ISO 23906-1:2020

Cigarettes — Determination of benzo[a]pyrene in cigarette mainstream smoke with an intense smoking regime using GC/MS — Part 1: Method using methanol as extraction solvent

This document specifies a method for the determination of benzo[a]pyrene (B[a]P) in the total particulate matter (TPM) of cigarette mainstream smoke using gas chromatography/mass spectrometry (GC-MS) with methanol as extraction solvent. This method is specified using ISO 20778 smoking parameters. NOTE An alternative method for the determination of B[a]P is specified in ISO 23906-2 with a different clean-up using cyclohexane solvent and a shorter total analytical time.

Cigarettes — Dosage par CG/SM du benzo[a]pyrène dans le courant principal de la fumée de cigarette avec un régime de fumage intense — Partie 1: Méthode utilisant du méthanol comme solvant d'extraction

General Information

• Status: Published
• Publication Date: 29-Sep-2020
• ICS: 65.160 - Tobacco, tobacco products and related equipment
• Technical Committee: ISO/TC 126 - Tobacco and tobacco products
• Drafting Committee: ISO/TC 126/WG 10 - Intense smoking regime
• Current Stage: 6060 - International Standard published
• Start Date: 30-Sep-2020
• Due Date: 16-Aug-2020
• Completion Date: 30-Sep-2020

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 23906-1
First edition
2020-09
Cigarettes — Determination
of benzo[a]pyrene in cigarette
mainstream smoke with an intense
smoking regime using GC/MS —
Part 1:
Method using methanol as extraction
solvent
Cigarettes — Dosage par CG/SM du benzo[a]pyrène dans le courant
principal de la fumée de cigarette avec un régime de fumage
intense —
Partie 1: Méthode utilisant du méthanol comme solvant d'extraction
Reference number
ISO 23906-1:2020(E)
©
ISO 2020

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ISO 23906-1:2020(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2020
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 the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address
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Phone: [phone]
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Published in Switzerland
ii © ISO 2020 – All rights reserved

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ISO 23906-1:2020(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Apparatus . 2
6 Reagents . 2
7 Standards . 3
7.1 General . 3
7.2 Primary B[a]P stock solution . 3
7.3 Secondary B[a]P stock solution . 3
7.4 B[a]P-d12 stock solution . 3
7.5 B[a]P-d12 spiking solution . 3
7.6 Working standard solutions . 3
7.7 Storage of standard solutions. 3
8 Preparation of sample . 4
8.1 Sampling . 4
8.2 Smoking . 4
8.3 Glass-fibre filter pad extraction . 4
8.4 Sample clean-up . 4
9 Determination . 5
9.1 GC-MS operating conditions . 5
9.2 Calibration . 6
9.3 Determination of B[a]P . 6
9.4 Calculation . 6
10 Repeatability and reproducibility . 6
11 Test report . 7
11.1 General . 7
11.2 Characteristic data about the cigarette . 8
11.3 Data about sampling . 8
11.4 Description of the test . 8
11.5 Test results. 8
Annex A (informative) Example of a chromatogram of a cigarette smoke extract .9
Bibliography .11
© ISO 2020 – All rights reserved iii

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ISO 23906-1:2020(E)

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 documents 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).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www .iso .org/ patents).
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.
This document was prepared by Technical Committee ISO/TC 126, Tobacco and tobacco products.
A list of all parts in the ISO 23906 series can be found on the ISO website.
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.
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ISO 23906-1:2020(E)

Introduction
Between 1999 and 2003, a task force composed of Cooperation Centre for Scientific Research Relative
to Tobacco (CORESTA) members studied the existing methodologies for the determination of benzo[a]
pyrene (B[a]P) in the mainstream smoke of cigarettes. Several methods have been proposed for this
determination, which are mainly based on two types of analytical methodology: high performance liquid
chromatography (HPLC) with fluorescence detection and gas chromatography/mass spectrometry
(GC-MS). In both cases, it is necessary to purify the total particulate matter (TPM) extract before
performing the chromatography in order to obtain a correct separation of the B[a]P peak.
The task force decided in the first instance to develop a method using HPLC with fluorescence detection.
However, after several collaborative experiments, it appeared that achieving a significant reduction
of the initially observed variability would be technically very difficult. The task force then decided
to investigate a GC-MS method as an alternative and was able to demonstrate, through collaborative
experiments, that a lower variability can be obtained with this methodology.
This document, produced through collaborative experiments conducted in 2012 involving 12
laboratories in 11 countries, provides a procedure for the determination of B[a]P in cigarette
[4]
mainstream smoke. The repeatability and reproducibility of this method have been assessed
according to ISO recommendations and are included.
No machine smoking regime can represent all human smoking behaviours.
— It is recommended that cigarettes also be tested under conditions of a different intensity of machine
smoking than those specified in this document.
— Machine smoking testing is useful to characterize cigarette emissions for design and regulatory
purposes, but communication of machine measurements to smokers can result in misunderstandings
about differences in exposure and risk across brands.
— Smoke emission data from machine measurements may be used as inputs for product hazard
assessment, but they are not intended to be nor are they valid measures of human exposure or
risks. Communicating differences between products in machine measurements as differences in
exposure or risk is a misuse of testing using International Standards.
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INTERNATIONAL STANDARD ISO 23906-1:2020(E)
Cigarettes — Determination of benzo[a]pyrene in
cigarette mainstream smoke with an intense smoking
regime using GC/MS —
Part 1:
Method using methanol as extraction solvent
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 document specifies a method for the determination of benzo[a]pyrene (B[a]P) in the total particulate
matter (TPM) of cigarette mainstream smoke using gas chromatography/mass spectrometry (GC-MS)
with methanol as extraction solvent.
This method is specified using ISO 20778 smoking parameters.
NOTE An alternative method for the determination of B[a]P is specified in ISO 23906-2 with a different
clean-up using cyclohexane solvent and a shorter total analytical time.
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 3402, Tobacco and tobacco products — Atmosphere for conditioning and testing
ISO 3696, Water for analytical laboratory use — Specification and test methods
ISO 8243, Cigarettes — Sampling
ISO 20778, Cigarettes — Routine analytical cigarette smoking machine — Definitions and standard
conditions with an intense smoking regime
ISO 20779, Cigarettes — Generation and collection of total particulate matter using a routine analytical
smoking machine with an intense smoking regime
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
© ISO 2020 – All rights reserved 1

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ISO 23906-1:2020(E)

4 Principle
— Sampling of the test cigarettes according to the sampling procedure specified in ISO 8243.
— Conditioning of the test cigarettes according to the conditioning procedure specified in ISO 3402.
— Smoking of the test cigarettes according to the smoking procedure specified in ISO 20779.
— Extraction of the TPM, collected on the appropriate glass-fibre filter pad, with methanol.
— Dilution of the methanol extract with water.
— Loading of the water/methanol solution onto a cyclohexyl solid phase extraction (CH SPE) cartridge,
followed by the elution of B[a]P with cyclohexane.
— Analytical determination of B[a]P by gas chromatography/mass spectrometry.
5 Apparatus
The usual laboratory apparatus and equipment and, in particular, the following.
5.1 Routine analytical cigarette-smoking machine, complying with the requirements of ISO 20778
and equipped for smoking in accordance with ISO 20779.
5.2 Gas chromatograph with a mass selective detector, equipped with its computerized control
and data acquisition and processing system. This system shall be able to pilot the mass spectrometer
in order to obtain chromatographic data under single ion monitoring (SIM) detection mode. The
gas chromatograph shall be configured to perform splitless injections on a capillary column. It is
recommended to equip the gas chromatograph with an autosampler for sample injection.
5.3 Fused silica capillary column, for example a (5 %-phenyl)-methylpolysiloxane stationary phase
and a 30 m length, 0,25 mm intern
...