prEN 16987
(Main)Foodstuffs - Determination of acrylamide in coffee and coffee products by HPLC-MS/MS and GC-MS
Foodstuffs - Determination of acrylamide in coffee and coffee products by HPLC-MS/MS and GC-MS
This standard specifies methods for the determination of acrylamide in coffee and coffee products by extraction with water, clean-up by solid-phase extraction and determination by HPLC-MS/MS and GC-MS. It was validated in a method validation study on roasted coffee, soluble coffee, coffee substitutes and coffee products with ranges from 53 μg/kg to 612,1 μg/kg.
Lebensmittel - Bestimmung von Acrylamid in Kaffee und Kaffee-Erzeugnissen mit HPLC-MS/MS und GC-MS
Diese Norm legt Verfahren zur Bestimmung von Acrylamid in Kaffee und Kaffee-Erzeugnissen durch Extraktion mit Wasser, Aufreinigung mit Festphasenextraktion und Bestimmung mittels HPLC-MS/MS bzw. GC-MS fest. Es wurde in einem Ringversuch für Röstkaffee, löslichen Kaffee, Kaffeesurrogate und Kaffee-Erzeugnisse in Bereichen von 53 μg/kg bis 612,1 μg/kg validiert.
Produits alimentaires - Détermination de la teneur en acrylamide dans le café et les produits du café par CLHP-SM/SM et CG-SM
La présente norme spécifie des méthodes de détermination de la teneur en acrylamide dans le café et les produits du café par extraction à l’eau, puis purification par extraction en phase solide et détermination par CLHP-SM/SM et CG-SM. Elle a été validée lors d’une étude de validation de méthode sur le café torréfié, le café soluble, les succédanés de café et les produits du café dans des gammes allant de 53 μg/kg à 612,1 μg/kg.
Živila - Določevanje akrilamida v kavi in proizvodih iz kave s HPLC-MS/MS in GC-MS
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2016
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Foodstuffs - Determination of acrylamide in coffee and coffee products by HPLC-MS/MS
and GC-MS
Lebensmittel - Bestimmung von Acrylamid in Kaffee und Kaffee-Erzeugnissen mit HPLC-
MS/MS und GC-MS
Produits alimentaires - Détermination de la teneur en acrylamide dans le café et les
produits du café par CLHP-SM/SM et CG-SM
Ta slovenski standard je istoveten z: prEN 16987
ICS:
67.140.20 Kava in kavni nadomestki Coffee and coffee substitutes
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
DRAFT
EUROPEAN STANDARD
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2016
ICS 67.140.20
English Version
Foodstuffs - Determination of acrylamide in coffee and
coffee products by HPLC-MS/MS and GC-MS
Lebensmittel - Bestimmung von Acrylamid in Kaffee
und Kaffee-Erzeugnissen mit HPLC-MS/MS und GC-MS
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 275.
If this draft becomes a European Standard, 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.
This draft European Standard was established by CEN 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.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2016 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 16987:2016 E
worldwide for CEN national Members.
Contents Page
Introduction . 3
European foreword . 3
1 Scope . 5
2 Normative references . 5
3 Principle . 5
4 Reagents . 5
5 Apparatus . 7
6 Procedure. 8
6.1 General . 8
6.2 Preparation of the sample extract . 8
6.3 Clean-up of the extracts . 8
6.3.1 Carrez precipitation . 8
6.3.2 Solid phase extraction . 8
6.4 HPLC-MS/MS measurement . 9
6.4.1 High performance liquid chromatography (HPLC). 9
6.4.2 Identification and quantification by mass spectrometry (HPLC-MS/MS) . 9
6.5 Measurement with GC-MS . 10
6.5.1 Derivatisation and sample preparation for gas chromatography . 10
6.5.2 Gas chromatography . 10
6.5.3 Identification and quantification by mass spectrometry . 10
7 Calibration . 11
7.1 General advice . 11
7.2 Determination of linearity and definition of the working range . 11
7.3 Calibration with internal standard . 11
7.4 Determination of the laboratory specific recovery . 11
8 Evaluation . 12
8.1 Criteria for identification . 12
8.2 Calculation and final results. 12
9 Precision data . 12
9.1 General . 12
9.2 Repeatability . 12
9.3 Reproducibility . 13
9.4 Recovery . 13
10 Measurement uncertainty . 13
11 Test report . 13
Annex A (informative) Performance characteristics . 14
Annex B (informative) Examples of absorber materials. 15
Annex C (informative) Examples for recommended columns and analysis conditions . 16
Bibliography . 23
European foreword
This document (prEN 16987:2016) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - horizontal methods”, the secretariat of which is held by DIN.
This document is currently submitted to the CEN Enquiry.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
Introduction
When applying this European Standard, all existing safety regulations should be followed.
Annexes A to C are informative.
WARNING — The use of this document 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.
1 Scope
This standard specifies methods for the determination of acrylamide in coffee and coffee products by
extraction with water, clean-up by solid-phase extraction and determination by HPLC-MS/MS and GC-
MS. It was validated in a method validation study on roasted coffee, soluble coffee, coffee substitutes
and coffee products with ranges from 53 μg/kg to 612,1 μg/kg.
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, Water for analytical laboratory use — Specification and test methods (ISO 3696)
3 Principle
The coffee sample is extracted with water or, in the case of soluble products, dissolved in water. A
clean-up by solid phase extraction is employed to remove interfering matrix compounds. Two
alternative methods can be used for the determination: high performance liquid chromatography with
mass spectrometric detection (HPLC-MS/MS) or, after a bromination of the acrylamide, gas
chromatography with mass spectrometric detection (GC-MS). In both cases isotopic labelled internal
standards are used.
4 Reagents
WARNING — In view of health risks when working with acrylamide, appropriate preventive and
protection measures shall be taken, such as using a fume cupboard, aspirating acrylamide-
containing solutions only with a pipette, and avoiding skin and eye contact or inhalation of
acrylamide-containing vapour
If available, reagents of “residue analysis grade” or “analytical reagent grade” shall be used. The level of
impurities in the reagents that contribute to the blank should be negligibly small. The blank shall be
checked regularly.
4.1 Water, of grade 1 according to EN ISO 3696, MS-grade is recommended.
4.2 Operating gases of high purity, suitable for GC and mass spectrometry according to the
instructions of the manufacturer of the apparatus.
4.3 Solvents, such as methanol, ethyl acetate, acetonitrile, n-hexane, MS-grade is recommended.
4.4 Acrylamide, C H NO, purity > 98 %, reference compound.
3 5
4.4.1 Acrylamide stock solution, mass concentration ρ = 1 000 μg/ml.
Weigh (0,10 ± 0,001) g of acrylamide into a 100 ml one-mark volumetric flask and swirl it in 30 ml of
water in order to dissolve the acrylamide. Fill up to the mark with water and mix well. The stock
solution is stable for at least three months when stored protected from light at a maximum of 6 °C.
Alternatively a commercially available solution with a mass concentration of ρ = 1 000 µg/ml may be
used. The information of the manufacturer regarding the stability of the solution shall be observed.
4.4.2 Acrylamide calibration solution, ρ = 10 μg/ml.
Using a pipette, transfer (1,0 ± 0,001) ml of the acrylamide stock solution (4.4.1) into a 100 ml one-
mark volumetric flask and fill up to the mark with water. This solution shall be stored protected from
light at a maximum of 6 °C and shall be freshly prepared every working day. Depending on the working
range more dilution steps might be necessary.
4.5 D3-acrylamide (acrylamide-2,3,3-d3) internal standard, C H D NO, purity > 98 %, reference
3 2 3
compound.
4.5.1 D3-acrylamide stock solution (internal standard),
Weigh (0,10 ± 0,001) g of D3-acrylamide into a 100 ml one-mark volumetric flask and swirl it in 30 ml
of water in order to dissolve the D3-acrylamide. Fill up to the mark with water and mix well. The stock
solution is stable for at least three months when stored protected from light at a maximum of 6 °C.
Alternatively a commercially available solution with a mass concentration of ρ = 1 000 µg/ml may be
used. The information of the manufacturer regarding the stability of the solution shall be observed.
4.5.2 D3-acrylamide internal standard solution.
Using a pipette, transfer (1,0 ± 0,001) ml of the D3-acrylamide stock solution (4.5.1) into a 100 ml one-
mark volumetric flask and fill up to the mark with water. This solution shall be stored protected from
light at a maximum of 6 °C and shall be freshly prepared every working day. Depending on the working
range more dilution steps might be necessary.
For HPLC-MS/MS the solutions according to 4.4.1 to 4.5.2 can be prepared using the HPLC eluent as a
solvent. The stability of these solutions depends on the solvent used and has
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