2015. Attention is drawn to the possibility that some of
20、 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. This document supersedes EN ISO 12228:1999. According to the CEN-CENELEC Internal Regulations, the national standards organizations of t
21、he 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, Nether
22、lands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12228-1:2014 has been approved by CEN as EN ISO 12228-1:2014 without any modification. BS EN ISO 12228-1:2014EN ISO 12228-1:2014 (E) 3 Foreword
23、This document (EN ISO 12228-1:2014) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oi s and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNO
24、R. 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 January 2015, and conflicting national standards shall be ithdrawn at the latest by January 2015. Attention is drawn to the possibility that some
25、 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. This document supersedes EN ISO 12228:1999. According to the CEN-CENELEC Internal Regulations, the national standards organizations o
26、f 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, Net
27、herlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12228-1:2014 has been approved by CEN as EN ISO 12228-1:2014 without any modification. ISO 12228-1:2014(E)Foreword iv1 Scope . 12 Normative r
28、eferences 13 Terms and definitions . 14 Principle 15 Reagents 26 Apparatus . 37 Sample . 37.1 Sampling . 37.2 Preparation of the test sample 38 Procedure. 48.1 Preparation of the aluminium oxide column 48.2 Test portion 48.3 Extraction of the unsaponifiable matter . 48.4 Thin-layer chromatography .
29、48.5 Isolation of the sterols 48.6 Preparation of sterol trimethylsilyl ethers . 58.7 Gas chromatography 59 Expression of results 59.1 Identification of sterols 59.2 Composition of sterols 59.3 Determination of the total sterol content 610 Precision . 710.1 Interlaboratory test. 710.2 Repeatability
30、limit, r . 710.3 Reproducibility limit, R 711 Test report . 7Annex A (informative) Figures 8Annex B (informative) Interlaboratory trial .15Bibliography .24 ISO 2014 All rights reserved iiiContents PageBS EN ISO 12228-1:2014ISO 12228-1:2014(E)ForewordISO (the International Organization for Standardiz
31、ation) 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
32、 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.The procedures used to
33、 develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of th
34、e ISO/IEC Directives, Part 2 (see www.iso.org/directives).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. Details of any patent rights identified dur
35、ing the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the mea
36、ning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 34,
37、 Food products, Subcommittee SC 11, Animal and vegetable fats and oils.This first edition of ISO 12228-1, together with ISO 12228-2, cancels and replaces ISO 12228:1999, which has been technically revised.ISO 12228 consists of the following parts, under the general title Determination of individual
38、and total sterols content Gas chromatographic method: Part 1: Animal and vegetable fats and oils Part 2: Olive oils and olive pomace oilsThis corrected version of ISO 12228-1:2014 incorporates the following corrections: In Annex B, the data in the tables have been revised and a new table for -Sitost
39、erol (Table B.11) has been added.iv ISO 2014 All rights reservedBS EN ISO 12228-1:2014Determination of individual and total sterols contents Gas chromatographic method Part 1: Animal and vegetable fats and oils1 ScopeThis part of ISO 12228 specifies a procedure for the gas chromatographic determinat
40、ion of the content and composition of sterols in animal and vegetable fats and oils. However, the determination of the contents and composition of sterols in olive and olive pomace oils is to be carried out using ISO 12228-2.2 Normative referencesThe following documents, in whole or in part, are nor
41、matively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 661, Animal and vegetable fats and oils Preparation of te
42、st sampleISO 3696, Water for analytical laboratory use Specification and test methods3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1composition of sterolscomposition of individual sterols in the sample, beginning with cholesterol and ending wit
43、h 7-avenasterol (see Table 1) under the conditions specified in this part of ISO 12228Note 1 to entry: The composition is expressed as a percentage of all peak areas, normalized to 100 %.3.2total sterol contentmass fraction of the sum of all individual sterols, as determined in accordance with the m
44、ethod specified in this part of ISO 12228, beginning with cholesterol and ending with 7-avenasterol (see Table 1), divided by the mass of the test portionNote 1 to entry: The content is expressed in milligrams per kilogram.4 PrincipleA test portion is saponified by boiling under reflux with an ethan
45、olic potassium hydroxide solution. The unsaponifiable matter is isolated by solid-phase extraction on an aluminium oxide column. The aluminium oxide column is used to retain the fatty acid anions; sterols pass through the column. The sterol fraction from the unsaponifiable matter is separated by thi
46、n-layer chromatography. The qualitative and quantitative compositions of the sterol fraction are determined by gas chromatography using cholestanol or betulin as the internal standard.INTERNATIONAL STANDARD ISO 12228-1:2014(E) ISO 2014 All rights reserved 1BS EN ISO 12228-1:2014ISO 12228-1:2014(E)5
47、ReagentsWARNING Attention is drawn to the regulations which specify the handling of hazardous substances. Technical, organizational, and personal safety measures shall be followed.Use only reagents of recognized analytical grade, unless otherwise stated, and water complying with grade 3 of ISO 36961
48、.5.1 Potassium hydroxide (KOH), ethanolic solution, molar concentration c(KOH) approximately 0,5 mol/l.Dissolve 3 g of potassium hydroxide in 5 ml of water and dilute to 100 ml with ethanol (5.3). The solution should be colourless or straw-coloured.5.2 Internal standard solution, cholestanol (5-chol
49、estan-3-ol) or betulin, volume fraction of 1,0 mg/ml solution in ethanol (see note to 5.10).NOTE In case of hydrogenated oils, which may contain cholestanol, the use of betulin (peak 17 in Table 1) is recommended.5.3 Ethanol, of minimum volume fraction = 95 %.5.4 Aluminium oxide, neutral, particle size 0,063 mm to 0,200 mm, activity grade I (water content = 0 %).5.5 Diethyl ether, freshly distilled, free from peroxides and residue.WARNING Diethyl ether is highly flammable and can form explosive peroxides. Explosive limits in air are
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