SIST-TS CEN/TS 16190:2012

SLOVENSKI STANDARD
SIST-TS CEN/TS 16190:2012
01-junij-2012
%ODWRREGHODQLELRORãNLRGSDGNLLQWOD'RORþHYDQMHGLRNVLQRYLQIXUDQRYLQ
GLRNVLQRPSRGREQLKSROLNORULUDQLKELIHQLORYVSOLQVNRNURPDWRJUDILMR]PDVQR
VHOHNWLYQLPGHWHNWRUMHPYLVRNHORþOMLYRVWL+5*&06
Sludge, treated biowaste and soil - Determination of dioxins and furans and dioxin-like
polychlorinated biphenyls by gas chromatography with high resolution mass selective
detection (HR GC-MS)
Schlamm, behandelter Bioabfall und Boden - Bestimmung von Dioxinen und Furanen
sowie Dioxin vergleichbaren polychlorierten Biphenylen mittels Gaschromatographie und
hochauflösender massenspektrometrischer Detektion (HR GC-MS)
Boue, biodéchet traité et sol - Détermination des dioxine et furane et biphényls
polychlorés de type dioxine par chromatographie en phase gazeuse-spectrométrie de
masse (HR GC-MS)
Ta slovenski standard je istoveten z: CEN/TS 16190:2012
ICS:
13.030.20 7HNRþLRGSDGNL%ODWR Liquid wastes. Sludge
13.080.10 .HPLMVNH]QDþLOQRVWLWDO Chemical characteristics of
soils
SIST-TS CEN/TS 16190:2012 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST-TS CEN/TS 16190:2012

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SIST-TS CEN/TS 16190:2012


TECHNICAL SPECIFICATION
CEN/TS 16190

SPÉCIFICATION TECHNIQUE

TECHNISCHE SPEZIFIKATION
February 2012
ICS 13.030.01
English Version
Sludge, treated biowaste and soil - Determination of dioxins and
furans and dioxin-like polychlorinated biphenyls by gas
chromatography with high resolution mass selective detection
(HR GC-MS)
Boues, biodéchets traités et sols - Détermination des Schlamm, behandelter Bioabfall und Boden - Bestimmung
dioxines et furanes et polychlorobiphényles de type dioxine von Dioxinen und Furanen sowie Dioxin vergleichbaren
par chromatographie en phase gazeuse avec spectrométrie polychlorierten Biphenylen mittels Gaschromatographie und
de masse à haute résolution (CG-SMHR) hochauflösender massenspektrometrischer Detektion (HR
GC-MS)
This Technical Specification (CEN/TS) was approved by CEN on 24 April 2011 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.

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, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16190:2012: E
worldwide for CEN national Members.

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Contents Page
Foreword .4
Introduction .5
1 Scope .6
2 Normative references .7
3 Abbreviations .7
4 Principle .7
5 Reagents .8
5.1 Chemicals .8
5.2 Standards .8
6 Apparatus and materials .8
6.1 General .8
6.2 Equipment for sample preparation .8
6.3 Soxhlet extractor.8
6.4 Clean-up apparatus .8
6.5 Concentration apparatus .9
6.6 Other equipment .9
7 Sample storage and sample pretreatment . 10
7.1 Sample storage . 10
7.2 Sample pretreatment . 10
8 Extraction and clean-up . 10
8.1 General . 10
8.2 Extraction . 10
8.3 Clean-up . 11
8.3.1 General . 11
8.3.2 Gel permeation chromatography . 12
8.3.3 Multilayer column . 12
8.3.4 Sulfuric acid treatment . 12
8.3.5 Activated carbon column . 12
8.3.6 Aluminium oxide column . 12
8.3.7 Removal of sulfur. 12
8.4 Final concentration of cleaned sample extract . 12
8.5 Addition of recovery standard . 13
9 HRGC/HRMS analysis . 13
9.1 General . 13
9.2 Gas chromatographic analysis . 13
9.3 Mass spectrometric detection . 13
9.4 Minimum requirements for identification of PCDF/PCDD and PCB . 15
9.5 Minimum requirements for quantification of PCDF/PCDD and PCB . 16
9.6 Calibration of the HRGC/HRMS system . 17
9.6.1 General . 17
9.6.2 Calibration for 2,3,7,8-congeners . 17
9.6.3 Calibration for sum of homologue groups . 17
9.7 Quantification of HRGC/HRMS results . 18
9.7.1 Quantification of concentrations of 2,3,7,8-congeners . 18
13
9.7.2 Quantification of recovery rates of C-labelled standards . 19
9.7.3 Quantification of sum of homologue groups . 19
9.7.4 Calculation of the toxic equivalent . 19
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9.7.5 Calculation of the limit of detection and the limit of quantification . 20
10 Precision. 20
11 Test report . 20
Annex A (informative) Toxic equivalent factors. 21
Annex B (informative) Repeatability and reproducibility data . 23
B.1 Materials used in the interlaboratory comparison study . 23
B.2 Interlaboratory comparison results . 24
Annex C (informative) Examples of extraction and clean-up methods . 27
C.1 Example A . 27
C.1.1 General . 27
C.1.2 Chemicals . 27
C.1.3 Procedure . 28
C.2 Example B: Approved clean-up methods . 33
Annex D (informative) Examples of operation of GC/HRMS determination . 35
D.1 Example . 35
D.1.1 General . 35
D.1.2 Gas chromatographic analysis . 35
D.1.3 Mass spectrometric detection . 36
Bibliography . 39

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Foreword
This document (CEN/TS 16190:2012) has been prepared by Technical Committee CEN/TC 400 “Project
Committee - Horizontal standards in the fields of sludge, biowaste and soil”, the secretariat of which is held by
DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
The preparation of this document by CEN is based on a mandate by the European Commission
(Mandate M/330), which assigned the development of standards on sampling and analytical methods for
hygienic and biological parameters as well as inorganic and organic determinants, aiming to make these
standards applicable to sludge, treated biowaste and soil as far as this is technically feasible.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: 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, Romania, Slovakia, Slovenia,
Spain, Sweden, Switzerland, Turkey and the United Kingdom.

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Introduction
Two groups of related chlorinated aromatic ethers are known as polychlorinated dibenzo-p-dioxins (PCDDs)
and polychlorinated dibenzofurans (PCDFs); they consist of a total of 210 individual substances (congeners):
75 PCDDs and 135 PCDFs.
A group of chlorinated aromatic compounds similar to polychlorinated dibenzo-p-dioxins (PCDDs) and
polychlorinated dibenzofurans (PCDFs) is known as polychlorinated biphenyls (PCBs) which consist of
209 individual substances.
PCDDs and PCDFs can form in the combustion of organic materials; they also occur as undesirable by-
products in the manufacture or further processing of chlorinated organic chemicals. PCDDs/PCDFs enter the
environment via these emission paths and through the use of contaminated materials. In fact, they are
universally present at very small concentrations. The 2,3,7,8-substituted congeners are toxicologically
significant. Toxicologically much less significant than the tetrachlorinated to octachlorinated dibenzo-p-
dioxins/dibenzofurans are the 74 monochlorinated to trichlorinated dibenzo-p-dioxins/dibenzofurans.
PCBs have been produced over a period of approximately 50 years until the end of the 1990s for the purpose
of different use in open and closed systems, e. g. as electrical insulators or dielectric fluids in capacitors and
transformers, as specialised hydraulic fluids, as a plasticizer in sealing material. Worldwide more than one
million tons of PCBs were produced.
PCDD/F as well as PCBs are emitted during thermal processes as e. g. waste incineration. In 1997 a group of
experts of the World Health Organisation (WHO) fixed toxicity equivalent factors (TEF) for PCDD and twelve
PCBs, known as dioxin-like PCBs (see Annex A). These twelve dioxin-like PCBs consist of four non-ortho
PCBs and eight mono-ortho PCBs (no or only one chlorine atoms in 2-, 2’-, 6- and 6’-position), having a planar
or mostly planar structure. Dioxin-like PCB can contribute considerably to the total WHO-TEQ.
Only skilled operators who are trained in handling highly toxic compounds should apply the method described
in this Technical Specification.
This Technical Specification is applicable for several types of matrices and validated for municipal sludge (see
also Annex B for the results of the validation).
WARNING — Persons using this Technical Specification should be familiar with usual laboratory
practice. This Technical Specification 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 Technical
Specification be carried out by suitably trained staff.
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1 Scope
This Technical Specification specifies a method for quantitative determination of 17 2,3,7,8-chlorine
substituted dibenzo-p-dioxins and dibenzofurans and dioxin-like polychlorinated biphenyls in sludge, treated
biowaste and soil using liquid column chromatographic clean-up methods and GC/HRMS.
The analytes to be determined with this Technical Specification are listed in Table 1.
Table 1 — Analytes and their abbreviations
Substance Abbreviation
Tetrachlorodibenzo-p-dioxin TCDD
Pentachlorodibenzo-p-dioxin PeCDD
Hexachlorodibenzo-p-dioxin HxCDD
Heptachlorodibenzo-p-dioxin HpCDD
Octachlorodibenzo-p-dioxin OCDD
Tetrachlorodibenzofuran TCDF
Pentachlorodibenzofuran PeCDF
Hexachlorodibenzofuran HxCDF
Heptachlorodibenzofuran HpCDF
Octachlorodibenzofuran OCDF
Polychlorinated biphenyl PCB
Trichlorobiphenyl TCB
Tetrachlorobiphenyl TeCB
Pentachlorobiphenyl PeCB
Hexachlorobiphenyl HxCB
Heptachlorobiphenyl HpCB
Decachlorobiphenyl DecaCB

The limit of detection depends on the kind of sample, the congener, the equipment used and the quality of
chemicals used for extraction and clean-up. Under the conditions specified in this Technical Specification,
limits of detection better than 1 ng/kg (expressed as dry matter) can be achieved.
This method is "performance based". It is permitted to modify the method if all performance criteria given in
this method are met.
NOTE In principle this method can also be applied for sediments, mineral wastes and for vegetation. It is the
responsibility of the user of this Technical Specification to validate the application for these matrices. For measurement in
complex matrices like fly ashes adsorbed on vegetation it can be necessary to further improve the clean up. This can also
apply to sediments and mineral wastes.
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2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
EN 16179, Sludge, treated biowaste and soil — Guidance for sample pretreatment
3 Abbreviations
PCB Polychlorinated biphenyls
PCDD/PCDF or PCDD/F Polychlorinated dibenzo-p-dioxins/dibenzofurans
I-TEF NATO/CCMS International toxic equivalent factor proposed by NATO-CCMS in 1988 (for
detailed description, see Annex A)
I-TEQ International toxic equivalent obtained by multiplying the mass determined with
the corresponding I-TEF including PCDDs and PCDFs (for detailed description,
see Annex A). Should only be used for comparison with older data
WHO-TEF Toxic equivalent factor proposed by WHO in 2005 (for detailed description, see
Annex A)
WHO-TEQ Toxic equivalent obtained by multiplying the mass determined with the
corresponding WHO-TEF including PCDD, PCDF and PCB (for detailed
description, see Annex A). WHO-TEQ , WHO-TEQ should be used to
PCB PCDD/F
distinguish different compound classes
4 Principle
This Technical Specification is based on the use of gas chromatography/mass spectrometry combined with
the isotope dilution technique to enable the separation, detection and quantification of PCDD/PCDF and
dioxin-like PCB in sludge, biowaste and soil. For the isotope dilution method 17 labelled PCDD/F and
12 labelled PCB internal standards are used. The extracts for the GC-MS measurements contain one or two
recovery standards. The gas chromatographic parameters offer information which enables the identification of
congeners (position of chlorine substitutes) whereas the mass spectrometric parameters enable the
differentiation between isomers with different numbers of chlorine substitutes and between dibenzo-p-dioxins,
furans and PCB.
13
C -labelled PCDD/F and PCB congeners are added to the sample prior to extraction and HRGC/HRMS
12
measurement. Losses during extraction and clean-up are detected and compensated by using these added
congeners as internal standards for quantification together with recovery standards which are added just
before the HRGC/HRMS analysis. For the determination of these substances it is necessary to separate
PCBs from PCDDs/PCDFs and vice versa.
The main purpose of the clean-up procedure of the raw sample extract is the removal of sample matrix
components, which may overload the separation method, disturb the quantification or otherwise severely
impact the performance of the identification and quantification method and the separation of PCDD/F from
dioxin-like PCB. Furthermore, the enrichment of the analytes in the final sample extract is achieved. Extraction
procedures are usually based on Soxhlet or equivalent extraction methods of dried, preferably freeze dried,
samples. Sample clean-up is usually carried out by multi-column liquid chromatographic techniques using
different adsorbents. The determination of PCDD/F and PCBs is based on quantification by the isotope-
dilution technique using HRGC/HRMS.
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5 Reagents
5.1 Chemicals
Solvents used for extraction and clean-up shall be of pesticide grade or equivalent quality and checked for
blanks. Adsorbents like aluminium oxide, silica gel, diatomaceous earth and others used for clean-up shall be
of analytical grade quality or better and pre-cleaned and activated if necessary.
NOTE See Annex C for a specific list of solvents and chemicals.
5.2 Standards
13
 C-spiking solution for PCDD/F (internal Standard);
13
 C-spiking solution for PCB (internal Standard);
 Calibration solutions PCDD/F;
 Calibration solutions PCB;
 Recovery standard PCDD/F;
 Recovery standard PCB.
NOTE See Annex C for examples of concentration of the standard solutions.
6 Apparatus and materials
6.1 General
The apparatus and materials listed below are meant as minimum requirements for "conventional" sample
treatment with Soxhlet extraction and column chromatographic clean-up. Additional apparatus and materials
may be necessary due to different methods of sample extraction and clean-up methods.
6.2 Equipment for sample preparation
6.2.1 Laboratory fume hood, of sufficient size to contain the sample preparation equipment listed below.
6.2.2 Desiccator.
6.2.3 Balances, consisting of an analytical type capable of weighing 0,1 mg and a top-loading type capable
of weighing 10 mg.
6.3 Soxhlet extractor
6.3.1 Soxhlet, 50 mm internal diameter, 150 ml or 250 ml capacity with 500 ml round bottom flask.
6.3.2 Thimble, 43 mm × 123 mm, to fit Soxhlet.
6.3.3 Hemispherical heating mantle, to fit 500 ml round-bottom flask.
6.4 Clean-up apparatus
6.4.1 Disposable pipettes, either disposable Pasteur pipettes, or disposable serological pipettes.
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6.4.2 Glass chromatographic columns, of the following sizes:
 150 mm length × 8 mm internal diameter, with coarse-glass frit or glass-wool plug, 250 ml reservoir and
glass or polytetrafluoroethylene (PTFE) stopcock;
 200 mm length × 15 mm internal diameter, with coarse-glass frit or glass-wool plug, 250 ml reservoir and
glass or PTFE stopcock;
 300 mm length × 25 mm internal diameter, with coarse-glass frit or glass-wool plug, 300 ml reservoir and
glass or PTFE stopcock.
6.4.3 Oven, capable of maintaining a constant temperature (± 5 °C) in the range of 105 °C to 450 °C for
baking and storage of adsorbents.
6.5 Concentration apparatus
6.5.1 Rotary evaporator, equipped with a variable temperature water bath and:
 vacuum source for rotary evaporator equipped with shutoff valve at the evaporator and vacuum gauge;
 recirculating water pump and chiller, providing cooling water of (9 ± 4) °C (use of tap water for cooling the
evaporator wastes large volumes of water and can lead to inconsistent performance as water
temperatures and pressures vary);
 round-bottom flask, 100 ml and 500 ml or larger, with ground-glass fitting compatible with the rotary
evaporator.
6.5.2 Nitrogen blowdown apparatus, equipped with either a water bath controlled in the range of 30 °C to
60 °C or a heated stream of nitrogen, installed in a fume hood.
1)
6.5.3 Kuderna-Danish concentrator.
6.5.4 Sample vials, of the following types:
 amber glass, nominated volume 2 ml to 5 ml, with PTFE-lined screw cap;
 glass, 0,3 ml, conical, with PTFE-lined screw or crimp cap.
6.6 Other equipment
6.6.1 Gas chromatograph, equipped with a splitless or on-column or temperature programmed injection
port for the use with capillary columns, and an oven temperature programme which enables isothermal hold.
6.6.2 GC column for PCDDs/PCDFs and for isomer specificity for 2,3,7,8-TCDD (e. g. 60 m length ×
0,32 mm internal diameter; 0,25 µm; 5 % phenyl, 94 % methyl, 1 % vinyl silicone bonded-phase fused-silica
capillary column).
6.6.3 Mass spectrometer, 28 eV to 80 eV electron impact ionization, capable of repetitively selectively
monitoring of twelve exact masses minimum at high resolution (> 10 000) during a period of approximately
1 s.
6.6.4 Data system, capable of collecting, recording, and storing mass spectrometric data.

1) Kuderna Danish is an example of a suitable product available commercially. This information is given for the
convenience of users of this Technical Specification and does not constitute an endorsement by CEN of this product.
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7 Sample storage and sample pretreatment
7.1 Sample storage
Samples should be stored in suitable containers with an appropriate closure material such as
polytetrafluoroethylene (PTFE). Samples to be frozen may be stored in aluminium containers pre-cleaned by
heating to 450 °C for minimum 4 h or by rinsing with a non-chlorinated solvent.
Samples should be kept cold (< 8 °C) and in the dark. The sample pretreatment should take place within
three days of sampling. Alternatively, samples may be frozen (–18 °C) directly after sampling and kept frozen
before sample pretreatment.
7.2 Sample pretreatment
Drying and homogenization should be carried out according to EN 16179, if not otherwise specified. Store the
ground material in a desiccator or a tightly closed glass container.
8 Extraction and clean-up
8.1 General
In this Technical Specification the minimum requirements for extraction and clean-up to be met are described
as well as examples of operation. The analyst may use any of the procedures given below and in Annex C or
any suitable alternative procedures.
The determination of PCDDs/PCDFs is based on quantification by the isotope-dilution technique using
13
HRGC/HRMS. C -labelled 2,3,7,8-chlorine substituted PCDD/PCDF congeners are added at different
12
stages of the whole method. Losses during extraction and clean-up can be detected and compensated by
using these added congeners as internal standards for quantification together with recovery standards which
are added just before the HRGC/HRMS analysis. However, due to possible differences in the binding and
13
adsorption characteristics between the native PCDDs/PCDFs and the C -labelled congeners, which are
12
added during analysis, complete substantiation of the extraction efficiency and compensation of losses during
clean-up is not ensured. Therefore, in addition the applied methods shall be validated thoroughly. Examples of
well-proven extraction and clean-up methods are given in Annex C.
The main purpose of the clea
...