DIN EN ISO 12872-2014 Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872 2010) German version EN ISO 12872 2014《橄榄油和橄榄果渣油 2-甘油单棕榈酸酯.pdf

1、July 2014Translation by DIN-Sprachendienst.English price group 11No 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!%2l“2159773www.din.deDDIN EN ISO 12872Olive oils and olive-pomace oils Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010);English version EN ISO 12872:2014,English translation of DIN EN ISO 12872:2014-07Olivenle und Oliventresterle Verfahren zur Bestimmung des Prozentante

3、iles von 2-Glycerylmonopalmitat in Olivenlen(ISO 12872:2010);Englische Fassung EN ISO 12872:2014,Englische bersetzung von DIN EN ISO 12872:2014-07Huiles dolive et huiles de grignons dolive Dtermination de la teneur en 2-glycryl monopalmitate (ISO 12872:2010);Version anglaise EN ISO 12872:2014,Traduc

4、tion anglaise de DIN EN ISO 12872:2014-07www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 19 pages06.14 DIN EN ISO 12872:2014-07 2 A comma is used as the decimal marker. National foreword The text of ISO 12872:2010 has been prepared by T

5、echnical Committee ISO/TC 34 “Food products” (Secretariat: AFNOR, France), Subcommittee SC 11 “Animal and vegetable fats and oils” (Secretariat: BSI, United Kingdom) and has been taken over as EN ISO 12872:2014 by Technical Committee CEN/TC 307 “Oil seeds, vegetable and animal fats and oils and thei

6、r 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 landwirtschaftliche Produkte (Food and Agricultu

7、ral 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 Standards corresponding to the Interna

8、tional Standards referred to in this document are as follows: ISO 661 DIN EN ISO 661 ISO 835 DIN EN ISO 835 ISO 4788 DIN EN ISO 4788 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

9、 oils Preparation of test sample DIN EN ISO 835, Laboratory glassware Graduated pipettes DIN EN ISO 4788, Laboratory glassware Graduated measuring cylinders DIN EN ISO 5555, Animal and vegetable fats and oils Sampling DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and resul

10、ts Part 1: General principles and definitions DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method EN ISO 12872April 2014 ICS 67.200.10 English Version Oli

11、ve oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010) Huiles dolive et huiles de grignons dolive - Dtermination de la teneur en 2-glycryl monopalmitate (ISO 12872:2010)Olivenle und Oliventresterle - Verfahren zur Bestimmung des Prozentanteiles von 2-G

12、lycerylmonopalmitat in Olivenlen (ISO 12872:2010) This European Standard was approved by CEN 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 alte

13、ration. Up-to-date lists and bibliographical 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 mad

14、e by translation under the responsibility 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, Eston

15、ia, 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, Sweden, Switzerland, Turkey and United Kingdom. CEN-CENELEC Management Centr

16、e: 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 12872:2014 EEUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGEUROPEAN STANDARDNORME EURO

17、PENNEEUROPISCHE NORMContents Page Foreword . 3 Introduction . 4 1 Scope . 5 2 Normative references. 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 8 9.1 Preparatory steps . 8 9.2 Column chromatography 8 9.3 Hydroly

18、sis with pancreatic lipase 8 9.4 Preparation of the silylated derivatives and gas chromatography . 9 9.5 Gas chromatography . 9 10 Expression of results . 9 11 Precision . 10 11.1 Interlaboratory test . 10 11.2 Repeatability . 10 11.3 Reproducibility . 10 12 Test report . 10 Annex A (informative) Ch

19、romatograms . 11 Annex B (informative) Results of an interlaboratory test 14 Annex C (informative) Preparation and activity of the lipase 15 Bibliography 17 2DIN EN ISO 12872:2014-07 EN ISO 12872:2014 (E) ForewordThe text of ISO 12872:2010 has been prepared by Technical Committee ISO/TC 34 “Food pro

20、ducts” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 12872:2014 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 Eu

21、ropean 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 2014, and conflicting national standards shall be withdrawn at the latest by October 2014. Attention is drawn to the possibility that some of the e

22、lements 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 this

23、 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, Slovakia

24、, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12872:2010 has been approved by CEN as EN ISO 12872:2014 without any modification. 3DIN EN ISO 12872:2014-07EN ISO 12872:2014 (E)Introduction As part of the Trade standard applying to olive oils

25、 and olive-pomace oils, the International Olive Council (IOC) published COI/T.20/Doc. 23:20066. COI/T.20/Doc. 23 was applicable to olive and olive-pomace oils and was used to distinguish between lampante virgin olive oils and crude olive-pomace oils. Olive pomace is the residual paste which still co

26、ntains 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. 4DIN EN ISO 12872:2014-07 EN ISO 12872:2014 (E) 1 Scope This International Standard specifies a procedure for the determinat

27、ion of the content, as a percentage mass fraction, of 2-glyceryl monopalmitate in olive oils and olive-pomace oils that are liquid at ambient temperature (20 C). NOTE This International Standard is based on COI/T.20/Doc. 23:20066. 2 Normative references The following referenced documents are indispe

28、nsable 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) applies. ISO 661, Animal and vegetable fats and oils Preparation of test sample 3 Terms and definitions

29、For the purposes of this document, the following terms and definitions apply. 3.1 2-glyceryl monopalmitate content mass fraction of 2-glyceryl monopalmitate in the monoacylglycerol fraction, determined according to the method specified in this International Standard NOTE The 2-glyceryl monopalmitate

30、 content is expressed as a percentage. 4 Principle The oil, after suitable preparation, is subjected to the action of pancreatic lipase. A partial hydrolysis takes place that is specific for positions 1 and 3 of the triacylglycerol molecule so that 2-monoacylglycerols are obtained as reaction produc

31、ts. The percentage of 2-glyceryl monopalmitate in the monoacylglycerol fraction is determined, after silylation, by capillary gas chromatography. 5 Reagents WARNING Comply with any local regulations which specify the handling of hazardous substances. Technical, organizational and personal safety mea

32、sures shall be followed. During the analysis, unless otherwise stated, use only reagents of recognized analytical grade, and distilled or demineralized water or water of equivalent purity. 5DIN EN ISO 12872:2014-07EN ISO 12872:2014 (E)5.1 Silica gel, with a particle size of 0,063 mm to 0,200 mm (70/

33、280 mesh), prepared as follows: put the silica gel into a porcelain cup, dry in an oven at 160 C for 4 h, then cool at ambient temperature in a desiccator. Add a volume of water equivalent to 5 % of the mass of the silica gel as follows: weigh 152 g of silica gel into a 500 ml Erlenmeyer flask, add

34、8 g of water, stopper and homogenize carefully. Leave to settle for at least 12 h before using. 5.2 n-Hexane, chromatography grade. 5.3 Isopropanol. 5.4 Isopropanol-water mixture, volume fractions 50 ml/100 ml. 5.5 Pancreatic lipase, activity between 2,0 and 10 lipase units per milligram (see Annex

35、C). 5.6 Buffer solution of tris(hydroxymethyl)aminomethane: prepare an aqueous solution (1 mol/l) with pH 8 and mix with concentrated HCl, volume fractions 50 ml/100 ml. 5.7 Sodium cholate, special enzyme grade, aqueous solution, mass fraction 0,1 g/100 g. Use this solution within 15 days of prepara

36、tion. 5.8 Calcium chloride, aqueous solution, mass fraction 22 g/100 g. 5.9 Diethyl ether, chromatography grade. 5.10 Elution solvent: mixture of n-hexane-diethyl ether, volume fraction of n-hexane 87 ml/100 ml and of diethyl ether 13 ml/100 ml. 5.11 Sodium hydroxide, aqueous solution, mass fraction

37、 12 g/100 g. 5.12 Phenolphthalein, ethanolic solution, mass concentration 1 g/100 ml. 5.13 Carrier gas: hydrogen or helium, gas chromatography grade. 5.14 Auxiliary gases: hydrogen, free from moisture and organic substances, and synthetic air, gas chromatography grade. 5.15 Silylation reagent: mixtu

38、re of pyridine, hexamethyldisilazane (HMDS) and trimethylchlorosilane (TMCS); volume fractions: 9 ml/13 ml, 3 ml/13 ml, and 1 ml/13 ml, respectively. 5.16 Reference samples: pure monoacylglycerols and mixtures of monoacylglycerols with a known composition similar to that of the sample. 6 Apparatus U

39、sual laboratory equipment and in particular the following. 6.1 Erlenmeyer flasks, of capacity 25 ml. 6.2 Beakers, of capacities 100 ml, 250 ml, and 300 ml. 6.3 Glass chromatography column, 21 mm to 23 mm internal diameter, 400 mm in length, with septum and stopcock. 6.4 Measuring cylinders, of capac

40、ities 10 ml, 50 ml, 100 ml, and 200 ml, ISO 47882class A. 6.5 Round-bottomed flasks, of capacities 100 ml and 250 ml. 6.6 Rotary evaporator. 6DIN EN ISO 12872:2014-07 EN ISO 12872:2014 (E) 6.7 Centrifuge tubes, conical bottom, of capacity 10 ml, with ground-glass stopper. 6.8 Centrifuge, suitable fo

41、r 10 ml and 100 ml tubes. 6.9 Water bath, capable of maintaining a temperature of (40 0,5) C. 6.10 Graduated pipettes, of capacities 1 ml and 2 ml, ISO 8351class A. 6.11 Hypodermic syringe, 1 ml. 6.12 Microsyringe, 100 l. 6.13 Separating funnel, 1 000 ml. 6.14 Gas chromatograph, suitable for use wit

42、h capillary columns, equipped with the components specified in 6.14.1 to 6.14.5. 6.14.1 Cold on-column injector. 6.14.2 Flame ionization detector. 6.14.3 Column oven, capable of maintaining the temperature to within 1 C. 6.14.4 Computer-based integration system. 6.14.5 Fused silica capillary column,

43、 of length 8 m to 12 m and internal diameter 0,25 mm to 0,32 mm, coated with methylpolysiloxane or 5 % phenyl methylpolysiloxane, with a film thickness of 0,10 m to 0,30 m, suitable for use at 370 C. 6.15 Microsyringe, 10 l, with hardened needle, of minimum length 7,5 cm in length, suitable for on-c

44、olumn injection. 7 Sampling Sampling is not part of the method specified in this International Standard. A recommended sampling method is given in ISO 55553. It is important that the laboratory receive a truly representative sample which has not been damaged or changed during transport or storage. 8

45、 Preparation of the test sample Prepare the test sample in accordance with ISO 661. Oils with a free acidity greater than 3 % require a neutralization step as specified here. Introduce 50 g of the oil into a 1 000 ml separating funnel (6.13) and dissolve it in 200 ml of n-hexane (5.2). Add 100 ml of

46、 isopropanol (5.3) and a volume of sodium hydroxide solution (5.11) corresponding to the free acidity of the oil plus an excess of 5 %. Shake vigorously for 1 min, add 100 ml of water, shake again and leave to settle. After separation, remove the lower soapy layer. Also remove any intermediate layer

47、 of mucilage and insoluble matter which often forms. Wash the n-hexane solution of the oil with successive 50 ml to 60 ml portions of the isopropanol-water mixture (5.4) until the washed phase is neutral to phenolphthalein (5.12). Remove most of the hexane by distillation under vacuum, e.g. by using

48、 a rotary evaporator (6.6), and transfer the oil to a 100 ml round-bottomed flask (6.5). Dry it under vacuum to complete solvent removal. By the end of this procedure, ensure that the acidity of the oil is below 0,5 %. 7DIN EN ISO 12872:2014-07EN ISO 12872:2014 (E)9 Procedure 9.1 Preparatory steps 9

49、.1.1 Introduce 1,0 g of the oil, pretreated (Clause 8) where necessary, into a 25 ml Erlenmeyer flask (6.1) and dissolve it in 10 ml of elution solvent (5.10). Leave the solution to settle for at least 15 min before starting the silica gel column chromatography procedure. If the solution is cloudy, centrifuge it to ensure optimal conditions for chromatography. Ready-to-use 500 mg silica gel solid phase extraction (SPE) cartridges may be us