ISO 16000-39:2019

INTERNATIONAL ISO
STANDARD 16000-39
First edition
2019-06
Indoor air —
Part 39:
Determination of amines —
Analysis of amines by (ultra-) high-
performance liquid chromatography
coupled to high resolution or tandem
mass spectrometry
Reference number
©
ISO 2019
© ISO 2019
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: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Amines in indoor air . 2
4.1 Properties of amines . 2
4.2 Origin and occurrence of amines . 2
5 Analytical procedure . 2
5.1 Preparation of sample solutions . 2
5.2 High-performance liquid chromatography . 2
5.3 Tandem mass spectrometry . 3
5.4 High resolution mass spectrometry . 3
5.5 Sample sequence and external calibration . 3
6 Testing of the suitability of the equipment and the instrumental analysis .3
6.1 General . 3
6.2 High-performance or ultra-performance liquid chromatography (HPLC or UHPLC) . 3
6.3 Tandem mass spectrometry (MS-MS) . 4
6.4 High resolution mass spectrometry (HRMS) . 4
7 Reporting requirements . 4
Annex A (informative) List of included amines . 6
Annex B (informative) Performance data of the analytical method . 9
Bibliography .12
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 146, Air quality, Subcommittee SC 6,
Indoor air.
A list of all parts in the ISO 16000 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 © ISO 2019 – All rights reserved

Introduction
ISO 16000 (all parts) describe general requirements relating to the measurement of indoor air pollutants
and the important conditions to be observed before or during the sampling of individual pollutants or
groups of pollutants, as well as the measurements procedures themselves.
The definition of indoor environment is given by ISO 16000-1. Dwellings [living rooms, bedrooms, do-
it-yourself (DIY) rooms, sport rooms and cellars, kitchens and bathrooms], workrooms or workplaces
in buildings which are not subject to health and safety inspections with respect to air pollutants (e.g.
offices, salesrooms), public buildings (e.g. restaurants, theatres, cinemas and other meeting rooms) and
passenger cabins of motor vehicles and public transportation are among the most important types of
indoor environment.
INTERNATIONAL STANDARD ISO 16000-39:2019(E)
Indoor air —
Part 39:
Determination of amines — Analysis of amines by (ultra-)
high-performance liquid chromatography coupled to high
resolution or tandem mass spectrometry
1 Scope
This document, along with ISO 16000-38, specifies the measurement method for determining the mass
concentration of primary, secondary and tertiary aliphatic and aromatic amines in indoor air using
accumulated sampling and high-performance liquid chromatography (HPLC) coupled with tandem
mass spectrometry (MS-MS) or high-resolution mass spectrometry (HRMS). The analytical procedure
is covered by this document. The sampling procedure and the manufacturing of the samplers are
covered by ISO 16000-38.
This document describes specifications for the chromatography and the mass spectroscopy for the
amines. Measurement results are expressed in µg/m .
Although primarily intended for the measurement of amines listed in Tables A.1 and A.2, it can also be
used for the measurement of other amines in indoor air. This document gives instructions and describes
procedures for the inclusion of other amines.
The range of application of this document concerning the concentrations of amines in indoor air
depends on the linear range of the calibration line and hence on the gas sample volume (here: from 5 l
up to 100 l), the eluate volume (from 1 ml up to 5 ml), the injection volume (from 1 µl up to 10 µl) and
the sensitivity of the analytical equipment (e.g. linear range from 2 pg up to 2 ng amine). The range
of application can be expected to be from approximately 0,002 µg/m (100 l sample) up to 2 000 µg/
m (5 l sample) for a common analytical equipment (e.g. Waters „TQD“) for the majority of the amines
listed in Tables A.1 and A.2. The analysis of derivatives of ethanolamine is usually about 10 times more
sensitive and the analysis of short-chained aliphatic amines is usually about 10 times less sensitive than
the analysis of an average amine.
The performance data of the analytical method is given in Annex B, particularly in Tables B.1 and B.2.
This document can be used also for the determination of amines in water if the detection limit is
sufficient.
This document does not cover the determination of isocyanates in indoor air (nor in water samples) as
corresponding amines (covered by ISO 17734-1 and ISO 17734-2).
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 16000-38, Indoor air — Part 38: Determination of amines in indoor and test chamber air — Active
sampling on samplers containing phosphoric acid impregnated filters
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardisation at the following addresses:
— ISO online browsing platform: available at http: //www .iso .org/obp
— IEC Electropedia: aivailable at http: //www .elctropedia .org/
3.1
amines
-3
nitrogen containing compounds with a sufficient vapor pressure (>10 Pa) and a free electron pair at
the nitrogen atom which can be protonated
4 Amines in indoor air
4.1 Properties of amines
Amines are basic and polar substances.
There are primary, secondary, tertiary, and quaternary amines.
Quaternary amines are not included in this document, as they have no free electron pair and therefore
have very different properties. In this document the term “amines” refers only to primary, secondary,
and tertiary amines.
Not protonated amines are oxidation sensitive.
Reaction of amines with acids results in ammonium salts of the amines. The ammonium salts are not
oxidation sensitive.
4.2 Origin and occurrence of amines
Amines are produced by technical chemical processes and processing and in addition by biotic or
abiotic decomposition of nitrogen compounds. Besides sources of biological origin, indoor air sources of
amines could be, for instance, products containing polyurethane, especially foams, e.g. in vehicle seats,
mattresses, pillows, and upholstered furniture or as thermal insulation or sound absorbing material.
Several amines, in particular aromatic amines, are known as harmful compounds. Furthermore, most
amines have an unacceptable odour in combination with a low odour threshold.
Further sources are for example food, e.g. fish (aliphatic amines), and cigarette smoke (aromatic
amines).
5 Analytical procedure
5.1 Preparation of sample solutions
ISO 16000-38 describes the procedure to get the aqueous eluate of the ammonium salts of the sampled
amines. The sample solution (which contains already approximately 0,02 µmol/µl up to 0,1 µmol/µl
phosphoric acid) can be used for the injection into the HPLC system without further treatment.
5.2 High-performance liquid chromatography
— stationary phase of separating column: pentafluorophenyl;
— common column dimensions: length 150 mm, inner diameter: 2,1 mm;
2 © ISO 2019 – All rights reserved

— common particle size: 3 μm (HPLC), 1,7 µm [Ultra Performance Liquid Chromatography (UHPLC)];
— column temperature: 40 °C;
— injection volume: 1 µl to 10 μl;
— mobile phase: acetonitrile and water with each 0,02 % formic acid (500 µl formic acid in 2,5 l
acetonitrile or 2,5 l water, respectively);
— mobile phase composition: 28 % acetonitrile/72 % water, isocratic.
5.3 Tandem mass spectrometry
+
— ionisation: electrospray ionisation, positive mode (ESI );
— source temperature: 120 °C;
— desolvation gas: nitrogen, 900 l/h;
— desolvation temperature: 420 °C;
— cone gas: nitrogen, 50 l/h;
— precursor ion: M+1;
— collision gas: argon;
— calibration: external and addi
...

INTERNATIONAL ISO
STANDARD 16000-39
First edition
2019-06
Indoor air —
Part 39:
Determination of amines —
Analysis of amines by (ultra-) high-
performance liquid chromatography
coupled to high resolution or tandem
mass spectrometry
Reference number
©
ISO 2019
© ISO 2019
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: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Amines in indoor air . 2
4.1 Properties of amines . 2
4.2 Origin and occurrence of amines . 2
5 Analytical procedure . 2
5.1 Preparation of sample solutions . 2
5.2 High-performance liquid chromatography . 2
5.3 Tandem mass spectrometry . 3
5.4 High resolution mass spectrometry . 3
5.5 Sample sequence and external calibration . 3
6 Testing of the suitability of the equipment and the instrumental analysis .3
6.1 General . 3
6.2 High-performance or ultra-performance liquid chromatography (HPLC or UHPLC) . 3
6.3 Tandem mass spectrometry (MS-MS) . 4
6.4 High resolution mass spectrometry (HRMS) . 4
7 Reporting requirements . 4
Annex A (informative) List of included amines . 6
Annex B (informative) Performance data of the analytical method . 9
Bibliography .12
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 146, Air quality, Subcommittee SC 6,
Indoor air.
A list of all parts in the ISO 16000 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 © ISO 2019 – All rights reserved

Introduction
ISO 16000 (all parts) describe general requirements relating to the measurement of indoor air pollutants
and the important conditions to be observed before or during the sampling of individual pollutants or
groups of pollutants, as well as the measurements procedures themselves.
The definition of indoor environment is given by ISO 16000-1. Dwellings [living rooms, bedrooms, do-
it-yourself (DIY) rooms, sport rooms and cellars, kitchens and bathrooms], workrooms or workplaces
in buildings which are not subject to health and safety inspections with respect to air pollutants (e.g.
offices, salesrooms), public buildings (e.g. restaurants, theatres, cinemas and other meeting rooms) and
passenger cabins of motor vehicles and public transportation are among the most important types of
indoor environment.
INTERNATIONAL STANDARD ISO 16000-39:2019(E)
Indoor air —
Part 39:
Determination of amines — Analysis of amines by (ultra-)
high-performance liquid chromatography coupled to high
resolution or tandem mass spectrometry
1 Scope
This document, along with ISO 16000-38, specifies the measurement method for determining the mass
concentration of primary, secondary and tertiary aliphatic and aromatic amines in indoor air using
accumulated sampling and high-performance liquid chromatography (HPLC) coupled with tandem
mass spectrometry (MS-MS) or high-resolution mass spectrometry (HRMS). The analytical procedure
is covered by this document. The sampling procedure and the manufacturing of the samplers are
covered by ISO 16000-38.
This document describes specifications for the chromatography and the mass spectroscopy for the
amines. Measurement results are expressed in µg/m .
Although primarily intended for the measurement of amines listed in Tables A.1 and A.2, it can also be
used for the measurement of other amines in indoor air. This document gives instructions and describes
procedures for the inclusion of other amines.
The range of application of this document concerning the concentrations of amines in indoor air
depends on the linear range of the calibration line and hence on the gas sample volume (here: from 5 l
up to 100 l), the eluate volume (from 1 ml up to 5 ml), the injection volume (from 1 µl up to 10 µl) and
the sensitivity of the analytical equipment (e.g. linear range from 2 pg up to 2 ng amine). The range
of application can be expected to be from approximately 0,002 µg/m (100 l sample) up to 2 000 µg/
m (5 l sample) for a common analytical equipment (e.g. Waters „TQD“) for the majority of the amines
listed in Tables A.1 and A.2. The analysis of derivatives of ethanolamine is usually about 10 times more
sensitive and the analysis of short-chained aliphatic amines is usually about 10 times less sensitive than
the analysis of an average amine.
The performance data of the analytical method is given in Annex B, particularly in Tables B.1 and B.2.
This document can be used also for the determination of amines in water if the detection limit is
sufficient.
This document does not cover the determination of isocyanates in indoor air (nor in water samples) as
corresponding amines (covered by ISO 17734-1 and ISO 17734-2).
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 16000-38, Indoor air — Part 38: Determination of amines in indoor and test chamber air — Active
sampling on samplers containing phosphoric acid impregnated filters
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardisation at the following addresses:
— ISO online browsing platform: available at http: //www .iso .org/obp
— IEC Electropedia: aivailable at http: //www .elctropedia .org/
3.1
amines
-3
nitrogen containing compounds with a sufficient vapor pressure (>10 Pa) and a free electron pair at
the nitrogen atom which can be protonated
4 Amines in indoor air
4.1 Properties of amines
Amines are basic and polar substances.
There are primary, secondary, tertiary, and quaternary amines.
Quaternary amines are not included in this document, as they have no free electron pair and therefore
have very different properties. In this document the term “amines” refers only to primary, secondary,
and tertiary amines.
Not protonated amines are oxidation sensitive.
Reaction of amines with acids results in ammonium salts of the amines. The ammonium salts are not
oxidation sensitive.
4.2 Origin and occurrence of amines
Amines are produced by technical chemical processes and processing and in addition by biotic or
abiotic decomposition of nitrogen compounds. Besides sources of biological origin, indoor air sources of
amines could be, for instance, products containing polyurethane, especially foams, e.g. in vehicle seats,
mattresses, pillows, and upholstered furniture or as thermal insulation or sound absorbing material.
Several amines, in particular aromatic amines, are known as harmful compounds. Furthermore, most
amines have an unacceptable odour in combination with a low odour threshold.
Further sources are for example food, e.g. fish (aliphatic amines), and cigarette smoke (aromatic
amines).
5 Analytical procedure
5.1 Preparation of sample solutions
ISO 16000-38 describes the procedure to get the aqueous eluate of the ammonium salts of the sampled
amines. The sample solution (which contains already approximately 0,02 µmol/µl up to 0,1 µmol/µl
phosphoric acid) can be used for the injection into the HPLC system without further treatment.
5.2 High-performance liquid chromatography
— stationary phase of separating column: pentafluorophenyl;
— common column dimensions: length 150 mm, inner diameter: 2,1 mm;
2 © ISO 2019 – All rights reserved

— common particle size: 3 μm (HPLC), 1,7 µm [Ultra Performance Liquid Chromatography (UHPLC)];
— column temperature: 40 °C;
— injection volume: 1 µl to 10 μl;
— mobile phase: acetonitrile and water with each 0,02 % formic acid (500 µl formic acid in 2,5 l
acetonitrile or 2,5 l water, respectively);
— mobile phase composition: 28 % acetonitrile/72 % water, isocratic.
5.3 Tandem mass spectrometry
+
— ionisation: electrospray ionisation, positive mode (ESI );
— source temperature: 120 °C;
— desolvation gas: nitrogen, 900 l/h;
— desolvation temperature: 420 °C;
— cone gas: nitrogen, 50 l/h;
— precursor ion: M+1;
— collision gas: argon;
— calibration: external and addi
...