Environmental Solid Matrices - Determination of polychlorinated biphenyls (PCB) by gas chromatography - mass selective detection (GC-MS) or electron-capture detection (GC-ECD)

This European Standard specifies a method for quantitative determination of seven selected polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in soil, sludge, sediment, treated biowaste and waste and using GC-MS and GC-ECD.
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this European Standard, lower limit of application from 1 μg/kg (expressed as dry matter) for soils, sludge and biowaste to 10 μg/kg (expressed as dry matter) for solid waste can be achieved. For some specific samples the limit of 10 μg/kg cannot be reached.
Sludge, waste and treated biowaste may differ in properties, as well as in the expected contamination levels of PCBs and presence of interfering substances. These differences make it impossible to describe one general procedure. This European Standard contains decision tables based on the properties of the sample and the extraction and clean-up procedure to be used.
NOTE   For the analysis of PCB in insulating liquids, petroleum products, used oils and aqueous samples is referred to EN 61619, EN 12766–1 and EN ISO 6468 respectively.
The method may be applied to the analysis of other PCB congeners not specified in the scope, provided suitability is proven by proper in-house validation experiments

Feststoffe in der Umwelt - Bestimmung von polychlorierten Biphenylen (PCB) mittels Gaschromatographie und massenspektrometrischer Detektion (GC-MS) oder Elektronen-Einfang-Detektion (GC-ECD)

Dieses Dokument legt Verfahren für die quantitative Bestimmung von sieben ausgewählten polychlorierten Biphenylen (PCB 28, PCB 52, PCB 101, PCB 118, PCB 138, PCB 153 und PCB 180) in Boden, Schlamm, Sediment, behandeltem Bioabfall und Abfall unter Anwendung der GC MS und GC ECD fest (siehe Tabelle 2).
[Tabelle 2]
Die Nachweisgrenze hängt von den zu bestimmenden Stoffen, den verwendeten Geräten, der Qualität der für die Extraktion der Probe verwendeten Chemikalien und der Reinigung des Extrakts ab.
Unter den in diesem Dokument festgelegten Bedingungen kann eine untere Anwendungsgrenze zwischen 1 μg/kg (angegeben als Trockenmasse) für Böden, Schlamm und Bioabfall und 10 μg/kg (angegeben als Trockenmasse) für Feststoffabfall erreicht werden. Bei einigen speziellen Proben kann die Grenze von 10 µg/kg nicht erreicht werden.
Schlamm, Abfall und behandelter Bioabfall können sich hinsichtlich ihrer Eigenschaften sowie hinsichtlich der erwarteten Kontaminationsniveaus von PCB und der Anwesenheit von störenden Stoffen unterscheiden. Aufgrund dieser Unterschiede ist es unmöglich, ein allgemeines Verfahren zu beschreiben. Dieses Dokument enthält Entscheidungstabellen, die auf den Eigenschaften der Probe und den anzuwendenden Extraktions- und Aufreinigungsverfahren basieren.
ANMERKUNG   Für die Analyse von PCB in Isolierflüssigkeiten, Mineralölerzeugnissen, Gebrauchtölen und wässrigen Proben wird jeweils auf EN 61619, EN 12766 1 bzw. EN ISO 6468 verwiesen.
Das Verfahren kann für die Analyse weiterer, nicht im Anwendungsbereich festgelegter PCB Kongenere angewendet werden, vorausgesetzt, seine Eignung wurde durch geeignete laborinterne Validierungs¬untersuchungen nachgewiesen.

Matrices solides environnementales - Dosage des polychlorobiphényles (PCB) par chromatographie en phase gazeuse-spectrométrie de masse (CG-SM) ou chromatographie en phase gazeuse avec détection par capture d’électrons (CG-ECD)

Le présent document spécifie les méthodes de dosage, par CG-SM et CG-ECD, de sept polychlorobiphényles choisis (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 et PCB180) dans les sols, les boues, les sédiments, les biodéchets traités et les déchets (voir Tableau 2).
La limite de détection dépend des composés à analyser, de l’équipement utilisé, de la qualité des réactifs chimiques utilisés pour l’extraction de l’échantillon et la purification de l’extrait.
Dans les conditions spécifiées dans le présent document, il est possible d’atteindre une limite inférieure d’application de 1 μg/kg (exprimée sur matière sèche) pour les sols, les boues et les biodéchets à 10 μg/kg (exprimée sur matière sèche) pour les déchets solides. Pour certains échantillons spécifiques, la limite de 10 μg/kg ne peut pas être atteinte.
Les boues, les déchets et les biodéchets traités peuvent présenter des différences en termes de propriétés, de niveaux prévus de contamination par les PCB et de présence de substances interférentes. En raison de ces différences, il est impossible de décrire un mode opératoire général. Le présent document contient des tables de décision basées sur les propriétés de l’échantillon ainsi que sur le mode opératoire d’extraction et de purification à utiliser.
NOTE   Pour l’analyse des PCB dans les liquides isolants, les produits pétroliers, les huiles usagées et les échantillons aqueux, se référer respectivement aux normes EN 61619, EN 12766-1 et EN ISO 6468.
La méthode peut être appliquée à l’analyse de congénères de PCB autres que ceux décrits dans le domaine d’application, à condition d’en définir l’applicabilité par des tests de validation internes adaptés.

Trdni matriksi z vidika okolja - Določevanje polikloriranih bifenilov (PCB) s plinsko kromatografijo z masno selektivnim detektorjem (GC-MS) ali s plinsko kromatografijo z detektorjem z zajetjem elektronov (GC-ECD)

General Information

Status
Published
Public Enquiry End Date
04-Dec-2018
Publication Date
23-Sep-2020
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
17-Aug-2020
Due Date
22-Oct-2020
Completion Date
24-Sep-2020

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SLOVENSKI STANDARD
SIST EN 17322:2020
01-november-2020
Nadomešča:
SIST EN 15308:2017
SIST EN 16167:2018+AC:2019

Trdni matriksi z vidika okolja - Določevanje polikloriranih bifenilov (PCB) s plinsko

kromatografijo z masno selektivnim detektorjem (GC-MS) ali s plinsko
kromatografijo z detektorjem z zajetjem elektronov (GC-ECD)

Environmental Solid Matrices - Determination of polychlorinated biphenyls (PCB) by gas

chromatography - mass selective detection (GC-MS) or electron-capture detection (GC-

ECD)

Feststoffe in der Umwelt - Bestimmung von polychlorierten Biphenylen (PCB) mittels

Gaschromatographie und massenspektrometrischer Detektion (GC-MS) oder
Elektronen-Einfang-Detektion (GC-ECD)
Matrices solides environnementales - Dosage des polychlorobiphényles (PCB) par
chromatographie en phase gazeuse-spectrométrie de masse (CG-SM) ou
chromatographie en phase gazeuse avec détection par capture d’électrons (CG-ECD)
Ta slovenski standard je istoveten z: EN 17322:2020
ICS:
13.030.10 Trdni odpadki Solid wastes
13.080.10 Kemijske značilnosti tal Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST EN 17322:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN 17322:2020
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SIST EN 17322:2020
EN 17322
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2020
EUROPÄISCHE NORM

ICS 13.030.01; 13.030.10; 13.030.20; 13.080.10 Supersedes EN 15308:2016, EN 16167:2018+AC:2019

English Version
Environmental Solid Matrices - Determination of
polychlorinated biphenyls (PCB) by gas chromatography -
mass selective detection (GC-MS) or electron-capture
detection (GC-ECD)

Matrices solides environnementales - Dosage des Feststoffe in der Umwelt - Bestimmung von

polychlorobiphényles (PCB) par chromatographie en polychlorierten Biphenylen (PCB) mittels

phase gazeuse-spectrométrie de masse (CG-SM) ou Gaschromatographie und massenspektrometrischer

chromatographie en phase gazeuse avec détection par Detektion (GC-MS) oder Elektronen-Einfang-Detektion

capture d'électrons (CG-ECD) (GC-ECD)
This European Standard was approved by CEN on 15 June 2020.

This European Standard was corrected and reissued by the CEN-CENELEC Management Centre on 9 September 2020.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,

Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATIO N
EUROPÄISCHES KOMITEE FÜR NORMUN G
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17322:2020 E

worldwide for CEN national Members.
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SIST EN 17322:2020
EN 17322:2020 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 6

4 Principle ............................................................................................................................................................. 7

5 Interferences .................................................................................................................................................... 8

5.1 Interference with sampling and extraction ........................................................................................... 8

5.2 Interference with GC ...................................................................................................................................... 8

6 Safety remarks ................................................................................................................................................. 8

7 Reagents ............................................................................................................................................................. 9

7.1 General ................................................................................................................................................................ 9

7.2 Reagents for extraction ................................................................................................................................. 9

7.3 Reagents for clean-up .................................................................................................................................... 9

7.4 Gas chromatographic analysis ................................................................................................................ 12

7.5 Standards ........................................................................................................................................................ 12

7.6 Preparation of standard solutions ......................................................................................................... 14

8 Apparatus ........................................................................................................................................................ 15

8.1 Extraction and clean-up procedures ..................................................................................................... 15

8.2 Gas chromatograph ..................................................................................................................................... 16

9 Sample storage and preservation .......................................................................................................... 16

9.1 Sample storage .............................................................................................................................................. 16

9.2 Sample pre-treatment ................................................................................................................................ 16

10 Procedure........................................................................................................................................................ 17

10.1 Blank test ........................................................................................................................................................ 17

10.2 Extraction ........................................................................................................................................................ 17

10.3 Concentration ................................................................................................................................................ 20

10.4 Clean-up of the extract ............................................................................................................................... 20

10.5 Addition of the injection standard ......................................................................................................... 24

10.6 Gas chromatographic analysis (GC) ....................................................................................................... 24

10.7 Mass spectrometry (MS) ............................................................................................................................ 24

10.8 Electron capture detection (ECD) ........................................................................................................... 28

11 Performance characteristics .................................................................................................................... 30

12 Precision .......................................................................................................................................................... 30

13 Test report ...................................................................................................................................................... 30

Annex A (informative) Repeatability and reproducibility data ............................................................... 31

A.1 Materials used in the inter-laboratory comparison study ............................................................ 31

A.2 Inter-laboratory comparison results .................................................................................................... 32

Annex B (informative) Examples for gas chromatographic conditions and retention times

of PCBs .............................................................................................................................................................. 36

Annex C (informative) Calculation method for the estimation of total PCB content ........................ 37

Bibliography ................................................................................................................................................................. 45

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SIST EN 17322:2020
EN 17322:2020 (E)
European foreword

This document (EN 17322:2020) has been prepared by Technical Committee CEN/TC 444

“Environmental characterization of solid matrices”, the secretariat of which is held by NEN.

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 January 2021, and conflicting national standards shall

be withdrawn at the latest by January 2021.

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 16167:2018+AC:2019 and EN 15308:2016.

This document is the result of the merging of EN 16167:2018+AC:2019 and EN 15308:2016, with minor

technical modifications.

According to the CEN-CENELEC Internal Regulations, the national standards organisations of the

following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia,

Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland,

Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of North

Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United

Kingdom.
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SIST EN 17322:2020
EN 17322:2020 (E)
Introduction

Polychlorinated biphenyls (PCB) have been widely used as additives in industrial applications where

chemical stability has been required. This stability on the other hand creates environmental problems

when PCB are eventually released into the environment. Since some of these PCB compounds are highly

toxic, their presence in the environment (air, water, soil, sediment and waste) is regularly monitored and

controlled. At present determination of PCB is carried out in these matrices in most of the routine

laboratories following the preceding steps for sampling, pre-treatment, extraction and clean-up, by

measurement of specific PCB by means of gas chromatography in combination with mass spectrometric

detection (GC-MS) or gas chromatography with electron capture detector (GC-ECD).

This document was developed by merging of EN 16167:2018+AC:2019, initially elaborated as a CEN

Technical Specification in the European project 'HORIZONTAL' and validated by CEN/TC 400 with the

support of BAM, with EN 15308, published by CEN/TC 292.

Considered the different matrices and possible interfering compounds, this document does not contain

one single possible way of working. Several choices are possible, in particular relating to clean-up.

Detection with both MS-detection and ECD-detection is possible. Two different extraction procedures are

described and 9 clean-up procedures. The use of internal and injection standards is described in order to

have an internal check on choice of the extraction and clean-up procedure. The method is as far as

possible in agreement with the method described for PAH (EN 16181:2018 and EN 15527:2008). It has

been tested for ruggedness.

This document is applicable and validated for several types of matrices as indicated in Table 1 (see also

Annex A for the results of the validation).
Table 1 — Matrices for which this European Standard is applicable and validated
Matrix Materials used for validation
Soil Sandy soil
Mix of soil from the vicinity of Berlin, Germany and PCB-free
German reference soil
Sludge Mix of municipal waste water treatment plant sludge from
North Rhine Westphalia, Germany
Biowaste Mix of compost from the vicinity of Berlin, Germany and
sludge from North Rhine Westphalia, Germany
Waste Contaminated soil, building debris, waste wood, sealant
waste, electronic waste, shredder light fraction, cable
shredder waste

WARNING — Persons using this document should be familiar with usual 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 and to ensure compliance

with any national regulatory conditions.

IMPORTANT — It is absolutely essential that tests conducted according to this document be carried out

by suitably trained staff.
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SIST EN 17322:2020
EN 17322:2020 (E)
1 Scope

This document specifies methods for quantitative determination of seven selected polychlorinated

biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in soil, sludge, sediment,

treated biowaste, and waste using GC-MS and GC-ECD (see Table 2).
Table 2 — Target analytes of this European Standard
Target analyte
CAS-RN
PCB28 2,4,4'-trichlorobiphenyl 7012–37–5
PCB52 2,2',5,5'-tetrachlorobiphenyl 35693–99–3
PCB101 2,2',4,5,5'-pentachlorobiphenyl 37680–73–2
PCB118 2,3',4,4',5-pentachlorobiphenyl 31508–00–6
PCB138 2,2',3,4,4',5'-hexachlorobiphenyl 35065–28–2
PCB153 2,2',4,4',5,5'-hexachlorobiphenyl 35065–27–1
PCB180 2,2',3,4,4',5,5'-heptachlorobiphenyl 35065–29–3
CAS-RN Chemical Abstracts Service Registry Number.

The limit of detection depends on the determinants, the equipment used, the quality of chemicals used

for the extraction of the sample and the clean-up of the extract.

Under the conditions specified in this document, lower limit of application from 1 μg/kg (expressed as

dry matter) for soils, sludge and biowaste to 10 μg/kg (expressed as dry matter) for solid waste can be

achieved. For some specific samples the limit of 10 μg/kg cannot be reached.

Sludge, waste and treated biowaste may differ in properties, as well as in the expected contamination

levels of PCB and presence of interfering substances. These differences make it impossible to describe

one general procedure. This document contains decision tables based on the properties of the sample and

the extraction and clean-up procedure to be used.

NOTE The analysis of PCB in insulating liquids, petroleum products, used oils and aqueous samples is referred

to in EN 61619, EN 12766-1 and EN ISO 6468 respectively.

The method can be applied to the analysis of other PCB congeners not specified in the scope, provided

suitability is proven by proper in-house validation experiments.
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.

EN ISO 5667-15, Water quality — Sampling — Part 15: Guidance on the preservation and handling of

sludge and sediment samples (ISO 5667-15)

EN ISO 16720, Soil quality — Pretreatment of samples by freeze-drying for subsequent analysis (ISO 16720)

EN ISO 22892, Soil quality — Guidelines for the identification of target compounds by gas chromatography

and mass spectrometry (ISO 22892)

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

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SIST EN 17322:2020
EN 17322:2020 (E)

ISO 18512, Soil quality — Guidance on long and short-term storage of soil samples

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

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at https://www.iso.org/obp/ui
3.1
critical pair

pair of congeners that shall be separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic

separation meets minimum quality criteria
∆ t
(1)
Rx2× ( )
Ya+ Yb
where
R resolution
Δt difference in retention times of the two peaks a and b in seconds (s)
Y peak width at the base of peak a in seconds (s)
Y peak width at the base of peak b in seconds (s)
Figure 1 — Example of a chromatogram of a critical pair
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SIST EN 17322:2020
EN 17322:2020 (E)
3.2
congener

member of the same kind, class or group of chemicals, e.g. anyone of the two hundred and nine individual

PCB

NOTE 1 to entry: The IUPAC congener numbers are for easy identification; they do not represent the order of

chromatographic elution.
3.3
injection standard

C -labelled PCB or other PCB that is unlikely to be present in samples, added to the sample extract

before injection into the gas chromatograph, to monitor variability of instrument response and the

recovery of the internal standards
3.4
internal standard

C -labelled PCB or other PCB that are unlikely to be present in samples, added to the sample before

extraction and used for quantification of PCB content
3.5
polychlorinated biphenyl
PCB
biphenyl substituted with one to ten chlorine atoms
3.6
sediment

solid material, both mineral and organic, deposited in the bottom of a water body

[SOURCE: ISO 5667-12:2017]
4 Principle

Due to the multi-matrix character of this European Standard, different procedures for different steps

(modules) are allowed. Which modules should be used depends on the sample. A recommendation is

given in this document. Performance criteria are described and it is the responsibility of the laboratories

applying this document to show that these criteria are met. Using of spiking standards (internal

standards) allows an overall check on the efficiency of a specific combination of modules for a specific

sample. But it does not necessarily give the information regarding the extensive extraction efficiency of

the native PCB bonded to the matrix.
After pre-treatment, the sample is extracted with a suitable solvent.

The extract is concentrated by evaporation. If necessary, interfering compounds are removed by a clean-

up method suitable for the specific matrix, before the concentration step.

The extract is analysed by gas chromatography. The various compounds are separated using a capillary

column with a stationary phase of low polarity. Detection occurs by mass spectrometry (MS) or an

electron capture detector (ECD).

PCB are identified and quantified by comparison of relative retention times and relative peak heights (or

peak areas) with respect to internal standards added. The efficiency of the procedure depends on the

composition of the matrix that is investigated.
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SIST EN 17322:2020
EN 17322:2020 (E)
5 Interferences
5.1 Interference with sampling and extraction

Use sampling containers of materials (preferably of steel, aluminium or glass) that do not affect the

sample during the contact time. Avoid plastics and other organic materials during sampling, sample

storage or extraction. Keep the samples from direct sunlight and prolonged exposure to light.

During storage of the samples, losses of PCB may occur due to adsorption on the walls of the containers.

The extent of the losses depends on the storage time.
5.2 Interference with GC

Substances that co-elute with the target PCB may interfere with the determination. These interferences

may lead to incompletely resolved signals and may, depending on their magnitude, affect accuracy and

precision of the analytical results. Peak overlap does not allow an interpretation of the result. Asymmetric

peaks and peaks being broader than the corresponding peaks of the reference substance suggest

interferences.
Chromatographic separation between the following pairs can be critical.
— PCB28 – PCB31
— PCB52 – PCB73
— PCB101 – PCB89 / PCB90
— PCB118 – PCB106
— PCB138 – PCB164 / PCB163

The critical pair PCB28 and PCB31 is used for selection of the capillary column (see 8.2.2). If molecular

mass differences are present, quantification can be made by mass selective detection. If not or using ECD,

the specific PCB is reported as the sum of all PCBs present in the peak. Typically, the concentrations of

the co-eluting congeners compared to those of the target congeners are low. When incomplete resolution

is encountered, peak integration shall be checked and, when necessary, corrected.

Presence of tetrachlorobenzyltoluene (TCBT)-mixtures or sulfur can disturb the determination of the

PCB with GC-ECD.
High mineral oil content can also disturb the determination of PCB with GC-MS.
6 Safety remarks

PCBs are highly toxic and shall be handled with extreme care. Avoid contact with solid materials, solvent

extracts and solutions of standard PCB. Contact of solutions of standard with the body should be

prevented. It is strongly advised that standard solutions are prepared centrally in suitably equipped

laboratories or are purchased from suppliers specialized in their preparation.

Solvent solutions containing PCB and samples shall be disposed of in a manner approved for disposal of

toxic wastes.

For the handling of hexane precautions shall be taken because of its neurotoxic properties.

National regulations enforcing locally stricter requirements are used with respect to all hazards

associated with this method.
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EN 17322:2020 (E)
7 Reagents
7.1 General

All reagents shall be of recognized analytical grade. The purity of the reagents used shall be checked by

running a blank test as described in 10.1. The blank shall be less than 50 % of the lowest reporting limit.

7.2 Reagents for extraction
7.2.1 Acetone (2-propanone), (CH ) CO.
3 2
7.2.2 n-heptane, C H .
7 16
7.2.3 Petroleum ether, boiling range 40 °C to 60 °C.
Hexane-like solvents with a boiling range between 30 °C and 98 °C are allowed.

7.2.4 Sodium sulfate, Na SO . The anhydrous sodium sulfate shall be kept carefully sealed.

2 4
7.2.5 Distilled water or water of equivalent quality, H O.
7.2.6 Sodium chloride, NaCl,
7.2.7 Keeper substance. High boiling compound, i.e. octane, nonane.
7.3 Reagents for clean-up
7.3.1 Clean-up A using aluminium oxide
7.3.1.1 Aluminium oxide, Al O
2 3
Basic or neutral, specific surface 200 m /g, activity Super I [13].
7.3.1.2 Deactivated aluminium oxide
Deactivated with approximately 10 % water.

Add approximately 10 g of water (7.2.5) to 90 g of aluminium oxide (7.3.1.1). Shake until all lumps have

disappeared. Allow the aluminium oxide to condition before use for some 16 h, sealed from the air, use it

for maximum two weeks.

NOTE 1 The activity depends on the water content. It can be necessary to adjust the water content.

NOTE 2 Commercially available aluminium oxides with 10 % mass fraction water can also be used.

7.3.2 Clean-up B using silica gel 60 for column chromatography
7.3.2.1 Silica gel 60, particle size 63 µm to 200 µm.
7.3.2.2 Silica gel 60, water content: mass fraction w(H O) = 10 %.

Silica gel 60 (7.3.2.1), heated for at least 3 h at 450 °C, cooled down and stored in a desiccator containing

magnesium perchlorate or a suitable drying agent. Before use heat at least for 5 h at 130 °C in a drying

oven. Then allow cooling in a desiccator and add 10 % water (mass fraction) (7.2.5) in a flask. Shake for

5 min intensively by hand until all lumps have disappeared and then for 2 h in a shaking device. Store the

deactivated silica gel in the absence of air, use it for maximum of two weeks.
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SIST EN 17322:2020
EN 17322:2020 (E)
7.3.3 Clean-up C using gel permeation chromatography (GPC)
7.3.3.1 Bio-Beads S-X3.
7.3.3.2 Ethyl acetate, C H O .
4 8 2
7.3.3.3 Cyclohexane, C H .
6 12

Preparation of GPC, for example: put 50 g Bio-Beads S-X3 (7.3.3.1) into a 500 ml Erlenmeyer flask and

add 300 ml elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 1:1 (volume

fraction) in order to allow the beads to swell; after swirling for a short time until no lumps are left,

maintain the flask closed for 24 h. Drain the slurry into the chromatography tube for GPC. After

approximately three days, push in the plungers of the column so that a filling level of

approximately 35 cm is obtained. To further compress the gel, pump approximately 2 l of elution mixture

through the column at a flow rate of 5 ml · min and push in the plungers to obtain a filling level of

approximately 33 cm.
7.3.4 Clean-up D using Florisil
7.3.4.1 Florisil , baked 2 h at 600 °C. Particle size 150 µm to 750 µm.
7.3.4.2 Iso-octane, C H .
8 18
7.3.4.3 Toluene, C H .
7 8
7.3.4.4 Iso-octane/Toluene 95/5 volumetric fraction
7.3.5 Clean-up E using silica H SO /silica NaOH
2 4

7.3.5.1 Silica, SiO , particle size 70 µm to 230 µm, baked at 180 °C for a minimum of 1 h, and stored

in a pre-cleaned glass bottle with screw cap that prevents moisture from entering.

7.3.5.2 Sulfuric acid H SO 95 – 97 % percent mass fraction
2 4
7.3.5.3 Silica, treated with sulfuric acid.
Mix 56 g silica (7.3.5.1) and 44 g sulfuric acid (7.3.5.2).
7.3.5.4 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.3.5.5 Silica, treated with sodium hydroxide.
Mix 33 g silica (7.3.5.1) and 17 g sodium hydroxide (7.3.5.4).
1 GPC is also known as SEC (size exclusion chromatography).

2 Bio-Beads is an example of a suitable product available commercially. This information is given for the convenience of users

of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products can be used if

they can be shown to lead to the same results.

3 Florisil is a trade name for a prepared diatomaceous substance, mainly consisting of anhydrous magnesium silicate. This

information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of

this product. Equivalent products can be used if they can be shown to lead to the same results.

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EN 17322:2020 (E)
7.3.5.6 n-hexane, C H
6 14
7.3.6 Clean-up F using benzenesulfonic acid/sulfuric acid
7.3.6.1 silica gel with particle size between 40 µm to 200 µm.
7.3.6.2 benzenesulfonic acid C H O S > 98 % percent mass fraction
6 6 3

Mix 500 mg of silica gel with sulfuric acid (7.3.5.2) or benzenesulfonic acid (7.3.6.2) and add it into a 3 ml

column
7.3.7 Clean-up G using TBA sulfite reagent

7.3.7.1 Tetrabutylammonium reagent (TBA sulfite reagent) 97 % percent mass fraction

7.3.7.2 2-Propanol, C H O.
3 8
7.3.7.3 Sodium sulfite, Na SO > 98 % percent mass fraction
2 3

Saturate a solution of tetrabutylammonium hydrogen sulphate in a mixture of equal volume of water and

2-propanol, c((C H ) NHSO ) = 0,1 mol/l, with sodium sulphite.
4 9 4 4
NOTE 25 g of sodium sulphite might be sufficient for 100 ml of solution.
7.3.8 Clean-up H using pyrogenic copper

WARNING — Pyrogenic copper is spontaneously inflammable. Suitable precautions shall be taken.

7.3.8.1 Copper(II)-s
...

SLOVENSKI STANDARD
SIST EN 17322:2020
01-november-2020
Nadomešča:
SIST EN 15308:2017
SIST EN 16167:2018+AC:2019

Trdni matriksi z vidika okolja - Določevanje polikloriranih bifenilov (PCB) s plinsko

kromatografijo z masno selektivnim detektorjem (GC-MS) ali s plinsko
kromatografijo z detektorjem z zajetjem elektronov (GC-ECD)

Environmental Solid Matrices - Determination of polychlorinated biphenyls (PCB) by gas

chromatography - mass selective detection (GC-MS) or electron-capture detection (GC-

ECD)

Feststoffe in der Umwelt - Bestimmung von polychlorierten Biphenylen (PCB) mittels

Gaschromatographie und massenspektrometrischer Detektion (GC-MS) oder
Elektronen-Einfang-Detektion (GC-ECD)
Matrices solides environnementales - Dosage des polychlorobiphényles (PCB) par
chromatographie en phase gazeuse-spectrométrie de masse (CG-SM) ou
chromatographie en phase gazeuse avec détection par capture d’électrons (CG-ECD)
Ta slovenski standard je istoveten z: EN 17322:2020
ICS:
13.030.10 Trdni odpadki Solid wastes
13.080.10 Kemijske značilnosti tal Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST EN 17322:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN 17322:2020
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SIST EN 17322:2020
EN 17322
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2020
EUROPÄISCHE NORM

ICS 13.030.01; 13.030.10; 13.030.20; 13.080.10 Supersedes EN 15308:2016, EN 16167:2018+AC:2019

English Version
Environmental Solid Matrices - Determination of
polychlorinated biphenyls (PCB) by gas chromatography -
mass selective detection (GC-MS) or electron-capture
detection (GC-ECD)

Matrices solides environnementales - Dosage des Feststoffe in der Umwelt - Bestimmung von

polychlorobiphényles (PCB) par chromatographie en polychlorierten Biphenylen (PCB) mittels

phase gazeuse-spectrométrie de masse (CG-SM) ou Gaschromatographie und massenspektrometrischer

chromatographie en phase gazeuse avec détection par Detektion (GC-MS) oder Elektronen-Einfang-Detektion

capture d'électrons (CG-ECD) (GC-ECD)
This European Standard was approved by CEN on 15 June 2020.

This European Standard was corrected and reissued by the CEN-CENELEC Management Centre on 9 September 2020.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,

Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATIO N
EUROPÄISCHES KOMITEE FÜR NORMUN G
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17322:2020 E

worldwide for CEN national Members.
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SIST EN 17322:2020
EN 17322:2020 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 6

4 Principle ............................................................................................................................................................. 7

5 Interferences .................................................................................................................................................... 8

5.1 Interference with sampling and extraction ........................................................................................... 8

5.2 Interference with GC ...................................................................................................................................... 8

6 Safety remarks ................................................................................................................................................. 8

7 Reagents ............................................................................................................................................................. 9

7.1 General ................................................................................................................................................................ 9

7.2 Reagents for extraction ................................................................................................................................. 9

7.3 Reagents for clean-up .................................................................................................................................... 9

7.4 Gas chromatographic analysis ................................................................................................................ 12

7.5 Standards ........................................................................................................................................................ 12

7.6 Preparation of standard solutions ......................................................................................................... 14

8 Apparatus ........................................................................................................................................................ 15

8.1 Extraction and clean-up procedures ..................................................................................................... 15

8.2 Gas chromatograph ..................................................................................................................................... 16

9 Sample storage and preservation .......................................................................................................... 16

9.1 Sample storage .............................................................................................................................................. 16

9.2 Sample pre-treatment ................................................................................................................................ 16

10 Procedure........................................................................................................................................................ 17

10.1 Blank test ........................................................................................................................................................ 17

10.2 Extraction ........................................................................................................................................................ 17

10.3 Concentration ................................................................................................................................................ 20

10.4 Clean-up of the extract ............................................................................................................................... 20

10.5 Addition of the injection standard ......................................................................................................... 24

10.6 Gas chromatographic analysis (GC) ....................................................................................................... 24

10.7 Mass spectrometry (MS) ............................................................................................................................ 24

10.8 Electron capture detection (ECD) ........................................................................................................... 28

11 Performance characteristics .................................................................................................................... 30

12 Precision .......................................................................................................................................................... 30

13 Test report ...................................................................................................................................................... 30

Annex A (informative) Repeatability and reproducibility data ............................................................... 31

A.1 Materials used in the inter-laboratory comparison study ............................................................ 31

A.2 Inter-laboratory comparison results .................................................................................................... 32

Annex B (informative) Examples for gas chromatographic conditions and retention times

of PCBs .............................................................................................................................................................. 36

Annex C (informative) Calculation method for the estimation of total PCB content ........................ 37

Bibliography ................................................................................................................................................................. 45

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SIST EN 17322:2020
EN 17322:2020 (E)
European foreword

This document (EN 17322:2020) has been prepared by Technical Committee CEN/TC 444

“Environmental characterization of solid matrices”, the secretariat of which is held by NEN.

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 January 2021, and conflicting national standards shall

be withdrawn at the latest by January 2021.

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 16167:2018+AC:2019 and EN 15308:2016.

This document is the result of the merging of EN 16167:2018+AC:2019 and EN 15308:2016, with minor

technical modifications.

According to the CEN-CENELEC Internal Regulations, the national standards organisations of the

following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia,

Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland,

Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of North

Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United

Kingdom.
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SIST EN 17322:2020
EN 17322:2020 (E)
Introduction

Polychlorinated biphenyls (PCB) have been widely used as additives in industrial applications where

chemical stability has been required. This stability on the other hand creates environmental problems

when PCB are eventually released into the environment. Since some of these PCB compounds are highly

toxic, their presence in the environment (air, water, soil, sediment and waste) is regularly monitored and

controlled. At present determination of PCB is carried out in these matrices in most of the routine

laboratories following the preceding steps for sampling, pre-treatment, extraction and clean-up, by

measurement of specific PCB by means of gas chromatography in combination with mass spectrometric

detection (GC-MS) or gas chromatography with electron capture detector (GC-ECD).

This document was developed by merging of EN 16167:2018+AC:2019, initially elaborated as a CEN

Technical Specification in the European project 'HORIZONTAL' and validated by CEN/TC 400 with the

support of BAM, with EN 15308, published by CEN/TC 292.

Considered the different matrices and possible interfering compounds, this document does not contain

one single possible way of working. Several choices are possible, in particular relating to clean-up.

Detection with both MS-detection and ECD-detection is possible. Two different extraction procedures are

described and 9 clean-up procedures. The use of internal and injection standards is described in order to

have an internal check on choice of the extraction and clean-up procedure. The method is as far as

possible in agreement with the method described for PAH (EN 16181:2018 and EN 15527:2008). It has

been tested for ruggedness.

This document is applicable and validated for several types of matrices as indicated in Table 1 (see also

Annex A for the results of the validation).
Table 1 — Matrices for which this European Standard is applicable and validated
Matrix Materials used for validation
Soil Sandy soil
Mix of soil from the vicinity of Berlin, Germany and PCB-free
German reference soil
Sludge Mix of municipal waste water treatment plant sludge from
North Rhine Westphalia, Germany
Biowaste Mix of compost from the vicinity of Berlin, Germany and
sludge from North Rhine Westphalia, Germany
Waste Contaminated soil, building debris, waste wood, sealant
waste, electronic waste, shredder light fraction, cable
shredder waste

WARNING — Persons using this document should be familiar with usual 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 and to ensure compliance

with any national regulatory conditions.

IMPORTANT — It is absolutely essential that tests conducted according to this document be carried out

by suitably trained staff.
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EN 17322:2020 (E)
1 Scope

This document specifies methods for quantitative determination of seven selected polychlorinated

biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in soil, sludge, sediment,

treated biowaste, and waste using GC-MS and GC-ECD (see Table 2).
Table 2 — Target analytes of this European Standard
Target analyte
CAS-RN
PCB28 2,4,4'-trichlorobiphenyl 7012–37–5
PCB52 2,2',5,5'-tetrachlorobiphenyl 35693–99–3
PCB101 2,2',4,5,5'-pentachlorobiphenyl 37680–73–2
PCB118 2,3',4,4',5-pentachlorobiphenyl 31508–00–6
PCB138 2,2',3,4,4',5'-hexachlorobiphenyl 35065–28–2
PCB153 2,2',4,4',5,5'-hexachlorobiphenyl 35065–27–1
PCB180 2,2',3,4,4',5,5'-heptachlorobiphenyl 35065–29–3
CAS-RN Chemical Abstracts Service Registry Number.

The limit of detection depends on the determinants, the equipment used, the quality of chemicals used

for the extraction of the sample and the clean-up of the extract.

Under the conditions specified in this document, lower limit of application from 1 μg/kg (expressed as

dry matter) for soils, sludge and biowaste to 10 μg/kg (expressed as dry matter) for solid waste can be

achieved. For some specific samples the limit of 10 μg/kg cannot be reached.

Sludge, waste and treated biowaste may differ in properties, as well as in the expected contamination

levels of PCB and presence of interfering substances. These differences make it impossible to describe

one general procedure. This document contains decision tables based on the properties of the sample and

the extraction and clean-up procedure to be used.

NOTE The analysis of PCB in insulating liquids, petroleum products, used oils and aqueous samples is referred

to in EN 61619, EN 12766-1 and EN ISO 6468 respectively.

The method can be applied to the analysis of other PCB congeners not specified in the scope, provided

suitability is proven by proper in-house validation experiments.
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.

EN ISO 5667-15, Water quality — Sampling — Part 15: Guidance on the preservation and handling of

sludge and sediment samples (ISO 5667-15)

EN ISO 16720, Soil quality — Pretreatment of samples by freeze-drying for subsequent analysis (ISO 16720)

EN ISO 22892, Soil quality — Guidelines for the identification of target compounds by gas chromatography

and mass spectrometry (ISO 22892)

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

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EN 17322:2020 (E)

ISO 18512, Soil quality — Guidance on long and short-term storage of soil samples

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

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at https://www.iso.org/obp/ui
3.1
critical pair

pair of congeners that shall be separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic

separation meets minimum quality criteria
∆ t
(1)
Rx2× ( )
Ya+ Yb
where
R resolution
Δt difference in retention times of the two peaks a and b in seconds (s)
Y peak width at the base of peak a in seconds (s)
Y peak width at the base of peak b in seconds (s)
Figure 1 — Example of a chromatogram of a critical pair
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EN 17322:2020 (E)
3.2
congener

member of the same kind, class or group of chemicals, e.g. anyone of the two hundred and nine individual

PCB

NOTE 1 to entry: The IUPAC congener numbers are for easy identification; they do not represent the order of

chromatographic elution.
3.3
injection standard

C -labelled PCB or other PCB that is unlikely to be present in samples, added to the sample extract

before injection into the gas chromatograph, to monitor variability of instrument response and the

recovery of the internal standards
3.4
internal standard

C -labelled PCB or other PCB that are unlikely to be present in samples, added to the sample before

extraction and used for quantification of PCB content
3.5
polychlorinated biphenyl
PCB
biphenyl substituted with one to ten chlorine atoms
3.6
sediment

solid material, both mineral and organic, deposited in the bottom of a water body

[SOURCE: ISO 5667-12:2017]
4 Principle

Due to the multi-matrix character of this European Standard, different procedures for different steps

(modules) are allowed. Which modules should be used depends on the sample. A recommendation is

given in this document. Performance criteria are described and it is the responsibility of the laboratories

applying this document to show that these criteria are met. Using of spiking standards (internal

standards) allows an overall check on the efficiency of a specific combination of modules for a specific

sample. But it does not necessarily give the information regarding the extensive extraction efficiency of

the native PCB bonded to the matrix.
After pre-treatment, the sample is extracted with a suitable solvent.

The extract is concentrated by evaporation. If necessary, interfering compounds are removed by a clean-

up method suitable for the specific matrix, before the concentration step.

The extract is analysed by gas chromatography. The various compounds are separated using a capillary

column with a stationary phase of low polarity. Detection occurs by mass spectrometry (MS) or an

electron capture detector (ECD).

PCB are identified and quantified by comparison of relative retention times and relative peak heights (or

peak areas) with respect to internal standards added. The efficiency of the procedure depends on the

composition of the matrix that is investigated.
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5 Interferences
5.1 Interference with sampling and extraction

Use sampling containers of materials (preferably of steel, aluminium or glass) that do not affect the

sample during the contact time. Avoid plastics and other organic materials during sampling, sample

storage or extraction. Keep the samples from direct sunlight and prolonged exposure to light.

During storage of the samples, losses of PCB may occur due to adsorption on the walls of the containers.

The extent of the losses depends on the storage time.
5.2 Interference with GC

Substances that co-elute with the target PCB may interfere with the determination. These interferences

may lead to incompletely resolved signals and may, depending on their magnitude, affect accuracy and

precision of the analytical results. Peak overlap does not allow an interpretation of the result. Asymmetric

peaks and peaks being broader than the corresponding peaks of the reference substance suggest

interferences.
Chromatographic separation between the following pairs can be critical.
— PCB28 – PCB31
— PCB52 – PCB73
— PCB101 – PCB89 / PCB90
— PCB118 – PCB106
— PCB138 – PCB164 / PCB163

The critical pair PCB28 and PCB31 is used for selection of the capillary column (see 8.2.2). If molecular

mass differences are present, quantification can be made by mass selective detection. If not or using ECD,

the specific PCB is reported as the sum of all PCBs present in the peak. Typically, the concentrations of

the co-eluting congeners compared to those of the target congeners are low. When incomplete resolution

is encountered, peak integration shall be checked and, when necessary, corrected.

Presence of tetrachlorobenzyltoluene (TCBT)-mixtures or sulfur can disturb the determination of the

PCB with GC-ECD.
High mineral oil content can also disturb the determination of PCB with GC-MS.
6 Safety remarks

PCBs are highly toxic and shall be handled with extreme care. Avoid contact with solid materials, solvent

extracts and solutions of standard PCB. Contact of solutions of standard with the body should be

prevented. It is strongly advised that standard solutions are prepared centrally in suitably equipped

laboratories or are purchased from suppliers specialized in their preparation.

Solvent solutions containing PCB and samples shall be disposed of in a manner approved for disposal of

toxic wastes.

For the handling of hexane precautions shall be taken because of its neurotoxic properties.

National regulations enforcing locally stricter requirements are used with respect to all hazards

associated with this method.
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7 Reagents
7.1 General

All reagents shall be of recognized analytical grade. The purity of the reagents used shall be checked by

running a blank test as described in 10.1. The blank shall be less than 50 % of the lowest reporting limit.

7.2 Reagents for extraction
7.2.1 Acetone (2-propanone), (CH ) CO.
3 2
7.2.2 n-heptane, C H .
7 16
7.2.3 Petroleum ether, boiling range 40 °C to 60 °C.
Hexane-like solvents with a boiling range between 30 °C and 98 °C are allowed.

7.2.4 Sodium sulfate, Na SO . The anhydrous sodium sulfate shall be kept carefully sealed.

2 4
7.2.5 Distilled water or water of equivalent quality, H O.
7.2.6 Sodium chloride, NaCl,
7.2.7 Keeper substance. High boiling compound, i.e. octane, nonane.
7.3 Reagents for clean-up
7.3.1 Clean-up A using aluminium oxide
7.3.1.1 Aluminium oxide, Al O
2 3
Basic or neutral, specific surface 200 m /g, activity Super I [13].
7.3.1.2 Deactivated aluminium oxide
Deactivated with approximately 10 % water.

Add approximately 10 g of water (7.2.5) to 90 g of aluminium oxide (7.3.1.1). Shake until all lumps have

disappeared. Allow the aluminium oxide to condition before use for some 16 h, sealed from the air, use it

for maximum two weeks.

NOTE 1 The activity depends on the water content. It can be necessary to adjust the water content.

NOTE 2 Commercially available aluminium oxides with 10 % mass fraction water can also be used.

7.3.2 Clean-up B using silica gel 60 for column chromatography
7.3.2.1 Silica gel 60, particle size 63 µm to 200 µm.
7.3.2.2 Silica gel 60, water content: mass fraction w(H O) = 10 %.

Silica gel 60 (7.3.2.1), heated for at least 3 h at 450 °C, cooled down and stored in a desiccator containing

magnesium perchlorate or a suitable drying agent. Before use heat at least for 5 h at 130 °C in a drying

oven. Then allow cooling in a desiccator and add 10 % water (mass fraction) (7.2.5) in a flask. Shake for

5 min intensively by hand until all lumps have disappeared and then for 2 h in a shaking device. Store the

deactivated silica gel in the absence of air, use it for maximum of two weeks.
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EN 17322:2020 (E)
7.3.3 Clean-up C using gel permeation chromatography (GPC)
7.3.3.1 Bio-Beads S-X3.
7.3.3.2 Ethyl acetate, C H O .
4 8 2
7.3.3.3 Cyclohexane, C H .
6 12

Preparation of GPC, for example: put 50 g Bio-Beads S-X3 (7.3.3.1) into a 500 ml Erlenmeyer flask and

add 300 ml elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 1:1 (volume

fraction) in order to allow the beads to swell; after swirling for a short time until no lumps are left,

maintain the flask closed for 24 h. Drain the slurry into the chromatography tube for GPC. After

approximately three days, push in the plungers of the column so that a filling level of

approximately 35 cm is obtained. To further compress the gel, pump approximately 2 l of elution mixture

through the column at a flow rate of 5 ml · min and push in the plungers to obtain a filling level of

approximately 33 cm.
7.3.4 Clean-up D using Florisil
7.3.4.1 Florisil , baked 2 h at 600 °C. Particle size 150 µm to 750 µm.
7.3.4.2 Iso-octane, C H .
8 18
7.3.4.3 Toluene, C H .
7 8
7.3.4.4 Iso-octane/Toluene 95/5 volumetric fraction
7.3.5 Clean-up E using silica H SO /silica NaOH
2 4

7.3.5.1 Silica, SiO , particle size 70 µm to 230 µm, baked at 180 °C for a minimum of 1 h, and stored

in a pre-cleaned glass bottle with screw cap that prevents moisture from entering.

7.3.5.2 Sulfuric acid H SO 95 – 97 % percent mass fraction
2 4
7.3.5.3 Silica, treated with sulfuric acid.
Mix 56 g silica (7.3.5.1) and 44 g sulfuric acid (7.3.5.2).
7.3.5.4 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.3.5.5 Silica, treated with sodium hydroxide.
Mix 33 g silica (7.3.5.1) and 17 g sodium hydroxide (7.3.5.4).
1 GPC is also known as SEC (size exclusion chromatography).

2 Bio-Beads is an example of a suitable product available commercially. This information is given for the convenience of users

of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products can be used if

they can be shown to lead to the same results.

3 Florisil is a trade name for a prepared diatomaceous substance, mainly consisting of anhydrous magnesium silicate. This

information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of

this product. Equivalent products can be used if they can be shown to lead to the same results.

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EN 17322:2020 (E)
7.3.5.6 n-hexane, C H
6 14
7.3.6 Clean-up F using benzenesulfonic acid/sulfuric acid
7.3.6.1 silica gel with particle size between 40 µm to 200 µm.
7.3.6.2 benzenesulfonic acid C H O S > 98 % percent mass fraction
6 6 3

Mix 500 mg of silica gel with sulfuric acid (7.3.5.2) or benzenesulfonic acid (7.3.6.2) and add it into a 3 ml

column
7.3.7 Clean-up G using TBA sulfite reagent

7.3.7.1 Tetrabutylammonium reagent (TBA sulfite reagent) 97 % percent mass fraction

7.3.7.2 2-Propanol, C H O.
3 8
7.3.7.3 Sodium sulfite, Na SO > 98 % percent mass fraction
2 3

Saturate a solution of tetrabutylammonium hydrogen sulphate in a mixture of equal volume of water and

2-propanol, c((C H ) NHSO ) = 0,1 mol/l, with sodium sulphite.
4 9 4 4
NOTE 25 g of sodium sulphite might be sufficient for 100 ml of solution.
7.3.8 Clean-up H using pyrogenic copper

WARNING — Pyrogenic copper is spontaneously inflammable. Suitable precautions shall be taken.

7.3.8.1 Copper(II)-s
...

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