Soil quality - Determination of polychlorinated biphenyls (PCB) by gas chromatography with mass selective detection (GC-MS) and gas chromatography with electron-capture detection (GC-ECD)

ISO 13876:2013 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.

Qualité du sol - Détermination des polychlorobiphényles (PCB) par chromatographie en phase gazeuse avec détection sélective de masse (GC-MS) et chromatographie en phase gazeuse avec détection par capture d'électrons (GC-ECD)

Kakovost tal - Določevanje polikloriranih bifenilov (PCB) s plinsko kromatografijo z masno selektivnim detektorjem (GC-MS) ali s plinsko kromatografijo z detektorjem z zajetjem elektronov (GC-ECD)

Ta mednarodni standard določa metodo za kvantitativno določevanje sedmih izbranih polikloriranih bifenilov (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 in PCB180) v blatu,
obdelanih bioloških odpadkih in tleh z uporabo metod GC-MS in GC-ECD (glej preglednico 2).
Meja zaznavanja je odvisna od determinant, uporabljene opreme, kakovosti kemikalij, uporabljenih
za ekstrakcijo vzorca, in očiščenja ekstrakta.
Pod pogoji, določenimi v tem mednarodnem standardu, je mogoče doseči mejo uporabe 1 μg/kg (izraženo
kot suho snov).
Blato in obdelani biološki odpadki se lahko razlikujejo glede lastnosti, predvidenih stopenj onesnaženja s polikloriranimi bifenili ter prisotnosti motečih snovi. Zaradi teh razlik ni možen opis enotnega splošnega postopka. Ta mednarodni standard vključuje tabele odločanja, ki temeljijo na lastnostih vzorca, ter postopek za ekstrakcijo in očiščenje, ki ga je treba uporabiti.

General Information

Status
Published
Public Enquiry End Date
24-Jun-2019
Publication Date
03-Sep-2019
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
30-Aug-2019
Due Date
04-Nov-2019
Completion Date
04-Sep-2019

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INTERNATIONAL ISO
STANDARD 13876
First edition
2013-11-15
Soil quality — Determination of
polychlorinated biphenyls (PCB)
by gas chromatography with mass
selective detection (GC-MS) and
gas chromatography with electron-
capture detection (GC-ECD)
Qualité du sol — Détermination des polychlorobiphényles (PCB) par
chromatographie en phase gazeuse avec détection sélective de masse
(GC-MS) et chromatographie en phase gazeuse avec détection par
capture d’électrons (GC-ECD)
Reference number
ISO 13876:2013(E)
ISO 2013
---------------------- Page: 1 ----------------------
ISO 13876:2013(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2013

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2013 – All rights reserved
---------------------- Page: 2 ----------------------
ISO 13876:2013(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 2

4 Principle ........................................................................................................................................................................................................................ 2

5 Interferences ............................................................................................................................................................................................................ 3

5.1 Interference with sampling and extraction .................................................................................................................... 3

5.2 Interference with GC .......................................................................................................................................................................... 3

6 Safety remarks ........................................................................................................................................................................................................ 3

7 Reagents ........................................................................................................................................................................................................................ 4

7.1 General ........................................................................................................................................................................................................... 4

7.2 Reagents for extraction .................................................................................................................................................................... 4

7.3 Reagents for clean-up ........................................................................................................................................................................ 4

7.4 Gas chromatographic analysis ................................................................................................................................................... 7

7.5 Standards ..................................................................................................................................................................................................... 7

7.6 Preparation of standard solutions ......................................................................................................................................... 9

8 Apparatus ..................................................................................................................................................................................................................10

9 Sample storage and preservation ....................................................................................................................................................11

9.1 Sample storage .....................................................................................................................................................................................11

9.2 Sample pretreatment ......................................................................................................................................................................11

10 Procedure..................................................................................................................................................................................................................12

10.1 Blank test ..................................................................................................................................................................................................12

10.2 Extraction .................................................................................................................................................................................................12

10.3 Concentration........................................................................................................................................................................................14

10.4 Clean-up of the extract ..................................................................................................................................................................14

10.5 Addition of the injection standard ......................................................................................................................................17

10.6 Gas chromatographic analysis (GC) ...................................................................................................................................18

10.7 Mass spectrometry (MS) .............................................................................................................................................................18

10.8 Electron capture detection (ECD) ........................................................................................................................................23

11 Performance characteristics .................................................................................................................................................................25

12 Precision ....................................................................................................................................................................................................................25

13 Test report ................................................................................................................................................................................................................25

Annex A (informative) Repeatability and reproducibility data .............................................................................................26

Annex B (informative) Examples for retention times of PCBs ................................................................................................28

Bibliography .............................................................................................................................................................................................................................29

© ISO 2013 – All rights reserved iii
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ISO 13876:2013(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the meaning of ISO specific terms and expressions related to conformity

assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers

to Trade (TBT) see the following URL: Foreword - Supplementary information

The committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 3, Chemical

methods and soil characteristics.
iv © ISO 2013 – All rights reserved
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ISO 13876:2013(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 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, 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).

The European Standard EN 16167:2012 on which this International Standard is based, 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 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 and 11 clean-up procedures are described. 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 ISO 13859). It has

been tested for ruggedness.

This International 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 International Standard is applicable and validated

Matrix Materials used for validation
Sludge Municipal sewage sludge
Biowaste Compost
© ISO 2013 – All rights reserved v
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INTERNATIONAL STANDARD ISO 13876:2013(E)
Soil quality — Determination of polychlorinated biphenyls
(PCB) by gas chromatography with mass selective
detection (GC-MS) and gas chromatography with electron-
capture detection (GC-ECD)

WARNING — Persons using this International Standard should be familiar with usual laboratory

practice. This International 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 International

Standard be carried out by suitably trained staff.
1 Scope

This International 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 International 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 37680-37-2
PCB118 2,3’,4,4’,5-pentachlorobiphenyl 31508-00-6
PCB138 2,2’,3,4,4’,5’-hexachlorobiphenyl 35056-28-2
PCB153 2,2’,4,4’,5,5’-hexachlorobiphenyl 35065-27-1
PCB180 2,2’,3,4,4’,5,5’-heptachlorobiphenyl 35065-29-3
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 International Standard, a limit of application of 1 µg/kg (expressed

as dry matter) can be achieved.

Sludge and treated biowaste can 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 International 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, 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.

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

and sediment samples
© ISO 2013 – All rights reserved 1
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ISO 13876:2013(E)

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 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method

ISO 14507, Soil quality — Pretreatment of samples for determination of organic contaminants

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

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

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

mass spectrometry
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
polychlorinated biphenyl
PCB
biphenyl substituted by one to ten chlorine atoms
[SOURCE: EN 15308:2008, 3.1]
3.2
congener

member of the same kind, class, or group of chemicals, e.g. anyone of the 209 individual PCBs

[SOURCE: EN 15308:2008, 3.2]

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

chromatographic elution.
3.3
critical pair

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

separation meets minimum quality criteria
[SOURCE: EN 15308:2008, 3.6]
4 Principle

Due to the horizontal character of this International Standard, different procedures for different steps

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

given in this International Standard. Performance criteria are described and it is the responsibility of

the laboratories applying this International Standard to show that these criteria are met. Use of spiking

standards (internal standards) allows an overall check on the efficiency of a specific combination of

modules for a specific sample. However, 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. The various compounds are separated using a capillary

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

electron capture detector (ECD) (8.2.1).
2 © ISO 2013 – All rights reserved
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ISO 13876:2013(E)

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 can 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 can interfere with the determination. These interferences

can lead to incompletely resolved signals and, depending on their magnitude, can 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 (8.2.1). 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 can interfere with the quantification of

PCB in GC-MS. In presence of mineral oil, GC-ECD can 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 can disturb the determination of the PCB with

GC-ECD.
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. 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.

© ISO 2013 – All rights reserved 3
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ISO 13876:2013(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 69 °C are allowed.
7.2.4 Anhydrous sodium sulfate, Na SO .
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
7.2.5 Distilled water, or water of equivalent quality, H O.
7.2.6 Sodium chloride, NaCl, anhydrous.
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 , basic or neutral, specific surface of 200 m /g, activity Super I

2 3
according to Brockmann.
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 The activity depends on the water content. It can be necessary to adjust the water content.

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) is heated for at least 3 h at 450 °C, cooled down in a desiccator and stored containing

magnesium perchlorate or a suitable drying agent. Before use, heat at for least 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.
4 © ISO 2013 – All rights reserved
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ISO 13876:2013(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 of elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 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

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 for 2 h at 600 °C, particle size of 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.
7.3.5 Clean-up E using silica H SO /silica NaOH
2 4

7.3.5.1 Silica, SiO , particle size of 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 Silica, treated with sulfuric acid.
Mix 56 g of silica (7.3.5.1) and 44 g of sulfuric acid (7.3.8.1).
7.3.5.3 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.3.5.4 Silica, treated with sodium hydroxide.
Mix 33 g of silica (7.3.5.1) and 17 g of sodium hydroxide (7.3.5.3).
7.3.5.5 n-hexane, C H .
6 14
7.3.6 Clean-up F using benzenesulfonic acid/sulfuric acid

7.3.6.1 3-ml silica gel column, of adsorbent mass 500 mg, particle size of 40 µm.

1) Bio-Beads is an example of a suitable product available commercially. This information is given for the

convenience of users of this International Standard and does not constitute an endorsement by ISO of this product.

Equivalent products may be used if they can be shown to lead to the same result.

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 International Standard and does not constitute

an endorsement by ISO of this product. Equivalent products may be used if they can be shown to lead to the same

results.
© ISO 2013 – All rights reserved 5
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ISO 13876:2013(E)

7.3.6.2 3-ml benzenesulfonic acid column, of adsorbent mass 500 mg, particle size of 40 µm.

7.3.7 Clean-up G using DMF/hexane partitioning
7.3.7.1 Dimethylformamide(DMF), C H NO.
3 7
7.3.8 Clean-up H using concentrated sulfuric acid
7.3.8.1 Sulfuric acid, H SO of purity 96 % to 98 % (mass fraction).
2 4
7.3.9 Clean-up I using TBA sulfite reagent
7.3.9.1 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
NOTE 25 g of sodium sulfite should be sufficient for 100 ml of solution.
7.3.9.2 2-Propanol, C H O.
3 8
7.3.9.3 Sodium sulfite, Na SO .
2 3
7.3.10 Clean-up J using pyrogenic copper

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

7.3.10.1 Copper(II)-sulfate pentahydrate, CuSO · 5 H O.
4 2
7.3.10.2 Hydrochloric acid, c(HCl) = 2 mol/l.
7.3.10.3 Zinc granules, Zn, particle size of 0,3 mm to 1,4 mm.

7.3.10.4 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
NOTE Other commercially available detergents can also be suitable.
7.3.10.5 Deoxygenated water.
7.3.10.6 Pyrogenic copper.

Dissolve 45 g of copper(II)-sulfate pentahydrate (7.3.10.1) in 480 ml of water containing 20 ml of

hydrochloric acid (7.3.10.2) in a 1 000-ml beaker.

Take 15 g of zinc granules size (7.3.10.3), add 25 ml of water and one drop of anionic detergent solution

(7.3.10.4) in another 1 000-ml beaker.

Stir with a magnetic stirrer at a high speed to form a slurry. Then while stirring at this high speed,

carefully add the copper(II)-sulfate solution drop by drop using a glass rod.

Hydrogen is liberated and elemental pyrogenic copper is precipitated (red 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

(7.3.10.5) three times to eliminate residual salts.
6 © ISO 2013 – All rights reserved
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ISO 13876:2013(E)

Then, the water is carefully replaced with 250 ml of acetone (7.2.1) (while 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 of hexane (7.3.5.5) 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 at 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.
7.3.11 Clean-up K using silica/silver nitrate
7.3.11.1 Silver nitrate, AgNO .
7.3.11.2 Silver nitrate/silica adsorbent.

Dissolve 10 g of AgNO (7.3.11.1) in 40 ml of water and add this mixture in portions to 90 g of silica

(7.3.5.1). 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, regularly 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.
7.4 Gas chromatographic analysis

Operating gases for gas chromatography/ECD or MS are of high purity and in accordance with the

manufacturer’s specifications.
7.5 Standards
7.5.1 General

Choose the internal standards substances whose physical and chemical properties (such as extraction

behaviour, r
...

SLOVENSKI STANDARD
SIST ISO 13876:2019
01-oktober-2019

Kakovost tal - Določevanje polikloriranih bifenilov (PCB) s plinsko kromatografijo

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

Soil quality - Determination of polychlorinated biphenyls (PCB) by gas chromatography

with mass selective detection (GC-MS) and gas chromatography with electron-capture

detection (GC-ECD)

Qualité du sol - Détermination des polychlorobiphényles (PCB) par chromatographie en

phase gazeuse avec détection sélective de masse (GC-MS) et chromatographie en
phase gazeuse avec détection par capture d'électrons (GC-ECD)
Ta slovenski standard je istoveten z: ISO 13876:2013
ICS:
13.080.10 Kemijske značilnosti tal Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST ISO 13876:2019 en

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

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SIST ISO 13876:2019
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SIST ISO 13876:2019
INTERNATIONAL ISO
STANDARD 13876
First edition
2013-11-15
Soil quality — Determination of
polychlorinated biphenyls (PCB)
by gas chromatography with mass
selective detection (GC-MS) and
gas chromatography with electron-
capture detection (GC-ECD)
Qualité du sol — Détermination des polychlorobiphényles (PCB) par
chromatographie en phase gazeuse avec détection sélective de masse
(GC-MS) et chromatographie en phase gazeuse avec détection par
capture d’électrons (GC-ECD)
Reference number
ISO 13876:2013(E)
ISO 2013
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SIST ISO 13876:2019
ISO 13876:2013(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2013

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2013 – All rights reserved
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SIST ISO 13876:2019
ISO 13876:2013(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 2

4 Principle ........................................................................................................................................................................................................................ 2

5 Interferences ............................................................................................................................................................................................................ 3

5.1 Interference with sampling and extraction .................................................................................................................... 3

5.2 Interference with GC .......................................................................................................................................................................... 3

6 Safety remarks ........................................................................................................................................................................................................ 3

7 Reagents ........................................................................................................................................................................................................................ 4

7.1 General ........................................................................................................................................................................................................... 4

7.2 Reagents for extraction .................................................................................................................................................................... 4

7.3 Reagents for clean-up ........................................................................................................................................................................ 4

7.4 Gas chromatographic analysis ................................................................................................................................................... 7

7.5 Standards ..................................................................................................................................................................................................... 7

7.6 Preparation of standard solutions ......................................................................................................................................... 9

8 Apparatus ..................................................................................................................................................................................................................10

9 Sample storage and preservation ....................................................................................................................................................11

9.1 Sample storage .....................................................................................................................................................................................11

9.2 Sample pretreatment ......................................................................................................................................................................11

10 Procedure..................................................................................................................................................................................................................12

10.1 Blank test ..................................................................................................................................................................................................12

10.2 Extraction .................................................................................................................................................................................................12

10.3 Concentration........................................................................................................................................................................................14

10.4 Clean-up of the extract ..................................................................................................................................................................14

10.5 Addition of the injection standard ......................................................................................................................................17

10.6 Gas chromatographic analysis (GC) ...................................................................................................................................18

10.7 Mass spectrometry (MS) .............................................................................................................................................................18

10.8 Electron capture detection (ECD) ........................................................................................................................................23

11 Performance characteristics .................................................................................................................................................................25

12 Precision ....................................................................................................................................................................................................................25

13 Test report ................................................................................................................................................................................................................25

Annex A (informative) Repeatability and reproducibility data .............................................................................................26

Annex B (informative) Examples for retention times of PCBs ................................................................................................28

Bibliography .............................................................................................................................................................................................................................29

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SIST ISO 13876:2019
ISO 13876:2013(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the meaning of ISO specific terms and expressions related to conformity

assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers

to Trade (TBT) see the following URL: Foreword - Supplementary information

The committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 3, Chemical

methods and soil characteristics.
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SIST ISO 13876:2019
ISO 13876:2013(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 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, 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).

The European Standard EN 16167:2012 on which this International Standard is based, 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 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 and 11 clean-up procedures are described. 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 ISO 13859). It has

been tested for ruggedness.

This International 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 International Standard is applicable and validated

Matrix Materials used for validation
Sludge Municipal sewage sludge
Biowaste Compost
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SIST ISO 13876:2019
INTERNATIONAL STANDARD ISO 13876:2013(E)
Soil quality — Determination of polychlorinated biphenyls
(PCB) by gas chromatography with mass selective
detection (GC-MS) and gas chromatography with electron-
capture detection (GC-ECD)

WARNING — Persons using this International Standard should be familiar with usual laboratory

practice. This International 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 International

Standard be carried out by suitably trained staff.
1 Scope

This International 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 International 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 37680-37-2
PCB118 2,3’,4,4’,5-pentachlorobiphenyl 31508-00-6
PCB138 2,2’,3,4,4’,5’-hexachlorobiphenyl 35056-28-2
PCB153 2,2’,4,4’,5,5’-hexachlorobiphenyl 35065-27-1
PCB180 2,2’,3,4,4’,5,5’-heptachlorobiphenyl 35065-29-3
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 International Standard, a limit of application of 1 µg/kg (expressed

as dry matter) can be achieved.

Sludge and treated biowaste can 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 International 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, 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.

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

and sediment samples
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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 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method

ISO 14507, Soil quality — Pretreatment of samples for determination of organic contaminants

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

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

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

mass spectrometry
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
polychlorinated biphenyl
PCB
biphenyl substituted by one to ten chlorine atoms
[SOURCE: EN 15308:2008, 3.1]
3.2
congener

member of the same kind, class, or group of chemicals, e.g. anyone of the 209 individual PCBs

[SOURCE: EN 15308:2008, 3.2]

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

chromatographic elution.
3.3
critical pair

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

separation meets minimum quality criteria
[SOURCE: EN 15308:2008, 3.6]
4 Principle

Due to the horizontal character of this International Standard, different procedures for different steps

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

given in this International Standard. Performance criteria are described and it is the responsibility of

the laboratories applying this International Standard to show that these criteria are met. Use of spiking

standards (internal standards) allows an overall check on the efficiency of a specific combination of

modules for a specific sample. However, 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. The various compounds are separated using a capillary

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

electron capture detector (ECD) (8.2.1).
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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 can 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 can interfere with the determination. These interferences

can lead to incompletely resolved signals and, depending on their magnitude, can 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 (8.2.1). 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 can interfere with the quantification of

PCB in GC-MS. In presence of mineral oil, GC-ECD can 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 can disturb the determination of the PCB with

GC-ECD.
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. 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.

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ISO 13876:2013(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 69 °C are allowed.
7.2.4 Anhydrous sodium sulfate, Na SO .
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
7.2.5 Distilled water, or water of equivalent quality, H O.
7.2.6 Sodium chloride, NaCl, anhydrous.
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 , basic or neutral, specific surface of 200 m /g, activity Super I

2 3
according to Brockmann.
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 The activity depends on the water content. It can be necessary to adjust the water content.

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) is heated for at least 3 h at 450 °C, cooled down in a desiccator and stored containing

magnesium perchlorate or a suitable drying agent. Before use, heat at for least 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.
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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 of elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 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

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 for 2 h at 600 °C, particle size of 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.
7.3.5 Clean-up E using silica H SO /silica NaOH
2 4

7.3.5.1 Silica, SiO , particle size of 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 Silica, treated with sulfuric acid.
Mix 56 g of silica (7.3.5.1) and 44 g of sulfuric acid (7.3.8.1).
7.3.5.3 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.3.5.4 Silica, treated with sodium hydroxide.
Mix 33 g of silica (7.3.5.1) and 17 g of sodium hydroxide (7.3.5.3).
7.3.5.5 n-hexane, C H .
6 14
7.3.6 Clean-up F using benzenesulfonic acid/sulfuric acid

7.3.6.1 3-ml silica gel column, of adsorbent mass 500 mg, particle size of 40 µm.

1) Bio-Beads is an example of a suitable product available commercially. This information is given for the

convenience of users of this International Standard and does not constitute an endorsement by ISO of this product.

Equivalent products may be used if they can be shown to lead to the same result.

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 International Standard and does not constitute

an endorsement by ISO of this product. Equivalent products may be used if they can be shown to lead to the same

results.
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SIST ISO 13876:2019
ISO 13876:2013(E)

7.3.6.2 3-ml benzenesulfonic acid column, of adsorbent mass 500 mg, particle size of 40 µm.

7.3.7 Clean-up G using DMF/hexane partitioning
7.3.7.1 Dimethylformamide(DMF), C H NO.
3 7
7.3.8 Clean-up H using concentrated sulfuric acid
7.3.8.1 Sulfuric acid, H SO of purity 96 % to 98 % (mass fraction).
2 4
7.3.9 Clean-up I using TBA sulfite reagent
7.3.9.1 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
NOTE 25 g of sodium sulfite should be sufficient for 100 ml of solution.
7.3.9.2 2-Propanol, C H O.
3 8
7.3.9.3 Sodium sulfite, Na SO .
2 3
7.3.10 Clean-up J using pyrogenic copper

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

7.3.10.1 Copper(II)-sulfate pentahydrate, CuSO · 5 H O.
4 2
7.3.10.2 Hydrochloric acid, c(HCl) = 2 mol/l.
7.3.10.3 Zinc granules, Zn, particle size of 0,3 mm to 1,4 mm.

7.3.10.4 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
NOTE Other commercially available detergents can also be suitable.
7.3.10.5 Deoxygenated water.
7.3.10.6 Pyrogenic copper.

Dissolve 45 g of copper(II)-sulfate pentahydrate (7.3.10.1) in 480 ml of water containing 20 ml of

hydrochloric acid (7.3.10.2) in a 1 000-ml beaker.

Take 15 g of zinc granules size (7.3.10.3), add 25 ml of water and one drop of anionic detergent solution

(7.3.10.4) in another 1 000-ml beaker.

Stir with a magnetic stirrer at a high speed to form a slurry. Then while stirring at this high speed,

carefully add the copper(II)-sulfate solution drop by drop using a glass rod.

Hydrogen is liberated and elemental pyrogenic copper is precipitated (red 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

(7.3.10.5) three times to eliminate residual salts.
6 © ISO 2013 – All rights reserved
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SIST ISO 13876:2019
ISO 13876:2013(E)
Then, the water is ca
...

SLOVENSKI STANDARD
oSIST ISO 13876:2019
01-junij-2019

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Soil quality - Determination of polychlorinated biphenyls (PCB) by gas chromatography

with mass selective detection (GC-MS) and gas chromatography with electron-capture

detection (GC-ECD)

Qualité du sol - Détermination des polychlorobiphényles (PCB) par chromatographie en

phase gazeuse avec détection sélective de masse (GC-MS) et chromatographie en
phase gazeuse avec détection par capture d'électrons (GC-ECD)
Ta slovenski standard je istoveten z: ISO 13876:2013
ICS:
13.080.10 .HPLMVNH]QDþLOQRVWLWDO Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
oSIST ISO 13876:2019 en

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

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oSIST ISO 13876:2019
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oSIST ISO 13876:2019
INTERNATIONAL ISO
STANDARD 13876
First edition
2013-11-15
Soil quality — Determination of
polychlorinated biphenyls (PCB)
by gas chromatography with mass
selective detection (GC-MS) and
gas chromatography with electron-
capture detection (GC-ECD)
Qualité du sol — Détermination des polychlorobiphényles (PCB) par
chromatographie en phase gazeuse avec détection sélective de masse
(GC-MS) et chromatographie en phase gazeuse avec détection par
capture d’électrons (GC-ECD)
Reference number
ISO 13876:2013(E)
ISO 2013
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oSIST ISO 13876:2019
ISO 13876:2013(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2013

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2013 – All rights reserved
---------------------- Page: 4 ----------------------
oSIST ISO 13876:2019
ISO 13876:2013(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 2

4 Principle ........................................................................................................................................................................................................................ 2

5 Interferences ............................................................................................................................................................................................................ 3

5.1 Interference with sampling and extraction .................................................................................................................... 3

5.2 Interference with GC .......................................................................................................................................................................... 3

6 Safety remarks ........................................................................................................................................................................................................ 3

7 Reagents ........................................................................................................................................................................................................................ 4

7.1 General ........................................................................................................................................................................................................... 4

7.2 Reagents for extraction .................................................................................................................................................................... 4

7.3 Reagents for clean-up ........................................................................................................................................................................ 4

7.4 Gas chromatographic analysis ................................................................................................................................................... 7

7.5 Standards ..................................................................................................................................................................................................... 7

7.6 Preparation of standard solutions ......................................................................................................................................... 9

8 Apparatus ..................................................................................................................................................................................................................10

9 Sample storage and preservation ....................................................................................................................................................11

9.1 Sample storage .....................................................................................................................................................................................11

9.2 Sample pretreatment ......................................................................................................................................................................11

10 Procedure..................................................................................................................................................................................................................12

10.1 Blank test ..................................................................................................................................................................................................12

10.2 Extraction .................................................................................................................................................................................................12

10.3 Concentration........................................................................................................................................................................................14

10.4 Clean-up of the extract ..................................................................................................................................................................14

10.5 Addition of the injection standard ......................................................................................................................................17

10.6 Gas chromatographic analysis (GC) ...................................................................................................................................18

10.7 Mass spectrometry (MS) .............................................................................................................................................................18

10.8 Electron capture detection (ECD) ........................................................................................................................................23

11 Performance characteristics .................................................................................................................................................................25

12 Precision ....................................................................................................................................................................................................................25

13 Test report ................................................................................................................................................................................................................25

Annex A (informative) Repeatability and reproducibility data .............................................................................................26

Annex B (informative) Examples for retention times of PCBs ................................................................................................28

Bibliography .............................................................................................................................................................................................................................29

© ISO 2013 – All rights reserved iii
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oSIST ISO 13876:2019
ISO 13876:2013(E)
Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards

bodies (ISO member bodies). The work of preparing International Standards is normally carried out

through ISO technical committees. Each member body interested in a subject for which a technical

committee has been established has the right to be represented on that committee. International

organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.

ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of

electrotechnical standardization.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www.iso.org/patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the meaning of ISO specific terms and expressions related to conformity

assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers

to Trade (TBT) see the following URL: Foreword - Supplementary information

The committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 3, Chemical

methods and soil characteristics.
iv © ISO 2013 – All rights reserved
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oSIST ISO 13876:2019
ISO 13876:2013(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 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, 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).

The European Standard EN 16167:2012 on which this International Standard is based, 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 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 and 11 clean-up procedures are described. 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 ISO 13859). It has

been tested for ruggedness.

This International 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 International Standard is applicable and validated

Matrix Materials used for validation
Sludge Municipal sewage sludge
Biowaste Compost
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INTERNATIONAL STANDARD ISO 13876:2013(E)
Soil quality — Determination of polychlorinated biphenyls
(PCB) by gas chromatography with mass selective
detection (GC-MS) and gas chromatography with electron-
capture detection (GC-ECD)

WARNING — Persons using this International Standard should be familiar with usual laboratory

practice. This International 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 International

Standard be carried out by suitably trained staff.
1 Scope

This International 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 International 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 37680-37-2
PCB118 2,3’,4,4’,5-pentachlorobiphenyl 31508-00-6
PCB138 2,2’,3,4,4’,5’-hexachlorobiphenyl 35056-28-2
PCB153 2,2’,4,4’,5,5’-hexachlorobiphenyl 35065-27-1
PCB180 2,2’,3,4,4’,5,5’-heptachlorobiphenyl 35065-29-3
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 International Standard, a limit of application of 1 µg/kg (expressed

as dry matter) can be achieved.

Sludge and treated biowaste can 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 International 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, 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.

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

and sediment samples
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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 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method

ISO 14507, Soil quality — Pretreatment of samples for determination of organic contaminants

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

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

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

mass spectrometry
3 Terms a nd definiti ons
For the purposes of this document, the following terms and definitions apply.
3.1
polychlorinated biphenyl
PCB
biphenyl substituted by one to ten chlorine atoms
[SOURCE: EN 15308:2008, 3.1]
3.2
congener

member of the same kind, class, or group of chemicals, e.g. anyone of the 209 individual PCBs

[SOURCE: EN 15308:2008, 3.2]

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

chromatographic elution.
3.3
critical pair

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

separation meets minimum quality criteria
[SOURCE: EN 15308:2008, 3.6]
4 Principle

Due to the horizontal character of this International Standard, different procedures for different steps

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

given in this International Standard. Performance criteria are described and it is the responsibility of

the laboratories applying this International Standard to show that these criteria are met. Use of spiking

standards (internal standards) allows an overall check on the efficiency of a specific combination of

modules for a specific sample. However, 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. The various compounds are separated using a capillary

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

electron capture detector (ECD) (8.2.1).
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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 can 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 can interfere with the determination. These interferences

can lead to incompletely resolved signals and, depending on their magnitude, can 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 (8.2.1). 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 can interfere with the quantification of

PCB in GC-MS. In presence of mineral oil, GC-ECD can 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 can disturb the determination of the PCB with

GC-ECD.
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. 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.

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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 69 °C are allowed.
7.2.4 Anhydrous sodium sulfate, Na SO .
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
7.2.5 Distilled water, or water of equivalent quality, H O.
7.2.6 Sodium chloride, NaCl, anhydrous.
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 , basic or neutral, specific surface of 200 m /g, activity Super I

2 3
according to Brockmann.
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 The activity depends on the water content. It can be necessary to adjust the water content.

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) is heated for at least 3 h at 450 °C, cooled down in a desiccator and stored containing

magnesium perchlorate or a suitable drying agent. Before use, heat at for least 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.
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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 of elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 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

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 for 2 h at 600 °C, particle size of 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.
7.3.5 Clean-up E using silica H SO /silica NaOH
2 4

7.3.5.1 Silica, SiO , particle size of 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 Silica, treated with sulfuric acid.
Mix 56 g of silica (7.3.5.1) and 44 g of sulfuric acid (7.3.8.1).
7.3.5.3 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.3.5.4 Silica, treated with sodium hydroxide.
Mix 33 g of silica (7.3.5.1) and 17 g of sodium hydroxide (7.3.5.3).
7.3.5.5 n-hexane, C H .
6 14
7.3.6 Clean-up F using benzenesulfonic acid/sulfuric acid

7.3.6.1 3-ml silica gel column, of adsorbent mass 500 mg, particle size of 40 µm.

1) Bio-Beads is an example of a suitable product available commercially. This information is given for the

convenience of users of this International Standard and does not constitute an endorsement by ISO of this product.

Equivalent products may be used if they can be shown to lead to the same result.

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 International Standard and does not constitute

an endorsement by ISO of this product. Equivalent products may be used if they can be shown to lead to the same

results.
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7.3.6.2 3-ml benzenesulfonic acid column, of adsorbent mass 500 mg, particle size of 40 µm.

7.3.7 Clean-up G using DMF/hexane partitioning
7.3.7.1 Dimethylformamide(DMF), C H NO.
3 7
7.3.8 Clean-up H using concentrated sulfuric acid
7.3.8.1 Sulfuric acid, H SO of purity 96 % to 98 % (mass fraction).
2 4
7.3.9 Clean-up I using TBA sulfite reagent
7.3.9.1 Tetrabutylammo nium 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
NOTE 25 g of sodium sulfite should be sufficient for 100 ml of solution.
7.3.9.2 2-Propanol, C H O.
3 8
7.3.9.3 S odium sulfite, Na SO .
2 3
7.3.10 Clean-up J using pyrogenic copper

WA R N I NG — P y r ogen ic copper i s s pont a ne ou sly i n f l a m m able . Su it able pr e c aut ion s sh a l l be t a ken .

7.3.10.1 Copper(II)-sulfate pentahydrate, CuSO · 5 H O.
4 2
7.3.10.2 Hydrochloric acid, c(HCl) = 2 mol/l.
7.3.10.3 Zinc granules, Zn, particle size of 0,3 mm to 1,4 mm.

7.3.10.4 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
NOTE Other commercially available detergents can also be suitable.
7.3.10.5 Deoxygenated water.
7.3.10.6 Pyrogenic copper.

Dissolve 45 g of copper(II)-sulfate pentahydrate (7.3.10.1) in 480 ml of water containing 20 ml of

hydrochloric acid (7.3.10.2) in a 1 000-ml beaker.

Take 15 g of zinc granules size (7.3.10.3), add 25 ml of water and one drop of anionic detergent solution

(7.3.10.4) in another 1 000-ml beaker.

Stir with a magnetic stirrer at a high speed to form a slurry. Then while stirring at this high speed,

carefully add the copper(II)-sulfate solution drop by drop using a glass rod.

Hydrogen is liberated and elemental pyrogenic copper is precipitated (red 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

(7.3.10.5) three times to eliminate residual salts.
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