ISO/FDIS 22032

ISO/DIS FDIS 22032:2025(en)
ISO/TC 147/SC 2
Secretariat: DIN
Date: 2026-02-04
Water quality — Determination of polybrominated diphenyl
ethers (PBDE) in sediment, suspended (particulate) matter and
biota — Method using gas chromatography -coupled with tandem
mass spectrometry (GC-MS/MS) or with high resolution mass
spectrometry (GC-MS/MS; HRMS)
Second edition
Date: 2025-06-26
This draft is submitted to a parallel vote in ISO, CEN.

Qualité de l'eau — Dosage d'éthers diphényliques polybromés (PBDE) dans les sédiments, les matières en
suspension (particules) et le biote — Méthode par chromatographie en phase gazeuse couplée à la
spectrométrie de masse en tandem (CG-SM/SM) ou à la spectrométrie de masse haute résolution (CG-SMHR)
FDIS stage
This draft is submitted to a parallel vote in ISO, CEN.

ISO/DIS 22032:2025(en)
DRAFT International Standard
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Contents
Foreword . v
Introduction . vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 2
5 Interferences . 4
6 Reagents and standards . 4
7 Apparatus . 7
8 Sampling and sample pretreatment . 8
9 Procedure . 9
9.1 Extraction of sediment or particulate matter samples by pressurized liquid extraction
(PLE) or Soxhlet . 9
9.2 Extraction of biota samples . 9
10 Clean-up of sample extracts . 10
11 Measurement and integration of the chromatogram . 13
12 Calibration . 13
12.1 General . 13
12.2 Estimation of the linear range . 13
12.3 Calibration of the measuring method using an internal standard, . 14
12.4 Injection standard . 14
13 Identification . 14
14 Quantification. 14
14.1 Quantification using internal standards, including quality checks of the recovery of the
internal standards . 14
14.2 Testing the validity of calibration . 15
15 Expression of results . 15
16 Test report . 15
Annex A (normative) Program for pressurized liquid extraction . 17
Annex B (normative) Clean-up methods . 21
Annex C (informative) Typical GC-MS conditions and m/z values for identification and
quantification . 26
Annex D (informative) Examples of linearity check and calibration working solutions . 6
Annex E (informative) Performance data . 8
Bibliography . 11

iv
ISO/DIS FDIS 22032:20252026(en)
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
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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 documentdocuments 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).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent rights
in respect thereof. As of the date of publication of this document, ISO had not received notice of (a) patent(s)
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Any trade name used in this document is information given for the convenience of users and does not
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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 147, Water quality, Subcommittee SC 2,
Physical, chemical and biochemical methods, in collaboration with the European Committee for
Standardization (CEN) Technical Committee CEN/TC 230, Water analysis, in accordance with the Agreement
on technical cooperation between ISO and CEN (Vienna Agreement).
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 .
This second edition cancels and replaces the first edition (ISO 22032:2006), which has been technically
revised.
The main changes are as follows:
— — the scope has been expanded to include biota;
— — GC-MS/MS has been included as a detection method;
— — a description of a clean-up set with manual and automated methods for sediment, for suspended
particulate matter and biota has been included.
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 .
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.
v
Introduction
This document enables the analysis of polybrominated diphenyl ethers (PBDE) related to 91/271/EEC - the
Urban Waste Water Treatment Directive, the European Union directive concerning urban waste waterthe
collection, treatment and discharge of urban waste water and the treatment and discharge of waste water
from certain industrial sectors.
vi
DRAFT International Standard ISO/DIS 22032:2025(en)

Water quality — Determination of polybrominated diphenyl ethers
(PBDE) in sediment, suspended (particulate) matter and biota —
Method using gas chromatography coupled with tandem mass
spectrometry (GC-MS/MS) or with high resolution mass
spectrometry (GC-MS/MS, GC-HRMS)
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 with this document be
carried out by suitably qualified staff.
1 Scope
This document specifies a method for the determination of selected polybrominated diphenylethers
(PBDE) (see Figure 1Figure 1 and Table 1Table 1)) in sediment, suspended particulate matter and biota
using gas chromatography/ or mass spectrometry (GC-MS/MS or GC-HRMS) in the electron impact (EI),
negative ion chemical ionization (NCI) or atmospheric pressure ionization (APCI) mode.
The method is applicable to sediment and suspended particulate matter samples with limits of
quantification of 0,2 µg/kg dry mass (dm) for brominated diphenylether (BDE) BDE-28 to BDE-183, of
2 µg/kg dry mass (dm) for BDE--209.
The method is applicable as well with lower limits of quantification (LOQ), if specific clean-up methods,
described in Clause 10, Table 3Clause 10, Table 3,, method 1 and method 2 in combination with
measurement methods GC-MS/MS or GC-HRMS after electron impact ionization (El) or negative ion
chemical ionization (NCI) for BDE-209 are used. Depending on the analytical capability of the instrument,
limits of quantification down to 0,003 µg/kg dm for BDE-28 to BDE-154 and 0,02 µg/kg dm for BDE-183
and 1 µg/kg dm for BDE-209 and lower are possible.
The method is applicable to biota samples with limits of quantification down to 0,000 2 µg/kg fresh mass
(fm) (BDE-28 to BDE-154) and 0,03 μg/kg fresh mass (fm) (BDE-183), if specific clean-up methods,
described in Table 4Table 4 in combination with measurement methods GC-MS/MS or GC-HRMS after
electron impact ionization (El) are used.
Performance data are listed in Annex EAnnex E.
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 5667-12, Water quality — Sampling — Part 12: Guidance on sampling of bottom sediments from rivers,
lakes and estuarine areas
ISO 5667-13, Water quality — Sampling — Part 13: Guidance on sampling of sludges
ISO 5667-17, Water quality — Sampling — Part 17: Guidance on sampling of bulk suspended solids
ISO 8466--1:2021, Water quality — Calibration and evaluation of analytical methods — Part 1: Linear
calibration function
ISO 10870, Water quality — Guidelines for the selection of sampling methods and devices for benthic
macroinvertebrates in fresh waters
ISO/TS 13530, Water quality — Guidance on analytical quality control for chemical and physicochemical
water analysis
EN 13946, Water quality — Guidance for the routine sampling and preparation of benthic diatoms from
rivers and lakes
EN 14011, Water quality — Sampling of fish with electricity
EN 14757, Water quality — Sampling of fish with multi-mesh gillnets
EN 16150, Water quality — Guidance on pro-rata Multi-Habitat sampling of benthic macro-invertebrates
from wadeable rivers
EN 16190, Soil, treated biowaste and sludge — Determination of dioxins and furans and dioxin-like
polychlorinated biphenyls by gas chromatography with high resolution mass selective detection
(HR GC--MS)
EN 17218, Water quality — Guidance on sampling of mesozooplankton from marine and brackish water
using mesh
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— — ISO Online browsing platform: available at https://www.iso.org/obp
— — IEC Electropedia: available at https://www.electropedia.org/
4 Principle
Brominated diphenyl ethers are extracted from dried sample material (sediment, suspended particulate
matter, biota) using organic solvents.
2 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
Figure 1 — General structure of polybrominated diphenyl ethers
Table 1 — PBDE congeners determined by this method
a b
No. Congener Formula Abbreviation CAS-RN Molar mass
g/mol
1 2,4,4'-Tribromodiphenyl ether C12H7Br3O BDE--28 41318-75-6 406,9
2 2,2',4,4'-Tetrabromodiphenyl ether C H Br O BDE--47 5436-43-1 485,8
12 6 4
3 2,2',4,4',5-Pentabromodiphenyl ether C12H5Br5O BDE--99 60348-60-9 564,7
4 2,2',4,4',6-Pentabromodiphenyl ether C12H5Br5O BDE--100 189084-64-8 564,7
5 2,2',4,4',5,5'-Hexabromodiphenyl C H Br O BDE--153 68631-49-2 643,6
12 4 6
ether
6 2,2',4,4',5,6'-Hexabromodiphenyl C12H4Br6O BDE--154 207122-15-4 643,6
ether
7 2,2',3,4,4',5',6-Heptabromodiphenyl C12H3Br7O BDE--183 207122-16-5 722,5
ether
8 Decabromodiphenyl ether C12Br10O BDE--209 1163-19-5 959,2
a Numbering analogous to IUPAC nomenclature for Polychlorinated Biphenylethers PCB.
b
CAS-RN Chemical Abstracts Services Registration Number.b Chemical Abstracts Service (CAS) Registry Number® is a
trademark of the American Chemical Society (ACS). This information is given for the convenience of users of this document
and does not constitute an endorsement by ISO of the product named. Equivalent products may be used if they can be shown
to lead to the same results.
Clean-up of the extract is carried out by various methods as specified in this document. Depending on the
matrix and the concentration of PBDE in the samples five different clean-up methods for sediment
samples and three methods for biota samples can be selected, e.g. column chromatography or gel
permeation chromatography. Options for avoiding dichloromethane and toluene in the sample clean-up
are available.
After clean-up and concentration, separation of the brominated diphenyl ethers is accomplished by
capillary gas chromatography. For detection, different types of GC-MS equipment can be used, applying
either mass spectrometry in multiple reaction mode, or high resolution mass spectrometry with different
ionisation techniques as electron impact (EI), negative ion chemical ionization (NCI) or atmospheric
pressure chemical ionisation (APCI).
For the determination of the analyte concentration in the sample, an internal standard calibration is used.
5 Interferences
Non--specific matrix interferences, as well as interferences from other environmental contaminants are
dealt with using the given clean-up procedure.
Sources of contamination of the samples can be the following: brominated diphenyl ethers used as flame-
retardants or for other purposes in organic polymers, used in vial covers, Pasteur pipette fillers, recycled
paper and are possibly transported also via air dust. Therefore, any contact of samples, reagents or any
material used with these organic polymers shall be avoided.
PCB-180 can interfere with BDE-47 if a short column is used. The application of MS/MS detection as well
as avoiding m/z 323,87 in the detection by GC-HRMS allows for separation the compounds via mass
spectrometry.
Interferences with other chlorinated substances can occur, if chromatographic separation is not sufficient
as described in Reference [13[1]:]:
— — BDE-47 at m/z 323,878 5 with Heptachlorobiphenyl at m/z 323,864 7 such as PCB-180;
— — BDE-100 and BDE-99 at m/z 403,787 0 with Octachloronaphthalene at m/z 403,745 0;
— — BDE-100 and BDE-99 at m/z 405,784 9 with Heptachlorodibenzofurans at m/z 405,784 7.
MS/MS detection at optional m/z values (see Table C.1Table C.1)) can solve the problem. Therefore,
careful evaluation of instrumentation, masses and mass transitions used in MS shall be done.
Further interferences from BDE-congeners not listed in this document, as well as by other brominated
compounds can be found in Reference [14[2].]. Coelutions can occur especially when using short columns
and short oven programs. Such potential interferences depending on the analytical column are:
— — BDE-16, BDE-33 interfering with BDE-28;
— — BDE-184, BDE-182 and BDE-175 interfering with BDE-183.
These and further interferences should be overcome by a sufficient chromatographic separation.
6 Reagents and standards
Use only reagents and materials with negligibly low concentrations of brominated diphenyl ethers, verify
this in every analysis series by blank determinations over the total procedure. The blank over the total
procedure mustshall be less than the reported limit of detection (see the definition of the limit of
detection in ISO/TS 13530). If necessary, traces of PBDE in solid materials can be reduced by heating at
400 °C.
6.1 6.1 Solvents for extraction, preparation of stock solutions and clean up.
6.1.1 6.1.1 n--Heptane, C H .
7 16
6.1.2 6.1.2 Toluene, C H .
7 8
6.1.3 6.1.3 Acetone (2-propanone), C H O.
3 6
4 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
6.1.4 6.1.4 Dichloromethane, CH Cl .
2 2
6.1.5 6.1.5 Cyclohexane, C H .
6 12
6.1.6 6.1.6 Dodecane, C H for automatized clean up or as a keeper.
12 26,
6.1.7 6.1.7 n-Hexane, C H .
6 14
6.1.8 6.1.8 Ethanol, C H O.
2 6
6.1.9 6.1.9 Ethyl acetate, C H O .
4 8 2
6.1.10
6.2 6.2 Reference stock solutions.
See Table 1Table 1. Solutions of reference substances are commercially available.
Store the prepared solutions in a refrigerator at (5 ± 3) °C, alternatively according to the manufacturer.
6.3 6.3 Internal standard stock solutions.
Solutions of C-labelled reference substances for use as internal standards are commercially available.
Store the prepared solutions in a refrigerator at (5 ± 3) °C, alternatively according to the manufacturer.
See Table 2Table 2.
Table 2 — List of C-labelled internal standards
No Name Formula Abbreviation Molar mass
g/mol
Internal standards
13 13 13
1 2,4,4'-Tribromo[ C ]diphenyl ether C H Br O C-BDE--28 418,8
12 12 7 3
13 13 13
2 2,2',4,4'-Tetrabromo[ C12]diphenyl ether C12H6Br4O C-BDE--47 497,7
13 13 13
3 2,2',4,4',5-Pentabromo[ C ]diphenyl ether C H Br O C-BDE--99 576,6
12 12 5 5
13 13 13
4 2,2',4,4',6-Pentabromo[ C12]diphenyl ether C12H5Br5O C-BDE--100 576,6
13 13 13
5 2,2',4,4',5,5'-Hexabromo[ C ]diphenyl ether C H Br O C-BDE--153 655,5
12 12 4 6
13 13 13
6 2,2',4,4',5,6'-Hexabromo[ C ]diphenyl ether C H Br O C-BDE--154 655,5
12 12 4 6
13 13 13
7 2,2',3,4,4',5',6-Heptabromo[ C12]diphenyl ether C12H3Br7O C-BDE--183 734,4
13 13 13
8 Decabromo[ C ]diphenyl ether C Br O C-BDE--209 971,1
12 12 10
6.4 6.4 Internal standard working solutions for calibration and addition to the samples, see
Annex DAnnex D.
6.5 6.5 Sodium sulfate, Na SO , anhydrous powdered baked in oven (7.5(7.5)) for 4 hours at
2 4
400 °C.
6.6 6.6 Water, free of blanks and potential interferences.
6.7 6.7 Extraction filters, e.g. cellulose for PLE-extractor.
6.8 6.8 Soxhlet thimbles, (e.g. 27 mm × 100 mm) (pre-cleaned).
6.9 6.9 Sand (blank free), e.g. Ottawa sand or sea sand for blank analysis over the total
procedure and as a filling of the extraction cells,; alternative: glass granulate.
6.10 6.10 Basic alumina, (Al O ), activity Super I, particle size 0,063 mm to 0,2 mm, pH 10 (100 g/l,
2 3
3 1)
H O, 20 °C) (in slurry) density 3,94 g/cm , e.g. MP Alumina B – Super I for Dioxin Analysis 1344-28-1
(mpbio.com), avoid long storage and contact with air humidity.
6.11 6.11 Basic alumina, (Al O ), Activityactivity I, particle size 0,063 mm to 0,2 mm, pH 8,5 to
2 3
pH 10,5 (100 g/l, H O, 20 °C) (in slurry) density 4 g/cm , (20 °C), pore size 9 nm, avoid long storage and
contact with air humidity.
6.12 6.12 Silica 60, (70 mesh to 230 mesh).
6.13 6.13 Silver nitrate, (AgNO ).
6.14 6.14 Sulfuric acid, H SO , 95 % to 97 %.
2 4
6.15 6.15 Hydrochloric acid, HCl, 2 mol/l.
6.16 6.16 Material for fixing the adsorbent materials in the clean-up columns, e.g. quartz wool
or glass wool, silanised glass wool.
6.17 6.17 Copper powder, grain size < 63 µm.
6.18 6.18 Silica-sulfuric acid.
Add 44 g sulfuric acid (6.14(6.14)) dropwise to 56 g silica (6.12(6.12).). Subsequently shake for 30 min.
Store tightly closed in brown glass bottles. The mixture is stable for at least 1 month.
6.19 6.19 Silica-silver nitrate.
Dissolve 10 g AgNO (6.13(6.13)) in 40 ml water (6.6(6.6),), add solution stepwise to 90 g silica
(6.12(6.12)) and stir, activate at 125 °C for 5 hours. Store tightly closed in brown glass bottles. The
mixture is stable at least for 1 month.
6.20 6.20 Operating gases, for GC--MS, of high purity and in accordance with manufacturer’s
specifications.
6.21 6.21 Nitrogen, N , for concentrating of extracts.
6.22 6.22 Linearity check and calibration working solutions.
Considering the working range and the sample amount, prepare calibration solutions using syringes
(7.17(7.17)) with the lowest point corresponding to the LOQ of the method, (example given in
Annex DAnnex D. ). The concentration of the internal standard should be between the medium and the
top level of the calibration range, (example given in Annex DAnnex D.).
6.23 6.23 Injection standards.

1)
This information is given for the convenience of users of this document and does not constitute an endorsement
by ISO of the product named. Equivalent products may be used if they can be shown to lead to the same results.

6 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
The following C-labelled injection standards are commercially available and have proven to be
practicable:
13 13
— — 3,3',4,4'-Tetrabromo( C )diphenyl ether ( C-BDE-77);
13 13
— — 3,3',4,5'-Tetrabromo( C )diphenyl ether ( C-BDE-79);
13 13
— — 2,3’,4,4’,5-Pentabromo( C )diphenyl ether ( C-BDE-118);
13 13
— — 2,2',3,4,4',5-Hexabromo( C )diphenyl ether ( C-BDE-137);
13 13
— — 2,2',3,4,4',5’-Hexabromo( C )diphenyl ether ( C-BDE-138);
13 13
— — 2,2',3,4,4',6-Hexabromo( C )diphenyl ether ( C-BDE-139);
13 13
— — 2,2',3,4,4',5,5'-Heptabromo( C )diphenyl ether ( C-BDE-180);
13 13
— — 2,3,3',4,4',5,6-Heptabromo( C )diphenyl ether ( C-BDE-190);
13 13
— — 2,2',3,4,4',5,5',6-Octabromo( C )diphenyl ether ( C-BDE-203);
13 13
— — 2,3,3',4,4',5,5',6-Octabromo( C )diphenyl ether ( C-BDE-205);
13 13
— — 2,2',3,3',4,4',5,5',6-Nonabromo( C )diphenyl ether ( C-BDE-206).
Use commercially available solutions (6.2, 6.3(6.2, 6.3)) (e.g. in nonane, toluene or iso-octane) or prepare
stock solutions, e.g. by dissolving 10 mg of each of the reference substances in n--heptane (6.1.1(6.1.1))
in an amber, 10--ml volumetric flask 7.157.15 and bring to volume (concentration: 1 mg/ml).
Examples for calibration working solutions are given in Annex DAnnex D. For the linearity check use at
least six concentration levels.
The solutions may be stored in a refrigerator at (5 ± 3) °C in the dark for at least 1 year. Check the
concentration of the calibration solutions against an independently prepared standard prior to use.
7 Apparatus
Clean all glassware at least by rinsing with acetone (2-propanone) (6.1.3(6.1.3).).
7.1 7.1 Wide-necked bottle, 1 000 ml up to 5 000 ml capacity, for wet sediment, particulate
matter or biota.
7.2 7.2 Freeze drying apparatus.
7.3 7.3 Deep freezer.
7.4 7.4 Mortar and pestle, or a grinding mill.
7.5 7.5 Drying ovens, capable of maintaining temperatures in the ranges of 100 °C to 400 °C for
baking of clean-up materials, for baking of glassware and for dry residue determination of samples.
7.6 7.6 Sieve shaker with appropriate sieve meshes (aperture size), 2 mm.
7.7 7.7 Desiccator.
7.8 7.8 Pressurized liquid extractor (PLE) and appropriate filter materials suited for the
device, alternatively Soxhlet extraction apparatus, consisting of round bottom flasks (e.g. 250 ml),
Soxhlet extractors and Soxhlet thimbles (e.g. 27 mm × 100 mm) see 6.86.8,, vertical condensers (e.g.
300 mm) and heating apparatus or Twisselmann extraction system.
7.9 7.9 Evaporation device, e.g. rotary evaporator or concentration device with suitable gases
6.216.21.
7.10 7.10 Automatic sample processing system, e.g. DexTech/DexTech Plus from LC-Tech,
12)
MiuraTM GO--2HT (Miura Co. Ltd).
7.11 7.11 Small glass columns for chromatographic clean--up, 1 cm inner diameter,
approximately 12 mm × 140 mm with a stopcock.
7.12 7.12 Big glass columns for chromatographic clean--up, > 2 cm inner diameter,
approximately 25 mm × 150 mm with a stopcock.
7.13 7.13 Ready to use columns for automatized clean-up from, e.g. LC-Tech or Miura-system:
see Clause B.7Clause B.7.
7.14 7.14 Gel Permeation Chromatographypermeation chromatography (GPC) clean--up
system (with modular design), pump, sampling injector, sample rack; fraction collector, column: e.g.
Ashahipak GF 310 HQ 7,5 mm × 300 mm, 5 µm particle size.
7.15 7.15 Volumetric flasks, 1 ml, 2 ml, 10 ml and 25 ml, preferable amber glass.
7.16 7.16 Pasteur pipettes, e.g. 2 ml.
7.17 7.17 Syringes, 2 µl, 5 µl, 10 µl and 50 µl, volume precision ±2 %.
7.18 7.18 GC-sample vials, e.g. 2 ml, amber glass with a micro insert and a fluoropolymer--lined
screw--cap is most suitable.
7.19 7.19 Gas chromatograph, with operating gases 6.206.20 with a splitless injection port or a
temperature programmable injection port, coupled to a tandem mass spectrometer (GC--MS/MS) or GC-
HRMS with electron impact or chemical ionization and appropriate reactant gas (e.g. CH ) or atmospheric
pressure ionization.
7.20 7.20 Analytical column, Fusedfused silica column with non-polar low bleed separating phase
(see Annex CAnnex C for examples); e.g. inner diameter < 0,25 mm, length 15 m, film thickness of
0,1 µm is recommended.
8 Sampling and sample pretreatment
Take samples as specified in , see e.g. ISO 5667--12, ISO 5667--13 or ISO 5667-17, in a bottle (7.1(7.1).).
Store and transport in the dark at approximately 4 °C. Pre--treat the samples immediately in the
laboratory by homogenizing and freeze--drying (7.2(7.2),), alternatively mix itthem with sodium sulfate

DexTech/DexTech Plus from LC-Tech, MiuraTM GO-2HT (Miura Co. Ltd) are automatic sample processing systems.
This information is given for the convenience of users of this document and does not constitute an endorsement by
ISO of the automatic sample processing systems. Equivalent systems may be used if they can be shown to lead to
the same results.”
2)
DexTech/DexTech Plus from LC-Tech, MiuraTM GO-2HT (Miura Co. Ltd) are automatic sample processing
systems. This information is given for the convenience of users of this document and does not constitute an
endorsement by ISO of the automatic sample processing systems. Equivalent systems may be used if they can be
shown to lead to the same results.

8 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
(6.5(6.5).). If an immediate pre-treating is not possible, samples can be frozen (7.3(7.3)) and stored in a
freezer at a temperarure below −15 °C. Deagglomerize the dried samples using apparatus e.g. (7.4(7.4))
and sieve it using device (7.6(7.6)) according to the analytical task.
Take and pretreat biota samples as specified in, see e.g. EN 14011, EN 14757, EN 16150, EN 17218,
ISO 10870 or EN 13946, immediately either before or after homogenizing by freezing. Freeze dry to
remove water and to enhance the surface for later extraction.
Store the dried samples protected against air humidity, e.g. in a desiccator (7.7(7.7).).
9 Procedure
9.1 Extraction of sediment or particulate matter samples by pressurized liquid
extraction (PLE) or Soxhlet
Place the filter (6.7(6.7)) or thimble (6.8(6.8)) and, if applicable, the sand (6.9(6.9)) in the extractor
according to the instructions for the extractor (PLE) (7.8(7.8)) or soxhletSoxhlet extractor (7.8(7.8).).
Transfer a suitable amount, e.g. 5 g of, of the pre--treated, dry sample into the prepared extractor cell.
Add a suitable amount of the internal standard working solution (6.4(6.4)) prepared from the internal
standard stock solutions (6.3(6.3)) to the sample and add 2 ml of the extraction solvent (6.1(6.1)) to avoid
losses of the internal standard. Add sand (6.9(6.9)) on the top.
Extract with n--heptane, or n--hexane (6.1or mixtures with acetone (6.1).).
AnThe extraction program inprograms for PLE and soxhlet areor Soxhlet given in Clauses A.1Clauses A.1
and A.3, respectively.
Make sure that exhaustive extraction is achieved.
A.3Extraction of BDE-209 requires specific attention and, sometimes, longer extraction times than for
other PBDE congeners. The solvent n-heptane allows for a complete extraction with an extraction
program given in Clauses A.1 and A.3 shall be applied.
Protect samples and extracts carefully from sunlight to avoid photodegradation of the PBDE, especially
BDE-209.
NOTE 1 Other extraction techniques, can be used after performing comparability exercise with PLE and the given
program or soxhletSoxhlet extraction.
Extraction of BDE-209 requires specific attention and, sometimes, longer extraction times than for other
PBDE congeners.
NOTE 2 Other extraction solvents, can be used after performing comparability exercise.
Concentrate the extract from the extractor gently (at a temperature of 40 °C) to 1 ml ± 0,5 ml using a
suitable evaporation device (7.9(7.9).).
9.2 Extraction of biota samples
9.2.1 PLE, Soxhlet- or Twisselmann -extraction
Place the filter (6.7(6.7)) or thimble (6.8(6.8)) and the sand (6.9(6.9)) in the extractor according to the
instructions for the extractor (PLE) or alternative extractor (7.8(7.8).). Transfer a suitable amount, 2 g to
5 g, of the dried biota sample, into the prepared extractor cell. Add the internal standard working solution
(6.4(6.4)) to the sample and add 2 ml of the extraction solvent to avoid losses of the internal standard.
Add sand on the top, if applicable. The internal standards can be added also after fat extraction, if an
appropriate validation of the quantitative extraction is given.
Extraction The extraction programs arefor PLE or Soxhlet given in Clauses A.2Clauses A.2 and A.4A.4.
Make sure that exhaustive extraction is achieved.
Extraction of BDE-209 requires specific attention and, sometimes, longer extraction times than other
PBDE congeners. The solvent n-heptane allows for a complete extraction with an extraction program
given in Clauses A.1 and A.3 shall be applied.
Protect samples and extracts carefully from sunlight to avoid photodegradation of the PBDE, especially
BDE-209.
NOTE 1 Other extraction techniques, likesuch as ultrasonic extraction, can be used after performing
comparability exercise with PLE and the given program or soxhletSoxhlet extraction.
Make sure that exhaustive extraction is achieved.
Extraction of BDE-209 requires specific attention and, sometimes, longer extraction times than other
PBDE congeners.
NOTE 2 Other extraction solvents, can be used after performing comparability exercise.
Concentrate the extract from the extractor gently (at a temperature of 40 °C) to 1 ml ± 0,5 ml using a
suitable evaporation device (7.9(7.9).).
9.2.2 Cold extraction alternative (Biotabiota)
This procedure describes a cold extraction of fat by means of dichloromethane/cyclohexane (a volume
fraction of 1:1) (6.1.4(6.1.4 and 6.1.56.1.5).).
A mixture of (30,0 ± 0,1) g of the wet sample, 70 g sodium sulfate (6.5(6.5),), 30 g glass granulate or as an
alternative sea sand (6.9(6.9)) and the internal standard solution (6.4(6.4)) are put in a mortar (7.4(7.4).).
The mixture is finely ground using the pestle to produce a powder. AboutApproximately 5 g of sodium
sulfate (6.5(6.5)) is filled into a chromatographic glass column (7.11(7.11)) sealed with a plug of silanised
glass wool (6.16(6.16).). Afterwards the finely ground sample powder is added to the column. Extract
with 350 ml of a dichloromethane/cyclohexane (a volume ratio of 1:1) mixture and collect the eluate in a
500 ml round bottom flask. This eluate is carefully concentrated by means of an evaporation device
(7.9(7.9)) at a temperature of (40 ± 5) °C.
Remove the solvent entirely.
10 Clean-up of sample extracts
Depending on the matrix and the concentration of PBDE in the samples, five different clean-up methods
for sediment samples (see Table 3Table 3)) and three methods for biota samples (see Table 4Table 4))
shall be selected, e.g. column chromatographic methods, or combined methods using GPC or avoiding
dichloromethane and toluene. One of the methods is identical to ISO 18635 for Short Chain
Polychlorinatedshort chain polychlorinated paraffins (SCCP) analysis in sediment and allows for
enhancement of laboratory efficiency.
Before applying the method, the elution volume shall be checked. The clean-up columns are always
prepared before solvent addition, hence do not prepare by the slurry technique.
10 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
Table 3 — Clean-up methods for sediments and suspended particle matter
(see Annex BAnnex B))
Clean-up
Method 1 Method 2 Method 3 Method 4 Method 7
method
Low Low
concentration of concentration of
High Medium High
matrix matrix
concentration of concentration concentration of
Matrix constituents, constituents,
matrix of matrix matrix
characterisation e.g. sometimes e.g. sometimes
constituents, constituents, constituents,
in suspended in suspended
possibly PCB possibly PCB lipids, PCB
particulate particulate
matter matter
No use of
dichloromethan
Additional Use of Use of
= clean-up in
e and toluene
Information dichloromethane Use of toluene dichloromethan
ISO 18635
information and toluene e and toluene
No sulfur
removal
Column Column
chromatographic chromatographi
clean-up with c clean-up with
sulfur removal sulfur removal
using layers of using layers of
Clean-up step silica-sulfuric  silica-sulfuric
acid and silica- acid and silica-
silver nitrate silver nitrate
(see (see
Clause B.1Clause Clause B.1Claus
B.1)) e B.1))
Column
chromatographi
c clean-up with
silica-sulfuric
acid in a small
Clean-up step
column
(see
Clause B.2Claus
e B.2))
Column
chromatographic
clean-up with
Alumina B Super
I (see
Clean-up step
Clause B.3Clause
B.3))
Use of
dichloromethane
Column
chromatograph
ic clean-up
Clean-up step
with Alumina
and activated
copper powder
(see
Clean-up
Method 1 Method 2 Method 3 Method 4 Method 7
method
Clause B.4Claus
e B.4))
Gel
chromatograph
Clean-up step  ic clean-up (see
Clause B.5Claus
e B.5))
Biota
Automatedauto
mated clean up
Clean-up step
(see
Clause B.7Claus
e B.7))
NOTE For samples containing high amounts of sulfur: Gelgel permeation chromatography is more efficient for
sulfur removal than using copper powder.
Concentrate (7.9(7.9)) the final eluate carefully to a volume of about 0,5 ml to 1 ml and dry it with sodium
sulfate (6.5(6.5),), if necessary. For further concentration a keeper can be added to avoid evaporation to
dryness, e.g. 10 µl dodecane (6.1.6(6.1.6)) for a final volume of e.g. 100 µl. Transfer the extract to a GC-
vial (7.18(7.18).).
Table 4 — Clean-up methods for biota
Clean-up
Method 5 Method 6 Method 7
method
High concentration of
Matrix High concentration of matrix High concentration of matrix
matrix constituents, lipids,
characterisation constituents, lipids, PCB constituents, lipids, PCB
PCB
Additional Use of dichloromethane and
Use of dichloromethane No use of dichloromethane
information toluene
Column chromatographic Column chromatographic
clean-up by modified silica clean-up by modified silica –
Clean-up step
– big column (see big column (see
Clause B.6Clause B.6)) Clause B.6Clause B.6))
Column chromatographic
clean-up by alumina B
Super I (see
Clean-up step
Clause B.3Clause B.3)
)
Column chromatographic
clean-up with Alumina and
Clean-up step
activated copper powder
(see Clause B.4Clause B.4))
Gel chromatographic clean-
Clean-up step  up (see Clause B.5Clause
B.5))
Biota automated clean up
Clean-up step
(see Clause B.7Clause B.7))
12 © ISO 2024 – All rights reserved

ISO/DIS 22032:2025(en)
Concentrate (7.9(7.9)) the final eluate carefully to a volume of about 0,5 ml to 1 ml and dry it with sodium
sulfate (6.5(6.5),), if necessary. For further concentration a keeper can be added to avoid evaporation to
dryness, e.g. 10 µl dodecane (6.1.6(6.1.6)) for a final volume of e.g. 100 µl. Transfer the extract to a GC-
vial (7.18(7.18).).
11 Measurement and integration of the chromatogram
Optimize the operating conditions of the GC--MS system (7.19(7.19 and 7.207.20)) according to the
manufacturer’s instructions. Examples of the gas chromatographic conditions are given in
Annex CAnnex C.
Prior to analysis, establish the operating conditions and verify the GC--MS system performance and the
calibration for all analytes and their internal standards by analysis of a calibration standard.
Add the injection standard (6.23(6.23)) to the final extract if applicable, and analyse the sample with
GC--MS.
Especially for the analysis of BDE--209, minimise the exposure time of the samples to high temperatures
during sample injection and separation stages, because of the thermal degradation of BDE--209 at
temperatures higher than 300 °C. Optimize the injection step, paying special attention to the peak height
of BDE--209.
The m/z transitions are examples for measured ions and integrated signals for quantification. It is also
possible to summarize more than one ion for quantification.
12 Calibration
12.1 General
See normative references for the generally applicable procedures to calibrate The calibration and to
quantitate concentrations of substancesquantification shall be performed in water analysis:accordance
with ISO 8466--1, ISO/TS 13530 or; the calibration and quantification in soil, treated biowaste and sludge
according to shall be performed in accordance with EN 16190.
The strategy of calibration of the measurement method using an internal standard (seeaccording to
ISO 8466--1) is selected. For that, matrix free calibration solutions with C-labelled internal standards
are used. Isotopic dilution analysis, as described in according to EN 16190 can be used for calibration and
quantification as well.
Use at minimum one C-labelled internal standard per degree of bromination.
Matrix effects will be corrected by addition of the internal standard solution to the sample prior to sample
preparation.
Further injection standards can be used to check the recovery rate of the internal standards.
Calibration and quantification in the linear range is recommended, although non-linear relationships
between concentration and response can be used according to ISO/TS 13530.
For appropriate working solutions, see 6.226.22.
12.2 Estimation of the linear range
Calibration working solutions should be used as described in Annex DAnnex D.
The estimation of the linear range isshall be performed by calculating and evaluating the point-to-point
slope. see in accordance with ISO 8466-1:2021, 5.3. After calculation of the section-wise slope of each two
consecutive measuring points, differences of slopes should be lower than 10 % of the median.
12.3 Calibration of the measuring method using an internal standard,
Use calibration working solutions described in Annex DAnnex D.
Modern software allows for calibration according to the equations in ISO 8466--1:2021, 6.4. The operator
may decide, if the calibration line should be forced through zero or not, depending on the significance of
intercept. The lowest calibration point shall be lower than or equal to the corresponding limit of
quantification in the solid sample.
12.4 Injection standard
An injection standard is also referred as a “syringe or volumetric or recovery standard”. It is a compound
of known chemical purity added to every sample, procedural blank or calibration standard at a known
c
...