1、BS ISO12873:2010ICS 67.200.10NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDOlive oils and olive-pomace oils Determination of waxcontent by capillarygas chromatographyBS EN ISO 12873:2014BS EN ISO 12873:2014ISBN 978 0 580 83970 2Amendments/corrigenda issued sin
2、ce publicationDate Comments30 April 2014 This corrigendum renumbers BS ISO 12873:2010 as BS EN ISO 12873:2014This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 June 2010 The British Standards Institution 2014. Published by BSI Standards Limit
3、ed 2014National forewordThis British Standard is the UK implementation of EN ISO 12873:2014. It is identical to ISO 12873:2010. It supersedes BS ISO 12873:2010, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee AW/307, Oilseeds, animal and vegetable fats
4、 and oils and their by-products.A list of organizations represented on this committee can be obtained on request to its secretary.The publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application.Compliance with a British Standa
5、rd cannot confer immunity from legal obligations.EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 12873 April 2014 ICS 67.200.10 English Version Olive oils and olive-pomace oils - Determination of wax content by capillary gas chromatography (ISO 12873:2010) Huiles dolive et huiles de grignon
6、s dolive - Dtermination de la teneur en cires par chromatographie en phase gazeuse sur colonne capillaire (ISO 12873:2010) Olivenle und Oliventresterle - Bestimmung des Wachsgehaltes in Olivenlen mittels Kapillarsulen-Gaschromatographie (ISO 12873:2010) This European Standard was approved by CEN on
7、11 April 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 bibliographical references concerning such national standards may b
8、e 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
9、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, I
10、reland, 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 EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management
11、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 12873:2014 EBS ISO 12873:2010ISO 12873:2010(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe
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14、has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. COPYRIGHT PROTECTED DOCUMENT ISO 2010 All rights reserved. Unless otherwise specified, no p
15、art of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISOs member body in the country of the requester. ISO copyright office Case postale
16、 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2010 All rights reservedEN ISO 12873:2014 (E) 3 Foreword The text of ISO 12873:2010 has been prepared by Technical Committee ISO/TC 34 “Food products” of the Int
17、ernational Organization for Standardization (ISO) and has been taken over as EN ISO 12873:2014 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 Standard s
18、hall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by October 2014, and conflicting national standards shall be withdrawn at the latest by October 2014. Attention is drawn to the possibility that some of the elements of this d
19、ocument 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 Standar
20、d: 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
21、, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12873:2010 has been approved by CEN as EN ISO 12873:2014 without any modification. BS EN ISO 12873:2014EN ISO 12873:2014 (E)BS ISO 12873:2010ISO 12873:2010(E) ISO 2010 All rights reserved iiiForeword ISO (the In
22、ternational 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 h
23、as 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
24、 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 bodi
25、es 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
26、or all such patent rights. ISO 12873 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils. BS ISO 12873:2010ISO 12873:2010(E) iv ISO 2010 All rights reservedIntroduction As part of the Trade standard applying to olive oils and olive-pom
27、ace oils, the International Olive Oil Council OOC) now known as International Olive Council (IOC) published COI/T.20/Doc. 18:20074. COI/T.20/Doc. 18 was applicable to olive and olive-pomace oils and was used to distinguish between oilsobtained by either pressing or centrifuging and olive-pomace oils
28、. Olive pomace is the residual paste which still contains a variable amount of water and oil after pressing or centrifuging. In 2008, the IOC submitted the document to ISO/TC 34/SC 11 for adoption as an International Standard. BS EN ISO 12873:2014ISO 12873:2010(E)blankBS ISO 12873:2010INTERNATIONAL
29、STANDARD ISO 12873:2010(E) ISO 2010 All rights reserved 1Olive oils and olive-pomace oils Determination of wax content by capillary gas chromatography 1 Scope This International Standard specifies the determination of the wax content, as a mass fraction expressed in milligrams per kilogram, of olive
30、 oils and olive-pomace oils. The individual waxes are separated according to the number of carbon atoms. The method is recommended for distinguishing between olive oil obtained by pressing or centrifuging and that obtained from olive pomace (olive-pomace oil). NOTE This International Standard is bas
31、ed on COI/T.20/Doc. 18/Rev. 2:20074. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments
32、) applies. ISO 661, Animal and vegetable fats and oils Preparation of test sample 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 wax content mass fraction of those substances in a sample determined in accordance with the method specified in
33、this International Standard NOTE The wax content is expressed in milligrams per kilogram. 4 Principle After a suitable internal standard has been added, the oil is fractionated by chromatography on a hydrated silica gel column. The fraction eluted under test conditions (which has a lower polarity th
34、an the triglycerides) is recovered and analysed by capillary gas chromatography. 5 Reagents WARNING Comply with any local regulations which specify the handling of hazardous substances. Technical, organizational and personal safety measures shall be followed. During the analysis, unless otherwise st
35、ated, use only reagents of recognized analytical grade, and distilled or demineralized water or water of equivalent purity. BS EN ISO 12873:2014BS ISO 12873:2010ISO 12873:2010(E) 2 ISO 2010 All rights reserved5.1 Silica gel, with a particle size of 60 m to 200 m, prepared as follows: place the gel i
36、n the muffle oven at 500 C for at least 4 h; allow to cool, then add 2 % of water in relation to the mass of silica gel used; shake well to homogenize the slurry. Store in darkness for at least 12 h prior to use. 5.2 n-Hexane, chromatography grade. 5.3 Diethyl ether, chromatography grade. 5.4 n-Hept
37、ane, chromatography grade. 5.5 Internal standard, solution of lauryl arachidate in hexane, mass concentration 0,1 g/100 ml. NOTE Palmityl palmitate or myristyl stearate may also be used. 5.6 Sudan I (1-phenylazo-2-naphthol). 5.7 Carrier gas: hydrogen or helium, gas chromatography grade. 5.8 Auxiliar
38、y gases: hydrogen, free from moisture and organic substances, and synthetic air, gas chromatography grade. 6 Apparatus Usual laboratory apparatus and, in particular, the following. 6.1 Erlenmeyer flask, 25 ml. 6.2 Glass column for liquid chromatography, of diameter 15,0 mm and length 30 cm to 40 cm,
39、 with stopcock. 6.3 Gas chromatograph, suitable for use with a capillary column, equipped with the components specified in 6.3.1 to 6.3.5. 6.3.1 Cold on-column injector. 6.3.2 Thermostat-controlled oven with temperature programming. 6.3.3 Flame-ionization detector. 6.3.4 Computer-based integration s
40、ystem. 6.3.5 Capillary column, fused silica, of length 8 m to 12 m and internal diameter 0,25 mm to 0,32 mm, with SE-52 or SE-541)liquid phase or equivalent, with a film thickness of 0,10 m to 0,30 m. 6.4 Microsyringe for on-column injection, of capacity 10 l, with a hardened needle. 6.5 Electric sh
41、aker. 6.6 Rotary evaporator. 6.7 Muffle oven. 6.8 Analytical balance, for weighing to an accuracy of 0,1 mg. 1) SE-52 and SE-54 are examples of suitable products available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by
42、ISO of these products. BS EN ISO 12873:2014ISO 12873:2010(E)BS ISO 12873:2010ISO 12873:2010(E) ISO 2010 All rights reserved 37 Sampling Sampling is not part of the method specified in this International Standard. A recommended sampling method is given in ISO 55551. It is important that the laborator
43、y receive a truly representative sample which has not been damaged or changed during transport or storage. 8 Preparation of the test sample Prepare the test sample in accordance with ISO 661. 9 Procedure 9.1 Preparation of the chromatography column Suspend 15 g of silica gel (5.1) in n-hexane (5.2)
44、and introduce into the chromatography column (6.2). Allow to settle spontaneously. Use an electric shaker (6.5) to assist in complete settling to make the chromatographic band more homogeneous. Percolate 30 ml n-hexane to remove any impurities. Weigh, to the nearest 0,1 mg, about 500 mg of the sampl
45、e into a 25 ml Erlenmeyer flask (6.1), add a suitable amount of internal standard (5.5) depending on the assumed wax content. Add 0,1 mg of lauryl arachidate in the internal standard solution (5.5) in the case of olive oil, and 0,25 mg to 0,50 mg in the case of olive-pomace oil. Transfer the test po
46、rtion to the chromatographic column with the aid of two 2 ml portions of n-hexane. Allow the solvent to drain off to 1 mm above the upper level of the absorbent. Percolate a further 70 ml of n-hexane to remove any n-alkanes naturally present. Then start chromatographic elution by collecting 180 ml o
47、f a mixture of 99 ml/100 ml n-hexane (5.2) and 1 ml/100 ml diethyl ether (5.3) at a rate of about 15 drops every 10 s. Perform the column chromatography at room temperature. Prepare the n-hexane-diethyl ether mixture freshly every day. To check visually that the waxes have been completely eluted, ad
48、d 100 l of Sudan I dye (5.6) solution at a concentration of 1 g/100 ml to the test portion solution. The dye is retained on the chromatographic column between the waxes and triglycerides. Hence, when the dye reaches the bottom of the column, suspend elution because all the waxes have been eluted. Ev
49、aporate the resultant fraction in a rotary evaporator (6.6) until the solvent is almost removed. Remove the last 2 ml of solvent under a weak stream of nitrogen, then add 2 ml to 4 ml of n-heptane. 9.2 Gas chromatographic analysis 9.2.1 Preliminary procedure If the column is being used for the first time, it is advisable to condition it. Run a light flow of gas through the column, then switch on the gas chromatograph. Heat gradually for approximately 4 h until a temperature of 350 C is reached. Maintain this temperature
