INTERNATIONAL ISO

STANDARD 28540

First edition
2011-08-01

Water quality — Determination of

16 polycyclic aromatic hydrocarbons
(PAH) in water — Method using gas
chromatography with mass spectrometric
detection (GC-MS)
Qualité de l'eau — Détermination de 16 hydrocarbures aromatiques
iTeh STANDARD
polycycliques (HAP)PREVIEW
dans l'eau — Méthode par chromatographie en
phase gazeuse avec détection par spectrométrie de masse (CG-SM)
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ISO 28540:2011
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Reference number
ISO 28540:2011(E)

© ISO 2011
ISO 28540:2011(E)

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ISO 28540:2011
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 ISO 28540:2011(E)

Contents Page

Foreword ............................................................................................................................................................ iv
Introduction ......................................................................................................................................................... v
1 Scope ...................................................................................................................................................... 1
2 Normative references ............................................................................................................................ 1
3 Terms and definitions ........................................................................................................................... 1
4 Principle ................................................................................................................................................. 2
5 Interferences .......................................................................................................................................... 4
6 Reagents ................................................................................................................................................ 4
7 Apparatus ............................................................................................................................................... 7
8 Sampling ................................................................................................................................................ 8
9 Procedure ............................................................................................................................................... 8
10 Calibration ............................................................................................................................................ 11
11 iTeh STANDARD PREVIEW
Measurement of samples.................................................................................................................... 12
12 (standards.iteh.ai)
Identification ........................................................................................................................................ 12
13 Calculation ........................................................................................................................................... 14
14
ISO 28540:2011
Expression of results .......................................................................................................................... 16
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15 Test report ............................................................................................................................................
01f50d98ce47/iso-28540-2011 17
Annex A (informative) Examples for GC-MS conditions ............................................................................... 18
Annex B (informative) Precision and accuracy.............................................................................................. 19
Annex C (informative) Examples of the construction of special apparatus ............................................... 22
Annex D (informative) Example of chromatograms ...................................................................................... 24
Annex E (informative) Extraction with extraction disks................................................................................ 26
Bibliography ...................................................................................................................................................... 27

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ISO 28540:2011(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.

International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.

The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.

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.

ISO 28540 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 2, Physical,
chemical and biochemical methods.
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 ISO 28540:2011(E)

Introduction
Polycyclic aromatic hydrocarbons (PAH) occur in nearly all types of water, these substances are adsorbed on
solids (sediments, suspended matter) as well as dissolved in the liquid phase.

ISO 17993[7] specifies methods for the determination of 15 PAH by high performance liquid chromatography
in drinking water, ground water, and surface water.

ISO 7981-1[3] and ISO 7981-2[4] specify methods for the determination of 6 PAH by high performance thin
layer chromatography or by high performance liquid chromatography in drinking water and ground water.

This International Standard describes a method for at least 16 PAH using gas chromatography with mass
spectrometric detection (GC-MS) in drinking water, ground water and surface water.

Some PAH are known or suspected to cause cancer. Maximum acceptable levels have been set in a number
of countries. For instance, the European Council Directive 98/83/EC on the quality of water intended for
human consumption (Reference [10]) set the maximum acceptable level for benzo[a]pyrene at 0,010 µg/l, and
for the sum of four specified PAH (benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[ghi]perylene,
indeno[1,2,3-cd]pyrene) at 0,100 µg/l.

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 iTeh STANDARD PREVIEW
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ISO 28540:2011
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01f50d98ce47/iso-28540-2011
INTERNATIONAL STANDARD ISO 28540:2011(E)

Water quality — Determination of 16 polycyclic aromatic

hydrocarbons (PAH) in water — Method using gas
chromatography with mass spectrometric detection (GC-MS)

WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This standard does not purport to address all of the safety problems, if any, associated with
its use. It is the responsibility of the user to establish appropriate safety and health practices and to
ensure compliance with any national regulatory conditions.

IMPORTANT — It is absolutely essential that tests conducted according to this International Standard
be carried out by suitably trained staff.

1 Scope
This International Standard specifies a method for the determination of at least 16 selected PAH (see Table 1)
in drinking water and ground water in mass concentrations above 0,005 µg/l and in surface water in mass
iTeh STANDARD PREVIEW
concentrations above 0,01 µg/l (for each individual compound).

This International Standard can be(standards.iteh.ai)

used for samples containing up to 150 mg/l of suspended matter.

This method is, with some modification, also suitable for the analysis of waste water. It is possible that this
method is applicable to other PAH, providedISO
the 28540:2011
method is validated for each case.
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2 Normative references
The following referenced documents are indispensable for the application 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 5667-1, Water quality — Sampling — Part 1: Guidance on the design of sampling programmes and
sampling techniques

ISO 5667-3, Water quality — Sampling — Part 3: Preservation and handling of water samples

ISO 8466-1, Water quality — Calibration and evaluation of analytical methods and estimation of performance
characteristics — Part 1: Statistical evaluation of the linear calibration function

3 Terms and definitions

For the purposes of this document, the following terms and definitions apply.

3.1
analyte
substance to be determined

[ISO 15089:2000[5], 3.2]

NOTE Substances determinable by this International Standard are listed in Table 1.

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ISO 28540:2011(E)

3.2
calibration solution
solution prepared from a secondary standard and/or stock solutions and used to calibrate the response of the
instrument with respect to analyte concentration

[ISO 18073:2004[8], 3.1.2]

3.3
GC-MS determination diagnostic ion
selected fragment or molecular ion of the target compound with the highest possible specificity

3.4
injection standard
standard mixture added to a sample before injection into the GC-MS apparatus, to monitor variability of
instrument response and to calculate internal standard recovery

NOTE In this International Standard, the injection standard mixture contains an isotopically labelled PAH.

3.5
internal standard
isotopically labelled PAH or PAH unlikely to be present in the sample, added to samples prior to extraction,
against which the concentrations of native substances are calculated

3.6
selected ion monitoring/recording mode
SIM/SIR iTeh STANDARD PREVIEW
measuring the intensity of selected diagnostic ions only

NOTE
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[9]
Adapted from ISO 22892:2006 , 3.8.

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4 Principle 01f50d98ce47/iso-28540-2011
The PAH (see Table 1) present in the aqueous sample are extracted from the water sample by liquid-liquid
extraction with hexane. An internal standard mixture is added to the sample prior to extraction. The extract is
concentrated by evaporation, and the residue taken up in a solvent appropriate for clean-up or GC analysis.

NOTE 1 Other volatile solvents can be used as well if it is proven that there is equal or better recovery (recovery mass
fraction between 70 % and 110 %).

The liquid-liquid extraction method shall not be used with samples containing more than 150 mg/l of
suspended matter.

WARNING — The use of this International Standard may involve hazardous materials, operations and
equipment. This International Standard does not purport to address all the safety problems associated
with its use. It is the responsibility of the user of this standard to establish appropriate safety and
health practices and determine the applicability of regulatory limitations prior to use.

NOTE 2 For samples that contain more than 150 mg/l suspended matter the procedure described in ISO 17858:2007[6],
4.1, 4.2 and 4.3, can be used.

If necessary, extracts of surface water samples can be cleaned by column chromatography prior to analysis.
Prior to injection, injection standards are added to each extract, and an aliquot of the extract is injected into
the gas chromatograph.

PAH are separated on a suitable fused silica capillary column, coated with a film of cross-linked non-polar
polysiloxane or slightly polar modified polysiloxane with an efficient separation. The column shall be suitable
for separating benzo[a]pyrene and benzo[e]pyrene. Identification and quantification is performed by means of
mass spectrometry using electron impact ionization (EI).

2 © ISO 2011 – All rights reserved

 ISO 28540:2011(E)

Table 1 — Polycyclic aromatic hydrocarbons determinable by this International Standard

Name Name
Chemical formula Chemical formula
Molar mass Structure Molar mass Structure
% carbon % carbon
CAS number CAS number
Naphthalene Fluorene
C10H8 C13H10
128,17 g/mol 166,22 g/mol
93,75 % C 93,59 % C
91-20-3 86-73-7
Acenaphthylene Acenaphthene
C12H8 C12H10
152,20 g/mol 154,21 g/mol
94,6 % C 93,05 % C
208-96-8 83-32-9
Anthracene Phenanthrene
C14H10 C14H10
178,23 g/mol 178,23 g/mol
94,05 % C 94,05 % C
120-12-7 85-01-8
Pyrene Fluoranthene
C16H10 C16H10
202,26 g/mol iTeh STANDARD PREVIEW 202,26 g/mol
95,0 % C 95,0 % C
129-00-0 (standards.iteh.ai) 206-44-0
Chrysene Benzo[a]anthracene
C18H12 ISO 28540:2011 C18H12
228,29 g/mol https://standards.iteh.ai/catalog/standards/sist/e97873a0-4177-45f1-b2f4-
228,29 g/mol
94,45 % C 01f50d98ce47/iso-28540-2011
94,45 % C
218-01-9 56-55-3
Benzo[k]fluoranthenea Benzo[b]fluoranthenea
C20H12 C20H12
252,32 g/mol 252,32 g/mol
95,2 % C 95,2 % C
207-08-9 205-99-2
Indeno[1,2,3-cd]pyrenea Benzo[a]pyrenea
C22H12 C20H12
276,34 g/mol 252,32 g/mol
95,6 % C 95,2 % C
193-39-5 50-32-8
Benzo[ghi]perylene a Dibenzo[a,h]anthracenea
C22H12 C22H14
276,34 g/mol 278,35 g/mol
95,6 % C 94,7 % C
191-24-2 53-70-3
a Compound specified by Council Directive 98/93/EC (Reference [10]).

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ISO 28540:2011(E)

5 Interferences

5.1 Interferences with sampling, extraction, and concentration

Use sampling containers of materials that do not affect the analyte content during the contact time (preferably
of stainless steel or glass). Avoid plastics and other organic materials during sampling, sample storage or
extraction. Care should be taken when using surfactants for cleaning sample containers, because they may
lead to the formation of emulsions during liquid-liquid extraction.

If automatic samplers are used, avoid the use of silicone or rubber material for the tubes. If these materials
are present, ensure that the contact time is minimized. Rinse the sampling line with the water to be sampled
before taking the test sample. Use ISO 5667-1 and ISO 5667-3 for guidance.

Keep the test samples away from direct sunlight and prolonged exposure to light. Store the samples in amber
containers. Clear glass bottles are suitable as well, but then the samples shall be kept in a dark place.

During storage of the test samples, losses of PAH may occur due to adsorption on to the walls of the
containers. The extent of the losses may depend on the storage time.

5.2 Interferences with GC-MS

Substances that co-elute with the target PAH may interfere with the determination. These interferences may
lead to incompletely resolved signals and may, depending on their magnitude, affect accuracy and precision
of the analytical results. Non-symmetrical peaks and peaks broader than the corresponding peaks of the
reference substance suggest interferences.
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Chromatographic separation between dibenzo[a,h]anthracene and indeno[1,2,3-cd]pyrene is most critical.
Due to their molecular mass differences, quantification can be made by mass selective detection. When
incomplete resolution is encountered, peak integration shall be checked and, when necessary, the baseline
ISO 28540:2011
corrected. Sufficient resolution (e.g. not less than R = 0,8) between the peaks of benzo[b]fluoranthene and
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benzo[k]fluoranthene as well as of benzo[a]pyrene and benzo[e]pyrene is to be set as quality criterion for the
capillary column. Benzo[j]fluoranthene 01f50d98ce47/iso-28540-2011
cannot be separated from benzo[k]fluoranthene and
benzo[b]fluoranthene. It is possible that triphenylene is not completely separated from benzo[a]anthracene
and chrysene. If this is the case, state this fact in the test report.

NOTE Benzo[j]fluoranthene, benzo[e]pyrene, and triphenylene are not part of the 16 target analytes.

6 Reagents

WARNING — The use of this International Standard may involve hazardous materials, operations and
equipment. This International Standard does not purport to address all the safety problems associated
with its use. It is the responsibility of the user of this standard to establish appropriate safety and
health practices and determine the applicability of regulatory limitations prior to use.

During the analysis, unless otherwise stated, use only reagents of recognized analytical grade, “for residue
analysis” or “for GC analysis”, where appropriate, and distilled or demineralized water or water of equivalent
purity. Otherwise, pay extra attention that each batch of solvents does not contain blank concentrations
affecting the results.

6.1 Sodium thiosulfate pentahydrate, Na2S2O3·5H2O, for dechlorination.

6.2 Solvents.

6.2.1 Hexane, C6H14.

6.2.2 Acetonitrile, CH3CN.

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 ISO 28540:2011(E)

6.2.3 Acetone, C3H6O.

6.2.4 Decane, C10H22.

6.2.5 Isooctane, C8H18.

6.2.6 Dichloromethane, CH2Cl2.

6.3 Sodium sulfate, Na2SO4, anhydrous, precleaned by heating to 500 °C for 4 h or free of interfering
compounds.

6.4 Gases.

6.4.1 Nitrogen, 99,999 % volume fraction, for the purpose of evaporating the extracts.

6.4.2 Helium, 99,999 % volume fraction, for gas chromatography.

6.5 Standards.

6.5.1 Reference substances (see Table 2) and internal standards.

Choose internal standards with physical and chemical properties (such as extraction behaviour, retention
time) that are similar to those of the compounds to be analysed.

Use a minimum of three internal standards, e.g. three deuterated PAH, for evaluation of results (see
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Clauses 11 and 12). Verify the stability of the internal standards regularly. Table 2 contains native PAH and a
number of deuterated PAH that can be used. The internal standards are added to the sample to be extracted
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and are therefore dissolved in a water-soluble solvent.

NOTE 1 13C isotopically labelled PAH standards

ISOcan also be used as internal standards.
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NOTE 2 Certified solutions of PAH and single solid PAH substances with certified purity are available from a limited
01f50d98ce47/iso-28540-2011
number of suppliers 1) or from other commercial providers.

Because of the dangerous nature of these substances, commercially available, preferably certified, standard
solutions should be used. Skin contact should be avoided.

6.5.2 Injection standard.

Add an isotopically labelled non-polar substance to the final extract and to the calibration solutions (6.8)
before GC-MS injection to check the recovery of the internal standards.

Prepare a stock solution of the injection standard in an appropriate solvent of mass concentration   10 µg/ml.

6.6 Single substance stock solutions.

Prepare solutions of the single substances (see Table 1) in an appropriate solvent, e.g. hexane (6.2.1) or
acetonitrile (6.2.2), with mass concentration   200 µg/ml.

These solutions can be used for confirmation and identification of single PAH in the chromatogram.

1) The Institute for Reference Materials and Measurements (IRMM), Geel, Belgium, the National Institute of Science and
Technology (NIST), Washington DC, USA, and the Cambridge Isotope Laboratory (CIL), Andover, MA, USA, are
examples of suitable product suppliers. This information is given for the convenience of users of this International
Standard and does not constitute an endorsement of these suppliers by ISO.

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ISO 28540:2011(E)

6.7 Multiple substance stock solution.

Dilute a sufficient volume (e.g. 5 ml) of the single substance stock solutions (6.6) in a one-mark volumetric
flask (7.14, e.g. 100 ml) with an appropriate solvent, e.g. hexane (6.2.1) or acetonitrile (6.2.2), to prepare a
solution of mass concentration   10 µg/ml.

Alternative commercially available (certified) combined and mixed solutions containing only one or some of
the reference substances (see Table 1) at an appropriate mass concentration of the relevant single substance,
e.g. 10 µg/ml in an appropriate solvent, such as acetonitrile (6.2.2) or hexane (6.2.1), may be used.

The solutions 6.5 to 6.7 are stable for at least 1 year when stored in the dark at room temperature and
protected from evaporation. Check the stability of the standard solution regularly. For that purpose,
independent solutions for quality control shall be available within a laboratory.

Table 2 — Native PAH and deuterated PAH

PAH reference substances Labelled internal standard substances deuterated PAH

Naphthalene (CAS No. 91-20-3) Naphthalene-d8 (CAS No. 1146-65-2)

Acenaphthene (CAS No. 83-32-9) Acenaphthene-d10 (CAS No. 15067-26-2)
Acenaphthylene (CAS No. 208-96-8) Acenaphthylene-d8 (CAS No. 93951-97-4)
Fluorene (CAS No. 86-73-7) Fluorene-d10 (CAS No. 81103-79-9)
Anthracene (CAS No. 120-12-7) Anthracene-d10 (CAS No. 1719-06-8)
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Phenanthrene (CAS No. 85-01-8) Phenanthrene-d10 (CAS No. 1517-22-2)
Fluoranthene (CAS No. 206-44-0)
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Fluoranthene-d10 (CAS No. 93951-69-0)
Pyrene (CAS No. 129-00-0) Pyrene-d10 (CAS No. 1718-52-1)
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Benzo[a]anthracene (CAS No. 56-55-3) Benzo[a]anthracene-d12 (CAS No. 1718-53-2)
01f50d98ce47/iso-28540-2011
Chrysene (CAS.No. 218-01-9) Chrysene-d12 (CAS No. 1719-03-5)
Benzo[b]fluoranthene (CAS No. 205-99-2) Benzo[b]fluoranthene-d12 (CAS No. 93951-98-5)
Benzo[j]fluoranthenea (CAS No. 205-82-3)
Triphenylenea (CAS No 217-59-4)
Benzo[k]fluoranthene (CAS No. 207-08-9) Benzo[k]fluoranthene-d12 (CAS No. 93952-01-3)
Benzo[a]pyrene (CAS No. 50-32-8) Benzo[a]pyrene-d12 (CAS No. 63466-71-7)
Benzo[e]pyrenea (CAS No. 192-97-2)
Indeno[1,2,3-cd]pyrene (CAS No. 193-39-5) Indeno[1,2,3-cd]pyrene-d12 (CAS No. 203578-33-0)
Dibenzo[a,h]anthracene (CAS No. 53-70-3) Dibenzo[a,h]anthracene-d14 (CAS No. 13250-98-1)
Benzo[ghi]perylene (CAS No. 191-24-2) Benzo[ghi]perylene-d12 (CAS No. 93951-66-7)
a Not one of the 16 target analytes; used only to check that resolution is sufficient.

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