1、BRITISH STANDAR BS EN ISO 23275-1:2008Animal and vegetable fats and oils Cocoa butter equivalents in cocoa butter and plain chocolate Part 1: Determination of the presence of cocoa butter equivalentsICS 67.190; 67.200.10nullnull nullnullnullnullnullnullnull nullnullnullnullnullnullnull nullnullnull
2、nullnullnullnullnullnullnullnullnullnull nullnullnullnullnullnull nullnull nullnullnullnullnullnullnullnullnull nullnull nullnullnullnullnullnullnullnullnull nullnullnullLicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSINational forewordThis British S
3、tandard is the UK implementation of EN ISO 23275-1:2008. It is identical with ISO 23275-1:2006. It supersedes BS ISO 23275-1:2006, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee AW/307, Oil seeds, animal and vegetable fats and oils and their by produc
4、ts.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 a contract. Users are responsible for its correct application.Compliance with a British Standard cannot confer immunity fr
5、om legal obligations.BS EN ISO 23275-1:2008This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 November 2006 BSI 2009ISBN 978 0 580 63597 7Amendments/corrigenda issued since publicationDate Comments 30 June 2009 This corrigendum renumbers BS I
6、SO 23275-1:2006 as BS EN ISO 23275-1:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSIEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 23275-1November 2008ICS 67.200.10; 67.190English VersionAnimal and vegetable fats and oils - Cocoa butter
7、equivalents incocoa butter and plain chocolate - Part 1: Determination of thepresence of cocoa butter equivalents (ISO 23275-1:2006)Corps gras dorigines animale et vgtale - quivalents aubeurre de cacao dans le beurre de cacao et dans lechocolat de mnage - Partie 1: Dtermination de laprsence dquivale
8、nts au beurre de cacao (ISO 23275-1:2006)Tierische und pflanzliche Fette und le - Kakaobutter-quivalente in Kakaobutter und Zartbitterschokolade - Teil1: Bestimmung der Prsenz von Kakaobutter-quivalenten(ISO 23275-1:2006)This European Standard was approved by CEN on 23 October 2008.CEN members are b
9、ound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the C
10、EN 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 translationunder the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same statu
11、s as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slo
12、vakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2008 CEN All rights of exploitation in any form and by any means reservedworldwide f
13、or CEN national Members.Ref. No. EN ISO 23275-1:2008: ELicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSIEN ISO 23275-1:2008 (E) ii Foreword The text of ISO 23275-1:2006 has been prepared by Technical Committee ISO/TC 34 “Agricultural food products” o
14、f the International Organization for Standardization (ISO) and has been taken over as EN ISO 23275-1:2008 by 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 European
15、 Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 2009, and conflicting national standards shall be withdrawn at the latest by May 2009. Attention is drawn to the possibility that some of the elements of thi
16、s 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 Stan
17、dard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom.
18、 Endorsement notice The text of ISO 23275-1:2006 has been approved by CEN as a EN ISO 23275-1:2008 without any modification. BS EN ISO 23275-1:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSIiiiContentsPageIntroduction v1Scope12Terms and definit
19、ions .13Principle14Reagents and materials15Apparatus.26Sampling.27Preparation of test sample37.1 Preparation of cocoa butter CRM for calibration purposes and system suitability check37.2 Preparation of chocolate sample.38Procedure.38.1Fat extraction .38.2 Separation of individual triacylglycerols by
20、 HR-GC38.3Identification.39Calculation49.1Determination of response factors49.2 Calculation of percentages of triacylglycerols.49.3 Decision if sample is pure cocoa butter410 Procedural requirements510.1 General considerations.510.2 System suitability511 Precision.511.1 Interlaboratory test511.2 Rep
21、eatability.611.3 Reproducibility.612 Test report6Annex A (informative) Results of interlaboratory test.7Bibliography12BS ISO 23275-1:2006iEN ISO 23275-1:2008 (E) BS EN ISO 23275-1:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSIIntroduction “Coc
22、oa butter equivalents” is the general term forfats used to replace cocoa butter in chocolate. Theyresemble the chemical composition and physic al properties of cocoa butter very close ly, making themtherefore extremelydifficult to quantify and even in some cases to detect. In principle, cocoa butter
23、equivalents must by definition be fats low in lauric acid, rich in symmetrical mono-unsaturated triacylglycerolsof the type 1,3-dipalmitoyl-2-oleoylglycerol, 1-palmitoyl-2-oleo yl-3-stearoylglycerol and 1,3-distearoyl-2-oleoylglycerol, miscible with cocoa butter, and obtained onlyby refining and fra
24、ctionation. Within the European Union, the following vegetable fats, obtained from the plants listed below, maybe used singlyor in blends, according to Directive 2000/36/EC 1: illip, Borneo tallow or tengkawang ( Shorea spp . ), palm oil ( Elaeis guineensis, Elaeis olifera ), sal( Shorea robusta), s
25、hea ( Butyrospermum parkii), kokum gurgi ( Garcinia indica ), and mango kernel ( Mangifera indica ). This part of ISO23275 specifies a procedure for the detection of these fats (restrictions are only made forpure illip fat samples) in cocoa butter and plain chocolate. ISO 23275-2 specifies a procedu
26、re allowing areliable quantification of these fats at the level of 5 %, complying with the statutorylimit laid down in Directive2000/36/EC 1 of the European Parliament and the Council. To facilitate the usage of both parts of ISO23275, an analytical toolbox named “CoCal-1” has beenestablished. “CoCa
27、l-1” contains the validated methods for detection (part 1) and quantification (part 2) ofCBEs in plain chocolate, and also a certified cocoa butter reference material(IRMM-801) to calibrate theanalysts instruments and an electronic evaluation sheet for Microsoft Excel to calculate the final result.
28、Ananalyst working on CBE detection and quantification has only to calibrate the gas chromatographic separation system using IRMM-801, separate the triglyceride fractions of the sample in question, and use the electronicevaluation sheet for subsequent data treatment to detect and quantify CBEs. Infor
29、mation on “CoCal-1” isavailable on thewebsite of the Institute for Reference Materials and Measurements: http:/www.irmm.jrc.be. EN ISO 23275-1:2008 (E) ivBS EN ISO 23275-1:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSI1Animal and vegetable fat
30、s and oils Cocoa butter equivalents in cocoa butter and plain chocolate Part 1: Determination of the presence of cocoa butter equivalents 1 Scope This part of ISO23275 specifies a procedure for the detection of cocoa butter equivalents (CBEs) in cocoa butter (CB)and plain chocolate by high-resolutio
31、n capillarygas liquid chromatography(HR-GC)of triacylglycerols and subsequent data evaluation by regression analysis. The method is applicable for the detection of2 % CBE admixture to cocoa butter,corresponding to about 0,6 % CBE in chocolate (i.e. the assumed fat content of chocolate is 30 %). 2 Te
32、rms and definitions For the purposes of this document, the following terms and definitions apply.2.1 cocoa butter equivalentsCBEs fats detected in cocoa butter and plain chocolate according the procedure specified in this part of ISO 23275 NOTE The result is expressed as a qualitative result, i.e. C
33、BEs present/CBEs not present (YES/NO). 3 Principle Cocoa butter, or the fat obtained by solvent extraction from plain chocolate, is separated by HR-GC into triacylglycerol fractions according to their molecular mass and degree of unsaturation. The presence of CBEsis detected bylinear regression anal
34、ysis applied to individual triacylglycerol fractions of the fat analysed. 4 Reagentsand materials Use only reagents of recognized analytical grade, unless otherwise stated. WARNING Attention is drawn to the regulations which specify the handling of dangerous matter.Technical, organizational and pers
35、onal safety measures shall be followed. 4.1 Cocoa butter Certified Reference Material (CRM) IRMM-801 2, for calibration purposes and systemsuitability check.4.2 Fat solvent , non-chlorinated solvents (e.g. diethyl ether, n -heptane, iso-octane). EN ISO 23275-1:2008 (E) BS EN ISO 23275-1:2008Licensed
36、 Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSI2 5 Apparatus 5.1 Analyticalbalance,with a readability of 0,1 mg. 5.2 Drying oven, maintained at 55 C. A dry heater blockmaybe used. 5.3 Food grater, i.e. a kitchen blender with a design featuring the motor ab
37、ove the receiving container toavoid melting the samples1). 5.4 Rotary evaporator. Alternative evaporation procedures maybe used. 5.5 Pipettes , of capacity 1 ml.5.6 Volumetric flasks , ofcapacity 20 ml. 5.7 Microsyringe, with maximum volume 10 l, graduated to 0,1 l, or automatic sampler. 5.8 Gas chr
38、omatograph (GC), fitted with a cold on-column injection system and a flame ionization detector(FID). Alternative injection systems e.g. a split injector, a programmed-temperature vaporizer (PTV) ora moving-needle injector may be used provided the same results are obtained as indicated in 1 0.2. The
39、separation and detection have been found to be satisfactory if the following experimental conditions are followed: GC column: 25 m to 30 m length, with 0,25 mm i.d., fused silica coated with thermostable 50 %phenylmethylpolysiloxane to a film thickness of 0,1 m to 0,15 m. temperature programme: 100
40、C (initial temperature), programme rate 30 C/min to 340 C (final temperature). carrier gas: helium or hydrogen (purity W 99,999 %). NOTE Suitable columns and alternative experimental conditions, used in an international collaborative study, arelisted in Annex A. Operating conditions may be changed t
41、o obtain optimumseparation of cocoa butter triacylglycerols. 5.9 Chromatographic datasystem. 6 Sampling A representative sample should have been sent to the laboratory. It should not have been damaged orchanged during transport or storage. Sampling is not part of the method specified in this part of
42、 ISO23275. A recommended sampling method isgiven in ISO5555. 1) Philips HR2833 is an example of suitable equipment available commercially. This information is given for the convenience of the users of this part of ISO23275 and does not constitute an endorsement by ISO of this product. EN ISO 23275-1
43、:2008 (E) BS EN ISO 23275-1:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 02/11/2009 06:58, Uncontrolled Copy, (c) BSI37 Preparation of test sample 7.1Preparation of cocoa butter CRM for calibration purposes and system suitability check Before opening and using the cocoa butter CRM (4.1
44、), the ampoule shall be warmed in a drying oven (5.2)until the contents have melted. When a clear so lutionis obtained, mix the contents by repeated inversion fornot less than 20 s. Then open and transfer the contents to a clean vial, which can be tightly sealed and preserved in a cool place for fut
45、ure usage. 7.2Preparation of chocolate sample Chill approx. 200 g of chocolate until hard, and grate to fine granular condition using a mechanica l device (5.3).Mix thoroughly and preserve in tightly stoppered bottle in a cool place. 8 Procedure 8.1 Fat extraction Separate the fat from 10 g to 40 g
46、of grated chocolate (as described in 7.2) byextracting with two or three 100 mlportionsofafat solvent (4.2). Centrifuge and decant. Combine the extracts and evaporate most of the fat solvent (5.4) then finallydry it under a stream of nitrogen. Alternative extraction procedures may be used (e.g. by S
47、oxhlet, by supercritical carbon dioxide or by using microwaves) provided that the same results are obtained. 8.2Separation of individual triacylglycerols by HR-GC The test samples cocoa butter, fat extracted from chocolate, cocoa butter CRM (4.1) shall be warmed in adrying oven (5.2) until completel
48、y melted. If the liquid sample contains sediment, filter the sample inside theoven to obtain a clear filtrate. Pipettes (or similar equipment) used for transferring the sample during weighingoperations should be brought to a temperature of ca. 55 C in a drying oven (5.2) in order to a void partial f
49、at fractionation. Weigh ca. 0,2 g of test sample in a 20 ml volumetric flask (5.6) and dilute to the mark with a suitable fat solvent (4.2). Pipette (5.5) 1 ml of the resulting solution into another 20 ml volumetric flask and dilute to the mark with the same solvent.Inject 0,5 l to 1,0 l of the final test solution ( fat = 0,5 mg/ml) into the HR-GC system using the cold on-column injection system. Alternative sample amounts and injectors may be used provided that the detection system employed gives a linear response
