EN ISO 17059-2009 en Oilseeds - Extraction of oil and preparation of methyl esters of triglyceride fatty acids for analysis by gas chromatography (Rapid method)《含油种子 气相色谱法分析用油的萃取和甘.pdf

1、BRITISH STANDARDBS EN ISO 17059:2009Oilseeds Extraction of oil and preparation of methyl esters of triglyceride fatty acids for analysis by gas chromatography (Rapid method)ICS 67.200.20g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g5

2、5g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58National forewordThis British Standard is the UK implementation of EN ISO 17059:2009. It is identical with ISO 17059:2007. It supersedes BS ISO 17059:2007 which is withdrawn.The UK participation in its preparation

3、was entrusted to Technical Committee AW/307, Oil seeds, animal and vegetable fats and oils and their by products.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.

4、Users are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN ISO 17059:2009This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 August 2007 BSI 2010Amendments/corrigend

5、a issued since publicationDate Comments 31 January 2010 This corrigendum renumbers BS ISO 17059:2007 as BS EN ISO 17059:2009ISBN 978 0 580 66795 4EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 17059 August 2009 ICS 67.200.20 English Version Oilseeds - Extraction of oil and preparation of m

6、ethyl esters of triglyceride fatty acids for analysis by gas chromatography (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) l

7、samen - Extraktion von l und Herstellung von Methylester aus den Fettsuren 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 Regulation

8、s 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 be obtained on application to the CEN Management Centre or to any CEN member. This Europ

9、ean 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 the CEN Management Centre has the same status as the official versions. CEN members are the nat

10、ional 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, Slovakia, Slovenia, Spain, Sweden, Switzerland and

11、 United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN 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 170

12、59:2009: EForeword The text of ISO 17059:2007 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 17059:2009 by Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils an

13、d 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 by endorsement, at the latest by February 2010, and conflicting national standards

14、 shall be withdrawn at the latest by February 2010. 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 for identifying any or all such patent rights. According to the CEN/CENELEC Inter

15、nal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Mal

16、ta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland 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. iiBS EN ISO 17059:2009EN ISO 17059:2009 (E)iiiIntroductio

17、n Chromatographic analysis of the fatty acid methyl esters (FAME) of oilseeds requires oil extraction 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

18、for the determination of oil content and are tedious or time consuming2, 3. Consequently, the 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 ste

19、p. This International Standard specifies a rapid and optimized method for a combined oil extraction 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

20、 in the Scope4, 5. The FAME are prepared according to the transesterification method described 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 Internati

21、onal Standard was compared to ISO 6592in an interlaboratory test6. Results showed very good agreement 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 %.

22、BS EN ISO 17059:2009EN ISO 17059:2009 (E)This page deliberately set blank1Oilseeds Extraction of oil and preparation of methyl esters of triglyceride fatty acids for analysis by gas chromatography (Rapid method) 1 Scope This International Standard specifies a rapid method for extraction of oil and f

23、or 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 rapid method to high erucic acid content rape

24、seed 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 edition cited applies. For undated references

25、, 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 oil is cold extracted from previously crus

26、hed 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 otherwise specified. 4.1 Iso-octane (2,2,4-trime

27、thylpentane) 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. BS EN ISO 17059:2009EN ISO 17059:2009 (E)2 5 Apparatus Usual laboratory apparatus and, in particular,

28、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 wick and anhydrous sodium sulfate up to a heig

29、ht 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. It should not have been damaged or changed duri

30、ng 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 crush a quantity of approximately 10 g using a bla

31、de 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 fatty acids. 7.2 Test portion 7.2.1 General The cr

32、ushed 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 linseed) Mix the crushed material and weigh, t

33、o 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 10 ml test tube (5.2). BS EN ISO 17059:2009EN

34、ISO 17059:2009 (E)37.3 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 extract is not clear, proceed with filtration and drying (7

35、.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 Filtration and drying of the extract Place the Pasteu

36、r 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 test tube. This extract is then ready for the preparati

37、on 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 5.3.3. Transfer (with dilution or not, according to

38、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 shall specify: all information necessary for the comp

39、lete 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 details of any incidents which may have influenced th

40、e test result(s). BS EN ISO 17059:2009EN ISO 17059:2009 (E)4 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 chromatography. Consequently, any new batch of reagent or s

41、olvent 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 should preferably be analysed as soon as possible. If neces

42、sary, 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 antioxidant in such a concentration as will not interfere with th

43、e subsequent analysis, for example a 0,05 g/l solution of BHT (2,6-di-t-butyl-4-methylphenol). BS EN ISO 17059:2009EN ISO 17059:2009 (E)5Bibliography 1 ISO 542:1990, Oilseeds Sampling 2 ISO 659:1998, Oilseeds Determination of oil content (Reference method) 3 XP V 03-030:1991 (French standard), Agric

44、ultural 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. Validation of a rapid method to extract oil in oleaginous seeds

45、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 precisio

46、n) of measurement methods and results Part 1: General principles and definitions 8 ISO 5725-2:1994, 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 BS EN ISO 17059:20

47、09EN ISO 17059:2009 (E)BSI GroupHeadquarters 389 Chiswick High Road, London, W4 4AL, UK Tel +44 (0)20 8996 9001 Fax +44 (0)20 8996 7001 standardsBSI - British Standards InstitutionBSI is the independent national body responsible for preparing British Standards. It presents the UK view on standards

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