SIST EN ISO 18862:2026

SLOVENSKI STANDARD
01-februar-2026
Nadomešča:
SIST EN ISO 18862:2020
Kava in proizvodi iz kave - Določevanje akrilamida - Metode z uporabo tekočinske
kromatografije visoke ločljivosti s tandemsko masno spektrometrijo (HPLC-
MS/MS) in plinske kromatografije z masno spektrometrijo (GC-MS) po derivatizaciji
(ISO 18862:2025)
Coffee and coffee products - Determination of acrylamide - Methods using high-
performance liquid chromatography with tandem mass spectrometric detection (HPLC-
MS/MS) and gas chromatography with mass spectrometric detection (GC-MS) after
derivatization (ISO 18862:2025)
Kaffee und Kaffee-Erzeugnisse - Bestimmung von Acrylamid - Verfahren mittels
Hochleistungs-Flüssigchromatographie mit massenspektrometrischer Detektion (HPLC-
MS/MS) und Gaschromatographie mit massenspektrometrischer Detektion (GC-MS)
nach Derivatisierung (ISO 18862:2025)
Café et dérivés du café - Dosage de l’acrylamide - Méthodes par chromatographie
liquide à haute performance avec détection par spectrométrie de masse en tandem
(CLHP–SM/SM) et chromatographie en phase gazeuse avec détection par spectrométrie
de masse (CG–SM) après dérivation (ISO 18862:2025)
Ta slovenski standard je istoveten z: EN ISO 18862:2025
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.

EN ISO 18862
EUROPEAN STANDARD
NORME EUROPÉENNE
December 2025
EUROPÄISCHE NORM
ICS 67.140.20 Supersedes EN ISO 18862:2019
English Version
Coffee and coffee products - Determination of acrylamide -
Methods using high-performance liquid chromatography
with tandem mass spectrometric detection (HPLC-MS/MS)
and gas chromatography with mass spectrometric
detection (GC-MS) after derivatization (ISO 18862:2025)
Café et dérivés du café - Dosage de l'acrylamide - Kaffee und Kaffee-Erzeugnisse - Bestimmung von
Méthodes par chromatographie liquide à haute Acrylamid - Verfahren mittels Hochleistungs-
performance avec détection par spectrométrie de Flüssigchromatographie mit massenspektrometrischer
masse en tandem (CLHP-SM/SM) et chromatographie Detektion (HPLC-MS/MS) und Gaschromatographie
en phase gazeuse avec détection par spectrométrie de mit massenspektrometrischer Detektion (GC-MS) nach
masse (CG-SM) après dérivation (ISO 18862:2025) Derivatisierung (ISO 18862:2025)
This European Standard was approved by CEN on 17 November 2025.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 18862:2025 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 18862:2025) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”
the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by June 2026, and conflicting national standards shall be
withdrawn at the latest by June 2026.
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 supersedes EN ISO 18862:2019.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 18862:2025 has been approved by CEN as EN ISO 18862:2025 without any modification.

International
Standard
ISO 18862
Second edition
Coffee and coffee products —
2025-11
Determination of acrylamide —
Methods using high-performance
liquid chromatography with
tandem mass spectrometric
detection (HPLC-MS/MS) and
gas chromatography with mass
spectrometric detection (GC-MS)
after derivatization
Café et dérivés du café — Dosage de l’acrylamide — Méthodes
par chromatographie liquide à haute performance avec
détection par spectrométrie de masse en tandem (CLHP–SM/
SM) et chromatographie en phase gazeuse avec détection par
spectrométrie de masse (CG–SM) après dérivation
Reference number
ISO 18862:2025(en) © ISO 2025
ISO 18862:2025(en)
© ISO 2025
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
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Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
ISO 18862:2025(en)
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Reagents . 2
6 Apparatus . 3
7 Sampling . 4
8 Procedure . 4
8.1 General .4
8.2 Preparation of the sample extract .4
8.3 Clean-up of the extracts .5
8.3.1 Carrez precipitation .5
8.3.2 Solid-phase extraction.5
8.4 HPLC-MS/MS measurement . .5
8.4.1 High-performance liquid chromatography (HPLC) .5
8.4.2 Identification and quantification by mass spectrometry (HPLC-MS/MS) .6
8.5 Measurement with GC-MS .6
8.5.1 Derivatization and sample preparation for gas chromatography .6
8.5.2 Gas chromatography .7
8.5.3 Identification and quantification by mass spectrometry .7
9 Calibration . 7
9.1 General advice .7
9.2 Determination of linearity and definition of the working range .7
9.3 Calibration with internal standard solution .7
9.4 Determination of the laboratory specific recovery .8
10 Evaluation . 8
10.1 Criteria for identification .8
10.2 Calculation and final results .8
11 Precision data . 9
11.1 General .9
11.2 Repeatability .9
11.3 Reproducibility .9
11.4 Recovery .9
12 Measurement uncertainty . 9
13 Test report . 9
Annex A (informative) Performance characteristics .11
Annex B (informative) Examples of absorber materials .12
Annex C (informative) Examples of columns and analysis conditions.13
Annex D (informative) Examples for sample preparation and chromatographic conditions
using HPLC-MS/MS . 19
Bibliography .25

iii
ISO 18862:2025(en)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO’s adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
The committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 15, Coffee, in
collaboration with the European Committee for Standardization (CEN) Technical Committee CEN/TC 275,
Food analysis - Horizontal methods, in accordance with the Agreement on technical cooperation between ISO
and CEN (Vienna Agreement).
This second edition cancels and replaces the first edition (ISO 18862:2016), which has been technically
revised.
The main changes are as follows:
— Annex D with examples for sample preparation and chromatographic conditions using HPLC-MS/MS has
been added.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
International Standard ISO 18862:2025(en)
Coffee and coffee products — Determination of acrylamide —
Methods using high-performance liquid chromatography with
tandem mass spectrometric detection (HPLC-MS/MS) and gas
chromatography with mass spectrometric detection (GC-MS)
after derivatization
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 take appropriate measures for ensuring the safety and
health of the personnel prior to application of this document and to fulfil statutory requirements for
this purpose.
1 Scope
This document specifies methods for the determination of acrylamide in coffee and coffee products by
extraction with water, clean-up by solid-phase extraction (SPE) and determination by high-performance
liquid chromatography with tandem mass spectrometric detection (HPLC-MS/MS) and gas chromatography
with mass spectrometric detection (GC-MS) after derivatization. The methods were validated in a validation
study for 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 are referred to in the text in such a way that some or all of their content constitutes
requirements of this document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 3696, Water for analytical laboratory use — Specification and test methods
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4 Principle
The coffee sample is extracted with water or, in the case of soluble products, dissolved in water. A clean-up
by SPE is employed to remove interfering matrix compounds. Two alternative methods can be used for the
determination: HPLC-MS/MS or, after a bromination of the acrylamide, GC-MS. In both cases, solutions of
isotopic labelled internal standard are used.

ISO 18862:2025(en)
5 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.
For HPLC-MS/MS, the solutions according to 5.4.1 to 5.5.2 can be prepared using the HPLC eluent as a solvent.
The stability of these solutions depends on the mobile phase used and shall be validated.
When using GC-MS, all standard solutions according to 5.4.2 and 5.5.2 shall be subjected to the derivatization
step according to 8.5.1.
Instead of D3-acrylamide, it is also possible to use C acrylamide for the preparation of the internal standard
solution. However, in Clauses 8 and 9, the procedure and calculation are described for D3-acrylamide only.
5.1 Water, of grade 1 according to ISO 3696
5.2 Operating gases of high purity, suitable for gas chromatography and mass spectrometry according
to the instructions of the manufacturer of the apparatus.
5.3 Solvents, such as methanol, ethyl acetate, acetonitrile and n-hexane. MS-grade.
5.4 Acrylamide, C H NO, purity > 98 %, reference substance.
3 5
5.4.1 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.
5.4.2 Acrylamide calibration solution, ρ = 10 μg/ml.
Using a pipette, transfer (1,0 ± 0,001) ml of the acrylamide stock solution (5.4.1) 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 can be necessary.
5.5 D3-acrylamide (acrylamide-2,3,3-d3) internal standard solution, C H D NO, purity > 98 %,
3 2 3
reference substance.
5.5.1 D3-acrylamide stock solution (internal standard solution).
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.

ISO 18862:2025(en)
5.5.2 D3-acrylamide internal standard solution.
Using a pipette, transfer (1,0 ± 0,001) ml of the D3-acrylamide stock solution (5.5.1) 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 can be necessary.
5.6 Saturated bromine water.
Saturate distilled water with bromine in a 100 ml one-mark volumetric flask (with a glass stopper) until a
phase of bromine is formed at the bottom of the flask (around 3,5 % of bromine at 4 °C). Acidify the bromine
water to a pH of about 1 using concentrated hydrobromic acid (ρ(HBr) = 1,48 g/cm ).
If stored at 4 °C and protected from light, the solution can be used for about four weeks.
5.7 Potassium bromide, KBr.
5.8 Sodium thiosulfate (pentahydrate), Na S O · 5 H O.
2 2 3 2
5.9 Triethylamine, (C H ) N.
2 5 3
5.10 Sodium sulfate, Na SO anhydrous, granular.
2 4,
5.11 Carrez solution I.
Dissolve 10,6 g of potassium hexacyanoferrate trihydrate (II) K [Fe(CN) ] · 3 H O in 100 ml of water. If
4 6 2
stored at 4 °C and protected from light, the solution is stable for six months.
5.12 Carrez solution II.
Dissolve 21,9 g of zinc acetate dihydrate Zn(CH COO) · 2 H O in 100 ml of water. If stored at 4 °C and
3 2 2
protected from light, the solution is stable for six months.
5.13 Borate buffer, pH 8,6.
Mix 68 ml of a 0,1 molar sodium borate solution (20,12 g Na B O per litre of water) and 32 ml of 0,1 molar
2 4 7
hydrochloric acid, c(HCl) = 0,1 mol/l, in a 100 ml one-mark volumetric flask.
6 Apparatus
The usual laboratory apparatus and, in particular, the following shall be used.
Apparatus and parts of the apparatus which come into contact with the sample and extract shall be
free of residues which can cause blank values. Glassware or equipment made of stainless steel or PTFE
(polytetrafluoroethylene) shall be used.
6.1 Analytical balance, capable of weighing to an accuracy of 0,1 mg.
6.2 Coffee mill, suitable for grinding roasted coffee beans.
6.3 Glassware, for collecting and storing the extracts, preferably made of amber glass, as sample vials for
manual or automatic use, equipped with an inert seal (e.g. vials with PTFE-coated septum).
6.4 Ultrasonic bath, capable of being maintained at 40 °C.

ISO 18862:2025(en)
6.5 Laboratory centrifuge, suitable for 15 ml and 50 ml centrifugal tubes and with a minimum g-force
of 2 000g.
6.6 Centrifuge tubes, of 15 ml and 50 ml.
6.7 One-mark volumetric flask, of 20 ml and 100 ml.
6.8 Pipettes, glass or automatic, suitable for measuring volume ranges of standard solutions and sample
extract dilutions.
6.9 Glass or polypropylene cartridges, with sorbents for the SPE, and for the clean-up of extracts in
8.3.2 and 8.5.1 (examples are given in Table B.1).
6.10 High performance liquid chromatograph (for the test procedure according to 8.4), equipped with
electrospray ionization (ESI) and a mass spectrometric detector (HPLC-MS/MS), and with a gas supply as
specified by the manufacturer.
6.11 HPLC column (for the test procedure according to 8.4), suitable for acrylamide chromatography
(examples are given in Table C.1).
6.12 Gas chromatograph, (for the test procedure according to 8.5) with mass spectrometric detector (GC-
MS) and operating gas supply (5.2) as specified by the manufacturer.
6.13 GC column, (for the test procedure according to 8.5) capillary column, suitable for acrylamide
chromatography (examples are given in Table C.2).
6.14 Membrane filter units, syringe filter (e.g. cellulose acetate filters 0,45 µm pore size) suitable for
filtration of sample eluate obtained by SPE before injection into the chromatographic system.
7 Sampling
Sampling is not part of the method specified in this document. The sampling procedure shall be subject to
agreement by the interested parties. A representative, thoroughly mixed sample shall be used, which has
not been damaged or adulterated during transport or storage.
In order to exclude changes in the acrylamide levels, the analysis shall be performed shortly after reception
of the sample. When stored for more than two weeks, samples should be kept at a temperature below 6 °C.
The date of receipt of the sample, as well as the date of roasting or the best-before date, shall be documented
along with the date of analysis.
8 Procedure
8.1 General
To avoid losses of the analyte, the samples shall be protected from light during extraction and further
preparation. For this reason, amber glassware shall always be used. Otherwise, the content of the vessels
and flasks shall be protected from incident light using aluminium foil.
Examples for sample preparation and chromatographic conditions using HPLC-MS/MS are given in Annex D.
8.2 Preparation of the sample extract
If necessary, grind the sample in a coffee mill (6.2) and homogenize thoroughly.

ISO 18862:2025(en)
Weigh 2 g of the homogenized sample of roasted coffee, soluble coffee or coffee substitute or 5 g of liquid
coffee beverage to the nearest 1 mg using an analytical balance (6.1) and transfer it into a 50 ml centrifuge
tube (6.6).
Add 2 ml of n-hexane to the test sample and shake briefly. Then spike the test sample with the internal
standard solution of D3-acrylamide in a concentration corresponding to the expected acrylamide level of the
sample.
EXAMPLE Weigh 2 g of coffee and add 100 µl internal standard solution (ρ = 10 µg/ml), which is equivalent to an
acrylamide mass fraction of 500 µg/kg in the coffee sample.
Add 20 ml of dist
...