SIST EN 16167:2018+AC:2019

SLOVENSKI STANDARD
01-marec-2019
1DGRPHãþD
SIST EN 16167:2018
7ODREGHODQLELRORãNLRGSDGNLLQEODWR'RORþHYDQMHSROLNORULUDQLKELIHQLORY3&%
VSOLQVNRNURPDWRJUDILMR]PDVQRVHOHNWLYQLPGHWHNWRUMHP*&06LQVSOLQVNR
NURPDWRJUDILMR]GHWHNWRUMHP]]DMHWMHPHOHNWURQRY*&(&'YNOMXþQRV
SRSUDYNRP$&
Soil, treated biowaste and sludge - Determination of polychlorinated biphenyls (PCB) by
gas chromatography with mass selective detection (GC-MS) and gas chromatography
with electron-capture detection (GC-ECD)
Boden, behandelter Bioabfall und Schlamm - Bestimmung von polychlorierten
Biphenylen (PCB) mittels Gaschromatographie mit Massenspektrometrie-Kopplung (GC-
MS) und Gaschromatographie mit Elektroneneinfangdetektion (GC-ECD)
Sols, biodéchets traités et boues - Dosage des polychlorobiphényles (PCBs) par
chromatographie en phase gazeuse-spectrométrie gazeuse couplée avec un détecteur
de masse (CG-SM) ou un détecteur par capture d'électrons (CG-ECD)
Ta slovenski standard je istoveten z: EN 16167:2018+AC:2019
ICS:
13.030.20 7HNRþLRGSDGNL%ODWR Liquid wastes. Sludge
13.080.10 .HPLMVNH]QDþLOQRVWLWDO Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN 16167:2018+AC
EUROPEAN STANDARD
NORME EUROPÉENNE
January 2019
EUROPÄISCHE NORM
ICS 13.030.01; 13.080.10 Supersedes EN 16167:2018
English Version
Soil, treated biowaste and sludge - Determination of
polychlorinated biphenyls (PCB) by gas chromatography
with mass selective detection (GC-MS) and gas
chromatography with electron-capture detection (GC-
ECD)
Sols, biodéchets traités et boues - Dosage des Boden, behandelter Bioabfall und Schlamm -
polychlorobiphényles (PCBs) par chromatographie en Bestimmung von polychlorierten Biphenylen (PCB)
phase gazeuse-spectrométrie gazeuse couplée avec un mittels Gaschromatographie mit
détecteur de masse (CG-SM) ou un détecteur par Massenspektrometrie-Kopplung (GC-MS) und
capture d'électrons (CG-ECD) Gaschromatographie mit Elektroneneinfangdetektion
(GC-ECD)
This European Standard was approved by CEN on 20 March 2018 and includes the Corrigendum issued by CEN on 30 January
2019.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.

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

EUROPÄISCHES KOMITEE FÜR NORMUN G

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 16167:2018+AC:2019 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 6
4 Principle . 7
5 Interferences . 7
5.1 Interference with sampling and extraction . 7
5.2 Interference with GC . 7
6 Safety remarks . 8
7 Reagents . 8
8 Apparatus . 14
9 Sample storage and preservation . 15
9.1 Sample storage . 15
9.2 Sample pretreatment . 15
10 Procedure. 16
10.1 Blank test . 16
10.2 Extraction . 16
10.3 Concentration . 18
10.4 Clean-up of the extract . 19
10.5 Addition of the injection standard . 22
10.6 Gas chromatographic analysis (GC) . 22
10.7 Mass spectrometry (MS) . 23
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
Annex B (informative) Examples for retention times of PCBs . 33
Annex C (informative) Calculation method for the estimation of total PCB content . 34
Bibliography . 39

European foreword
This document (EN 16167:2018+AC:2019) has been prepared by Technical Committee CEN/TC 444
“Test methods for 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 July 2019, and conflicting national standards shall be
withdrawn at the latest by July 2019.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes ˜EN 16167:2018™.
This document includes the corrigendum 1 which deletes the mandate information.
The start and finish of text introduced or altered by corrigendum is indicated in the text by tags ˜™
˜deleted text™.
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, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
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 PCBs 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, pretreatment, extraction, 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 European Standard was developed in the European project 'HORIZONTAL'. It is the result of a desk
study “3-12 PCB” and aims at evaluation of the latest developments in assessing PCBs in sludge, soil,
treated biowaste and neighbouring fields. Taken into account the different matrices and possible
interfering compounds, this European Standard 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. Three different extraction procedures are described and 11 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 PAHs (see EN 16181). It has been tested for ruggedness.
This European Standard 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
Sludge Municipal sewage sludge
Biowaste Compost
Soil Sandy soil
WARNING — Persons using this European Standard should be familiar with usual laboratory practice.
This European Standard 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 European
Standard be carried out by suitably trained staff.
1 Scope
This draft European Standard specifies a method for quantitative determination of seven selected
polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in sludge,
treated biowaste and soil using GC-MS and GC-ECD (see Table 2).
Table 2 — Target analytes of this European Standard
Target analyte a
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
a
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 European Standard, limit of application of 1 µg/kg (expressed as
dry matter) can be achieved.
Sludge and treated biowaste may differ in properties and also 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.
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 15934, Sludge, treated biowaste, soil and waste — Calculation of dry matter fraction after
determination of dry residue or water content
EN 16179, Sludge, treated biowaste and soil — Guidance for sample pretreatment
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
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 http://www.iso.org/obp
3.1
polychlorinated biphenyl
PCB
biphenyl substituted by one to ten chlorine atoms
[SOURCE: EN 15308:2016, 3.1]
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.
[SOURCE: EN 15308:2016, 3.2]
3.3
critical pair
pair of congeners that will be separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic
separation meets minimum quality criteria
[SOURCE: EN 15308:2016, 3.6]
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
[SOURCE: EN 15308:2016, 3.4, modified – "waste samples" is replaced here with "samples".]
3.5
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
[SOURCE: EN 15308:2016, 3.5, modified – "waste samples" is replaced here with "samples".]
4 Principle
Due to the horizontal 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 European Standard. Performance criteria are described and it is the responsibility of the
laboratories applying this European Standard 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 upon the extensive
extraction efficiency of the native PCB bonded to the matrix.
After pretreatment according to the methods referred to in 9.2, the test 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. The eluate is concentrated by evaporation.
The extract is analysed by gas chromatography with either mass spectrometric (GC-MS) or electron
capture detection (GC-ECD). Applying GC-MS the various compounds are separated using a capillary
column with a stationary phase of low polarity. In case of GC-ECD, extracts are analysed using two
columns of different polarity (see 8.2.1).
PCBs 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.
5 Interferences
5.1 Interference with sampling and extraction
Use sampling containers of materials (preferably of steel, aluminium or glass) that do not change 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 PCBs 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. 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.
— PCB28 – PCB31
— PCB52 – PCB73
— PCB101 – PCB89 / PCB90
— PCB118 – PCB106
— PCB138 – PCB164 / PCB163
Presence of considerable amounts of mineral oil in the sample may interfere with the quantification of
PCB in GC-MS. In presence of mineral oil, GC-ECD may be preferred or mineral oil can be removed using
clean-up procedure G (see 10.4.8) using DMF/n-hexane.
Presence of tetrachlorobenzyltoluene (TCBT)-mixtures may disturb the determination of the PCB with
GC-ECD.
6 Safety remarks
PCBs are highly toxic and shall be handled with extreme care. Contact between the body and solid
materials, solvent extracts and solutions of standard PCB shall not be allowed to occur. 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 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 shall be followed with respect to all hazards associated with this method.
7 Reagents
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.
7.1 Reagents for extraction
7.1.1 Propanone (Acetone), (CH ) CO.
3 2
7.1.2 n-heptane, C H .
7 16
7.1.3 Petroleum ether, boiling range 40 °C to 60 °C.
Hexane-like solvents with a boiling range between 40 °C and 89 °C are allowed.
7.1.4 Anhydrous sodium sulfate, Na SO :
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
7.1.5 Distilled water or water of equivalent quality, H O.
7.1.6 Sodium chloride, NaCl, anhydrous.
7.1.7 Keeper substance. High boiling compound, i.e. octane, nonane.
7.2 Reagents for clean-up:
7.2.1 Clean-up A using aluminium oxide:
7.2.1.1 Aluminium oxide, Al O .
2 3
Basic or neutral, specific surface 200 m /g, activity Super I according to Brockmann.
7.2.1.2 Deactivated aluminium oxide
Deactivated with approximately 10 % water.
Add approximately 10 g of water (7.1.5) to 90 g of aluminium oxide (7.2.1.1). 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 The activity depends on the water content. It can be necessary to adjust the water content.
7.2.2 Clean-up B using silica gel 60 for column chromatography
7.2.2.1 Silica gel 60, particle size 63 µm to 200 µm.
7.2.2.2 Silica gel 60, water content: mass fraction w(H O) = 10 %.
Silica gel 60 (7.2.2.1), 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) 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.
7.2.3 Clean-up C using gel permeation chromatography (GPC)
® 1)
7.2.3.1 Bio-Beads S-X3 .
7.2.3.2 Ethyl acetate, C H O .
4 8 2
7.2.3.3 Cyclohexane, C H .
6 12 ®
Preparation of GPC, for example: Put 50 g Bio-Beads S-X3 (7.2.3.1) into a 500 ml Erlenmeyer flask and
add 300 ml elution mixture made up of cyclohexane (7.2.3.3) and ethyl acetate (7.2.3.2) 1:1 (volume) 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
−1
rate of 5 ml · min and push in the plungers to obtain a filling level of approximately 33 cm.
®2)
7.2.4 Clean-up D using Florisil . ®
7.2.4.1 Florisil , baked 2 h at 600 °C. Particle size 150 µm to 750 µm.
7.2.4.2 Iso-octane, C H .
8 18 ®
1) Bio-Beads is an example of a suitable product available commercially. This information is given for the
convenience of users of this document and does not constitute an endorsement by CEN of this product. Equivalent
products may be used if they can be shown to lead to the same results. ®
2) 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 may be used if they can be shown to lead to the same
results.
7.2.4.3 Toluene, C H .
7 8
7.2.4.4 Iso-octane/Toluene 95/5 (v/v).
7.2.5 Clean-up E using silica H SO /silica NaOH.
2 4
7.2.5.1 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.
7.2.5.2 Silica, treated with sulfuric acid.
Mix 56 g silica (7.2.5.1) and 44 g sulfuric acid (7.2.8.1).
7.2.5.3 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.2.5.4 Silica, treated with sodium hydroxide.
Mix 33 g silica (7.2.5.1) and 17 g sodium hydroxide (7.2.5.3).
7.2.5.5 n-hexane, C H .
6 14
7.2.6 Clean-up F using benzenesulfonic acid/sulfuric acid
7.2.6.1 3 ml silica gel column, of adsorbent mass 500 mg, particle size 40 µm.
7.2.6.2 3 ml benzenesulfonic acid/sulfuric acid column, of adsorbent mass 500 mg, particle size
40 µm.
7.2.7 Clean-up G using DMF/hexane partitioning
7.2.7.1 Dimethylformamide(DMF), C H NO.
3 7
7.2.8 Clean-up H using concentrated sulfuric acid
7.2.8.1 Sulfuric acid, H SO of purity 96 % to 98 % (mass fraction).
2 4
7.2.9 Clean-up I using TBA sulfite reagent
7.2.9.1 Tetrabutylammonium reagent (TBA sulfite reagent)
Saturate a solution of tetrabutylammonium hydrogen sulfate in a mixture of equal volume of water and
2-propanol, c((C H ) NHSO ) = 0,1 mol/l, with sodium sulfite.
4 9 4 4
25 g of sodium sulfite should be sufficient for 100 ml of solution.
7.2.9.2 2-Propanol, C H O.
3 8
7.2.9.3 Sodium sulfite, Na SO .
2 3
7.2.10 Clean-up J using pyrogenic copper
WARNING — Pyrogenic copper is spontaneously inflammable. Suitable precautions shall be taken.
7.2.10.1 Copper(II)-sulfate pentahydrate, CuSO · 5 H O.
4 2
7.2.10.2 Hydrochloric acid, c(HCl) = 2 mol/l.
7.2.10.3 Zinc granules, Zn, particle size 0,3 mm to 1,4 mm.
7.2.10.4 Anionic detergent aqueous solution (e.g. 35 g/100 ml, n-dodecane-1-sulfonic acid sodium
salt (CH (CH ) SO Na).
3 2 11 3
Other commercially available detergents may also be suitable.
7.2.10.5 Deoxygenated water
Water from which dissolved O has been removed, e.g. by stripping with an inert gas, or by adequate
membrane filtration.
7.2.10.6 Pyrogenic copper
Dissolve 45 g copper(II)-sulphate pentahydrate (7.2.10.1) in 480 ml water containing 20 ml
hydrochloric acid (7.2.10.2) in a 1 000 ml beaker.
Take 15 g of zinc granules size (7.2.10.3), add 25 ml water and one drop of anionic detergent solution
(7.2.10.4) in another 1 000 ml beaker.
Stir with a magnetic stirrer at a high speed to form a slurry. Then whilst stirring at this high speed,
carefully add the copper(II)-sulphate solution drop by drop using a glass rod.
Hydrogen is liberated and elemental pyrogenic copper is precipitated (red coloured precipitate).
Stirring is continued until the hydrogen generation almost ceases. Then the precipitated copper is
allowed to settle. The supernatant water is carefully removed and the product washed with
deoxygenated water (7.2.10.5) three times, to eliminate residual salts.
Then the water is carefully replaced with 250 ml acetone (7.1.1) (whilst continuously stirring the
mixture). This operation is repeated twice more to ensure elimination of water.
Then the above procedure is repeated three times with 250 ml hexane (7.2.5.5), to ensure elimination
of the acetone.
Carefully transfer the copper with hexane into an Erlenmeyer flask and store under hexane. The flask
shall be sealed to prevent ingress of air and stored in an explosion-proof refrigerator 2 °C to 8 °C.
The shelf life of the pyrogenic copper is at least two months. The clean-up efficiency then declines. The
copper changes colour as the clean-up efficiency decreases.
7.2.11 Clean-up K using silica/silver nitrate
7.2.11.1 Silver nitrate, AgNO .
7.2.11.2 Silver nitrate/silica adsorbent
Dissolve 10 g of AgNO (7.2.11.1) in 40 ml water and add this mixture in portions to 90 g of silica
(7.2.5.1). Shake the mixture until it is homogenous and leave it for 30 min. Put the mixture into a drying
oven at (70 ± 5) °C. Within 5 h regular increase the temperature from 70 °C to 125 °C. Activate the
mixture for 15 h at 125 °C. Store the mixture in brown glass bottles.
7.3 Gas chromatographic analysis
Operating gases for gas chromatography/ECD or MS, of high purity and in accordance with the
manufacturer’s specifications.
7.4 Standards
7.4.1 General
Choose the internal standards substances whose physical and chemical properties (such as extraction
behaviour, retention time) are similar to those of the compounds to be analysed. C -PCBs should be
used as internal standards for the GC-MS method for evaluation of results. Verify the stability of the
internal standards regularly.
NOTE Certified solutions of PCB, and single solid PCB substances with certified purity are available from a
limited number of suppliers.
7.4.2 Calibration standards:
The calibration standard should contain the following compounds:
PCB28 2,4,4'-trichlorobiphenyl (CAS-RN 7012-37-5)
PCB52 2,2',5,5'-tetrachlorobiphenyl (CAS-RN 35693-99-3)
PCB101 2,2',4,5,5'-pentachlorobiphenyl (CAS-RN 37680-73-2)
PCB118 2,3',4,4',5-pentachlorobiphenyl (CAS-RN 31508-00-6)
PCB138 2,2',3,4,4',5'-hexachlorobiphenyl (CAS-RN 35065-28-2)
PCB153 2,2',4,4',5,5'-hexachlorobiphenyl (CAS-RN 35065-27-1)
PCB180 2,2',3,4,4',5,5'-heptachlorobiphenyl (CAS-RN 35065-29-3)
NOTE The numbers 28, 52, etc. correspond to the sequential numbers of chlorobiphenyls according to the
IUPAC rules for the nomenclature of organic compounds.
7.4.3 Internal and injection standards:
7.4.3.1 General
The PCB congeners to be considered as internal and injection standards are listed below. The internal
standard shall be added to the sample. For MS-detection labelled PCB congeners are advised.
When highly contaminated samples are analysed, an aliquot of the extract is often used for further
clean-up. This makes the costs of analyses caused by the use of labelled standard very high. In these
cases, it is allowed to add the internal standard in two steps. Step 1 addition of unlabelled internal
standards to the sample. Step 2 addition of labelled congeners to the aliquot of the extract used for
clean-up.
At least three congeners, covering the chromatogram shall be used as internal standard.
Other PCB not present in the sample, or C -labelled PCBs not used as internal standard, can be used
as injection standard.
NOTE Some PCB mixtures contain up to 2,5 % of PCB155.
PCB30, PCB143 and PCB207 are recommended as internal standards.
PCB198 or PCB209 are recommended as injection standards for ECD-detection because of lesser
interferences.
7.4.3.2 Labelled PCB congeners:
C -2,4,4’-trichlorobiphenyl
PCB28 12 (CAS RN 208263 76 7)
C -2,2‘,5,5‘-tetrachlorobiphenyl
PCB52 12 (CAS RN 208263 80 3)
C -2,2‘,4,5,5‘-pentachlorobiphenyl
PCB101 12 (CAS RN 104130-39-4)
C -2,3‘,4,4‘,5-pentachlorobiphenyl
PCB118 12 (CAS RN 104130-40-7)
C -2,2‘,3,4,4‘,5‘-hexachlorobiphenyl
PCB138 12 (CAS-RN 208263-66-5)
C -2,2‘,4,4‘,5,5‘-hexachlorobiphenyl
PCB153 12 (CAS RN 1855376-58-3)
C -2,2‘,3,4,4‘,5,5‘-heptachlorobiphenyl
PCB180 12
7.4.3.3 Non-labelled PCB congeners:
PCB29 2,4,5-trichlorobiphenyl (CAS-RN 15862-07-4)
PCB30 2,4,6-trichlorobiphenyl (CAS-RN 35693-92-6)
PCB143 2,2',3,4,5,6'-hexachlorobiphenyl (CAS-RN 68194-15-0)
PCB155 2,2',4,4',6,6'-hexachlorobiphenyl (CAS-RN 33979-03-2)
PCB198 2,2',3,3',4,5,5’,6,-octachlorobiphenyl (CAS-RN 68194-17-2)
PCB207 2,2',3,3',4,4',5,6,6'-nonachlorobiphenyl (CAS-RN 52663-79-3)
PCB209 2,2',3,3',4,4',5,5’,6,6'-decachlorobiphenyl (CAS-RN 2051-24-3)
7.4.3.4 PCB congeners for resolution check:
PCB28 2,4,4'-trichlorobiphenyl (CAS-RN 7012-37-5)
PCB31 2,4',5-trichlorobiphenyl (CAS-RN 16606-02-3)

7.5 Preparation of standard solutions:
7.5.1 Preparation of calibration standard solutions of PCBs:
Prepare individual concentrated primary standard solutions of about 0,4 mg/ml in n-heptane (7.1.2) by
weighing approximately 10 mg of each of the calibration standards (7.4.2) to the nearest 0,1 mg and
dissolving them in 25 ml of n-heptane.
Combine small quantities (2 ml to 10 ml) of these individual primary standard solutions into a mixed
standard solution of PCB.
Because of the dangerous nature of the substances to be used, commercially available - preferably
certified - standard solutions or mixed standard solutions are preferred. Avoid skin contact.
The working standard solutions shall be in the same solvent like the extract.
Store the primary and diluted standard solutions in a dark place at a temperature of (5 ± 3) °C. The
solutions are stable for at least one year, provided that evaporation of solvent is negligible.
Components present in mixed standard solutions should be completely separated by the gas
chromatographic columns used.
7.5.2 Preparation of internal standard solution:
Prepare a concentrated primary internal standard solution, containing at least three different
components (7.4.3), of about 0,4 mg/ml in n-heptane (7.1.2) by weighing approximately 10 mg of each
of the chosen internal standards to the nearest 0,1 mg and dissolving them in 25 ml of n-heptane.
Prepare from this a secondary internal solution with such a concentration that the added amount gives
a peak with measurable peak area or peak height in the chromatogram (at least 10 times the detection
limit).
If the two-step procedure for GC-MS is used, make two different internal standard solutions, one
containing the non-labelled compounds. At least two unlabelled congeners shall be used in the first
internal standard solution and at least three labelled congeners in the second solution.
7.5.3 Preparation of injection standard solution:
Prepare a concentrated primary injection standard solution, containing at least two different
components (7.4.3), of about 0,4 mg/ml in an appropriate solvent by weighing approximately 10 mg of
each of the chosen injection standards to the nearest 0,1 mg and dissolving them in 25 ml. Prepare from
this a secondary internal solution which such a concentration that the added amount gives a peak with
measurable peak area or peak surface in the chromatogram (at least 10 times the detection limit).
8 Apparatus
8.1 Extraction and clean-up procedures:
Usual laboratory glassware.
All glassware and material that comes into contact with the sample or extract shall be thoroughly
cleaned.
8.1.1 Sample bottles, made of glass, stainless steel or aluminium, with glass stopper or screw top and
polytetrafluoroethylene (PTFE) seal of appropriate volume.
Glass is not appropriate for sludge samples.
WARNING — For safety reasons, biologically active sludge samples shall not be stored in a sealed
container.
8.1.2 Shaking device, with horizontal movement (200 strokes to 300 strokes per min).
8.1.3 Water bath, adjustable up to 100 °C.
8.1.4 Separating funnels of appropriate volume.
8.1.5 Conical flasks of appropriate volume.
8.1.6 Soxhlet extraction apparatus, consisting of round bottom flask, e.g. 100 ml, Soxhlet extractors
and Soxhlet thimbles, e.g. 27 mm × 100 mm, vertical condensers, e.g. 300 mm, heating device.
8.1.7 Concentrator, Kuderna Danish type.
Other evaporators, e.g. a rotary evaporator, may be used if found to be equally suitable.
8.1.8 Boiling chips, glass or porcelain beads.
8.1.9 Quartz wool or silanized glass wool.
WARNING — Working with quartz wool imposes a risk to health through the release of fine quartz
particles. Inhalation of these should be prevented by using a fume cupboard and wearing a dust mask.
8.1.10 Calibrated test tubes, with a nominal capacity of 10 ml to 15 ml and ground glass stopper.
8.1.11 Chromatography tubes
Chromatography column of glass, 5 mm to 10 mm inside diameter, length e.g. 600 mm.
8.2 Gas chromatograph:
8.2.1 General:
Use a gas chromatograph equipped with a capillary column and mass spectrometric detection (MS) or
electron capture detector (ECD).
NOTE ECD, based on Ni and working with an encapsulated radioactive source, requires a licence according
to the appropriate national regulations.
Using ECD, gas chromatographs equipped with two detectors and with facilities for connecting two
capillary columns to the same injection system are very well suited for this analysis; with such
apparatus, the confirmatory analysis can be performed simultaneously.
8.2.2 Capillary columns, each comprising a 5 % phenyl-methyl silicone stationary phase coated onto
fused silica capillary column or an equivalent chemically bonded phase column. The chromatographic
peaks of PCB28 and PCB31 shall be resolved sufficiently (resolution at least 0,5) for integrating the
PCB28 peak. In general column length should be 25 m to 60 m. Internal diameter 0,18 mm to 0,32 mm
and film thickness 0,1 µm to 0,5 µm.
3)
Using ECD-detection, a second column, coated with a moderate polar phase (e.g. CP-Sil 19, OV 1701 ,
etc.), shall be used to confirm the result obtained.
NOTE The retention times for the PCB on different capillary columns are given in Annex B.
9 Sample storage and preservation
9.1 Sample storage
The samples shall be analysed as soon as possible after sampling. This applies in particular to the
examination of microbiologically active solids.
If necessary, sludge samples shall be stored according to EN ISO 5667-15.
Dried samples can be stored at room temperature in a dark place up to one month. Soil samples shall be
stored according to ISO 18512.
9.2 Sample pretreatment
Pretreat samples according to EN 16179, if not otherwise specified, and considering the specific drying
procedures as specified in Table 3 to obtain a test sample.

3) CP-Sil 19, OV 1701 are examples of suitable products available commercially. This information is given for the

convenience of users of this document and does not constitute an endorsement by CEN of these products.
Equivalent products may be used if they can be shown to lead to the same results.

Pretreatment is necessary to reduce the moisture content to enable extraction of the PCBs and to
increase the homogeneity.
Complete drying of the sample is essential if Soxhlet is used for extraction or to increase the
homogeneity.
Complete drying is also recommended if the sample shall be stored for a long period.
Table 3 — Drying techniques for samples of different matrices for subsequent analyses of PCB
Matrix Drying technique
Air drying Freeze drying Na SO No drying
2 4
(EN ISO 16720)
Sludge a
x x
x
Biowaste
x x x
(compost, mixed waste)
Soil
x x x x
(e.g. sand, clay)
a
Na2SO4 can be used for the preservation of hygroscopic dried sludge.
10 Procedure
10.1 Blank test
Perform a blank test following the applied procedure (selected extraction and clean-up procedure)
using the same amount of reagents that are used for the pretreatment, extraction, clean-up and analysis
of a sample. Analyse the blank immediately prior to analysis of the samples to demonstrate sufficient
freedom from contamination. The blank shall be less than 50 % of the lowest reporting limit.
10.2 Extraction
10.2.1 General
Depending on the test sample (matrix and moisture content), choose a suitable extraction method
(see Table 4). Extraction method 1 (10.2.2) or 3 (10.2.4) are recommended if it is important to break up
aggregates in the sample to access the PCBs. With wet samples these methods shall be applied in order
to eliminate the presence of water. If dissolving of the PCBs is the most important step (waste and
organic rich materials) and the sample is dry, extraction method 2 (10.2.3) using Soxhlet is
recommended. For sludge it has been shown that Soxhlet is applicable. In presence of plastics, use of
acetone shall be avoided, because the use of acetone leads to a high amount of co-extractives. However,
a general rule cannot be given, because samples may contain all: aggregates, organic matter and
(plastic) waste.
Other extraction procedures, e.g. ultrasonic extraction, microwave or high pressure liquid extraction
may be used provided:
— the laboratory can show that the extraction efficiency is equivalent to one of the extraction
procedures 1, 2 or 3 as described in this European Standard, or
— the sample requires another approach as shown by the laboratory and the results of the procedures
are in agreement with the performance criteria as described in 10.7.4 and 10.8.6.
NOTE For application of this European Standard for some types of waste, the addition of acetone with Soxhlet
extraction has been shown to be effective.
Extraction procedures described in this European standard are suitable to extract up to 20 g of dry
sample. If the test sample has a low density (i.e. some wastes) or the sample is homogeneous,
depending on the expected PCB content and on the homogeneity of the sample, less sample can be used.
In general the following amounts of dry sample can be used: 2 g to 10 g of sewage sludge, 2 to 20 g of
soils, 5 g to 20 g of compost or 2 g to 20 g of biowaste. The amount of sample shall be weighed with an
accuracy of at least 1 %.
Table 4 — Extraction procedures to be used for different matrices
Moisture Matrix Extraction Extraction Extraction Remark
status of solvent technique procedure
the test
sample
Dry Soil-like materials, Acetone/ Agitation Extraction
sludge, biowaste, petroleum procedure 1
compost ether (see 10.2.2)
Sludge, biowaste, Petroleum Soxhlet, Extraction
compost, ether pressurized procedure 2
liquid (see 10.2.3)
extraction
Wet Soil-like materials, Acetone/ Agitation Extraction Also applicable for
biowaste, compost petroleum procedure 1 field moist samples
ether (see 10.2.2) with dry matter
content > 75 %
Soil-like material Acet
...