DIN EN ISO 17059-2009 Oilseeds - Extraction of oil and preparation of methyl esters of triglyceride fatty acids for analysis by gas chromatography (Rapid method) (ISO 17059 2007) E.pdf

1、November 2009DEUTSCHE NORM English price group 8No part of this standard 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 67.200.20!$ZqE“1557834

2、www.din.deDDIN EN ISO 17059Oilseeds Extraction of oil and preparation of methyl esters of triglyceride fattyacids for analysis by gas chromatography (Rapid method)(ISO 17059:2007)English version of DIN EN ISO 17059:2009-11lsamen Extraktion von l und Herstellung von Methylester aus den Fettsuren der

3、Triglyceride frdie Analyse durch Gaschromatographie (Schnellverfahren) (ISO 17059:2007)Englische Fassung DIN EN ISO 17059:2009-11www.beuth.deDocument comprises pages11DIN EN ISO 17059:2009-11 National foreword This standard has been prepared by Technical Committee ISO/TC 34 “Food products” (Secretar

4、iat: AFNOR, France), Subcommittee SC 2 “Oleaginous seeds and fruits and oilseed meals” (Secretariat: AFNOR, France) in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products Methods of sampling and analysis” (Secretariat: AFNOR, France).

5、 The responsible German bodies involved in its preparation were the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee) and the Deutsche Gesellschaft fr Fettwissenschaft e.V. (German Fat Research Society), Joint Committee Gemeinschafts-a

6、usschuss fr die Analytik von Fetten, len, Fettprodukten, verwandten Stoffen und Rohstoffen. The DIN Standards corresponding to the International Standards referred to in the EN are as follows: ISO 542 DIN EN ISO 542 ISO 659 DIN EN ISO 659 ISO 664 DIN EN ISO 664 ISO 5509 DIN EN ISO 5509 ISO 5725-1 DI

7、N ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 National Annex NA (informative) Bibliography DIN EN ISO 542, Oilseeds Sampling DIN EN ISO 659, Oilseeds Determination of oil content (Reference method) DIN EN ISO 664, Oilseeds Reduction of laboratory sample to test sample DIN EN ISO 5509, Animal and vegetable

8、fats and oils Preparation of methyl esters of fatty acids 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: Basic method for the

9、determination of repeatability and reproducibility of a standard measurement method 2 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 17059 August 2009 ICS 67.200.20 English Version Oilseeds Extraction of oil and preparation of methyl esters of triglyceride fatty acids for analysis by gas c

10、hromatography (Rapid method) (ISO 17059:2007) Graines olagineuses Extraction de lhuile et prparation des esters mthyliques dacides gras de triglycrides pour analyse par chromatographie en phase gazeuse (Mthode rapide) (ISO 17059:2007) lsamen Extraktion von l und Herstellung von Methylester aus den F

11、ettsuren der Triglyceride fr die Analyse durch Gaschromatographie (Schnellverfahren) (ISO 17059:2007) This European Standard was approved by CEN on 23 July 2009. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard

12、the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN Management Centre or to any CEN member. This European Standard exists in three official versions (English, French, Ge

13、rman). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as theofficial versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech

14、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 United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN

15、DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 17059:2009: EContents Page 2EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-1

16、1 Foreword .3 Introduction.4 1 Scope 5 2 Normative references 5 3 Principle5 4 Reagents.5 5 Apparatus .6 6 Sampling.6 7 Procedure .6 8 Test report 7 Annex A (normative) General analytical procedure 8 Bibliography9 Foreword The text of ISO 17059:2007 has been prepared by Technical Committee ISO/TC 34

17、 “Food products” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 17059:2009 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 AFNO

18、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 February 2010, and conflicting national standards shall be withdrawn at the latest by February 2010. Attention is drawn to the possibility that s

19、ome 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. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to im

20、plement this European Standard: 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, Switzerla

21、nd and the United Kingdom. Endorsement notice The text of ISO 17059:2007 has been approved by CEN as a EN ISO 17059:2009 without any modification. 3EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 Introduction Chromatographic analysis of the fatty acid methyl esters (FAME) of oilseeds requires oil ext

22、raction from the oilseeds. To date, no International Standard has specified a method for extracting oil from oilseeds for FAME analysis. The methods usually performed in laboratories involve oil extraction for the determination of oil content and are tedious or time consuming2, 3. Consequently, the

23、total duration and cost of the analysis of triglyceride fatty acids in oilseeds, including oil extraction, preparation and gas chromatography of the FAME are considerably increased by the oil extraction step. This International Standard specifies a rapid and optimized method for a combined oil extra

24、ction and FAME preparation. The oil is only partially extracted from the seeds and the extracted fraction remains representative enough of the total content when the method is applied to the seeds specified in the Scope4, 5. The FAME are prepared according to the transesterification method described

25、 in ISO 5509 and slightly modified to be applied to iso-octane solutions of oil. Taking into account that no reference method for oil extraction exists, the oil extraction method specified in this International Standard was compared to ISO 6592in an interlaboratory test6. Results showed very good ag

26、reement between the two methods except when applied to rapeseed with high erucic acid content. In this case, this method led to values of erucic acid content higher by approximately a mass fraction of 1 %. 4EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 1 Scope This International Standard specifies

27、a rapid method for extraction of oil and for preparation of the methyl esters of fatty acids. The methyl esters thus obtained can be used for gas chromatography. This International Standard is applicable to the following oilseeds: rape, sunflower, soya beans, mustard, linseed. NOTE Applying this rap

28、id method to high erucic acid content rapeseed leads to an overestimation of erucic acid content by approximately a mass fraction of 1 %. 2 Normative references The following referenced documents are indispensable for the application of this International Standard. For dated references, only the edi

29、tion cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 664, Oilseeds Reduction of laboratory sample to test sample ISO 5509:2000, Animal and vegetable fats and oils Preparation of methyl esters of fatty acids 3 Principle The

30、oil is cold extracted from previously crushed grains by shaking in iso-octane. After filtration, the triglyceride fatty acids present in the iso-octane solution are transesterified with potassium hydroxide into methyl esters. 4 Reagents Use only reagents of recognized analytical grade, unless otherw

31、ise specified. 4.1 Iso-octane (2,2,4-trimethylpentane) of chromatographic quality. See Annex A. 4.2 Anhydrous sodium sulfate. 4.3 Other reagents used for the preparation of the methyl esters are specified in ISO 5509:2000, 5.3.1 and 5.3.3. 5EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 5 Apparatus

32、Usual laboratory apparatus and, in particular, the following. 5.1 Blade crusher, coffee grinder type. 5.2 Test tubes, of glass, of capacity 10 ml, with ground or screw type stopper and PTFE cap. 5.3 Graduated pipette, of capacity 5 ml. 5.4 Pasteur pipettes, of length 150 mm, filled with a glass wool

33、 wick and anhydrous sodium sulfate up to a height of 20 mm. 5.5 Test tubes, of glass, of capacity 5 ml, with ground or screw type stopper and PTFE cap. 5.6 Glass vial, of capacity 2 ml, with screw type stopper and PTFE cap. 6 Sampling A representative sample should have been sent to the laboratory.

34、It should not have been damaged or changed during transport or storage. Sampling is not part of the method specified in this International Standard. A recommended sampling method is given in ISO 5421. 7 Procedure 7.1 Preparation of the test sample Reduce the sample in accordance with ISO 664 and cru

35、sh a quantity of approximately 10 g using a blade crusher (5.1) during 15 s. NOTE For samples that are non-homogeneous in nature, i.e. contain significant quantities of unseparable seeds (such as Sinapis arvensis in canola) a larger sample (25 g) may be required to ensure an accurate estimate of fat

36、ty acids. 7.2 Test portion 7.2.1 General The crushed material test portion shall be adapted as a function of the oil content of the sample in order to permit the extraction of approximately 100 mg of oil. 7.2.2 Case of grains having an oil content exceeding 30 % by mass (rape, mustard, sunflower and

37、 linseed) Mix the crushed material and weigh, to the nearest 0,02 g, approximately 0,40 g of it in a 10 ml test tube (5.2). 7.2.3 Case of grains having an oil content between 15 % and 30 % by mass (soya beans) Mix the crushed material and weigh, to the nearest 0,04 g, approximately 0,80 g of it in a

38、 10 ml test tube (5.2). 6EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 7.3 Extraction of the oil Using a pipette (5.3), add 5 ml of iso-octane (4.1) to the tube containing the crushed material and stopper. Shake for 2 min, leave to settle or centrifuge if necessary. If the supernatant of the extrac

39、t is not clear, proceed with filtration and drying (7.4). If the supernatant of the extract is clear, filtration and drying are not necessary and may be omitted. Transfer 3 ml of the supernatant to a 5 ml test tube (5.5). The extract is then ready for the preparation of the methyl esters (7.5). 7.4

40、Filtration and drying of the extract Place the Pasteur pipette (5.4) containing anhydrous sodium sulfate (4.2) above a 5 ml test tube (5.5). Transfer the supernatant of the extract (7.3) to the Pasteur pipette and allow to drain off to obtain a volume of approximately 3 ml of clear extract in the te

41、st tube. This extract is then ready for the preparation of the methyl esters (7.5). 7.5 Preparation of the methyl esters Proceed according to ISO 5509:2000, from 5.6.2.2 to 5.6.2.4 on the extract prepared in the 5 ml test tube (7.3 or 7.4). NOTE The reagents are specified in ISO 5509:2000, 5.3.1 and

42、 5.3.3. Transfer (with dilution or not, according to ISO 5509:2000, 5.6.2.4 and the injection mode in gas chromatography) the supernatant containing the methyl esters into a 2 ml glass vial (5.6) with stopper. The solution is ready to inject into the gas chromatograph. 8 Test report The test report

43、shall specify: all information necessary for the complete identification of the sample; the sampling method used, if known; the test method used, with reference to this International Standard; all operating details not specified in this International Standard, or regarded as optional, together with

44、details of any incidents which may have influenced the test result(s). 7EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 Annex A (normative) General analytical procedures A.1 Reagents The reagents shall not produce peaks that interfere with those of the methyl esters of fatty acids during gas-liquid c

45、hromatography. Consequently, any new batch of reagent or solvent should be checked by using it to prepare the methyl esters of pure oleic acid and analysing them by gas-liquid chromatography. If any extra peaks appear, the reagent should be rejected. A.2 Storage of methyl ester solution The esters s

46、hould preferably be analysed as soon as possible. If necessary, the iso-octane solution containing the methyl esters may be stored under inert gas in a refrigerator. For a longer period of storage, it is advisable to protect the methyl esters against autoxidation by adding to the solution an antioxi

47、dant in such a concentration as will not interfere with the subsequent analysis, for example a 0,05 g/l solution of BHT (2,6-di-t-butyl-4-methylphenol). 8EN ISO 17059:2009 (E) DIN EN ISO 17059:2009-11 Bibliography 1 ISO 542:1990, Oilseeds Sampling 2 ISO 659:1998, Oilseeds Determination of oil conten

48、t (Reference method) 3 XP V 03-030:1991 (French standard), Agricultural products and foodstuffs Extraction of the crude fat with a view to its characterization 4 BLIN, F, 1998. Rapid extraction of oil from oilseeds with a view to the analysis of fatty acids. Report-CETIOM 5 QUINSAC, A. and BLIN, F.

49、Validation of a rapid method to extract oil in oleaginous seeds for fatty acid analysis. Proceedings of the 10th International Rapeseed Congress, Camberra, Australia, September 26 to 29, 1999, 122 6 CETIOM, 2002, Report of the Ring-Test (January - April 2001): Comparison between ISO/WD 17059 and ISO 659 ISO 5509 7 ISO 5725-1:1994, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions 8 ISO 5725-2:1994, Accuracy (trueness and pre