1、July 2014Translation 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 67
2、.200.10!%2x#“2158500www.din.deDDIN EN ISO 12873Olive oils and olive-pomace oils Determination of wax content by capillary gas chromatography(ISO 12873:2010);English version EN ISO 12873:2014,English translation of DIN EN ISO 12873:2014-07Olivenle und Oliventresterle Bestimmung des Wachsgehaltes mitt
3、els Kapillarsulen-Gaschromatographie(ISO 12873:2010);Englische Fassung EN ISO 12873:2014,Englische bersetzung von DIN EN ISO 12873:2014-07Huiles dolive et huiles de grignons dolive Dtermination de la teneur en cires par chromatographie en phase gazeuse sur colonnecapillaire (ISO 12873:2010);Version
4、anglaise EN ISO 12873:2014,Traduction anglaise de DIN EN ISO 12873:2014-07www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 15 pages06.14 DIN EN ISO 12873:2014-07 2 A comma is used as the decimal marker. National foreword The text of ISO
5、12873:2010 has been prepared by Technical Committee ISO/TC 34 “Food products” (Secre-tariat: AFNOR, France), Subcommittee SC 11 “Animal and vegetable fats and oils” (Secretariat: BSI, United Kingdom) and has been taken over as EN ISO 12873:2014 by Technical Committee CEN/TC 307 “Oil seeds, vegetable
6、 and animal fats and oils and their by-products Methods of sampling and analysis” (Secretariat: AFNOR, France). The document has been submitted to the Unique Acceptance Procedure (UAP). The responsible German bodies involved in its preparation were the Normenausschuss Lebensmittel und landwirtschaft
7、liche Produkte (Food and Agricultural Products Standards Committee) and the Deutsche Gesellschaft fr Fettwissenschaft (German Fat Research Society), Joint Committee NA 057-05-05 AA Gemeinschaftsausschuss fr die Analytik von Fetten, len, Fettprodukten, verwandten Stoffen und Roh-stoffen. The DIN Stan
8、dards corresponding to the International Standards referred to in this document are as follows: ISO 661 DIN EN ISO 661 ISO 5555 DIN EN ISO 5555 ISO 5725-1 DIN ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 National Annex NA (informative) Bibliography DIN EN ISO 661, Animal and vegetable fats and oils Preparat
9、ion of test sample DIN EN ISO 5555, Animal and vegetable fats and oils Sampling DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2:
10、 Basic method for the determination of repeatability and reproducibility of a standard measurement method EN ISO12873April 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 gr
11、ignons 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 CE
12、N on 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
13、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 of a CEN member into its own language and notifie
14、d 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 Macedonia, France, Germany, Greece, Hungary, Icela
15、nd, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by
16、 any means reserved worldwide for CEN national Members. Ref. No. EN ISO 12873:2014 EEUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMContents Page Foreword . 3 Introduction . 4 1 Scope . 5 2 Normative ref
17、erences. 5 3 Terms and definitions 5 4 Principle 5 5 Reagents . 5 6 Apparatus 6 7 Sampling . 7 8 Preparation of the test sample 7 9 Procedure 7 9.1 Preparation of the chromatography column . 7 9.2 Gas chromatographic analysis . 7 9.3 Performance of the analysis . 8 9.4 Peak identification 8 10 Quant
18、itative analysis and expression of results 8 11 Precision . 9 11.1 Interlaboratory test . 9 11.2 Repeatability . 9 11.3 Reproducibility . 9 12 Test report . 9 Annex A (informative) Chromatogram . 10 Annex B (informative) Results of an interlaboratory test 11 Annex C (informative) Determination of th
19、e linear velocity of the gas 12 Bibliography 13 2DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E) ForewordThe text of ISO 12873:2010 has been prepared by Technical Committee ISO/TC 34 “Food products” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 12873:201
20、4 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 shall be given the status of a national standard, either by publication of an identical text or
21、 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 document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible
22、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 Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, For
23、mer Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12873
24、:2010 has been approved by CEN as EN ISO 12873:2014 without any modification. 3DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E)Introduction As part of the Trade standard applying to olive oils and olive-pomace oils, the International Olive Oil Council (IOOC) now known as International Olive Council (IO
25、C) 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 oils obtained by either pressing or centrifuging and olive-pomace oils. Olive pomace is the residual paste which still contains a variable amount of water and oil a
26、fter pressing or centrifuging. In 2008, the IOC submitted the document to ISO/TC 34/SC 11 for adoption as an International Standard. 4DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E) 1 Scope This International Standard specifies the determination of the wax content, as a mass fraction expressed in mill
27、igrams per kilogram, of olive 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
28、International Standard is based 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 docum
29、ent (including any amendments) 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
30、with the method specified in 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 (
31、which has a lower polarity than 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
32、analysis, unless otherwise stated, use only reagents of recognized analytical grade, and distilled or demineralized water or water of equivalent purity. 5DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E)5.1 Silica gel, with a particle size of 60 m to 200 m, prepared as follows: place the gel in the muff
33、le 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-Heptane, chrom
34、atography 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 Auxiliary gases: h
35、ydrogen, 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, with stop
36、cock. 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 system. 6.3
37、.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 shaker. 6.6
38、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 ISO of the
39、se products. 6DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E) 7 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 laboratory receive a truly representative sample which has not been damage
40、d 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) and introduce into the chromatography column (6.2). Allow to sett
41、le 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 sample into a 25 ml Erlenmeyer flask (6.1), add a suitable amount of i
42、nternal 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 portion to the chromatographic column with the aid of two 2 ml port
43、ions 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 of a mixture of 99 ml/100 ml n-hexane (5.2) and 1 ml/100 ml diethy
44、l 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, add 100 l of Sudan I dye (5.6) solution at a concentration of 1 g/1
45、00 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. Evaporate the resultant fraction in a rotary evaporator (6.6) until
46、 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 t
47、hrough 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 for at least 2 h, then regulate the apparatus to the operating conditions (gas flow, light flame, oven temperature for column, detector, et
48、c.). Record the signal at a sensitivity that is at least twice as high as that required for the analysis. The baseline should be linear, with no peaks of any kind, and shall not drift. Negative straight-line drift indicates incorrect column connections, while positive drift is symptomatic of imprope
49、r column conditioning. 7DIN EN ISO 12873:2014-07EN ISO 12873:2014 (E)9.2.2 Operating conditions The following operating conditions are recommended. Column temperature: isothermal at 80 C for 1 min, increase at a rate of 20 C/min to 240 C, increase at a rate of 5 C/min to 325 C; 5 min isothermal at 325 C, increase at a rate of 20 C/min to 340 C; 10 min isothermal at 340 C; detector temperature: 350 C; injection volume: 1 l of
