1、Foodstuffs Determination of acrylamide in food and coffee by gas chromatographymass spectrometry (GCMS)PD CEN/TS 17083:2017BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06TECHNICAL SPECIFICATION SPCIFICATION TECHNIQUE TECHNISCHE SPEZIFIKATION CEN/TS 17083 June 2017 I
2、CS 67.050; 67.060; 67.140.20; 67.080.20 English Version Foodstuffs - Determination of acrylamide in food and coffee by gas chromatography-mass spectrometry (GC-MS)Produits alimentaires - Dosage par CG-SM des produits alimentaires et caf avec GC-MS Lebensmittel - Bestimmung von Acrylamid in Lebensmit
3、teln und Kaffee mit Gaschromatographie-Massenspektrometrie (GC-MS) This Technical Specification (CEN/TS) was approved by CEN on 14 May 2017 for provisional application. The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
4、submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard. CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available promptly at national level in an appropriate form. It is p
5、ermissible to keep conflicting national standards in force (in parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Es
6、tonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTE
7、E 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 form and by any means reserved worldwide for CEN national Members. Ref. No. CEN/TS 17083:2017 ENational fore
8、wordThis Published Document is the UK implementation of CEN/TS 17083:2017.The UK participation in its preparation was entrusted to Technical Committee AW/275, Food analysis - Horizontal methods.A list of organizations represented on this committee can be obtained on request to its secretary.This pub
9、lication 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 Limited 2017ISBN 978 0 580 80854 8ICS 67.080.20; 67.060; 67.050; 67.140.20Compliance with a British St
10、andard cannot confer immunity from legal obligations. This Published Document was published under the authority of the Standards Policy and Strategy Committee on 31 August 2017.Amendments/corrigenda issued since publicationDate Text affectedPUBLISHED DOCUMENTPD CEN/TS 17083:2017TECHNICAL SPECIFICATI
11、ON SPCIFICATION TECHNIQUE TECHNISCHE SPEZIFIKATION CEN/TS 17083 June 2017 ICS 67.050; 67.060; 67.140.20; 67.080.20 English Version Foodstuffs - Determination of acrylamide in food and coffee by gas chromatography-mass spectrometry (GC-MS)Produits alimentaires - Dosage par CG-SM des produits alimenta
12、ires et caf avec GC-MS Lebensmittel - Bestimmung von Acrylamid in Lebensmitteln und Kaffee mit Gaschromatographie-Massenspektrometrie (GC-MS) This Technical Specification (CEN/TS) was approved by CEN on 14 May 2017 for provisional application. The period of validity of this CEN/TS is limited initial
13、ly to three years. After two years the members of CEN will be requested to submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard. CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the
14、CEN/TS available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached. CEN members are the national standards bodies
15、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, Romania, Serbia, Slovakia, Sloveni
16、a, 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 form and by any means reserved wo
17、rldwide for CEN national Members. Ref. No. CEN/TS 17083:2017 EPD CEN/TS 17083:2017CEN/TS 17083:2017 (E) 2 Contents Page European foreword . 3 1 Scope 4 2 Normative references 4 3 Principle . 4 4 Reagents . 4 5 Standard solution preparation 6 6 Apparatus . 7 7 Procedure. 8 8 GC-MS analysis . 10 9 Cal
18、culating and reporting . 12 10 Test report 12 11 Precision data 13 Annex A (informative) Typical chromatograms 14 Annex B (informative) GC-MS troubleshooting 16 Annex C (informative) Single laboratory validation 18 Annex D (informative) Interlaboratory trial precision data . 20 Bibliography . 22 PD
19、CEN/TS 17083:2017CEN/TS 17083:2017 (E) 3 European foreword This document (CEN/TS 17083:2017) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN. Attention is drawn to the possibility that some of the elements of this docu
20、ment may be the subject of patent rights. CEN 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. Annexes A, B, C and D are informative. WARNING 1
21、 The use of this Technical Specification can involve hazardous materials, operations 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 an
22、d determine the applicability of regulatory limitations prior to use. WARNING 2 Some precaution is required when using polyacrylamide-based plastics because acrylamide may leach from these materials. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the follo
23、wing countries are bound to announce this Technical Specification: 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
24、ds, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. PD CEN/TS 17083:2017CEN/TS 17083:2017 (E) 4 1 Scope This Technical Specification specifies a method for the determination of acrylamide in cereal-based products, potato-based
25、 products and coffee by gas-chromatography mass spectrometry (GC-MS). The method has been single-laboratory validated via the analysis of spiked samples (French fries (uncooked), bread, water biscuit, infant cereal, biscuit, green coffee, roast coffee and instant coffee), ranging from 30 g/kg to 1 5
26、00 g/kg acrylamide. The results from the single laboratory validation were obtained by a laboratory with significant experience in acrylamide analysis. In addition, this method has also been studied by inter laboratory trial via the analysis of samples containing incurred acrylamide, ranging from ap
27、proximately 200 g/kg to 2 000 g/kg. Critical points of the method are identified in 7.5 and Clause 8. 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 cit
28、ed applies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN ISO 3696:1995, Water for analytical laboratory use - Specification and test methods (ISO 3696:1987) 3 Principle The test portion is extracted with hot water and isotopically label
29、led acrylamide is added as internal standard. High-fat samples are defatted with hexane, cleared with Carrez solution and centrifuged. Sample extracts are brominated and extracted with ethyl acetate. Following removal of the ethyl acetate by evaporation, triethylamine is added to partially debromina
30、te, after which a portion of the sample extract is injected onto a GC-MS system for quantification. The chromatographic separation is obtained on a mid-polarity capillary GC column. The acrylamide derivative is ionized at 70 eV and recorded in selected ion monitoring (SIM) mode, and quantified by co
31、mparison with the stable isotopically labelled analogue. 4 Reagents Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995 (electrical conductivity below 0,1 S/cm at 25 C), unless specified otherwise. Standard solutions are preferably prepared gravimetr
32、ically. An analytical balance (6.1) is used for the preparation of both native and stable isotope labelled acrylamide. WARNING Acrylamide has been classified by the International Agency for Research on Cancer (IARC) as probably carcinogenic to humans (see 2). Bromine is very toxic and corrosive and
33、hydrobromic acid is corrosive and hazardous. The prepared brominating solution (see 4.17) shall be considered as very toxic and corrosive. Protective equipment such as laboratory coat, disposable gloves and safety glasses shall be used. All handlings of acrylamide, bromine, hydrobromic acid, bromina
34、ting solution and organic solvents shall be performed in a fume cupboard with adequate air flow. IMPORTANT Dispose chemical waste according to applicable environmental rules and regulations. Bromine is very toxic to aquatic organisms hence discharge into the environment shall be avoided. Waste shall
35、 be disposed of appropriately. 4.1 Acrylamide, purity 99 %. PD CEN/TS 17083:2017CEN/TS 17083:2017 (E) 5 4.2 1,2,3-13C3-acrylamide, (acrylamide-13C3) isotopic 13C purity 99 %, supplied as 1 000 g/ml solution1)It is permissible to use deuterated acrylamide (acrylamide-d3) as an alternative internal st
36、andard. In the following sections of the procedure, MS detection and calculation are prescribed for acrylamide-13C3only. 4.3 Bromine, purity 99 %. 4.4 Ethyl acetate, MS grade. 4.5 Helium purified compressed gas, (purity equivalent to 99,995 % or better). 4.6 n-Hexane, MS grade. 4.7 Hydrobromic acid,
37、 48 % in water, assay 99,9 %. 4.8 Ice, crushed. 4.9 Magnesium sulfate, anhydrous powder. 4.10 Potassium bromide, purity 98,5 %. 4.11 Potassium hexacyanoferrate trihydrate (II), K4Fe(CN)6.3H2O, purity 98,5 %. 4.12 Sodium chloride. 4.13 Sodium thiosulfate pentahydrate, Na2S2O35 H2O, purity 99,5 %. 4.1
38、4 Sodium thiosulfate solution, substance concentration c = 1 mol/l. Dissolve 24,82 g of sodium thiosulfate pentahydrate in 100 ml of water. 4.15 Triethylamine, N(C2H5)3, purity 99 %. 4.16 Zinc acetate dihydrate, Zn(CH3COO)22 H2O, purity 98,5 %. 4.17 Brominating solution. Wear gloves and carry out th
39、e following steps in a fume cupboard. Prepare fresh reagent every three months. For preparation of bromine water (saturated), place a 1 l glass bottle containing ca. 400 ml of water in a crushed ice bath and stir with a magnetic stirrer for ca. 10 min. Using a measuring cylinder, add 15 ml 1 ml of b
40、romine (4.3), stopper the flask and continue stirring for at least 60 min. Cap securely and store at 4 C to 6 C. Weigh 400 g 2 g of potassium bromide (4.10) into a 2 l screw-capped glass bottle and add 1 000 ml 5 ml of water. Shake or stir to dissolve then add 20 ml 1 ml of hydrobromic acid (4.7) us
41、ing a measuring cylinder. 1) This is an example of a suitable product available commercially, manufactured by Cambridge Isotope Laboratories Inc, Andover MA, USA. CLM-813-1.2 or similar. This information is given for the convenience of users of this Technical Specification and does not constitute an
42、 endorsement of these products by CEN. Equivalent products may be used if they provide similar results. PD CEN/TS 17083:2017CEN/TS 17083:2017 (E) 6 Using measuring cylinders, add 320 ml 5 ml of saturated bromine water followed by 660 ml 5 ml of water to the glass bottle (6.3) and mix well. Stopper a
43、nd store at 4 C to 6 C. Discard after 3 months. To dispose of excess saturated bromine water, add solid sodium thiosulfate pentahydrate (4.13) and swirl until the brown colour disappears. 4.18 Carrez solution I. Dissolve 10,6 g of potassium hexacyanoferrate trihydrate (II) (4.11) in 100 ml of water.
44、 Discard after 6 months. 4.19 Carrez solution II. Dissolve 21,9 g of zinc acetate dihydrate (4.16) in 100 ml of water. Discard after 6 months. 5 Standard solution preparation 5.1 General Prepare all standard solutions preferably gravimetrically. The tare masses of all recipients and the masses after
45、 each preparation step are recorded and used for calculation of the standard concentrations. Volumetric standard preparation may also be applied provided that the volumetric glassware used complies with EN ISO 1042 (see 1). 5.2 Acrylamide stock standard solution, mass concentration approximately 1 0
46、00 g/ml. Weigh 0,1 g 0,001 g of acrylamide into a 100 ml volumetric flask. Add 30 ml of water, swirl to dissolve and dilute to the mark with water and mix well. The stock solution is stable for 1 month when stored in a refrigerator (4 C to 6 C) and protected from light. Alternatively, commercially-a
47、vailable certified standard solutions may be used if available. 5.3 Acrylamide working standard solution, approximately 1 g/ml. Pipette 100 l of acrylamide stock standard solution (5.2) into a 100 ml volumetric flask, dilute to 100 ml with water and mix well. The solution is stable for 2 weeks when
48、stored in a refrigerator (4 C to 6 C) and protected from light. 5.4 Acrylamide spiking solution, approximately 100 g/ml. Pipette 10 ml of acrylamide stock solution (5.2) into a 100 ml volumetric flask, dilute to 100 ml with water and mix well. The solution is stable for 2 weeks when stored in a refr
49、igerator (4 C to 6 C) and protected from light. 5.5 13C3-Acrylamide internal standard solution, approximately 20 g/ml. Pipette 200 l of 13C3-acrylamide stock standard solution (4.2) into a 10 ml volumetric flask dilute to 10 ml with water. The solution is stable for 2 weeks when stored in a refrigerator (4 C to 6 C) and protected from light. 5.6 Calibration standards Prepare calibration standards of approximately 0 g/l, 1 g/l, 3 g/l, 5 g/l, 10 g/l, 20 g/l and 100 g/l according to the fo
