BS EN 14103-2011 Fat and oil derivatives Fatty acid methyl esters (FAME) Determination of ester and linolenic acid methyl ester contents《油脂衍生物 脂肪酸甲酯(FAME) 酯和亚油酸甲酯含量的测定》.pdf

1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 14103:2011Fat and oil derivatives Fatty Acid Methyl Esters (FAME) Determination of ester and linolenic acid methyl ester contentsBS EN 14103:2011 BRITISH STANDARDNational f

2、orewordThis British Standard is the UK implementation of EN 14103:2011. It supersedes BS EN 14103:2003 which is withdrawn.The UK participation in its preparation was entrusted to T e c h n i c a l C o m m i t t e e A W / 3 0 7 , O i l s e e d s , a n i m a l a n d v e g e t a b l e fats and oils and

3、 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. Users are responsible for its correct application. BSI 2011 ISBN 978 0 580 68565 1 ICS 67.200.

4、10 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 May 2011.Amendments issued since publicationDate T e x t a f f e c t e dBS EN 14103:2011EUROPEAN STANDARD N

5、ORME EUROPENNE EUROPISCHE NORM EN 14103 April 2011 ICS 67.200.10 Supersedes EN 14103:2003English Version Fat and oil derivatives - Fatty Acid Methyl Esters (FAME) - Determination of ester and linolenic acid methyl ester contentsProduits drivs des corps gras - Esters mthyliques dacides gras (EMAG) -

6、Dtermination de la teneur en ester et en ester mthylique de lacide linolnique Erzeugnisse aus pflanzlichen und tierischen Fetten und len - Fettsure-Methylester (FAME) - Bestimmung des Ester-Gehaltes und des Gehaltes an Linolensure-Methylester This European Standard was approved by CEN on 10 March 20

7、11. 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 be obtained

8、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 the CEN-CEN

9、ELEC 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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malt

10、a, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of explo

11、itation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 14103:2011: EBS EN 14103:2011EN 14103:2011 (E) 2 Contents Page Foreword 31 Scope 42 Principle 43 Reagents . 44 Apparatus . 45 Sampling . 56 Procedure . 57 Expression of results 68 Precision . 79 Test report

12、 8Bibliography 13BS EN 14103:2011EN 14103:2011 (E) 3 Foreword This document (EN 14103:2011) has been prepared 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 Eur

13、opean 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 2011, and conflicting national standards shall be withdrawn at the latest by October 2011. Attention is drawn to the possibility that some of the el

14、ements 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. This document supersedes EN 14103:2003. The main modifications of the standard are intended: - to enlarge the scope of the method to FAME from

15、animal fat, by changing the internal standard (FAME C19 instead of FAME C17); - to verify the purity of the internal standard as it was proven that some lots may have a purity not good enough; - to improve the precision data by modifying the preparation of the test portion. According to the CEN/CENE

16、LEC 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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuani

17、a, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. BS EN 14103:2011EN 14103:2011 (E) 4 1 Scope The purpose of this document is to describe a procedure for the determination of the ester content in fatty acid methyl

18、 esters (FAME) intended for incorporation into diesel oil. It also allows determining the linolenic acid methyl ester content. It allows verifying that the ester content of FAME is greater than 90 % (m/m) and that the linolenic acid content is between 1 % (m/m) and 15 % (m/m). This method is suitabl

19、e for FAME which contains methyl esters between C6 and C24. NOTE For the purposes of this European Standard, the terms “% (m/m)” and “%(v/v)” are used to represent respectively the mass and volume fractions. WARNING The use of this method may involve hazardous equipment, materials and operations. Th

20、is method does not purport to address to all of the safety problems associated with its use, but it is the responsibility of the user to search and establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 2 Principle Determination of t

21、he percentage of total methyl esters of fatty acids and the percentage of linolenic acid methyl ester present in the sample, by gas chromatography according to a procedure using internal calibration (nonadecanoic acid methyl ester). 3 Reagents Use only reagents of recognized analytical grade, unless

22、 otherwise specified. 3.1 Toluene, analytical grade. 3.2 Nonadecanoic acid methyl ester (FAME C19), purity, min. 99,5 % (m/m)1). NOTE Standard should be kept in a dry storage in order to limit its water absorption. Its water content should be verified by Karl-Fischer when a new lot of standard is op

23、en. 3.3 Carrier gas, hydrogen or helium. 3.4 Auxiliary gases: air; hydrogen. 4 Apparatus Usual laboratory apparatus and, in particular, the following. 1) Nonadecanoic FAME with acceptable quality is available from Fluka - Ref: 74208 () or from Nu Chek Prep - Ref: N-19-M (www.nu-) or Dr. Ehrenstorfer

24、 GmbH ref 15 622 360 (www.analytical-). This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of these products. BS EN 14103:2011EN 14103:2011 (E) 5 4.1 Gas chromatograph, equipped with a variable split flow injector or equival

25、ent device, a temperature programmable oven and a flame ionization detector. 4.2 Capillary column, coated with polyethylene glycol (Carbowax 20M) stationary phase, the following characteristics have been found suitable: length: 30 m, internal diameter: 0,25 mm, film thickness: 0,25 m. NOTE Stationar

26、y phase other than polyethylene glycol should be tested first before being selected as co-elution between the internal standard (FAME C19) and other fatty acid methyl esters may exist. Indeed, there is a co-elution between FAME C19 and linoleic acid methyl ester (FAME C18:2) when using a column with

27、 a stationary phase such as 70 % cyanopropyl-polysilphenyl-siloxane. 4.3 Glass mono-use tubes equipped with plastic mono-use stopper, 10 ml capacity. 4.4 Pipette, 10 ml capacity. 4.5 Analytical balance, accuracy 0,1 mg. 5 Sampling Sampling is not part of the method specified in this European Standar

28、d. A recommended sampling method is given in EN ISO 5555 1. 6 Procedure 6.1 Operating conditions The chromatographic analysis conditions will be chosen taking into account the characteristics of the column being used and the type of carrier gas (hydrogen or helium). By way of indication, an example

29、of analysis conditions is described below: column temperature: 60 C hold for 2 min, programmed at 10 C.min-1up to 200 C, programmed at 5 C.min-1up to 240 C, final temperature hold for 7 min; injector temperature carrier gas flow rate 1-2 ml.min-1, a minimum flow rate of 1 ml.min-1shall be warranted

30、when operating at the maximum temperature; injected volume: 1 l; hydrogen pressure = 70 KPa; split flow = 100 ml.min-1. 6.2 Internal standard purity determination Prepare a solution of nonadecanoic acid methyl ester (3.2) in toluene (3.1) approximately at 10 mg.ml-1. Analyse 1 l of this solution by

31、gas chromatography according to the conditions described in (6.1), but the final temperature should be held for 20 min at 240 C, instead of 7 min. Calculate the purity of the nonadecanoic acid methyl ester taking into account all the peak eluted in the chromatogram, excepted the solvent peak. The pu

32、rity of nonadecanoic acid methyl ester should be at least 99,5 % (m/m). If the purity is lower than 99,5 % (m/m) do not use this standard for this determination. NOTE No correction of the purity of the internal standard is performed for the calculation of the FAME content. BS EN 14103:2011EN 14103:2

33、011 (E) 6 6.3 FAME sample preparation and analysis Accurately weigh approximately 100 mg (accuracy 0,1 mg) of homogenized sample in a 10 ml tube (4.3), and approximately 100 mg (accuracy 0,1 mg) of nonadecanoic acid methyl ester (3.2), and dilute with 10 ml of toluene (3.1). NOTE Standard and sample

34、s should be let at ambient temperature, in their container closed, at least 3 h prior being weighed, in order to limit the water absorption during weighting. Analyse 1 l of this solution by gas chromatography according to the conditions described in (6.1). For each sample, two test portions are prep

35、ared and give rise, each one, to two chromatographic analyses. 6.4 Identification The chromatographic conditions (injected quantity, oven temperature, carrier gas pressure and split flow rate) shall be adjusted so as to correctly visualize the methyl ester peaks of the lignoceric (C24:0) and nervoni

36、c (C24:1) acids. The integration shall be carried out as from the hexanoic acid methyl ester (C6:0) peak up to that of the nervonic acid methyl ester (C24:1) taking all the peaks identified as fatty acid methyl esters into consideration. In order to identify properly the fatty acid methyl esters, so

37、me commercial solutions may be used 2). NOTE If some unknown thin peaks are found (others than saturated and mono-unsaturated FAME) between the linolenic acid methyl ester (C18:3) and the nervonic acid methyl ester (C24:1), presence of fish oil in the sample can be suspected. As a general rule, the

38、separation is done according to carbon atom chain length, unsaturated FAME are eluted after the corresponding saturated ones. 7 Expression of results 7.1 Determination of ester content The ester C content, expressed as a mass percentage, is calculated using the following equation: 100EIEIEI=WWAAAC (

39、1) where A is the total peak area from the methyl ester in C6:0 to that in C24:1; AEIis the peak area corresponding to nonadecanoic acid methyl ester; WEIis the weight, in milligrams, of the nonadecanoic acid methyl ester being used as internal standard; W is the weight, in milligrams, of the sample

40、. NOTE 1 In the case of vegetable oils, the result of the calculation based on relative areas is considered to represent a percentage by mass. 2) Commercial solutions of mixtures of fatty acid methyl esters are available from Sigma Ref. 18918-1AMP () or from Nu Chek Prep Ref.17a (www.nu-). This info

41、rmation is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of these products. BS EN 14103:2011EN 14103:2011 (E) 7 NOTE 2 If the average of two determinations is higher than 100 % then discard the results and verify the experimental condition

42、s as well as the purity of the internal standard by using this method to determine the ester content of a commercial or prepared mixture, and the water content of the internal standard by Karl Fischer determination. 7.2 Determination of linolenic acid methyl ester The linolenic acid methyl ester con

43、tent L, expressed as a mass percentage, is calculated using the following equation: 100EIEIL=WWAAL (2) where ALis the peak area corresponding to linolenic acid methyl ester; AEIis the peak area corresponding to nonadecanoic acid methyl ester; WEIis the weight, in milligrams, of the nonadecanoic acid

44、 methyl ester being used as internal standard; W is the weight, in milligrams, of the sample. 7.3 Expression of results Ester content and linolenic acid methyl ester content are expressed in percentage (m/m), to the nearest 0,1 %. 8 Precision 8.1 Interlaboratory test An interlaboratory test organize

45、d in 2009 at European level with the participation of 18 laboratories, each having carried out two determinations on each sample, gave the statistical results indicated in Annex B. 8.2 Repeatability The absolute difference between two independent single test results, obtained using the same method o

46、n identical test material in the same laboratory by the same operator using the same equipment within a short time interval, shall not be greater more than once out of 20 determinations than the values given in Table 1: Table 1 For ester content 1,01 % (m/m) For linolenic acid methyl ester r = 0,028

47、3 + 0,0175 . X X being the mean value of the two results in question r in % (m/m). 8.3 Reproducibility The absolute difference between two single test results, obtained using the same method on identical test material in different laboratories by different operators using different equipment, shall

48、not be greater more than once out of 20 determinations than the values given in Table 2: BS EN 14103:2011EN 14103:2011 (E) 8 Table 2 For ester content 4,16 % (m/m) For linolenic acid methyl ester R = 0,3872 + 0,0285 . X X being the mean value of the two results in question R in % (m/m). 9 Test repor

49、t The test report 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 European Standard; all operating details not specified in this European Standard, or regarded as optional, together with details of any incidents which may have influenced the test result(s); the test result(s)