CEN/TS 17083:2017

Foodstuffs - Determination of acrylamide in food and coffee by gas chromatography-mass spectrometry (GC-MS)

This Technical Specification specifies a method for the determination of acrylamide in cereal-based products, potato-based 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 μg/kg to 1 500 μg/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 approximately 200 μg/kg to 2 000 μg/kg. Critical points of the method are identified in 7.5 and Clause 8.

Lebensmittel - Bestimmung von Acrylamid in Lebensmitteln und Kaffee mit Gaschromatographie-Massenspektrometrie (GC-MS)

Diese Technische Spezifikation legt ein Verfahren zur Bestimmung von Acrylamid in Getreideerzeugnissen, Kartoffelerzeugnissen und Kaffee durch Gaschromatographie gekoppelt mit Massen¬spektrometrie (GC MS) fest.
Dieses Verfahren wurde in einer Einzellaborvalidierung durch die Analyse von aufgestockten Proben (Pommes frites (ungegart), Brot, Kräcker, Getreidebeikost, Kekse, Rohkaffee, Röstkaffee und löslicher Kaffee) im Bereich von 30 μg/kg bis 1 500 μg/kg Acrylamid untersucht.
Die Ergebnisse der Einzellaborvalidierung wurden von einem Labor mit umfangreicher Erfahrung in der Acrylamid Analytik erzielt. Außerdem wurde dieses Verfahren auch in einem Ringversuch durch Untersuchung von Proben mit einem Anteil an entstandenem Acrylamid von etwa 200 μg/kg bis 2 000 μg/kg untersucht. Kritische Punkte des Verfahrens werden in 7.5 und Abschnitt 8 benannt.

Produits alimentaires - Dosage par CG-SM des produits alimentaires et café avec GC-MS

Živila - Določevanje akrilamida v živilih in kavi s plinsko kromatografijo/masno spektrometrijo (GC-MS)

Ta tehnična specifikacija opredeljuje metodo za določanje akrilamida v žitnih proizvodih, izdelkih iz krompirja in kavi s plinsko kromatografijo/masno spektrometrijo (GC-MS).
Metoda je bila validirana v laboratoriju z analizo primešanih vzorcev (krompirček, (nekuhan), kruh, krekerji, žita za dojenčke, piškoti, surova kava, pražena in instant kava) v razponu 30–1500 μg/kg akrilamida.
Rezultati validacije laboratorija so bili pridobljeni v laboratoriju z bogatimi izkušnjami z analizo akrilamida. Poleg tega je bila ta metoda raziskana tudi z medlaboratorijskim preskusom z analizo vzorcev, ki so vsebovali akrilamid v razponu približno 200–2000 μg/kg. Kritične točke metode so navedene v točkah 7.5 in 8.

General Information

• Status: Published
• Publication Date: 06-Jun-2017
• ICS: 67.050 - General methods of tests and analysis for food products
• ICS: 67.060 - Cereals, pulses and derived products
• ICS: 67.080.20 - Vegetables and derived products
• ICS: 67.140.20 - Coffee and coffee substitutes
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 13 - Neoformed contaminants
• Current Stage: 9060 - Closure of 2 Year Review Enquiry - Review Enquiry
• Start Date: 02-Sep-2024
• Completion Date: 02-Sep-2024
• Directive: 882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
• Mandate: M/463 - Mandate for Standardisation addressed to CEN for the methods of analysis for food contaminants
• Ref Project: SIST-TS CEN/TS 17083:2017 - Foodstuffs - Determination of acrylamide in food and coffee by gas chromatography-mass spectrometry (GC-MS)

Standards Content (Sample)

SLOVENSKI STANDARD
01-september-2017
äLYLOD'RORþHYDQMHDNULODPLGDYåLYLOLKLQNDYLVSOLQVNRNURPDWRJUDILMRPDVQR
VSHNWURPHWULMR*&06
Foodstuffs - Determination of acrylamide in food and coffee by gas chromatography-
mass spectrometry (GC-MS)
Lebensmittel - Bestimmung von Acrylamid in Lebensmitteln und Kaffee mit
Gaschromatographie-Massenspektrometrie (GC-MS)
Produits alimentaires - Dosage par CG-SM des produits alimentaires et café avec GC-
MS
Ta slovenski standard je istoveten z: CEN/TS 17083:2017
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN/TS 17083
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
June 2017
TECHNISCHE SPEZIFIKATION
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 Lebensmittel - Bestimmung von Acrylamid in
alimentaires et café avec GC-MS 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 initially 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 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 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, 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
© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 17083:2017 E
worldwide for CEN national Members.

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 Calculating 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
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 document 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 — 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 and 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
following 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,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
1 Scope
This Technical Specification specifies a method for the determination of acrylamide in cereal-based
products, potato-based 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 μg/kg to 1 500 μg/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 approximately 200 μg/kg
to 2 000 μg/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 cited 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 labelled 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 debrominate, 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 comparison 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 µS/cm at 25 °C), unless specified otherwise.
Standard solutions are preferably prepared gravimetrically. 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 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, brominating 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 be disposed of appropriately.
4.1 Acrylamide, purity ≥ 99 %.
13 13 13
4.2 [1,2,3- C ]-acrylamide, (acrylamide- C ) isotopic C purity 99 %, supplied as 1 000 µg/ml
3 3
1)
solution
It is permissible to use deuterated acrylamide (acrylamide-d ) as an alternative internal standard. In
the following sections of the procedure, MS detection and calculation are prescribed for acrylamide- C
only.
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, 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), K Fe(CN) .3H O, purity ≥ 98,5 %.
4 6 2
4.12 Sodium chloride.
4.13 Sodium thiosulfate pentahydrate, Na S O ·5 H O, purity ≥ 99,5 %.
2 2 3 2
4.14 Sodium thiosulfate solution, substance concentration c = 1 mol/l. Dissolve 24,82 g of sodium
thiosulfate pentahydrate in 100 ml of water.
H ) , purity ≥ 99 %.
4.15 Triethylamine, N(C2 5 3
4.16 Zinc acetate dihydrate, Zn(CH COO) •2 H O, purity ≥ 98,5 %.
3 2 2
4.17 Brominating solution.
Wear gloves and carry out the 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 bromine (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) using
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 endorsement of these products by CEN. Equivalent products may be used if they
provide similar results.
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 and 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. 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 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 ρ approximately 1 000 µg/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-available certified standard solutions may be used if available.
5.3 Acrylamide working standard solution, ρ approximately 1 µg/ml.
Pipette 100 µl of acrylamide
...