DIN EN 14105-2011 Fat and oil derivatives - Fatty Acid Methyl Esters (FAME) - Determination of free and total glycerol and mono- di- triglyceride contents German version EN 14105 2.pdf

1、July 2011 Translation by DIN-Sprachendienst.English price group 12No 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 6

2、7.200.10!$s3“1805816www.din.deDDIN EN 14105Fat and oil derivatives Fatty Acid Methyl Esters (FAME) Determination of free and total glycerol and mono-, di-, triglyceridecontentsEnglish translation of DIN EN 14105:2011-07Erzeugnisse aus pflanzlichen und tierischen Fetten und len Fettsure-Methylester (

3、FAME) Bestimmung des Gehaltes an freiem und Gesamtglycerin und Mono-, Di- undTriglyceridenEnglische bersetzung von DIN EN 14105:2011-07Produits drivs des corps gras Esters mthyliques dacides gras (EMAG) Dtermination de la teneur en glycrols libre et total et en mono-, di- et triglycridesTraduction a

4、nglaise de DIN EN 14105:2011-07SupersedesDIN EN 14105:2003-10www.beuth.deDocument comprises pagesIn case of doubt, the German-language original shall be considered authoritative.2307.11 DIN EN 14105:2011-07 A comma is used as the decimal marker. National foreword This standard has been prepared by T

5、echnical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products Methods of sampling and analysis” (Secretariat: AFNOR, France) in collaboration with Technical Committee CEN/TC 19 “Gaseous and liquid fuels, lubricants and related products of petroleum, synthetic and

6、biological origin” (Secretariat: NEN, Netherlands). The responsible German body involved in its preparation was the Normenausschuss Materialprfung (Materials Testing Standards Committee), Working Committee NA 062-06-32-01 UA Prfung von FAME of the Fachausschuss Minerall- und Brennstoffnormung in col

7、laboration with the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-05-05 AA Gemeinschaftsausschuss fr die Analytik von Fetten, len, Fettprodukten, verwandten Stoffen und Rohstoffen. Amendments This standard

8、 differs from DIN EN 14105:2003-10 as follows: a) the method has been updated to obtain better precision; b) a number of separate internal standards for mono- (C19), di- (C38) and triglycerides (C57) have been included; c) representative internal standards for monoglycerides, diglycerides and trigly

9、cerides are now used in order to avoid using calibration solutions for these families of compounds; d) the integration of the peaks has been improved; e) some evaluation of interference with minor components (i.e. dimers) has been done; f) performance criteria for the gas chromatography column calcu

10、lated with the response factors for the diglyceride and triglyceride internal standards have been introduced. Previous editions DIN EN 14105: 2003-10 2 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 14105 April 2011 ICS 67.200.10 Supersedes EN 14105:2003English Version Fat and oil derivatives

11、- Fatty Acid Methyl Esters (FAME) - Determination of free and total glycerol and mono-, di-, triglyceride contents Produits drivs des corps gras - Esters mthyliques dacides gras (EMAG) - Dtermination de la teneur en glycrols libre et total et en mono-, di- et triglycrides Erzeugnisse aus pflanzliche

12、n und tierischen Fetten und len - Fettsure-Methylester (FAME) - Bestimmung des Gehaltes an freiem und Gesamtglycerin und Mono-, Di- und Triglyceriden This European Standard was approved by CEN on 10 March 2011. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate

13、 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-CENELEC Management Centre or to any CEN member. This European Stand

14、ard 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-CENELEC Management Centre has the same status as the official versions. CEN members are the nati

15、onal standards bodies of Austria, Belgium, Bulgaria, Croatia, 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, Switzer

16、land 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 exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN

17、 14105:2011: EEN 14105:2011 (E) 2 Contents Page Foreword 31 Scope 42 Principle 43 Reagents .44 Apparatus .55 Preparation of solutions .65.1 1,2,4-Butanetriol stock solution, 1 mg/ml .65.2 Glycerol stock solution, 0,5 mg/ml 65.3 Standard glycerides stock solution, 2,5 mg/ml 65.4 Commercial mixture of

18、 monoglycerides .65.5 Calibration solutions .66 Sampling .77 Procedure .77.1 Operating conditions .77.2 Analysis of the calibration solutions .77.3 Analysis of the commercial mixture of monoglycerides .77.4 Preparation and analysis of the samples 87.5 Identification .87.6 Calibration 87.7 Column per

19、formance control .88 Determination of results .98.1 Integration of the peaks 98.2 Glycerol calibration function 98.3 Free glycerol .98.4 Glycerides 108.5 Total glycerol . 109 Expression of results . 1010 Precision 1010.1 Interlaboratory test . 1010.2 Repeatability 1110.3 Reproducibility 1111 Test re

20、port . 11Annex A (informative) Sample chromatogram 12Annex B (normative) Calibration function calculation . 16Annex C (informative) Worked example. 18Annex D (informative) Results of the interlaboratory trial . 19Bibliography . 21DIN EN 14105:2011-07 EN 14105:2011 (E) 3 Foreword This document (EN 14

21、105:2011) 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 European Standard shall be given the status of a national standard, either by publica

22、tion 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 elements of this document may be the subject of patent rights. CEN and/or CENELEC sh

23、all not be held responsible for identifying any or all such patent rights. This document supersedes EN 14105:2003. The main modifications of the standard are: - the utilization of representative internal standards for monoglycrides, diglycerides and triglycerides in order to avoid using calibration

24、solutions for these families of compounds; - the introduction of a performance criteria for the gas chromatography column calculated with the response factors for the diglyceride and triglyceride internal standards. The method has been updated to obtain better precision in general, needed for the li

25、mits required by European FAME specifications for automotive use 1. This has been done by introducing separate internal standards for mono- (C19), di- (C38) and triglycerides (C57). Next an improvement of the integration has been incorporated and some evaluation of interference with minor components

26、 (i.e. dimers) has been done. Via a new Round Robin study, improvement of the precision of free glycerol and diglyceride measurement has been proven. The precision statement of the former standard could be confirmed for triglyceride determination, but no improvement was made. According to the CEN/CE

27、NELEC 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, Lithua

28、nia, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. DIN EN 14105:2011-07 EN 14105:2011 (E) 4 1 Scope The purpose of this European Standard is to determine the free glycerol and residual mono-, di- and triglyceride

29、 contents in fatty acid methyl esters (FAME) intended for addition to mineral oils. The total glycerol content is then calculated from the obtained results. Under the conditions described, the quantification limits are 0,001 % (m/m) for free glycerol, 0,10 % (m/m) for all glycerides (mono-, di- and

30、tri-). This method is suitable for FAME prepared from rapeseed, sunflower, soybean, palm, animal oils and fats and mixture of them. It is not suitable for FAME produced from or containing coconut and palm kernel oils derivatives because of overlapping of different glyceride peaks. NOTE For the purpo

31、ses of this European Standard, the term “% (m/m)” is used to represent respectively the mass fraction. WARNING The use of this method may involve hazardous equipment, materials and operations. This method does not purport to address to all of the safety problems associated with its use, but it is th

32、e 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 Transformation of the glycerol and of the mono- and diglycerides into more volatile and stable silyl derivatives in presenc

33、e of pyridine and of N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA). Analysis of the sample after silylation, by gas chromatography on a short capillary column with thin film thickness, with an on-column injector or equivalent device, and flame ionization detection. After a calibration procedur

34、e, the quantification of glycerol is carried out in presence of the internal standard 1,2,4-butanetriol. Mono-, di- and triglycerides are directly evaluated in presence of an internal standard for each glyceride category: glyceryl monononadecanoate (Mono C19) for monoglycerides; glyceryl dinonadecan

35、oate (Di C38) for diglycerides; glyceryl trinonadecanoate (Tri C57) for triglycerides. 3 Reagents Use only reagents of recognized analytical grade, unless otherwise specified. 3.1 N-methyl-N-trimethysilyltrifluoroacetamide (MSTFA) 3.2 Pyridine, max. 0,1 % water, stored on molecular sieve NOTE Pyridi

36、ne silyl grade (3.10) can also be used. 3.3 Tetrahydrofurane (THF) 3.4 n-Heptane 3.5 Glycerol DIN EN 14105:2011-07 EN 14105:2011 (E) 5 3.6 1,2,4-Butanetriol 3.7 1 - Glyceryl monononadecanoate (Mono C19)1)3.8 1-3 Glyceryl dinonadecanoate (Di C38)2)3.9 Glyceryl trinonadecanoate (Tri C57)3)3.10 Pyridin

37、e, silyl grade 4 Apparatus Usual laboratory apparatus and, in particular, the following. 4.1 Gas chromatograph, equipped with an on-column injector or equivalent device, a temperature-programmable oven and a flame ionization detector. 4.2 Capillary column, capable of being programmed up to 400 C (“h

38、igh temperature“ type) for which the following characteristics are advised: 100 % dimethylpolysiloxane or 95 % dimethyl-5 % diphenylpolysiloxane stationary phase; length 15 m; internal diameter 0,32 mm; film thickness 0,1 m. 4.3 Volumetric flask, 50 ml capacity 4.4 Volumetric flasks, 20 ml capacity

39、4.5 Volumetric flasks, 10 ml capacity 4.6 Screw-cap vials with PTFE-faced septa, 10 ml capacity 4.7 Precision pipette, 1 ml capacity 4.8 Microsyringe, 100 l capacity 4.9 Microsyringe, 500 l capacity 4.10 Microsyringe, 5 l or 10 l capacity specially designed for on-column operation 4.11 Graduated cyl

40、inder, 10 ml capacity 4.12 Analytical balance, with an accuracy of 0,1 mg 1)Monononadecaoin available from Larodan, ref. 31-1900-11 (www.larodan.se) 2)1,3-dinonadecaoin available from Larodan, ref. 32-1903-8 (www.larodan.se) 3)Trinonadecaoin available from Larodan, ref. 33-1900-13 (www.larodan.se),

41、or from Sigma , ref. T4632-1G () DIN EN 14105:2011-07 EN 14105:2011 (E) 6 4.13 Carrier gas, hydrogen or helium 4.14 Auxiliary gases, such as air, hydrogen and nitrogen 5 Preparation of solutions 5.1 1,2,4-Butanetriol stock solution, 1 mg/ml Accurately weigh approximately 50 mg (accuracy 0,1 mg) of 1

42、,2,4-butanetriol (3.6) in a 50 ml volumetric flask (4.3) and make up to the mark with pyridine (3.2). 5.2 Glycerol stock solution, 0,5 mg/ml Accurately weigh approximately 50 mg (accuracy 0,1 mg) of glycerol (3.5) in a 10 ml volumetric flask (4.5) and make up to the mark with pyridine (3.2). Using a

43、 pipette (4.7), transfer 1 ml of this solution into a 10 ml volumetric flask (4.5) and make up to the mark with pyridine (3.2). 5.3 Standard glycerides stock solution, 2,5 mg/ml For each reference glyceride, monononadecanoate (3.7), dinonadecanoate (3.8) and trinonadecanoate (3.9), accurately weigh

44、approximately 50 mg (accuracy 0,1 mg) in a unique 20 ml volumetric flask (4.4) and make up to the mark with tetrahydrofurane (3.3). The solution shall be perfectly limpid at ambient temperature. After storage in refrigerator at 4 C the solution might show a precipitate that must re-dissolve spontane

45、ously when restored at ambient temperature, without any external heating. NOTE If stored at 4 C the solution is stable for almost 3 months. 5.4 Commercial mixture of monoglycerides Made up of mono-palmitoylglycerol (monopalmitin), mono-stearoylglycerol (monostearin) and of mono-oleoylglycerol (monoo

46、lein), present in quantities having an identical mass. Prepare a stock solution of this mixture by weighing approximately 100 mg in a 10 ml volumetric flask (4.5) and make up to the mark with pyridine (3.2). This solution may be used to locate the relevant peaks in GC paths. 5.5 Calibration solution

47、s Prepare four calibration solutions by transferring into a series of vials (4.6) the volumes of stock solutions of glycerol (5.2) and of 1,2,4-butanetriol (5.1) given in Table 1, using the 100 l microsyringes (4.8). Do not use syringe at maximum capacity, but dispense the half volume twice (i.e.: i

48、n case of 100 I dosing using a 100 I syringe, load 50 I twice). Make sure that needle and body of the syringe are free from air bubbles, and measure volumes only by difference (i.e.: when dispensing 80 I, fill syringe up to 100 I and supply solution up to the 20 I mark). DIN EN 14105:2011-07 EN 1410

49、5:2011 (E) 7 Table 1 Preparation of calibration solutions Stock solution 1 2 3 4 Syringe l l l l l glycerol solution (5.2) 10 40 70 100 100 internal butanetriol sol. (5.1) 80 80 80 80 100 6 Sampling Samples shall be taken in accordance with the requirements of national standards or regulations for the sampling of the product under test. A recommended sampling method is given in EN ISO 5555 2 or EN ISO 3170