ISO 23906-2:2023

Cigarettes - Determination of benzo[a]pyrene in cigarette mainstream smoke with an intense smoking regime using GC/MS - Part 2: Method using cyclohexane as extraction solvent

This document specifies a method for the determination of benzo[a]pyrene (B[a]P) in the total particulate matter of cigarette mainstream smoke with an intense smoking regime using gas chromatography/mass spectrometry (GC/MS) with cyclohexane as extraction solvent. This method is specified using ISO 20778 smoking parameters. An alternative method for the determination of B[a]P is specified in ISO 23906‑1, with a different clean-up using methanol 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 2: Méthode utilisant du cyclohexane comme solvant d'extraction

General Information

• Status: Published
• Publication Date: 09-Feb-2023
• 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: 10-Feb-2023
• Due Date: 04-Mar-2023
• Completion Date: 10-Feb-2023

Relations

• Consolidated By: ISO 6134:2024 - Rubber hoses and hose assemblies for saturated steam - Specification
• Effective Date: 28-Jan-2023

Overview

ISO 23906-2:2023 specifies a laboratory method for the determination of benzo[a]pyrene (B[a]P) in the total particulate matter (TPM) of cigarette mainstream smoke generated under an intense machine smoking regime. The method uses gas chromatography/mass spectrometry (GC/MS) for analysis and cyclohexane as the extraction solvent. Procedures are aligned with the ISO 20778 smoking parameters and ISO 20779 collection methods. Part 2 (this document) provides a cyclohexane extraction and solid-phase extraction (SPE) clean-up alternative to ISO 23906-1 (methanol-based clean-up).

Key topics and technical requirements

• Scope and intent: Determination of B[a]P in cigarette mainstream smoke TPM under an intense smoking regime; not a measure of human exposure or risk.
• Sampling & conditioning: Follows ISO 8243 (sampling) and ISO 3402 (conditioning).
• Machine smoking & collection: Uses ISO 20778 and ISO 20779 routines for smoking and TPM capture on glass-fibre filter pads (44 mm and 92 mm sizes; TPM capacity guidance provided).
• Extraction & clean-up: Extraction of filter pads with cyclohexane, followed by SPE (NH-bonded silica cartridges recommended) for sample clean-up.
• Analytical instrumentation: GC/MS with single ion monitoring (SIM) and splitless injections; example column: 50% phenyl/50% methyl polysiloxane, 30 m × 0.25 mm × 0.25 μm.
• Standards & calibration: Detailed preparation of internal standard (B[a]P-d12) and B[a]P stock/working solutions (e.g., 100 μg/ml primary stocks, 40 ng/ml spiking solution), calibration across relevant concentration ranges, and storage guidance (stable up to ~4 months at ≤4 °C).
• Quality & reporting: Procedures for calibration, calculation, repeatability/reproducibility assessment, blank controls, and required test-report elements.

Applications

• Regulatory and compliance testing of cigarette emissions for B[a]P content.
• Product characterization and comparative analysis of mainstream smoke polycyclic aromatic hydrocarbons (PAHs).
• Analytical method implementation in tobacco testing laboratories implementing GC/MS, SPE, and standardized smoking regimes. Note: The standard highlights that machine-derived emissions are inputs for product hazard assessment but are not measures of human exposure or risk.

Who should use this standard

• Analytical chemists in tobacco product testing laboratories
• Regulatory agencies and conformity-assessment bodies
• Tobacco manufacturers and contract testing labs
• Quality assurance personnel implementing ISO smoking regimes and GC/MS analyses

Related standards

• ISO 23906-1 (B[a]P determination - methanol clean-up alternative)
• ISO 20778, ISO 20779 (smoking machine parameters and TPM generation)
• ISO 3402 (conditioning) and ISO 8243 (sampling)

Keywords: ISO 23906-2:2023, benzo[a]pyrene, B[a]P, cigarette mainstream smoke, GC/MS, cyclohexane, SPE, ISO 20778, TPM, smoking regime.

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 23906-2
First edition
2023-02
Cigarettes — Determination
of benzo[a]pyrene in cigarette
mainstream smoke with an intense
smoking regime using GC/MS —
Part 2:
Method using cyclohexane 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 2: Méthode utilisant du cyclohexane comme solvant
d'extraction
Reference number
© ISO 2023
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 below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principles . 1
5 Apparatus . 2
6 Reagents . 3
7 Standards . 3
7.1 General . 3
7.2 Primary B[a]P-d12 stock solution: 100 µg/ml . 3
7.3 Secondary B[a]P-d12 spiking solution: 40 ng/ml . 3
7.4 Primary B[a]P stock solution: 100 µg/ml . 3
7.5 Secondary B[a]P stock solution: 1 000 ng/ml . 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
8.5 Blank solution . 5
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 Test results . 7
Annex A (informative) Example of a chromatogram of a cigarette smoke extract .8
Bibliography . 9
iii
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.
iv
Introduction
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 ISO standards.
v
INTERNATIONAL STANDARD ISO 23906-2:2023(E)
Cigarettes — Determination of benzo[a]pyrene in cigarette
mainstream smoke with an intense smoking regime using
GC/MS —
Part 2:
Method using cyclohexane 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 any other restrictions 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 of cigarette mainstream smoke with an intense smoking regime using gas
chromatography/mass spectrometry (GC/MS) with cyclohexane as extraction solvent.
This method is specified using ISO 20778 smoking parameters. An alternative method for the
determination of B[a]P is specified in ISO 23906-1, with a different clean-up using methanol solvent.
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 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 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 Principles
— 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 total particulate matter, collected on the glass-fibre filter pad, with cyclohexane.
— Clean-up procedure using solid phase extraction (SPE).
— 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, in accordance 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 an
...