DIN EN 16619-2015 Food analysis - Determination of benzo[a]pyrene benz[a]anthracene chrysene and benzo[b]fluoranthene in foodstuffs by gas chromatography mass spectrometry (GC-MS) .pdf

1、June 2015 Translation by DIN-Sprachendienst.English price group 16No 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、7.050!%CZb“2325563www.din.deDDIN EN 16619Food analysis Determination of benzoapyrene, benzaanthracene, chrysene andbenzobfluoranthene in foodstuffs by gas chromatography massspectrometry (GC-MS);English version EN 16619:2015,English translation of DIN EN 16619:2015-06Lebensmittelanalytik Bestimmung

3、von Benzoapyren, Benzaanthracen, Chrysen und Benzobfluoranthen inLebensmitteln mit Gaschromatographie und Massenspektrometrie (GC-MS);Englische Fassung EN 16619:2015,Englische bersetzung von DIN EN 16619:2015-06Analyse des produits alimentaires Dosage du benzo(a)pyrne, benzo(a)anthracne, chrysne et

4、benzo(b)fluoranthne dansles denres alimentaires par chromatographie en phase gazeuse couple laspectromtrie de masse (CG-SM);Version anglaise EN 16619:2015,Traduction anglaise de DIN EN 16619:2015-06www.beuth.deDocument comprises 39 pagesIn case of doubt, the German-language original shall be conside

5、red authoritative.06.15DIN EN 16619:2015-06 2 A comma is used as the decimal marker. National foreword This document (EN 16619:2015) has been prepared by Technical Committee CEN/TC 275 “Food analysis Horizontal methods” (Secretariat: DIN, Germany). The responsible German body involved in its prepara

6、tion was the DIN-Normenausschuss Lebensmittel und landwirtschaftliche Produkte (DIN Standards Committee Food and Agricultural Products), Working Committee NA 057-01-14 AA Prozesskontaminanten. EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16619 April 2015 ICS 67.050 English Version Food analy

7、sis - Determination of benzoapyrene, benzaanthracene, chrysene and benzobfluoranthene in foodstuffs by gas chromatography mass spectrometry (GC-MS)Analyse des produits alimentaires - Dosage du benzo(a)pyrne, benzo(a)anthracne, chrysne et benzo(b)fluoranthne dans les denres alimentaires par chromatog

8、raphie en phase gazeuse couple la spectromtrie de masse (CG-SM) Lebensmittelanalytik - Bestimmung von Benzoapyren, Benzaanthracen, Chrysen und Benzobfluoranthen in Lebensmitteln mit Gaschromatographie und Massenspektrometrie (GC-MS) This European Standard was approved by CEN on 7 February 2015. CEN

9、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 on appli

10、cation 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-CENELEC Man

11、agement 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latv

12、ia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, 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 Marn

13、ix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16619:2015 EEN 16619:2015 (E) 2 Contents Page Foreword 3 1 Scope 4 2 Normative references 4 3 Principle 4 4 Reagents .5 5 Standard preparation 9 6 Apparatu

14、s 12 7 Procedure 15 8 GC-MS analysis . 17 9 Calculation and reporting 22 10 Quality control . 22 11 Precision data . 24 12 Test report . 27 Annex A (informative) Typical chromatograms 28 Annex B (informative) Precision data . 30 Annex C (informative) Precision data from single laboratory validation

15、. 35 Bibliography . 37 DIN EN 16619:2015-06 EN 16619:2015 (E) 3 Foreword This document (EN 16619:2015) 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 sta

16、ndard, either by publication of an identical text or by endorsement, at the latest by October 2015 and conflicting national standards shall be withdrawn at the latest by October 2015. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights

17、. CEN and/or CENELEC shall not 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 document can involve hazardous materials, operati

18、ons and equipment. This document 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 t

19、o the CEN-CENELEC 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, Former Yugoslav Republic of Macedonia, France, Germany, Greec

20、e, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 16619:2015-06 EN 16619:2015 (E) 4 1 Scope This European Standard specifies a method for the

21、 determination of 4 of the 16 EU priority polycyclic aromatic hydrocarbons (PAHs), identified as target PAHs. They are benzaanthracene (BaA), benzoapyrene (BaP), benzobfluoranthene (BbF) and chrysene (CHR). The method allows their quantification in the presence of the other 12 EU priority PAHs (benz

22、ojfluoranthene (BjF), cyclopentacdpyrene (CPP), benzokfluoranthene (BkF), dibenza,hanthracene (DhA), benzocfluorene (BcL), dibenzoa,epyrene (DeP), benzoghiperylene (BgP), dibenzoa,hpyrene (DhP), dibenzoa,ipyrene (DiP), dibenzoa,lpyrene (DlP), indeno1,2,3-cdpyrene (IcP), 5-methylchrysene (5MC) in ext

23、ruded wheat flour, smoked fish, dry infant formula, sausage meat, freeze-dried mussels, edible oil and wheat flour, by gas-chromatography mass-spectrometry (GC-MS). The extraction of PAHs from solid samples is performed by pressurized liquid extraction (PLE). Soxhlet extraction was applied by some p

24、articipants in the method validation study by collaborative trial as alternative to PLE. The sample cleanup is performed by applying the following techniques in the reported sequence: size exclusion chromatography (SEC), and solid phase extraction (SPE). This method complies with the performance cha

25、racteristics specified in Commission Regulation (EU) No 836/2011 (see 1). In particular the specifications for the limit of detection (LOD) and of the limit of quantification (LOQ) (0,30 g/kg and 0,90 g/kg respectively) were met. The method has been validated in an interlaboratory study via the anal

26、ysis of both naturally contaminated and spiked samples, ranging from 0,5 g/kg to 11,9 g/kg. However, linearity of the instrument response was proven for the concentration range 0,5 g/kg to 20 g/kg. For the determination of PAHs in edible fats and oils, two other standards are also available, EN ISO

27、22959 and EN ISO 15753, for more information see 2 and 3. 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 undated references, the late

28、st edition of the referenced document (including any amendments) applies. EN ISO 1042:1999, Laboratory glassware - One-mark volumetric flasks (ISO 1042:1998) EN ISO 3696:1995, Water for analytical laboratory use - Specification and test methods (ISO 3696:1987) 3 Principle The sample is homogenized.

29、A test portion is mixed with desiccant, sand and the stable isotope labelled internal standard solution. It is then extracted with n-hexane or cyclohexane by pressurized liquid extraction, or alternatively by Soxhlet extraction. If applicable, co-extracted water is separated from the organic phase o

30、f the extract. The organic extract is evaporated to a small volume, filtered and purified by SEC, using a mixture of ethyl acetate and cyclohexane as eluent. After SEC, 200 l of toluene are added as a keeper to the collected SEC fraction. The SEC fraction is evaporated to about 200 l, and cleaned up

31、 by SPE on silica, using cyclohexane as eluent. The cleaned up sample extract is evaporated again to about 200 l. Finally, an injection standard solution is added to the sample prior to measurement by GC-MS. The injection is performed with a PTV, or split/splitless injection port. The chromatographi

32、c separation is obtained on a mid-polar capillary column with high selectivity for PAHs. The analytes are ionised by electron ionization (EI) at 70 eV. The target PAHs are recorded in Single Ion Monitoring (SIM) mode, and quantified by comparison with the stable isotope labelled analogues. DIN EN 16

33、619:2015-06 EN 16619:2015 (E) 5 4 Reagents 4.1 General Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specified. All reagents and standard solutions shall be stored according to the specifications given by the supplier. The spe

34、cifications given in this procedure for opened commercial solutions or for in-house prepared solutions aim to minimize solvent evaporation and to protect the analytes (PAHs) from degradation. Standard solutions are preferably prepared gravimetrically. Depending on the handled amount of substance a m

35、icro-balance (6.4) and/or an analytical balance (6.5) are used for the preparation of solutions of both native and stable isotope labelled PAHs. All concentrations are expressed as mass per mass. If necessary, the concentrations expressed as mass per volume could be obtained applying the density equ

36、ation (Formula (1). vm= (1) where density (in g/ml); m measured mass of the substance (in g); v volume of the solution (in ml). The density of toluene at 20 C is 0,8669 g/ml. Comprehensive information on the density of solvents at various temperatures is given in 4. All solutions and substances are

37、used at room temperature. WARNING 1 Some PAHs are considered carcinogenic. Persons using this document should be familiar with normal laboratory practices. It is the responsibility of the user of this document to apply practices which are in agreement with applicable occupational safety and health p

38、ractices. WARNING 2 Dispose chemical waste according to applicable environmental rules and regulations. WARNING 3 PAHs are degraded by UV light. Protect PAHs solutions from light (keep in the dark, use aluminium foil or amber glassware). WARNING 4 Some precaution is needed when using plastics as pol

39、ypropylene or PTFE because the analytes may be absorbed onto these materials. 4.2 Helium purified compressed gas (purity equivalent to 99,995 % or better). 4.3 Nitrogen purified compressed gas (purity equivalent to 99,995 % or better). 4.4 Disodium sulfate, (Na2SO4), anhydrous, granular. 4.5 Poly(ac

40、rylic acid), partial sodium salt-graft-poly(ethylene oxide) granular, 90 m to 850 m particle size. 4.6 Sand, 50 mesh to 70 mesh particle size. 4.7 n-Hexane. DIN EN 16619:2015-06 EN 16619:2015 (E) 6 4.8 Acetone. 4.9 Cyclohexane. 4.10 Toluene. 4.11 Ethyl acetate. 4.12 SEC eluent Mix 1 part per volume

41、of cyclohexane (4.9) with 1 part per volume of ethyl acetate (4.11). 4.13 SPE column For the solid phase extraction cleanup, a silica SPE column is used. Commercial cartridges of 500 mg 4 ml or self-filled cartridges of the same size containing 500 mg activated silica are used. The surface area of t

42、he silica should be around 500 m2/g. NOTE Commercial SPE columns made of polypropylene were used in the method validation study by collaborative trial. 4.14 Reference material for quality control A certified reference material, or any other suitable quality control material (e.g left over proficienc

43、y test material) may be applied for this purpose. The CITAC/Eurachem Guide to Quality in Analytical Chemistry may be consulted for guidance, see 5. Analyse this material with every sample batch and use it to control the method performances along time (see 10.4). 4.15 Native reference substances - co

44、mmercially available neat material or solutions of PAHs The list of native substances analysed with this method is provided in Table 1. The target analytes are given in bold font. Commercially available, preferably certified, standard solutions are preferred due to the higher level of safety in hand

45、ling. Triphenylene, benzojfluoranthene, and benzokfluoranthene are potentially interfering with the target analytes and are therefore used for evaluation of selectivity. DIN EN 16619:2015-06 EN 16619:2015 (E) 7 Table 1 Names and structures of the native PAHs NameaCAS number Structure Name aCAS numbe

46、r Structure Benzaanthracene (BaA) 56553 Figure 1 Benzobfluoranthene (BbF) 205992 Figure 2 Benzoapyrene (BaP) 50328 Figure 3 Chrysene (CHR) 218019 Figure 4 Triphenylene (TRP) 217594 Figure 5 Benzojfluoranthene (BjF) 205823 Figure 6 Benzokfluoranthene (BkF) 207089 Figure 7 a The acronym is given in pa

47、renthesis, the target analytes are given in bold. 4.16 Stable isotope labelled reference standards (in the form of commercially available stable isotope labelled PAH solutions) The stable isotope labelled analogues, applied for the quantification of the target PAHs are listed in Table 2. The commerc

48、ial solutions used in the method validation study by collaborative trial contained the stable isotope labelled PAHs at a level of about 100 g/kg in nonane. Preference is given to 13C labelled analogues as their chemical properties best match those of the native analytes. However, alternatively to 13

49、C labelled substances, deuterated analogues of the target analytes may be applied. The concentration levels of these solutions should be similar to the levels specified for the 13C labelled PAH solutions. NOTE 1 Highly deuterated PAHs are separated on the specified GC-column at least partially from their native analogues. NOTE 2 Both forms of benzaanthracene-13C6, which are displayed in Table 2, are equally suitable for the purpose of