BS EN 16995-2017 Foodstuffs Vegetable oils and foodstuff on basis of vegetable oils Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons.pdf

1、Foodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) with on-line HPLC-GC-FID analysisBS EN 16995:2017BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013

2、15:06EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16995 June 2017 ICS 67.200.10 English Version Foodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) with on-line HPLC-GC-

3、FID analysisProduits alimentaires - Huiles vgtales et produits alimentaires base dhuiles vgtales - Dosage des hydrocarbures saturs dhuile minrale (MOSH) et des hydrocarbures aromatiques dhuile minrale (MOAH) par analyse par CLHP-CG-FID en ligne Lebensmittel - Pflanzliche le und Lebensmittel auf Basi

4、s pflanzlicher le - Bestimmung von gesttigten Minerall-Kohlenwasserstoffen (MOSH) und aromatischen Minerall-Kohlenwasserstoffen (MOAH) mit on-line HPLC-GC-FID This European Standard was approved by CEN on 10 March 2017. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which

5、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-CENELEC Management Centre or to any CEN member. This Europ

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

7、 the national standards bodies of 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, Rom

8、ania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2017 CEN All rights of exploitation in any fo

9、rm and by any means reserved worldwide for CEN national Members. Ref. No. EN 16995:2017 ENational forewordThis British Standard is the UK implementation of EN 16995:2017.The UK participation in its preparation was entrusted to Technical Committee AW/275, Food analysis - Horizontal methods.A list of

10、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. The British Standards Institution 2017 Published by BSI Standards Limite

11、d 2017ISBN 978 0 580 80862 3ICS 67.200.10Compliance 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 July 2017.Amendments/corrigenda issued since publicationDate Text af

12、fectedBRITISH STANDARDBS EN 16995:2017EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16995 June 2017 ICS 67.200.10 English Version Foodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocar

13、bons (MOAH) with on-line HPLC-GC-FID analysisProduits alimentaires - Huiles vgtales et produits alimentaires base dhuiles vgtales - Dosage des hydrocarbures saturs dhuile minrale (MOSH) et des hydrocarbures aromatiques dhuile minrale (MOAH) par analyse par CLHP-CG-FID en ligne Lebensmittel - Pflanzl

14、iche le und Lebensmittel auf Basis pflanzlicher le - Bestimmung von gesttigten Minerall-Kohlenwasserstoffen (MOSH) und aromatischen Minerall-Kohlenwasserstoffen (MOAH) mit on-line HPLC-GC-FID This European Standard was approved by CEN on 10 March 2017. CEN members are bound to comply with the CEN/CE

15、NELEC 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 on application to the CEN-CENELEC Management Centre

16、 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-CENELEC Management Centre has the same status as the o

17、fficial versions. CEN members are the national standards bodies of 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, Netherlan

18、ds, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2017 CEN All

19、 rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16995:2017 EBS EN 16995:2017EN 16995:2017 (E) 2 Contents Page European foreword . 3 1 Scope 4 2 Normative references 4 3 Terms and definitions . 4 4 Principle . 4 5 Reagents . 5 6 Apparatus

20、. 8 7 Sample storage . 9 8 Preparation of the test sample 10 9 Preparation of the analytical sample 10 10 Liquid chromatography and gas chromatography 12 11 Precision 17 12 Test report 17 Annex A (informative) Examples of chromatograms . 18 Annex B (informative) Precision data . 32 Bibliography . 35

21、 BS EN 16995:2017EN 16995:2017 (E) 3 European foreword This document (EN 16995:2017) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by p

22、ublication of an identical text or by endorsement, at the latest by December 2017, and conflicting national standards shall be withdrawn at the latest by December 2017. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN shall not

23、 be held responsible for identifying any or all such patent rights. This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association. WARNING The use of this standard can involve hazardous materials, operations and equipment. This standa

24、rd does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this document to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. According to the CEN-CENELEC Internal Reg

25、ulations, the national standards organisations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland,

26、Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 16995:2017EN 16995:2017 (E) 4 1 Scope This European Standard specifies a highly efficient method for the determina

27、tion of saturated and aromatic hydrocarbons (from C10 to C50) in vegetable fats and oils and foodstuff on basis of vegetable oils for which it has been interlaboratory validated, with online-HPLC-GC-FID 1, 2 and 3. This standard is not intended to be applied to other matrices. The method can be used

28、 for the analysis of mineral oil saturated hydrocarbons (MOSH) and/or mineral oil aromatic hydrocarbons (MOAH). The method has been tested in an interlaboratory study via the analysis of both naturally contaminated and spiked vegetable oil samples and mayonnaise and margarine samples, ranging from 4

29、 mg/kg to 197 mg/kg for MOSH, and from 2 mg/kg to 51 mg/kg for MOAH. According to the results of the interlaboratory studies, the method has been proven suitable for MOSH and MOAH mass concentrations each above 10 mg/kg. In case of suspected interferences from natural sources, the fossil origin of t

30、he MOSH and MOAH fraction can be verified by examination of the pattern by GC-MS. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For unda

31、ted references, the latest edition of the referenced document (including any amendments) applies. EN ISO 661, Animal and vegetable fats and oils - Preparation of test sample (ISO 661) 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 mineral oi

32、l saturated hydrocarbons MOSH paraffinic (open-chain, usually branched) and naphthenic (cyclic, alkylated) hydrocarbons 3.2 mineral oil aromatic hydrocarbons MOAH aromatic mainly alkylated hydrocarbons 3.3 unresolved complex mixture UCM complex mixture of saturated or aromatic hydrocarbons not resol

33、ved by gas chromatography such as branched paraffins, alkylated naphthenes and alkylated aromatics 4 Principle The fractions of MOSH and MOAH are isolated and separated by an HPLC-GC-FID system. MOSH and MOAH fractions are separated on a silica gel column using a n-hexane/dichloromethane gradient an

34、d each transferred as 450 l fractions to GC using the Y-interface 4, while triglycerides are kept on the HPLC column. Solvent vapours are discharged via a solvent vapour exit located between the uncoated pre-column and the GC separation column. Volatile components are retained by solvent trapping BS

35、 EN 16995:2017EN 16995:2017 (E) 5 applying partially concurrent eluent evaporation. High boiling components spread over the entire length of the flooded zone and are refocused by the retention gap technique 2. The area attributed to mineral oil is calculated by subtraction of sharp peaks due to n-al

36、kanes (naturally occurring hydrocarbons), terpenes, squalene and its isomerization products, sterenes and olefins with the structure of carotenoids. MOSH and MOAH are quantitated by internal standard added before analysis. Verification standards are added for monitoring proper HPLC fractionation and

37、 GC transfer conditions. Some vegetable oils contain odd-numbered n-alkanes in the range of C21-C33 in such quantities that the chromatograms of the MOSH fraction are severely overloaded and that they might overlap with the mineral oil hump. In this case, it is recommended to use an additional clean

38、-up technique. Activated aluminium oxide strongly retains long chain n-alkanes. Mineral oil which contaminates edible oil almost exclusively consists of branched and cyclic components which are not retained by activated aluminium oxide. Therefore, the use of activated aluminium oxide enables the rem

39、oval of plant paraffins. Epoxidation is a purification step that is necessary for the quantification of MOAH. This purification step allows the elimination of olefins like squalene, which elute within the MOAH fraction and interfere with quantification (e.g. olive oil, palm oil). Epoxidation also re

40、moves certain olefins co-eluting with the MOSH fraction, therefore epoxidation also may be used as a purification step for the MOSH fraction. Since now, the epoxidation step is the best compromise to remove olefins even though it is not fully quantitative and the efficiency may be sample dependent.

41、Depending on the sample, this reaction may induce the epoxidation of a part of the MOAH or incomplete removal of the interfering olefins. 5 Reagents Unless otherwise specified, use only reagents of recognized analytical grade. 5.1 Silica Gel 601), extra pure for column chromatography with particle s

42、ize from 60 m to 200 m (70 mesh to 230 mesh) in glass bottle to prevent contamination. 5.2 Silica Gel 60, activated. Condition silica gel (5.1) in an oven for at least 16 h at 400 C. 5.3 Demineralized water. 5.4 Anhydrous sodium sulfate, analytical grade, purity 99 %. 5.5 n-Hexane, trace organic ana

43、lysis grade, for pesticide residue analysis. n-Hexane purity can be checked by concentrating 30 ml of n-hexane mixed with 25 l of internal standard solution (5.21) and 2 drops of bis(2-ethylhexyl) maleate (5.29) using a rotary evaporator, dissolving the residue in 0,2 ml of n-hexane and the analysis

44、 of 50 l by online-HPLC-GC-FID (6.8). Take care that in the evaporation step the residue is not evaporated to dryness to avoid loss of volatile hydrocarbons. The signal abundance of the residue after evaporation should not exceed a tenth of the signal abundance obtained at the quantification limit.

45、1) Silica gel is available from Merck, reference 7754 or 7734 (www.merck-). It is an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent produc

46、ts may be used if they can be shown to lead to the same results. BS EN 16995:2017EN 16995:2017 (E) 6 5.6 Dichloromethane (DCM), trace organic analysis grade, purity 99 %. DCM purity can be checked by concentrating 50 ml of DCM mixed with 25 l of internal standard solution (5.21) and 2 drops of bis(2

47、-ethylhexyl) maleate (5.29) using a rotary evaporator, dissolving the residue in 0,2 ml of n-hexane and the analysis of 50 l by online-HPLC-GC-FID (6.8). Take care that in the evaporation step the residue is not evaporated to dryness to avoid loss of volatile hydrocarbons. The signal abundance of th

48、e residue after evaporation should not exceed a fifth of the signal abundance obtained at the quantification limit. 5.7 Dichloromethane solution. Mix 30 ml DCM (5.6) with n-hexane (5.5) up to a volume of 100 ml. The solution should be freshly prepared daily. 5.8 Toluene. 5.9 1,1,2-Trichloroethane. 5

49、.10 Perylene (Per), purity 99 %. 5.11 5-Cholestane (Cho), purity 97 %. 5.12 n-Undecane (n-C11), purity 98 %. 5.13 n-Tridecane (n-C13), purity 97 %. 5.14 Tri-tert-butylbenzene (TBB). 5.15 Bicyclohexyl (CyCy), purity 99 %. 5.16 1-Methylnaphthalene (1-MN), purity 95 %. 5.17 2-Methylnaphthalene (2-MN), purity 97 %. 5.18 Pentylbenzene (PB), purity 96 %. 5.19 Stock solutions, mass concentration = 10 mg/ml. Prepare individual stock solutions by weighing, to the nearest 1 mg, 100 mg of n-C11 (