1、BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06Foodstuffs Determination of benzene in soft drinks, other beverages and vegetable-based infant foods by headspace gas chromatography mass spectrometry (HS-GC-MS)BS EN 16857:2017EUROPEAN STANDARD NORME EUROPENNE EUROPISC
2、HE NORM EN 16857 May 2017 ICS 67.160.20; 67.230 English Version Foodstuffs - Determination of benzene in soft drinks, other beverages and vegetable-based infant foods by headspace gas chromatography mass spectrometry (HS-GC-MS) Produits alimentaires - Dtermination de la teneur en benzne dans les boi
3、ssons non alcoolises, les autres boissons et les aliments pour nourrissons base de lgumes par chromatographie en phase gazeuse avec espace de tte couple la spectromtrie de masse (HS CG-SM) Lebensmittel - Bestimmung von Benzol in Erfrischungsgetrnken, anderen Getrnken und in Babynahrung auf Gemsebasi
4、s mit Headspace-Gaschromatographie/Massenspektrometrie (HS-GC-MS) This European Standard was approved by CEN on 20 February 2017. 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 standa
5、rd 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 Standard exists in three official versions (English, French, German). A version in any
6、 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 national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Repub
7、lic, 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, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. E
8、UROPEAN 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 worldwide for CEN national Members. Ref. No. EN 16857:2017 EN
9、ational forewordThis British Standard is the UK implementation of EN 16857:2017.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.Th
10、is 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 Limited 2017ISBN 978 0 580 80858 6ICS 67.230; 67.160.20Compliance with a British Standard cannot
11、 confer immunity from legal obligations. This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 June 2017.Amendments/corrigenda issued since publicationDate Text affected BRITISH STANDARDBS EN 16857:2017EUROPEAN STANDARD NORME EUROPENNE EUROPISCH
12、E NORM EN 16857 May 2017 ICS 67.160.20; 67.230 English Version Foodstuffs - Determination of benzene in soft drinks, other beverages and vegetable-based infant foods by headspace gas chromatography mass spectrometry (HS-GC-MS) Produits alimentaires - Dtermination de la teneur en benzne dans les bois
13、sons non alcoolises, les autres boissons et les aliments pour nourrissons base de lgumes par chromatographie en phase gazeuse avec espace de tte couple la spectromtrie de masse (HS CG-SM) Lebensmittel - Bestimmung von Benzol in Erfrischungsgetrnken, anderen Getrnken und in Babynahrung auf Gemsebasis
14、 mit Headspace-Gaschromatographie/Massenspektrometrie (HS-GC-MS) This European Standard was approved by CEN on 20 February 2017. 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 standar
15、d 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 Standard exists in three official versions (English, French, German). A version in any
16、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 national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republ
17、ic, 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, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EU
18、ROPEAN 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 worldwide for CEN national Members. Ref. No. EN 16857:2017 EBS
19、 EN 16857:2017EN 16857:2017 (E) 2 Contents Page European foreword . 3 1 Scope 4 2 Normative references 4 3 Principle . 4 4 Reagents . 4 5 Apparatus . 6 6 Procedure. 8 7 HS-GC-MS analysis . 8 8 Calculation 11 9 Precision data 11 10 Test report 12 Annex A (informative) Typical chromatograms 13 Annex B
20、 (informative) Precision data . 14 Bibliography . 15 BS EN 16857:2017EN 16857:2017 (E) 3 European foreword This document (EN 16857:2017) 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
21、given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by November 2017, and conflicting national standards shall be withdrawn at the latest by November 2017. Attention is drawn to the possibility that some of the elements of this documen
22、t 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 Europan Free Trade Association. WARNING The use of this standard can involve hazardo
23、us materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to take appropriate measures for ensuring the safety and health of the personnel prior to application of the standa
24、rd and to fulfil statutory requirements for this purpose. Benzene has been classified by IARC as carcinogenic to humans (see 1). According to the CEN-CENELEC Internal Regulations, the national standards organisations of the following countries are bound to implement this European Standard: Austria,
25、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, Slovenia, Spain, Sw
26、eden, Switzerland, Turkey and the United Kingdom. BS EN 16857:2017EN 16857:2017 (E) 4 1 Scope This European Standard specifies a method for the determination of benzene in soft drinks, other beverages and vegetable-based infant foods, by headspace gas chromatography mass spectrometry (HS-GC-MS). The
27、 method has been validated in an interlaboratory study via the analysis of spiked samples of carbonated soft drink, still fruit-based drink, carbonated fruit-based drink, vegetable and fruit juice containing carrot, infant food vegetable based and infant food containing meat, ranging from 1,9 g/kg t
28、o 18,6 g/kg. However, linearity of the instrument response was proven for the concentration range from 0,5 g/kg to 20 g/kg. The limit of quantification (LOQ) depends on the instrument but can generally be expected to be in the range from 0,5 g/kg to 1,0 g/kg. 2 Normative references The following doc
29、uments, 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 latest edition of the referenced document (including any amendments) applies. EN ISO 3696:1995, Water
30、for analytical laboratory use - Specification and test methods (ISO 3696:1987) 3 Principle The sample is homogenized, a test portion is heated in a closed system with isotopically-labelled benzene added as internal standard. A portion of the headspace is injected into a GC-MS system for identificati
31、on and quantification. The injection is performed with a split-splitless injection port. The chromatographic separation is obtained on a mid-polarity capillary GC column. The benzene is ionized at 70 eV, recorded in selected ion monitoring (SIM) mode, and quantified by comparison with the stable iso
32、tope labelled analogue. 4 Reagents Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specified. Prepare standard solutions preferably gravimetrically. Use an analytical balance (5.1) for the preparation of both native and stable i
33、sotope labelled benzene standard solutions. 4.1 Benzene, C6H6, purity is 99,0 % (CAS 71-43-2). While not necessary, it is recommended to store at 4 C to 6 C to prevent evaporation. Bring to room temperature before use. 4.2 Benzene-d6, purity is 99, 6 atom % D (CAS 1076-43-3). While not necessary, it
34、 is recommended to store at 4 C to 6 C to prevent evaporation. Bring to room temperature before use. 4.3 Helium purified compressed gas, purity equivalent to 99,995 % or better. 4.4 Methanol, suitable for headspace GC analysis (CAS 67-56-1). 4.5 Preparation of stock and standard solutions 4.5.1 Gene
35、ral Prepare all standard solutions preferably gravimetrically. Record the tare masses of all recipients and the masses after each preparation step and use for calculation of the mass concentrations of standard solutions. BS EN 16857:2017EN 16857:2017 (E) 5 Volumetric preparation of standard solution
36、s may also be applied provided that the volumetric glassware used complies with EN ISO 1042 (see 2). 4.5.2 Benzene stock solution, mass concentration approximately 2,20 mg/ml Pipette 20 ml 0,1 ml of methanol into a 20 ml headspace vial (5.17) and seal the vial with a crimp cap. Weigh the sealed vial
37、 to the nearest 0,1 mg and record the weight as W1. Using a microsyringe, transfer 50 l of benzene (4.1) through the septum of the vial containing the methanol and shake vigorously or vortex to mix. Reweigh the sealed vial and record the weight (W2) to the nearest 0,1 mg. Subtract W1from W2to determ
38、ine the weight of benzene transferred (W3). Calculate the mass concentration of stock solution from W3divided by the total volume (20,05 ml). Alternatively, commercially available certified standard solutions may be used if available. Store the benzene stock solution at 4 C to 6 C but allow it to at
39、tain ambient temperature before use. Discard after 2 weeks. Once the septum on the stock solution has been pierced, the cap shall be replaced. 4.5.3 Benzene intermediate standard solution, approximately 54 g/ml Using a microsyringe, transfer 500 l of benzene stock solution (4.5.2) to a sealed headsp
40、ace vial containing 20 ml 0,1 ml of methanol and shake vigorously. Prepare fresh daily. Calculate the exact mass concentration from the stock solution (4.5.2). 4.5.4 Benzene standard solution, approximately 0,5 g/ml Using a microsyringe, transfer 200 l of benzene intermediate standard solution (4.5.
41、3) to a sealed headspace vial containing 20 ml 0,1 ml of water and shake vigorously. Prepare fresh daily. Calculate the exact concentration from the stock solution (4.5.2). Use this standard solution to prepare a 6-point calibration from 0,5 g/kg to 20 g/kg (see 4.6). 4.5.5 Benzene-d6internal standa
42、rd (IS) stock solution, approximately 2,36 mg/ml Pipette 20 ml 0,1 ml of methanol into a 20 ml headspace vial (5.17) and seal the vial with a crimp cap. Weigh the sealed vial to the nearest 0,1 mg and record the weight as W1. Using a microsyringe, transfer 50 l of benzene-d6(4.2) through the septum
43、of the vial containing the methanol and shake vigorously or vortex. Reweigh the sealed vial and record the weight (W2) to the nearest 0,1 mg. Subtract W1from W2to determine the weight of benzene-d6transferred (W3). Calculate the mass concentration of IS stock solution from W3divided by the total vol
44、ume (20,05 ml). Alternatively, commercially available certified standard solutions may be used if available. Store the benzene-d6IS stock solution at 4 C to 6 C but allow it to attain ambient temperature before use. Discard after 2 weeks. Once the septum on the IS stock solution has been pierced, th
45、e cap shall be replaced. 4.5.6 Benzene-d6internal standard (IS) solution, approximately 4,2 g/ml Using a microsyringe, transfer 36 l of benzene-d6IS stock solution (4.5.5) to a sealed headspace vial containing 20 ml 0,1 ml of water and shake vigorously. Prepare fresh daily. Calculate the exact mass
46、concentration from the benzene-d6IS stock solution (4.5.5). 4.6 Preparation of the calibration solutions Prepare calibration solutions of approximately 0 g/kg, 0,5 g/kg, 1 g/kg, 2,5 g/kg, 5 g/kg, 10 g/kg and 20 g/kg benzene and approximately 10 g/kg IS according to the following scheme. Add 10 g 0,0
47、5 g of water to each of seven 20 ml headspace vials and transfer using a positive displacement pipette benzene standard solution (4.5.4), IS solution (4.5.6)and water according to the values given in Table 1. To avoid loss of benzene, seal the vials immediately following addition of the entire benze
48、ne and benzene-d6solutions and water. BS EN 16857:2017EN 16857:2017 (E) 6 Table 1 Preparation of calibration solutions Mass of water g Volume of 0,5 g/ml benzene (4.5.4) l Volume of 4,2 g/ml benzene-d6(4.5.6) l Volume of water l Benzene mass concentration g/kg Benzene d6mass concentration g/kg 10 0
49、25 400 0 10,1 10 10 25 390 0,5 10,1 10 20 25 380 1,0 10,1 10 50 25 350 2,5 10,1 10 100 25 300 5,0 10,1 10 200 25 200 10 10,1 10 400 25 0 20 10,1 5 Apparatus WARNING All glassware shall be meticulously cleaned (except disposable glassware). Usual laboratory glassware and equipment and, in particular, the following: 5.1 Analytical balance, capable of weighing to the nearest of 0,0 001 g. 5.2 Top pan balance, capable of weighing to the nearest 0,001 g. 5.3 Headspace gas chro
