ISO 23905:2020

Cigarettes — Determination of selected phenolic compounds in cigarette mainstream smoke using HPLC-FLD

This document specifies a method for the quantification of selected phenolic compounds by high performance liquid chromatography with fluorescence detection (HPLC-FLD) using ISO 3308 smoking parameters. The selected phenolic compounds are: hydroquinone, resorcinol, catechol, phenol, p-Cresol, m-Cresol and o-Cresol. This method is applicable to cigarettes with total particulate matter (TPM) yields between 1 mg/cigarette and 16 mg/cigarette.

Cigarettes — Dosage de composés phénoliques sélectionnés dans le courant principal de la fumée de cigarette par CLHP-FLD

General Information

• Status: Published
• Publication Date: 29-Jun-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-Jun-2020
• Due Date: 30-Sep-2020
• Completion Date: 30-Jun-2020

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 23905
First edition
2020-06
Cigarettes — Determination of
selected phenolic compounds in
cigarette mainstream smoke using
HPLC-FLD
Cigarettes — Dosage de composés phénoliques sélectionnés dans le
courant principal de la fumée de cigarette par CLHP-FLD
Reference number
ISO 23905:2020(E)
©
ISO 2020

---------------------- Page: 1 ----------------------
ISO 23905:2020(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
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 © ISO 2020 – All rights reserved

---------------------- Page: 2 ----------------------
ISO 23905:2020(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Apparatus . 2
6 Reagents . 2
7 Preparation . 3
7.1 General . 3
7.2 Preparation of solutions — Acetic acid, with a volume fraction of 1 % solution . 3
7.3 Preparation of standards. 3
7.3.1 Primary phenolic compounds stock solutions . 3
7.3.2 Secondary phenolic compounds stock solutions . 3
7.3.3 Phenolic compounds working standards . 4
8 Sampling . 4
9 Tobacco product preparation . 4
10 Sample generation — Smoking of cigarettes . 4
10.1 General . 4
10.2 Smoking machine setup . 4
10.3 Smoking . 4
11 Sample analysis . 5
11.1 Preparation of sample. 5
11.2 Determination . 5
11.2.1 HPLC-FLD operating conditions . 5
11.2.2 Calibration . 6
11.2.3 Calculation . 6
12 Repeatability and reproducibility . 6
12.1 General . 6
12.2 Results of the 2013 collaborative study . 7
13 Report .10
Annex A (informative) Examples of chromatograms .11
Bibliography .13
© ISO 2020 – All rights reserved iii

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ISO 23905:2020(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.
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 © ISO 2020 – All rights reserved

---------------------- Page: 4 ----------------------
ISO 23905:2020(E)

Introduction
1)
The CORESTA Smoke Analytes Sub-Group selected one method using reversed phase high performance
liquid chromatography with fluorescence detection (HPLC-FLD) for the determination of selected
phenolic compounds in cigarette mainstream smoke. Smoke was collected on a glass fibre filter pad and
extracted with a 1 % acetic acid solution.
[1]
A CORESTA recommended method (CRM) was written on the basis of the results obtained in a
collaborative study conducted in 2013 involving 18 laboratories using cigarettes manufactured from a
range of blend styles.
This document is based upon the CRM 78 and includes statistical evaluations carried out according to
[2] [3]
ISO 5725-1 and ISO 5725-2 .
No machine smoking regime can represent all human smoking behaviour.
— 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 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 as 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.
1) Available at: www .coresta .org.
© ISO 2020 – All rights reserved v

---------------------- Page: 5 ----------------------
INTERNATIONAL STANDARD ISO 23905:2020(E)
Cigarettes — Determination of selected phenolic
compounds in cigarette mainstream smoke using HPLC-FLD
WARNING — The use of this document involves 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 quantification of selected phenolic compounds by high
performance liquid chromatography with fluorescence detection (HPLC-FLD) using ISO 3308 smoking
parameters. The selected phenolic compounds are: hydroquinone, resorcinol, catechol, phenol, p-Cresol,
m-Cresol and o-Cresol.
This method is applicable to cigarettes with total particulate matter (TPM) yields between 1 mg/
cigarette and 16 mg/cigarette.
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 3308, Routine analytical cigarette-smoking machine — Definitions and standard conditions
ISO 3402, Tobacco and tobacco products — Atmosphere for conditioning and testing
ISO 4387, Cigarettes — Determination of total and nicotine-free dry particulate matter using a routine
analytical smoking machine
ISO 8243, Cigarettes — Sampling
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/
4 Principle
Selected phenolic compounds are collected by passing the mainstream smoke of cigarettes through a
glass fibre filter pad as specified in ISO 3308 (e.g. Cambridge filer pad, CFP).
The CFP is extracted by a 1 % acetic acid solution. The obtained filtered solution is diluted and analysed
by HPLC-FLD.
© ISO 2020 – All rights reserved 1

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ISO 23905:2020(E)

5 Apparatus
The usual laboratory apparatus for use in preparation of samples, solutions and standards and, in
particular, the following.
5.1 Routine analytical cigarette-smoking machine, complying with the requirements of ISO 3308.
5.2 High performance liquid chromatography system, consisting of a binary gradient pump, an
auto sampler with sampling loop and cooling unit, a fluorescence detector, a data collection system.
5.3 HPLC column, with pentafluorphenylpropyl (PFP) stationary phase (e.g. 3 µm, 150 mm × 4,6 mm
or equivalent).
5.4 Disposable guard column, such as PFP cartridge (e.g. 4 mm × 3,00 mm or equivalent).
5.5 Wrist action shaker, or equivalent.
5.6 Analytical balance, suitable for measuring to the nearest 0,1 mg.
5.7 Glassware, actinic red Erlenmeyer flasks of appropriate volumes with ground glass stoppers,
actinic red volumetric flasks (10 ml, 25 ml and 50 ml).
5.8 Mechanical pipettes with disposable plastic tips.
6 Reagents
All reagents shall be at least of analytical reagent grade.
6.1 Methanol, HPLC grade.
6.2 Acetic acid, HPLC grade.
6.3 Hydroquinone, > 99 %.
6.4 Resorcinol,
...

FINAL
INTERNATIONAL ISO/FDIS
DRAFT
STANDARD 23905
ISO/TC 126
Cigarettes — Determination of
Secretariat: DIN
selected phenolic compounds in
Voting begins on:
2020­04­01 cigarette mainstream smoke using
HPLC-FLD
Voting terminates on:
2020­05­27
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 SUPPOR TING
DOCUMENTATION.
IN ADDITION TO THEIR EVALUATION AS
Reference number
BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO­
ISO/FDIS 23905:2020(E)
LOGICAL, COMMERCIAL AND USER PURPOSES,
DRAFT INTERNATIONAL STANDARDS MAY ON
OCCASION HAVE TO BE CONSIDERED IN THE
LIGHT OF THEIR POTENTIAL TO BECOME STAN­
DARDS TO WHICH REFERENCE MAY BE MADE IN
©
NATIONAL REGULATIONS. ISO 2020

---------------------- Page: 1 ----------------------
ISO/FDIS 23905:2020(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
below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH­1214 Vernier, Geneva
Phone: [phone]
Fax: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii © ISO 2020 – All rights reserved

---------------------- Page: 2 ----------------------
ISO/FDIS 23905:2020(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Apparatus . 2
6 Reagents . 2
7 Preparation . 3
7.1 General . 3
7.2 Preparation of solutions — Acetic acid, with a volume fraction of 1 % solution . 3
7.3 Preparation of standards. 3
7.3.1 Primary phenolic compounds stock solutions . 3
7.3.2 Secondary phenolic compounds stock solutions . 3
7.3.3 Phenolic compounds working standards . 4
8 Sampling . 4
9 Tobacco product preparation . 4
10 Sample generation — Smoking of cigarettes . 4
10.1 General . 4
10.2 Smoking machine setup . 4
10.3 Smoking . 4
11 Sample analysis . 5
11.1 Preparation of sample. 5
11.2 Determination . 5
11.2.1 HPLC­FLD operating conditions . 5
11.2.2 Calibration . 6
11.2.3 Calculation . 6
12 Repeatability and reproducibility . 6
12.1 General . 6
12.2 Results of the 2013 collaborative study . 7
13 Report .10
Annex A (informative) Examples of chromatograms .11
Bibliography .13
© ISO 2020 – All rights reserved iii

---------------------- Page: 3 ----------------------
ISO/FDIS 23905:2020(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.
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 © ISO 2020 – All rights reserved

---------------------- Page: 4 ----------------------
ISO/FDIS 23905:2020(E)

Introduction
1)
The CORESTA Smoke Analytes Sub-Group selected one method using reversed phase high performance
liquid chromatography with fluorescence detection (HPLC-FLD) for the determination of selected
phenolic compounds in cigarette mainstream smoke. Smoke was collected on a glass fibre filter pad and
extracted with a 1 % acetic acid solution.
[3]
A CORESTA recommended method (CRM) was written on the basis of the results obtained in a
collaborative study conducted in 2013 involving 18 laboratories using cigarettes manufactured from a
range of blend styles.
This document is based upon the CRM 78 and includes statistical evaluations carried out according to
ISO 5725­1 and ISO 5725­2.
No machine smoking regime can represent all human smoking behaviour.
— 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 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 as 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.
1) Available at: www .coresta .org.
© ISO 2020 – All rights reserved v

---------------------- Page: 5 ----------------------
FINAL DRAFT INTERNATIONAL STANDARD ISO/FDIS 23905:2020(E)
Cigarettes — Determination of selected phenolic
compounds in cigarette mainstream smoke using HPLC-FLD
WARNING — The use of this document involves 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 quantification of selected phenolic compounds by high
performance liquid chromatography with fluorescence detection (HPLC-FLD) using ISO 3308 smoking
parameters. The selected phenolic compounds are: hydroquinone, resorcinol, catechol, phenol, p-cresol,
m­cresol and o­cresol.
This method is applicable to cigarettes with total particulate matter (TPM) yields between 1 mg/
cigarette and 16 mg/cigarette.
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 3308, Routine analytical cigarette-smoking machine — Definitions and standard conditions
ISO 3402, Tobacco and tobacco products — Atmosphere for conditioning and testing
ISO 4387, Cigarettes — Determination of total and nicotine-free dry particulate matter using a routine
analytical smoking machine
ISO 8243, Cigarettes — Sampling
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/
4 Principle
Selected phenolic compounds are collected by passing the mainstream smoke of cigarettes through a
glass fibre filter pad as specified in ISO 3308 (e.g. Cambridge filer pad, CFP).
The CFP is extracted by a 1 % acetic acid solution. The obtained filtered solution is diluted and analysed
by HPLC-FLD.
© ISO 2020 – All rights reserved 1

---------------------- Page: 6 ----------------------
ISO/FDIS 23905:2020(E)

5 Apparatus
The usual laboratory apparatus for use in preparation of samples, solutions and standards and, in
particular, the following.
5.1 Routine analytical cigarette-smoking machine, complying with the requirements of ISO 3308.
5.2 High performance liquid chromatography system, consisting of a binary gradient pump, an
auto sampler with sampling loop and cooling unit, a fluorescence detector, a data collection system.
5.3 HPLC column, with pentafluorphenylpropyl (PFP) stationary phase (e.g. 3 µm, 150 mm × 4,6 mm
or equivalent).
5.4 Disposable guard column, such as PFP cartridge (e.g. 4 mm × 3,00 mm or equivalent).
5.5 Wrist action shaker, or equivalent.
5.6 Analytical balance, suitable for measuring to the nearest 0,1 mg.
5.7 Glassware, actinic red Erlenmeyer flasks of appropriate volumes with ground glass stoppers,
actinic red volumetric flasks (10 ml, 25 ml and 50
...