SIST EN ISO 12010:2019

SLOVENSKI STANDARD
01-junij-2019
Nadomešča:
SIST EN ISO 12010:2014
Kakovost vode - Določevanje polikloriranih alkanov s kratko verigo (SCCP) v vodi
- Metoda s plinsko kromatografijo z masno selektivnim detektorjem (GC-MS) in
negativno kemijsko ionizacijo (NCI) (ISO 12010:2019)
Water quality - Determination of short-chain polychlorinated alkanes (SCCP) in water -
Method using gas chromatography-mass spectrometry (GC-MS) and negative-ion
chemical ionization (NCI) (ISO 12010:2019)
Wasserbeschaffenheit - Bestimmung von kurzkettigen Chloralkanen (SCCP) in Wasser -
Verfahren mittels Gaschromatographie-Massenspektrometrie (GC-MS) und negativer
chemischer Ionisation (NCI) (ISO 12010:2019)
Qualité de l'eau - Détermination des alcanes polychlorés à chaîne courte (SCCP) dans
l'eau - Méthode par chromatographie gazeuse-spectrométrie de masse (CG-SM) avec
ionisation chimique négative (ICN) (ISO 12010:2019)
Ta slovenski standard je istoveten z: EN ISO 12010:2019
ICS:
13.060.50 Preiskava vode na kemične Examination of water for
snovi chemical substances
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 12010
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2019
EUROPÄISCHE NORM
ICS 13.060.50 Supersedes EN ISO 12010:2014
English Version
Water quality - Determination of short-chain
polychlorinated alkanes (SCCP) in water - Method using
gas chromatography-mass spectrometry (GC-MS) and
negative-ion chemical ionization (NCI) (ISO 12010:2019)
Qualité de l'eau - Détermination des alcanes Wasserbeschaffenheit - Bestimmung von kurzkettigen
polychlorés à chaîne courte (SCCP) dans l'eau - Chloralkanen (SCCP) in Wasser - Verfahren mittels
Méthode par chromatographie gazeuse-spectrométrie Gaschromatographie-Massenspektrometrie (GC-MS)
de masse (CG-SM) avec ionisation chimique négative und negativer chemischer Ionisation (NCI) (ISO
(ICN) (ISO 12010:2019) 12010:2019)
This European Standard was approved by CEN on 18 February 2019.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 12010:2019 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 12010:2019) has been prepared by Technical Committee ISO/TC 147 "Water
quality" in collaboration with Technical Committee CEN/TC 230 “Water analysis” the secretariat of
which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by October 2019, and conflicting national standards shall
be withdrawn at the latest by October 2019.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 12010:2014.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO 12010:2019 has been approved by CEN as EN ISO 12010:2019 without any modification.

INTERNATIONAL ISO
STANDARD 12010
Second edition
2019-03
Water quality — Determination of
short-chain polychlorinated alkanes
(SCCP) in water — Method using gas
chromatography-mass spectrometry
(GC-MS) and negative-ion chemical
ionization (NCI)
Qualité de l'eau — Détermination des alcanes polychlorés à
chaîne courte (SCCP) dans l'eau — Méthode par chromatographie
gazeuse-spectrométrie de masse (CG-SM) avec ionisation chimique
négative (ICN)
Reference number
ISO 12010:2019(E)
©
ISO 2019
ISO 12010:2019(E)
© ISO 2019
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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below or ISO’s member body in the country of the requester.
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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

ISO 12010:2019(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Interferences . 2
6 Reagents and standards . 3
6.1 Solvents for extraction and preparation of stock solutions . 3
6.2 Reference SCCP stock solutions . 3
6.3 Internal standard stock solutions from individual congeners . 4
6.4 Calibration solutions . 5
6.5 Extraction auxiliary and clean-up materials . 5
7 Apparatus . 6
8 Sampling and sample pretreatment . 7
9 Procedure. 7
9.1 Extraction with liquid-liquid extraction . 7
9.2 Extraction with higher content of suspended matter . 7
9.3 Extract clean-up . 8
9.4 Measurement and integration of the chromatogram. 8
9.5 Calibration . 9
9.5.1 General. 9
9.5.2 Basic calibration .10
9.5.3 Identification and quantification with mass fragment combinations .11
9.5.4 Calculation of the results .11
9.5.5 Quality checks for internal standardization .12
10 Expression of results .12
11 Test report .12
Annex A (normative) Independent quality control check solutions .13
Annex B (informative) Explanation of the calibration of the sum of SCCPs with multiple
linear regression .15
Annex C (informative) Typical GC-MS conditions .21
Annex D (informative) Typical chromatograms of standard solutions and quality control
check solutions 1 µg/ml .24
Annex E (informative) Presentation of goodness of fit .31
Annex F (normative) Alternative clean-up with column chromatography .32
Annex G (informative) Alternative clean-up with gel chromatography .35
Annex H (informative) Chromatograms of real SPM samples .36
Annex I (informative) Performance data .40
Bibliography .42
ISO 12010:2019(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 on 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 the following
URL: www .iso .org/iso/foreword .html.
This document was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 2,
Physical, chemical and biochemical methods.
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.
This second edition cancels and replaces the first edition (ISO 12010:2012), which has been technically
revised. The main changes compared to the previous edition are:
— the m/z values (mass/charge ratios) for quantification and identification;
— the calibration mixtures;
— the clean up procedure by gel chromatography;
— reduced interferences.
iv © ISO 2019 – All rights reserved

ISO 12010:2019(E)
Introduction
The user should be aware that particular problems might require the specifications of additional
marginal conditions.
This document achieves synergetic effects in the practical laboratory work. The following points
partially allow a combination of water and sediment analysis:
[2]
1) same mass combination as for sediment analysis (see ISO 18635 );
2) same calibration mixtures as for sediment analysis (see ISO 18635);
3) same GPC-clean up as for sediment analysis (see ISO 18635).
INTERNATIONAL STANDARD ISO 12010:2019(E)
Water quality — Determination of short-chain
polychlorinated alkanes (SCCP) in water — Method using
gas chromatography-mass spectrometry (GC-MS) and
negative-ion chemical ionization (NCI)
WARNING — Persons using this document should be familiar with normal laboratory practice.
This document 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.
IMPORTANT — It is absolutely essential that tests conducted in accordance to this document be
carried out by suitably qualified staff.
1 Scope
This document specifies a method for the quantitative determination of the sum of short-chain
polychlorinated n-alkanes also known as short-chain polychlorinated paraffins (SCCPs) in the carbon
bond range n-C to n-C inclusive, in mixtures with chlorine mass fractions (“contents”) between
10 13
50 % and 67 %, including approximately 6 000 of approximately 8 000 congeners.
This method is applicable to the determination of the sum of SCCPs in unfiltered surface water, ground
water, drinking water and waste water using gas chromatography-mass spectrometry with electron
capture negative ionization (GC-ECNI-MS).
Depending on the capability of the GC-ECNI-MS instrument, the concentration range of the method
is from 0,1 µg/l or lower to 10 µg/l. Depending on the waste water matrix, the lowest detectable
concentration is estimated to be > 0,1 µg/l. The data of the interlaboratory trial concerning this method
are given in Annex I.
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 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
ISO/TS 13530, Water quality — Guidance on analytical quality control for chemical and physicochemical
water analysis
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 12010:2019(E)
4 Principle
Determination of the sum of SCCPs in the carbon bond range n-C to n-C inclusive, in technical and
10 13
environmental transposed mixtures with chlorine mass fractions (“contents”) between 50 % and 67 %
(for example a mean of approximately 4 to 10 chlorine atoms per molecule) and independent of the
C-number distribution pattern of the congeners. No recognition of the chlorine content is necessary.
The analysed sum of SCCPs includes the variety of SCCPs with their differing chlorine content
and C-number distribution patterns as found in technical mixtures as well as compositions in the
environment (References [5] to [9]).
SCCPs in whole water samples are fortified with an internal standard and extracted using liquid-
liquid extraction with an organic solvent. The sample enrichment procedure is followed by a clean-
up procedure to eliminate interfering compounds. Gas chromatography (GC) is undertaken using a
short capillary column within a short retention time range. The detection of selected mass fragments
is carried out using mass spectrometry (MS) in selected ion-monitoring mode using electron capture
negative ionization mode (ECNI). The mass fragments and the compositions of the calibration solutions
used in this document are essential for the analysis of the sum of SCCPs (see References [3] and [4]).
The selected ion chromatogram is integrated over the full retention time range of the SCCPs. The
quantification of the sum of SCCPs is carried out after establishing a calibration by a multiple linear
regression. The calibration requires the specified three differently composed standard mixtures
fortified with an internal standard.
These standard mixtures mimick different mixtures found in the environment. In this method, only
the multiple linear regression quantification with these specific mixtures enables the quantification of
the variety of observed mixtures of SCCP in the environment and in technical compositions, described
in Clause 1 and in References [3] and [4]. It is not possible to use only one reference mixture for that
complex task.
The method allows for a quantification of the sum of SCCPs expected to be within an expanded
measurement uncertainty of less than 50 %.
5 Interferences
Non-specific matrix interferences, as well as interferences from other environmental situations, are
dealt with using the given clean-up procedure. A further reduction of matrix effects can be achieved by
reducing the mass spectrometric resolution power to, for example, 0,4 u, which is often possible with
a quadrupole mass spectrometer. The exact m/z values are 374,958 8; 410,916 9; 422,935 5; 448,810 6
(see Reference [8]).
Applying the entire procedure using the clean up procedure given in 9.3, a selection of chlorinated
pollutants has been tested and found not to cause interferences below the concentrations given in
Table 1.
Table 1 — Highest concentration level which causes no interferences higher than the limit of
quantification of 0,1 µg/l
Highest concentration level which
causes no interferences higher
Potential interfering compounds
than the limit of quantification of
0,1 µg/l
a
Aroclor 1262 1,25 µg/l
a
Aroclor 1242 10 µg/l
a
Aroclor 1221 10 µg/l
a
Aroclor 1262, Aroclor 1242, Aroclor 1221, Halowax 1014 and Halowax 1051 are examples of suitable products
available commercially. These examples are given for the convenience of users of this document and do not constitute an
endorsement by ISO of these products.
2 © ISO 2019 – All rights reserved

ISO 12010:2019(E)
Table 1 (continued)
Highest concentration level which
causes no interferences higher
Potential interfering compounds
than the limit of quantification of
0,1 µg/l
Campheclor (toxaphene) 1,75 µg/l
a
Halowax 1014 10 µg/l
a
Halowax 1051 0,4 µg/l
MCCP (medium-chain chlorinated n-alkanes C -C ) 42 % chlorine 10 µg/l
14 17
MCCP (medium-chain chlorinated n-alkanes C -C ) 52 % chlorine 6 µg/l
14 17
MCCP (medium-chain chlorinated n-alkanes C -C ) 57 % chlorine 10 µg/l
14 17
a
Aroclor 1262, Aroclor 1242, Aroclor 1221, Halowax 1014 and Halowax 1051 are examples of suitable products
available commercially. These examples are given for the convenience of users of this document and do not constitute an
endorsement by ISO of these products.
6 Reagents and standards
Use solvents and reagents of sufficient purity, i.e. with negligibly low concentrations of SCCPs, e.g. lower
than the limit of detection of the method. Check blanks regularly over the entire procedure to ensure
they are suitable and establish proper analytical control.
6.1 Solvents for extraction and preparation of stock solutions
The solvent for extraction is n-heptane. Other non-polar solvents, e.g. n-hexane (C H ), cyclohexane
6 14
(C H ), can be used if the extraction efficiency is comparable with those of n-heptane.
6 12
Use 2,2,4-trimethylpentane (C H , isooctane) for conditioning of the glass bottles (7.1).
8 18
For preparation of the stock solution and dilutions of the internal standard, use propanone
(acetone), C H O.
3 6
For conditioning of the clean-up columns, use mixtures of n-heptane and propanone (acetone).
For the first elution step of the filtrated suspended matter, use methanol (CH OH).
6.2 Reference SCCP stock solutions
Use commercially available solutions, such as in cyclohexane or n-hexane, of the single mixtures of SCCP
congeners with defined carbon chain length and with different defined chlorine contents (see Table 2,
first two columns). Alternatively, use commercially available ready mixed solutions with the same
composition.
Mixtures of synthetic solutions were used to simulate environmentally occurring SCCPs or technical
products of SCCPs. For example, the synthetic mixed calibration stock solution “Lake Ontario water” is
mixed to resemble a Lake Ontario water as reported in Reference [6]. Its characteristic is a relatively
high content of C to C , especially C and a low chlorine content as partly reported in water samples
10 12 12
too. The synthetic mixed calibration stock solution “Perch” simulates a C-number distribution found in
a perch (see Reference [7]). The standard mixture “Sediment Drevnice” simulates a natural mixture
reported about a sediment of the river Drevnice (see Reference [8]) with a high content of C and a
higher chlorine content.
The compositions of the calibration mixtures as well as of the independent quality assurance solutions
are mandatory to achieve the quantification of the variety of SCCP-mixtures.
Prepare the solutions “Lake Ontario water”, ”Perch”, and “Sediment Drevnice” according to Table 2.
ISO 12010:2019(E)
Table 2 — Reference substances stock solutions
Reference substances stock solutions in
accordance with 6.2
Commercially available standard solutions
(Synthetic mixed standard solutions, which
resemble environmental mixtures composi-
tion, ng/ml)
“Lake Ontario “Sediment
“Perch”
Chlorine content
water” Drevnice”
n-alkane
% of the individual Mean number of chlorines in
chain
Chlorine Chlorine Chlorine
C-number mixtures as the molecules (calculated)
length a a a
content content content
certified
50,2 % 60,6 % 65,0 %
C 50,18 3,97 1 000
C 55,00 4,79 1 000
C 60,09 5,86 500
C 65,02 7,16 1 100 280
C 45,50 3,63 1 000
C 50,21 4,37 1 000
C 55,20 5,31 600
C 60,53 6,55 1 000 500
C 65,25 7,94 3 000 660
C 45,32 3,93 2 000
C 50,18 4,76 2 000 800
C 55,00 5,74 2 000 2 000
C 65,08 8,59 900 1 000
C 69,98 10,62 830
C 59,98 7,56 100 730
C 65,18 9,34 6 000
Sum of SCCP (ng/ml) 10 000 10 000 10 000
a
The chlorine content of the mixtures is calculated as the weighted mean.
Store the prepared solutions in a refrigerator at 2 °C to 8 °C. Avoiding losses of the solvent by evaporation,
solutions can be used for five years.
Use as well commercially available solutions, e.g. in cyclohexane or n-hexane, of the reference substances
stock solutions (see Table 2, last three columns) of SCCP. See Reference [8].
1)
An example is DRE-ZS22102105HP . See Reference [8].
6.3 Internal standard stock solutions from individual congeners
Use commercially available individual congener standard solutions and prepare a stock solution in
propanone (acetone) (6.1) at a concentration of, for example, 1 µg/ml.
Individual SCCP congeners with chlorine contents of between 50 % and 67 % are suitable as internal
standards, for example:
— 1,1,1,3,10,11-hexachloroundecane, with e.g. 0,1 µg/ml;
— 1,1,1,3,11,13,13,13-octachlorotridecane, with e.g. 0,1 µg/ml;
— 1,2,5,5,6,9,10-heptachlorodecane, with e.g. 0,01 µg/ml.
1) DRE-ZS22102105HP is an example of a suitable product available commercially. These examples are given for
the convenience of users of this document and do not constitute an endorsement by ISO of these products.
4 © ISO 2019 – All rights reserved

ISO 12010:2019(E)
NOTE 1 The different individual SCCP congeners used as internal standard substances contribute in
environmental samples to the sum of SCCPs. Nevertheless, the contribution is approximately <1 %, which means
that the enhancement of the measurement uncertainty is negligible.
NOTE 2 Different individual SCCP congeners can produce different response factors, hence it can be necessary
for different concentrations to be used.
If validated, other individual SCCP congeners can be used as the internal standard if the congener shows
the same properties over the entire analytical process as the SCCPs being determined.
The solutions can be stored in a refrigerator at 2 °C to 8 °C.
6.4 Calibration solutions
Use the standard mixtures according to Table 2. Prepare a minimum of nine calibration solutions (see
Table 3) with concentrations according to the detection capability of the mass spectrometer. Combine
and dilute the solutions (6.2) and the internal standard solution (6.3) with n-heptane to produce
solutions for the calibration range.
Table 3 — Calibration solutions
Internal standard
“Lake Ontario water” “Perch” “Sediment Drevnice”
e.g. 1,1,1,3,11,13,13,13-octa-
chlorotridecane
Sum of SCCPs, µg/ml µg/ml µg/ml µg/ml
µg/ml
0,15 0,15 0,1
0,15 0,15 0,1
0,15 0,15 0,1
0,3 0,3 0,1
0,3 0,3 0,1
0,3 0,3 0,1
0,6 0,6 0,1
0,6 0,6 0,1
0,6 0,6 0,1
The solutions may be stored in a refrigerator at 2 °C to 8 °C at least for six months. Check the
concentration of the calibration solutions against an independently prepared standard prior to use.
Quality control check solutions shall be prepared to check the calibration independently. To do so, use
the mixtures as given in Annex A.
6.5 Extraction auxiliary and clean-up materials
6.5.1 Copper powder, grain size < 63 µm. Copper powder is used in the clean-up procedure to remove
sulfur and sulfur-containing matrix components.
6.5.2 Hydrochloric acid, 2 mol/l, used for copper activation in the clean-up column.
6.5.3 Aluminium oxide, Al O , neutral, high activity (10 % water).
2 3
6.5.4 Glass wool.
6.6 Operating gases, for GC-MS, of high purity and in accordance with the manufacturer’s
specifications.
ISO 12010:2019(E)
6.7 Nitrogen, N , purity ≥ 99,996 % volume fraction, for concentrating the solutions.
6.8 Sodium sulfate, anhydrous, Na SO , powdered.
2 4
6.9 Test solution for check of linearity of the internal standards.
Prepare solutions of the internal standard used at concentrations of 0,1 µg/ml, 0,5 µg/ml, and 1 µg/ml.
7 Apparatus
Glassware and equipment which may come into contact with water samples or their extracts should be
free from interfering compounds.
Clean all glassware by rinsing with propanone (acetone) (6.1).
7.1 Flat-bottomed glass bottles, conical shoulder, 1 000 ml capacity, for collecting water samples,
preferably with glass stoppers.
The sample bottle shall enable direct extraction of the sample to be undertaken.
Before use, condition it by rinsing the dry sample bottle with, for example, 2 ml of isooctane (6.1). Then,
invert it and allow the solvent to drain and evaporate from it.
7.2 Evaporation device, e.g. rotary evaporator, or nitrogen evaporating system.
[1]
7.3 Separator, for example micro-separator in accordance with ISO 6468 , separation funnel or
other suitable device for phase separation.
7.4 Vials, compatible with the GC-autosampler (e.g. with a capacity of 1,5 ml).
7.5 Chromatographic column, internal diameter (ID) 10 mm (empty) for clean-up.
7.6 Gas chromatograph, temperature-programmable, with all required accessories, including gases,
capillary column, split/splitless injector and mass spectrometer detector with negative-ion chemical
ionization option and appropriate reactant gas (CH ).
7.7 Volumetric flasks, 1 ml, 2 ml, 10 ml and 25 ml.
7.8 Disposable glass Pasteur pipettes, e.g. 150 mm or 250 mm.
7.9 Syringes, 2 µl, 5 µl, 10 µl and 50 µl.
7.10 Analytical column
Fused silica column with medium or non-polar low bleed separating phase (see Annex C for examples);
e.g. ID < 0,25 mm, length 15 m and film thickness 0,1 µm.
7.11 Glass fibre filter, binderless, fine porosity (<0,45 µm particle retention).
7.12 Vacuum filtration device, volume 1 l.
7.13 Shaking device or magnetic stirrer device (with a magnetic stir bar).
6 © ISO 2019 – All rights reserved

ISO 12010:2019(E)
7.14 GPC clean-up system (with modular design).
7.14.1 Pump, sampling injector, sample rack; fraction collector.
2)
7.14.2 GPC-Column: Shodex CLNpakPAE 800 AC® , Maximum pore size 40 nm, column size 80 mm
(inner diameter) × 300 mm (length).
8 Sampling and sample pretreatment
Take samples as specified in ISO 5667-1 and ISO 5667-3. To collect water samples (1 l per sample), use
conditioned glass bottles (7.1). Do not fill the sample bottle completely (e.g. fill to the shoulder) in order
to allow the addition of the extracting solvent.
Samples are extracted without filtering the sample and suspended solids are not removed prior to
analysis.
Weigh, to the nearest gram, the sample bottle with its contents and cap, and record the mass for
subsequent use. Thoroughly shake the bottle to homogenize the water sample. Add the internal
standard solution (6.3) to achieve a concentration of, for example, 0,1 µg/l in the water sample. Record
the mass, in micrograms, of internal standard added. Shake the bottle thoroughly.
9 Procedure
9.1 Extraction with liquid-liquid extraction
Add 10 ml of extraction solvent, n-heptane (6.1), to the bottle and shake it or stir (7.13) thoroughly for
about 2 h to carry out the extraction directly in the sample bottle. Allow the phases to separate and
use the separator (7.3) to collect the organic extract in a separate tube. If an emulsion forms, break
it by centrifuging and/or by adding sodium sulfate (6.8) to the tube. Discard the remaining water to
waste. Transfer the solvent from the tube to the evaporating device (7.2) or, using a gentle stream
of nitrogen (6.7), carefully evaporate the solvent (at a temperature of 40 °C) to about 1 ml. Weigh, to
the nearest gram, the empty sample bottle and cap. Calculate the volume of water extracted and the
concentration of internal standard in the water.
Proceed as in 9.3.
9.2 Extraction with higher content of suspended matter
If the content of suspended matter is higher than approximately 200 mg/l, filter the sample through a
glass fibre filter (7.11) and collect the filtrate in the bottle (7.1).
Weigh, to the nearest gram, the empty sample bottle and cap. Calculate the volume of water extracted
and the concentration of internal standard in the water.
Add 10 ml of methanol to the filter (without vacuum) separately to extract the suspended matter. Allow
to soak for 5 min, then use vacuum to add methanol to the sample filtrate collected before.
Add 10 ml n-heptane (without vacuum) to the filter and allow to soak for another 5 min, then use weak
vacuum to add n-heptane also to the sample filtrate collected before.
Shake or stir (7.13) the mixture thoroughly for about 2 h to carry out the extraction directly in the
sample bottle. Allow the phases to separate and use the separator (7.3) to collect the organic extract in
a separate tube. If an emulsion forms, break it by centrifuging and/or by adding sodium sulfate (6.8) to
the tube. Discard the remaining water to waste. Transfer the solvent from the tube to the evaporating
2) Shodex CLNpakPAE 800 AC is an example of a suitable product available commercially. This example is given for
the convenience of users of this document and does not constitute an endorsement by ISO of this product.
ISO 12010:2019(E)
device (7.2) or, using a gentle stream of nitrogen (6.7), carefully evaporate the solvent (at a temperature
of 40 °C) to about 1 ml.
9.3 Extract clean-up
Often, matrix contents can be cleaned by procedure b) only.
A two-step clean-up procedure [a) and b)] for high matrix content shall be carried out.
Begin the cleanup procedure using the extract concentrated to approximately 1 ml in n-heptane:
a) column chromatographic clean-up with 2 g activated copper powder and (6.5.1) 2 g Al O , neutral,
2 3
high activity (6.5.3);
3)
b) gel chromatographic clean-up (7.14). with Shodex CLNpakPAE 800 AC® , 8,0 mm × 300 mm and
0,5 ml/min propanone (acetone) as the eluent.
Fill the copper powder in a glass column with a glass wool plug. Activate the copper by adding 10 ml
of 2 mol/l hydrochloric acid (6.5.2). Allow all of the hydrochloric acid (6.5.2) to soak into the copper
powder before washing the column, first with 25 ml of water and subsequently with 20 ml of acetone to
remove acid and water from the column. Then, a stopcock may be attached to the bottom of the column
for controlling the elution progress. After the solvent level has reached the upper level of the copper
powder, wash the copper layer with 3 × 2 ml n-heptane. Then, 2 g of aluminium oxide (10 % water)
and about 10 ml of n-heptane are added for obtaining a copper/aluminium oxide - sandwich column.
The wet column is used for applying the sample extract. The SCCPs are eluted by 10 ml of a mixture
of n-heptane/acetone (98:2) which is concentrated to approx. 1,2 ml. It shall be noted that the column
shall never run dry.
This concentrate is used for the subsequently following GPC clean-up. Inject e.g 1 ml of the concentrate
and elute in a fraction between 12 ml and 13,5 ml. This fraction is concentrated again to, for example, 1 ml,
dried by sodium sulfate (6.8) and transferred into a sample vial for injection into the GC-MS.
When using a new column for the GPC clean-up, verify the eluent volume for complete elution of the
analytes of interest by analysing an appropriate standard solution and/or spiked sample extract.
Recoveries of SCCPs should be > 50 % and no interfering peak should appear in the gas chromatogram.
If necessary, GPC conditions need to be modified to meet these requirements.
NOTE 1 Alternative clean-up procedures, an extended column chromatographic clean-up (see Annex F) and
a modified gel chromatographic clean-up (see Annex G) can be used. The interferences quantified in Clause 4 of
this document apply only to the conditions described in this clause.
NOTE 2 Due to the physico-chemical properties of SCCP’s results in water higher than the LOD (limit of
detection) are very rare. The absence of SCCP can be proved by testing without the described clean up steps
only after drying the extract. If the peak areas of the specific mass ions are below the peak areas of the LOD, a
result < LOD can be reported.
9.4 Measurement and integration of the chromatogram
Optimize the operating conditions of the GC-ECNI-MS system, e.g. according to the manufacturer’s
instructions. Examples of the gas chromatographic conditions are given in Annex C.
Prior to analysis, verify the performance of the GC-ECNI-MS system by analysis of calibration standards.
Use as a minimum the calibration solutions “Lake Ontario water” and “Sediment Drevnice” to optimize
the GC-ECNI-MS system.
3) Shodex CLNpakPAE 800 AC® is an example of a suitable product available commercially. This example is given
only as information for the convenience of users of this document and does not constitute an endorsement by ISO of
this product.
8 © ISO 2019 – All rights reserved

ISO 12010:2019(E)
Check the GC-ECNI-MS system performance regularly, for example between every 10 to 20 samples,
by independently prepared calibration solutions (see Table 3) with a concentration of e.g. 1 µg/ml
sum of SCCP.
The measurement is performed in the selected ion mode with four selected mass ion fragments (mass
to charge values, m/z), i.e. m/z 375, m/z 411, and m/z 423, m/z 449. The peak areas of the two masses,
m/z 375 and m/z 423, are used in the multiple linear regression calculation of the sum of SCCP’s. The
peak areas of m/z 411 and m/z 449 are used for an additional identification. This specific selection has
been carried out by data analysis to analyse the sum of SCCPs of a large variety of environmentally
occurring SCCP-mixtures. For an explanation of this selection, see Reference [4].
The integration of the different m/z values should be carried out within different retention time ranges
that are established from calibration solutions. Retention time ranges of chromatograms in Annex D
are given in Table 4 as an example.
Table 4 — Typical retention time ranges
Approximate retention time range Approximate retention time range of the
m/z value
a
response maximum
min min
375 6,6 to 9,0 7,1 to 8,2
411 7,0 to 8,8 7,4 to 8,2
423 7,0 to 9,0 7,5 to 8,4
449 7,0 to 8,7 7,5 to 8,2
a
This represents the major portion of the SCCPs for the mass ion fragment monitored and is represented by an
unresolved complex mixture of peaks.
An example for integration of a real sample is given in Annex H.
Use selected ion mode measurements for detecting the internal standard. Integrate the response of the
internal standard as a single peak with the following m/z values (see Table 5).
Table 5 — m/z values of internal standard
Internal standard substance m/z for quantification m/z optional
1,1,1,3,10,11-Hexachloroundecane 364 362
1,1,1,3,11,13,13,13-Octachlorotridecane 460 458
1,2,5,5,6,9,10-Heptachlorodecane 348 346
Examples with chromatograms of the mixtures are given in Annex D.
9.5 Calibration
9.5.1 General
Short-chain polychlorinated n-alkanes with 50 % to 67 % chlorine content are mixtures containing
approximately 6 000 congeners. SCCP compounds of different chlorine contents exhibit different
response factors in ECNI-MS. Interferences occur in the mass spectra because individual compounds
cannot be separated by GC.
Using multiple linear regression techniques quantification can be carried out to a large extent
independent of chlorine content. See Annex B and Reference [4].
While modern mass spectrometric software frequently does not enable multiple linear regression
techniques to be carried out, commercial software that does is available. See also the ready to use Excel
sheet accessible through the following link: http: //standards .iso .org/iso/12010/ed -2/en
ISO 12010:2019(E)
9.5.2 Basic calibration
Analyse the calibration solutions (6.4) and integrate th
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