EN 16618:2015
(Main)Food analysis - Determination of acrylamide in food by liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS)
Food analysis - Determination of acrylamide in food by liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS)
This European Standard specifies a method for the determination of acrylamide in bakery ware such as bread, toasted bread, crisp bread, butter cookies, and biscuits, as well as potato products such as potato chips, potato crisps, and potato pan cake and roasted coffee, by liquid chromatography in combination with electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS). This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples, ranging from 14,3 µg/kg to 9 083 µg/kg. It was developed at the Swedish National Food Administration and validated in a study organized by the Directorate General Joint Research Centre (DG JRC), Swedish National Food Administration and the Nordic Committee on Food Analysis (NMKL), see [1] and [2].
The limit of quantification (LOQ) depends on the type of instrument used and on the actual performance of the instrument. The majority of the laboratories participating in the validation study were able to determine acrylamide in a butter cookie sample at a level of 14,3 µg/kg. Thus, the validation by interlaboratory study showed that LOQ can be expected to be in the range between below 15 µg/kg and 30 µg/kg.
Lebensmittelanalytik - Bestimmung von Acrylamid in Lebensmitteln mit Flüssigchromatographie und Tandem-Massenspektrometrie (LC-ESI-MS/MS)
Diese Europäische Norm legt ein Verfahren zur Bestimmung von Acrylamid in Backwaren, wie z. B. Brot, getoastetem (geröstetem) Brot, Knäckebrot, Butterkeksen und Keksen, sowie in Kartoffelprodukten, wie z. B. Pommes frites, Kartoffelchips und Kartoffelpuffern, und Röstkaffee durch Flüssigchromatographie in Kombination mit Elektrospray-Ionisation und Tandem-Massenspektrometrie (LC-ESI-MS/MS) fest. Dieses Verfahren wurde in einem Ringversuch mit sowohl natürlich kontaminierten als auch aufgestockten Proben im Bereich von 14,3 µg/kg bis 9 083 µg/kg validiert. Es wurde von der Schwedischen Behörde für Lebensmittelsicherheit (Swedish National Food Administration) entwickelt und in einer Studie validiert, die von der Generaldirektion Gemeinsame Forschungsstelle (Directorate General Joint Research Centre, DG JRC), der Schwedischen Behörde für Lebensmittelsicherheit und dem Nordischen Komitee für Lebensmittelanalytik (Nordic Committee on Food Analysis, NMKL) organisiert wurde, siehe [1] und [2].
Die Bestimmungsgrenze (en: LOQ, limit of quantification) hängt von der Art des eingesetzten Gerätes und dessen tatsächlichem Leistungsvermögen ab. Die Mehrheit der an der Validierungsstudie teilnehmenden Laboratorien war in der Lage, Acrylamid in einer Butterkeksprobe bei einer Konzentration von 14,3 µg/kg zu bestimmen. Die Validierung durch den Ringversuch zeigte daher, dass die Bestimmungsgrenze im Bereich von weniger als 15 µg/kg bis 30 µg/kg erwartet werden kann.
Analyse des produits alimentaires - Dosage de l'acrylamide dans les produits alimentaires par chromatographie en phase liquide couplée à la spectrométrie de masse en tandem (CL-ESI-SM-SM)
Le présent document spécifie une méthode de dosage, par chromatographie en phase liquide en mode ionisation par électrovaporisation couplée à la spectrométrie de masse en tandem (CL-ESI-SM/SM), de l’acrylamide dans les produits de boulangerie tels que le pain, le pain grillé, le pain suédois, les biscuits au beurre et les biscuits épicés, ainsi que dans les produits à base de pomme de terre tels que les frites, les chips et les galettes de pomme de terre et le café torréfié. Cette méthode a été validée lors d’un essai interlaboratoires impliquant l’analyse d’échantillons naturellement contaminés et d’échantillons enrichis, sur une plage de 14,3 µg/kg à 9 083 µg/kg. Elle a été développée au sein de l’Agence suédoise des aliments et validée lors d’un essai organisé par le Centre commun de recherche de la Direction générale (DG JRC), l’Agence suédoise des aliments et le Comité nordique sur l’analyse des aliments (NMKL), voir [1] et [2].La limite de quantification (LOQ) dépend du type d’instrument utilisé et de la performance réelle de l’instrument. La majorité des laboratoires participant à l’essai de validation a pu doser l’acrylamide dans un échantillon de biscuit au beurre à un niveau de 14,3 µg/kg. La validation effectuée lors de l’essai interlaboratoires a montré que ce niveau devait se situer entre une valeur inférieure à 15 µg/kg et 30 µg/kg.
Analize živil - Določevanje akrilamida v živilih s tekočinsko kromatografijo s tandemsko masno spektrometrijo (LC-ESI-MS-MS)
General Information
Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittelanalytik - Bestimmung von Acrylamid in Lebensmitteln mit Flüssigkeitschromatographie und Tandem-Massenspektrometrie (LC-ESI-MS/MS)Analyse des produits alimentaires - Dosage de l'acrylamide dans les produits alimentaires par chromatographie en phase liquide couplée à la spectrométrie de masse en tandem (CL-ESI-SM-SM)Food analysis - Determination of acrylamide in food by liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS)67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 16618:2015SIST EN 16618:2015en,fr,de01-julij-2015SIST EN 16618:2015SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16618
April 2015 ICS 67.050; 67.060; 67.080.20; 67.140.20 English Version
Food analysis - Determination of acrylamide in food by liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS)
Analyse des produits alimentaires - Dosage de l'acrylamide dans les produits alimentaires par chromatographie en phase liquide couplée à la spectrométrie de masse en tandem (CL-ESI-SM-SM)
Lebensmittelanalytik - Bestimmung von Acrylamid in Lebensmitteln mit Flüssigchromatographie und Tandem-Massenspektrometrie (LC-ESI-MS/MS) This European Standard was approved by CEN on 7 February 2015.
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 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 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 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, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre:
Avenue Marnix 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 16618:2015 ESIST EN 16618:2015
Typical chromatograms . 18 Annex B (informative)
Precision data . 20 Bibliography . 24
Figure 1 — Acrylamide WARNING — Acrylamide has been classified by the International Agency for Research on Cancer (IARC) as probably carcinogenic to humans. Protective equipment as laboratory coat, disposable gloves and safety glasses shall be used. All handlings of acrylamide and organic solvents shall be performed in a fume cupboard with adequate air flow. 4.2 Deuterium-labelled acrylamide – acrylamide-2,3,3-D3 (CAS 122775-19-3). The chemical structure is:
Figure 2 — Deuterium-labelled acrylamide Alternatively, 13C-labelled acrylamide (acrylamide-13C3, CAS 287399-26-2) may be used. 4.3 Methanol (CAS 67-56-1). 4.4 Glacial acetic acid (CAS 64-19-7). 4.5 n-Hexane (CAS 110-54-3). Alternatively, cyclohexane (CAS 110-82-7) may be used. 4.6 Eluent for SPE column 2 (5.2.3) Mix 60 parts per volume of methanol (4.3) with 40 parts per volume of water. 4.7 HPLC mobile phase Mix 1 part per volume of glacial acetic acid (4.4) with 1 000 parts per volume of water. 4.8 Stock solutions of acrylamide and acrylamide-2,3,3-D3, mass concentration
= 1 000
Weigh, to the nearest 0,05 mg, approximately 100 mg of acrylamide and acrylamide-2,3,3-D3 respectively into separate 100 ml volumetric flasks, dissolve in water and dilute to 100 ml. Solutions can be stored at 4 °C for at least 3 months. 4.9 Internal standard solution 1,
= 10
Transfer 1 000 -2,3,3-D3 (4.8) to a 100 ml volumetric flask and dilute to the mark with water. SIST EN 16618:2015
= 1 000 ng/ml. Transfer 5 000
internal standard solution 1 (4.9) to a 50 ml volumetric flask and dilute to the mark with water. 4.11 Acrylamide standard solution 1,
= 100
Transfer 5 000 4.8) to a 50 ml volumetric flask and dilute to the mark with water. 4.12 Acrylamide standard solution 2,
= 10
Transfer 5 000
1 (4.11) to a 50 ml volumetric flask and dilute to the mark with water. 4.13 Acrylamide standard solution 3,
= 100 ng/ml. Transfer 1 000
2 (4.12) to a 100 ml volumetric flask and dilute to the mark with water. 4.14 LC-MS calibration solutions Dilute aliquots from standard solutions (4.9), (4.11), (4.12) and (4.13) with water to give calibration solutions of e.g. 0 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml, 50 ng/ml, 100 ng/ml, 250 ng/ml, 500 ng/ml, 1 000 ng/ml, 2 000 ng/ml, 5 000 ng/ml and 10 000 ng/ml respectively of acrylamide, all containing 400 ng/ml of acrylamide-2,3,3-D3. Examples for the preparation of calibration solutions are given in Table 1. Table 2 indicates the relation between calibration solution concentrations and acrylamide contents of food samples. Calibration shall be performed on at least six concentration levels distributed properly over the working range. The analysis of an even higher number of calibration solutions should be analysed if such a broad range of concentrations (0
Table 1 — Preparation of LC-MS calibration solutions Calibration solution ng/ml Volumetric flask ml Internal standard solution (4.9)
Acrylamide standard solution
0 100 4 000 0 5 100 4 000 5 000 of (4.13) 10 100 4 000 10 000 of (4.13) 20 100 4 000 200 of (4.12) 50 100 4 000 500 of (4.12) 100 100 4 000 1 000 of (4.12) 250 100 4 000 2 500 of (4.12) 500 100 4 000 5 000 of (4.12) 1 000 100 4 000 1 000 of (4.11) 2 000 100 4 000 2 000 of (4.11) 5 000 100 4 000 5 000 of (4.11) 10 000 50 4 000 5 000 of (4.11) SIST EN 16618:2015
Roasted coffee
10 10 50 5 Apparatus Usual laboratory glassware and equipment and, in particular, the following: 5.1 LC-MS/MS system 5.1.1 HPLC apparatus, comprising the following: 5.1.1.1 Thermostated column compartment. 5.1.1.2 Injection system, capable of injecting 10
5.1.1.3 Mobile phase pump, capable of maintaining a mobile phase flow of 0,4 ml/min. 5.1.2 HPLC column The stationary phase of the column is graphitized carbon1a), particle size 5
mm x 2,1 mm with a guard column1a), particle size 5
mm x 2 mm. Alternative columns/stationary phases may be applied provided that similar performance to the graphitized carbon column can be demonstrated. 5.1.3 Mass spectrometer Triple quadrupole mass spectrometer operating in positive electrospray and, selected reaction monitoring mode (SRM), set to obtain unit resolution. 5.1.4 Data acquisition and analysis system Suitable data collection and evaluation software. 5.1.5 Divert valve (optional) HPLC valve installed between HPLC column and mass spectrometer in order to direct the HPLC effluent either to waste or to the mass spectrometer, see 7.1.1. 5.2 Solid phase extraction system 5.2.1 Vacuum manifold for solid phase extraction
1) a) Hypercarb™ column, Thermo Hypersil-Keystone® column, b) ISOLUTE ® Multimode SPE column and c
...
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