oSIST prEN ISO 22032:2025

SLOVENSKI STANDARD
01-junij-2025
Kakovost vode - Določanje polibromiranih difenil etrov (PBDE) v sedimentu,
suspendiranih (trdnih) delcih in bioti - Metoda s plinsko kromatografijo-tandemsko
masno spektrometrijo ali masno spektrometrijo visoke ločljivosti (GC-MS/MS;
HRMS) (ISO/DIS 22032:2025)
Water quality - Determination of polybrominated diphenyl ethers (PBDE) in sediment,
suspended (particulate) matter and biota - Method using gas chromatography-tandem
mass spectrometry or high resolution mass spectrometry (GC-MS/MS; HRMS) (ISO/DIS
22032:2025)
Wasserbeschaffenheit - Bestimmung von polybromierten Diphenylethern (PBDE) in
Sedimenten, Schwebstoffen und Biota - Verfahren mittels
Gaschromatographie/Massenspektrometrie und der hochauflösenden
Massenspektrometrie (GC-MS/MS; GC-HRMS) (ISO/DIS 22032:2025)
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 ou
à la spectrométrie de masse haute résolution (CG-SM/SM ; CG-SMHR) (ISO/DIS
22032:2025)
Ta slovenski standard je istoveten z: prEN ISO 22032
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.

DRAFT
International
Standard
ISO/DIS 22032
ISO/TC 147/SC 2
Water quality - Determination of
Secretariat: DIN
polybrominated diphenyl ethers
Voting begins on:
(PBDE) in sediment, suspended
2025-04-07
(particulate) matter and biota -
Voting terminates on:
Method using gas chromatography-
2025-06-30
tandem mass spectrometry or high
resolution mass spectrometry (GC-
MS/MS; HRMS)
ICS: 13.060.50
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
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USER PURPOSES, DRAFT INTERNATIONAL
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NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
PROVIDE SUPPORTING DOCUMENTATION.
Reference number
ISO/DIS 22032:2025(en)
DRAFT
ISO/DIS 22032:2025(en)
International
Standard
ISO/DIS 22032
ISO/TC 147/SC 2
Water quality - Determination of
Secretariat: DIN
polybrominated diphenyl ethers
Voting begins on:
(PBDE) in sediment, suspended
(particulate) matter and biota -
Voting terminates on:
Method using gas chromatography-
tandem mass spectrometry or high
resolution mass spectrometry (GC-
MS/MS; HRMS)
ICS: 13.060.50
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2025
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
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Published in Switzerland Reference number
ISO/DIS 22032:2025(en)
ii
ISO/DIS 22032:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 2
3 Terms and definitions . 2
4 Principle . 3
5 Interferences . 3
6 Reagents and standards . . 4
7 Apparatus . 6
8 Sampling and sample pretreatment. 7
9 Procedure . 7
9.1 Extraction of sediment/particulate matter samples by PLE or Soxhlet .7
9.2 Extraction of biota samples .8
9.2.1 PLE, Soxhlet- or Twisselmann-extraction .8
9.2.2 Cold extraction alternative (Biota) .8
10 Clean-up of sample extracts . 9
11 Measurement and integration of the chromatogram .10
12 Calibration .11
12.1 General .11
12.2 Estimation of the linear range see ISO 8466-1:2021, 5.3 .11
12.3 Calibration of the measuring method using an internal standard, see ISO 8466-1:2021,
6.4 .11
12.4 Injection standard .11
13 Identification .11
14 Quantification .12
14.1 Quantification using internal standards, including quality checks of the recovery of the
internal standards, see ISO 8466-1, 6.4 . 12
14.2 Testing the validity of calibration see ISO 8466-1, Clause 7 . 12
15 Expression of results .12
16 Test report .12
Annex A (normative) Program for pressurized liquid extraction . 14
Annex B (normative) Clean-up Methods .16
Annex C (informative) Typical GC-MS conditions and m/z values for Identification and
quantification .20
Annex D (informative) Examples for linearity check and calibration working solutions .26
Annex E (informative) Performance data .28
Bibliography .31

iii
ISO/DIS 22032:2025(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 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 can be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent
rights identified during the development of the document will be in the Introduction and/or on the ISO list of
patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO's adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 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).
This second edition cancels and replaces the first edition (ISO 22032:2006), which has been technically
revised.
The main changes are as follows:
— expansion of the scope to biota;
— including GC-MS/MS as detection method;
— including a description of a clean up set with manual and automated methods for sediment, for suspended
particulate matter and biota.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
ISO/DIS 22032:2025(en)
Introduction
This document is related to 91/271/EEC - the Urban Waste Water Treatment Directive, the European Union
directive concerning urban waste water collection, treatment and discharge of urban waste water and the
treatment and discharge of waste water from certain industrial sectors.

v
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-tandem mass
spectrometry or high resolution mass spectrometry (GC- MS/
MS; 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 1 and Table 1) in sediment, suspended particulate matter and biota using gas chromatography/
mass spectrometry (GC-MS/MS or GC-HRMS) in the electron impact (EI), negative ion chemical ionization
(NCI) or atmospheric pressure ionization (APCI) mode.
Figure 1 — General structure of polybrominated diphenyl ethers
The method is applicable to sediment and suspended particulate matter samples with limits of quantification
of 0,2 µg/kg dry weight (dw) for BDE-28 to BDE-183, of 2 µg/kg dry weight (dw) 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 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 dw for BDE-28 to BDE-154 and 0,02 µg/kg dw for BDE-183 and 1 µg/kg dw 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 weight
(fw) (BDE-28 to BDE-154) and 0,03 μg/kg fresh weight (fw) (BDE-183), if specific clean-up methods,
described in Table 4 in combination with measurement methods GC-MS/MS or GC-HRMS after electron
impact ionization (El) are used.

ISO/DIS 22032:2025(en)
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 C H Br O BDE-28 41318-75-6 406,9
12 7 3
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 C H Br O BDE‑99 60348-60-9 564,7
12 5 5
4 2,2',4,4',6-Pentabromodiphenyl ether C H Br O BDE‑100 189084-64-8 564,7
12 5 5
5 2,2',4,4',5,5'-Hexabromodiphenyl ether C H Br O BDE-153 68631-49-2 643,6
12 4 6
6 2,2',4,4',5,6'-Hexabromodiphenyl ether C H Br O BDE‑154 207122‑15‑4 643,6
12 4 6
7 2,2',3,4,4',5',6-Heptabromodiphenyl ether C H Br O BDE-183 207122-16-5 722,5
12 3 7
8 Decabromodiphenyl ether C Br O BDE‑209 1163-19-5 959,2
12 10
a
Numbering analogous to IUPAC nomenclature for PCB.
b
CAS-RN Chemical Abstracts Services Registration Number.
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/DIS 22032:2025(en)
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. 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 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
[1]
described in (Eljarrat, 2003):
— 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.1 —) can solve the problem. Therefore, careful
evaluation of instrumentation, masses and mass transitions used in MS is necessary.
Further interferences from BDE-congeners not listed in this document, as well as by other brominated
[2]
compounds can be found in (Korytar, 2005). 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.

ISO/DIS 22032:2025(en)
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 must be less than the reported limit of detection (see ISO/TS 13530). If necessary, traces of PBDE
in solid materials can be reduced by heating at 400 °C.
6.1 Solvents for extraction, preparation of stock solutions and clean up
6.1.1 n-Heptane, C H .
7 16
6.1.2 Toluene, C H .
7 8
6.1.3 Acetone (2-propanone), C H O.
3 6
6.1.4 Dichloromethane, CH Cl .
2 2
6.1.5 Cyclohexane, C H .
6 12
6.1.6 Dodecane, C H for automatized clean up or as a keeper.
12 26,
6.1.7 n-Hexane, C H .
6 14
6.1.8 Ethanol, C H O.
2 6
6.2 Reference stock solutions
See Table 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 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 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[ C ]diphenyl ether C H Br O C‑BDE‑47 497,7
12 12 6 4
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[ C ]diphenyl ether C H Br O C‑BDE‑100 576,6
12 12 5 5
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[ C ]diphenyl ether C H Br O C-BDE-183 734,4
12 12 3 7
13 13 13
8 Decabromo[ C ]diphenyl ether C Br O C‑BDE‑209 971,1
12 12 10
6.4 Internal standard working solutions for calibration and addition to the samples see Annex D.

ISO/DIS 22032:2025(en)
6.5 Sodium sulfate, Na SO , anhydrous powdered baked in oven (7.5) for 4 hours at 400 °C.
2 4
6.6 Water, free of blanks and potential interferences.
6.7 Extraction filters, e.g. Cellulose for PLE-Extractor.
6.8 Soxhlet thimbles, (e.g. 27 mm × 100 mm) (pre-cleaned).
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 Basic alumina, (Al O ), activity Super I, particle size 0,063 mm to 0,2 mm, pH 10 (100 g/l, H O, 20 °C)
2 3 2
(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 Basic alumina, (Al O ), Activity I, particle size 0,063 mm to 0,2 mm, pH 8,5 to pH 10,5 (100 g/l, H O,
2 3 2
20 °C) (in slurry) density 4 g/cm , (20 °C), pore size 9 nm, avoid long storage and contact with air humidity.
6.12 Silica 60, (70 mesh to 230 mesh).
6.13 Silver nitrate, (AgNO ).
6.14 Sulfuric acid, H SO , 95 % to 97 %.
2 4
6.15 Hydrochloric acid, HCl, 2 mol/l.
6.16 Material for fixing the adsorbent materials in the clean up columns, e.g. quarz wool or glass wool,
silanised glass wool.
6.17 Copper powder, grain size < 63 µm.
6.18 silica/sulfuric acid: add 44 g sulfuric acid (6.14) dropwise to 56 g silica (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 silica/silver nitrate: dissolve 10 g AgNO (6.13) in 40 ml water (6.6), add solution stepwise to 90 g
silica (6.12) and stir, activate at 125 °C for 5 hours. Store tightly closed in brown glass bottles. The mixture is
at least stable for 1 month.
6.20 Operating gases, for GC-MS, of high purity and in accordance with manufacturer’s specifications.
6.21 Nitrogen, N , for concentrating of extracts.
6.22 Linearity check and calibration working solutions
Considering the working range and the sample amount prepare calibration solutions with the lowest point
corresponding to the LOQ of the method, e. g. for BDE-28 to BDE-154 0,002 µg/kg; 0,005 µg/kg; 0,01 µg/kg;
0,02 µg/kg; 0,04 µg/kg; 0,1 µg/kg; 0,2 µg/kg; 0,4 µg/kg; 1 µg/kg and for BDE-183 and BDE-209 0,02 µg/
kg; 0,05 µg/kg; 0,1 µg/kg; 0,5 µg/kg; 1 µg/kg; 2 µg/kg; 5 µg/kg; 10 µg/kg; 20 µg/kg; 50 µg/kg; 100 µg/
kg; 200 µg/kg; 400 µg/kg using syringes (7.17). The concentration of the internal standard should be at
13 13 13 13
medium level, e.g. 0,2 µg/kg for C-BDE-28 to C-BDE-154 and 60 µg/kg for C-BDE-183 and C‑BDE‑209.
See also Annex D.
ISO/DIS 22032:2025(en)
6.23 Injection standards
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) (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) in an amber, 10-ml
volumetric flask 7.15 and bring to volume (concentration: 1 mg/ml).
Examples for calibration working solutions are given in Annex 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).
7.1 Wide-necked bottle, 1 000 ml up to 5 000 ml capacity, for wet sediment, particulate matter or biota.
7.2 Freeze drying apparatus.
7.3 Deep freezer.
7.4 Mortar and pestle, or a grinding mill.
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 Sieve shaker with appropriate sieve meshes (aperture size), 2 mm.
7.7 Desiccator.
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 6.8, vertical condensers (e.g. 300 mm) and
heating apparatus or Twisselmann extraction system.

ISO/DIS 22032:2025(en)
7.9 Evaporation device, e.g. rotary evaporator or concentration device with suitable gases 6.21.
7.10 Automatic sample processing system, e.g. DexTech/DexTech Plus from LC-Tech, MiuraTM
GO-2HT(Miura Co. Ltd).
7.11 Small glass columns for chromatographic clean-up, 1 cm inner diameter, approx. 12 mm × 140 mm
with a stopcock.
7.12 Big glass columns for chromatographic clean-up, > 2 cm inner diameter, approx. 25 mm × 150 mm
with a stopcock.
7.13 Ready to use columns for automatized clean-up from, e.g. LC-Tech or Miura-system: see B.7.
7.14 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 Volumetric flasks, 1 ml, 2 ml, 10 ml, and 25 ml, preferable amber glass.
7.16 Pasteur pipettes, e.g. 2 ml.
7.17 Syringes, 2 µl, 5 µl, 10 µl and 50 µl, volume precision ±2 %.
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 Gas chromatograph, with operating gases 6.20 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 Analytical column, Fused silica column with non-polar low bleed separating phase (see Annex 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 ISO 5667-12, ISO 5667-13 or ISO 5667-17 in a bottle (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), alternatively mix it with sodium sulfate (6.5). If an immediate pre-treating is not
possible, samples can be frozen (7.3) and stored in a freezer at below −15 °C. Deagglomerize the dried
samples using apparatus e.g. (7.4) and sieve it using device (7.6) according to the analytical task.
Pretreat biota samples as specified e.g. in EN 14011, EN 14757, EN 16150, EN 17218, ISO 10870 and 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).
9 Procedure
9.1 Extraction of sediment/particulate matter samples by PLE or Soxhlet
Place the filter (6.7) or thimble (6.8) and, if applicable, the sand (6.9) in the extractor according to the
instructions for the extractor (PLE) (7.8) or soxhlet extractor (7.8). Transfer a suitable amount, e.g. 5 g of
suspended particulate matter of the pre-treated, dry sample into the prepared extractor cell. Add a suitable

ISO/DIS 22032:2025(en)
amount of the internal standard working solution (6.4) prepared from the internal standard stock solutions
(6.3) to the sample and add 2 ml of the extraction solvent (6.1) to avoid losses of the internal standard. Add
sand (6.9) on the top.
Extract with n-heptane, n-hexane or mixtures with acetone (6.1).
Examples for extraction programs in PLE and soxhlet are given in A.1 and A.3 , respectively.
Make sure that exhaustive extraction is achieved.
Extraction 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 A.1 and A.3 .
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 soxhlet extraction.
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).
9.2 Extraction of biota samples
9.2.1 PLE, Soxhlet- or Twisselmann-extraction
Place the filter (6.7) or thimble (6.8) and the sand (6.9) in the extractor according to the instructions for
the extractor (PLE) or alternative extractor (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) 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 programs are given in Annex A.2 and A.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 A.1 and A.3.
Protect samples and extracts carefully from sunlight to avoid photodegradation of the PBDE, especially
BDE‑209.
NOTE 1 Other extraction techniques, like ultrasonic extraction can be used after performing comparability exercise
with PLE and the given program or soxhlet extraction.
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).
9.2.2 Cold extraction alternative (Biota)
This procedure describes a cold extraction of fat by means of dichloromethane/cyclohexane (1/1, v/v) (6.1.4
and 6.1.5).
A mixture of 30,0 ± 0,1 g of the wet sample, 70 g sodium sulfate (6.5), 30 g glass granulate or as an alternative
sea sand (6.9) and the internal standard solution (6.4) are put in a mortar (7.4). The mixture is finely ground

ISO/DIS 22032:2025(en)
using the pestle to produce a powder. About 5 g of sodium sulfate (6.5) is filled into a chromatographic glass
column (7.11) sealed with a plug of silanised glass wool (6.16). Afterwards the finely ground sample powder
is added to the column. Extract with 350 ml of a dichloromethane/cyclohexane (1:1 v/v) 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) 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 (Table 3) and three methods for biota samples (Table 4)can 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 SCCP analysis in sediment and allows for enhancement of
laboratory efficiency.
Before applying the method, the elution volume should be checked. The clean-up columns are always
prepared before solvent addition, hence do not prepare by the slurry technique.
Table 3 — Clean-up methods for sediments and suspended particle matter
(see Annex B)
Clean-up method Method 1 Method 2 Method 3 Method 4 Method 7
Matrix character‑ High concentration Medium concen‑ Low concentra‑ Low concentra‑ High concentra‑
isation of matrix constitu‑ tration of matrix tion of matrix tion of matrix tion of matrix
ents, possibly PCB constituents, constituents, e.g. constituents, e.g. constituents,
possibly PCB sometimes in sometimes in lipids, PCB
suspended partic‑ suspended partic‑
ulate matter ulate matter
Additional Infor‑ Use of dichlo‑ = clean-up in Use of toluene No use of dichlo‑ Use of dichlo‑
mation romethane and ISO 18635 romethane and romethane and
toluene toluene toluene
No sulfur removal
Clean-up step B.1 Column B.1 Column
chromatographic chromatographic
clean-up with sul‑ clean-up with
fur removal using sulfur removal
layers of silica sul‑ using layers of
furic acid and silica silica sulfuric acid
silver nitrate and silica silver
nitrate
Clean-up step B.2 Column
chromato-graphic
clean-up with sili‑
ca-sulfuric acid in
a small column
Clean-up step B.3 Column
chromatographic
clean-up with Alu‑
mina B Super I
Use of dichlo‑
romethane
ISO/DIS 22032:2025(en)
TTabablele 3 3 ((ccoonnttiinnueuedd))
Clean-up method Method 1 Method 2 Method 3 Method 4 Method 7
Clean-up step B.4 Column
chromatographic
clean-up with
Alumina and
activated copper
powder
Clean-up step B.5 Gel chro‑
matographic
clean-up
B.7 Biota Auto‑
mated clean up
NOTE For samples containing high amounts of sulfur: Gel permeation chromatography is more efficient for sulfur
removal than using copper powder.
Concentrate (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), if necessary. For further concentration a keeper can be added to avoid evaporation to dryness, e.g.
10 µl dodecane (6.1.6) for a final volume of e.g. 100 µl. Transfer the extract to a GC-vial (7.18).
Table 4 — Clean-up methods for biota
Clean-up method Method 5 Method 6 Method 7
Matrix characteri‑ High concentration of matrix High concentration of matrix High concentration of matrix
zation constituents, lipids, PCB constituents, lipids, PCB constituents, lipids, PCB
Additional informa‑ Use of dichloromethane and
Use of dichloromethane No use of dichloromethane
tion toluene
B.6 column chromatographic B.6 column chromatographic
Clean-up step clean-up by modified silica – clean-up by modified silica –
big column big column
B.3 column chromatographic
Clean-up step
clean-up by alumina B Super I
B.4 Column chromatographic
Clean-up step clean-up with Alumina and
activated copper powder
B.5 Gelchromatographic clean-
Clean-up step
up
Clean-up step B.7 Biota Automated clean up
Concentrate (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), if necessary. For further concentration a keeper can be added to avoid evaporation to dryness, e.g.
10 µl dodecane (6.1.6) for a final volume of e.g. 100 µl. Transfer the extract to a GC-vial (7.18).
11 Measurement and integration of the chromatogram
Optimize the operating conditions of the GC-MS system (7.19 and 7.20) according to the manufacturer’s
instructions. Examples of the gas chromatographic conditions are given in Annex 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), if necessary, 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.
ISO/DIS 22032:2025(en)
12 Calibration
12.1 General
See normative references for the generally applicable procedures to calibrate and to quantitate
concentrations of substances in water analysis according to ISO 8466-1, ISO/TS 13530 or in soil, treated
biowaste and sludge according to EN 16190.
The strategy of calibration of the measurement method using an internal standard (ISO 8466-1, 6.1 b)) is
selected. For that, matrix free calibration solutions with C-labelled internal standards are used. Isotopic
dilution analysis, as described in 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 6.22.
12.2 Estimation of the linear range see ISO 8466-1:2021, 5.3
Use calibration working solutions described in Annex D.
The estimation of the linear range is performed by calculating and evaluating the point-to-point slope. After
calculation of the section-wise slope of each two consecutive measuring points, differences of slopes should
be lower than 10 % of t
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