Animal feeding stuffs: Methods of sampling and analysis - Determination of OCPs and PCBs by GC/MS

This document specifies a gas chromatographic mass spectrometric (GC/MS) method for the determination of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in animal feeding stuffs and oil.
The method is applicable to animal feeding stuffs consisting of less than 20 % by mass and oil/fatty samples containing residues of one or more of the following OCPs and PCBs and some of their isomers and degradation products:
   aldrin;
   dieldrin;
   chlordane, as the sum of chlordane isomers and oxychlordane;
   dichlorodiphenyltrichloroethane (DDT), as the sum of isomers op'-DDT, pp'-DDT, pp'-TDE (pp'-DDD), and pp'-DDE;
   endosulfan, as the sum of α-/β-isomers and endosulfan-sulphate;
   endrin, as the sum of endrin and delta-keto-endrin;
   heptachlor, as the sum of heptachlor and heptachlor epoxide;
   hexachlorobenzene (HCB);
   hexachlorocyclohexane isomers α-HCH (α-BHC), β-HCH (β-BHC), γ-HCH (γ-BHC or lindane);
   photo heptachlor;
   cis- and trans-nonachlor;
   non dioxin-like PCBs (ndl-PCBs), as the sum of PCB 28, 52, 101, 138, 153 and 180.
The method has been fully validated by a collaborative trial for the substances and corresponding ranges (ng/g) noted in Table 1.
The method has not been fully validated for oxychlordane, endrin ketone, cis- and trans-nonachlor and photo heptachlor in all matrices.
The method is not applicable to chlorocamphene (toxaphene), a complex mixture of polychlorinated camphenes. Chlorocamphene has a very distinctive chromatographic profile and is easily recognisable by GC/ECD. Positive identification of the toxaphene isomers can be performed by negative chemical ionisation mass spectrometry (NCI-MS), electron impact tandem mass spectrometry (EI MS × MS) or electron impact high resolution mass spectrometry (EI-HRMS), which is not within the scope of this method.
A limit of quantification (LOQ) for the mentioned organochlorine pesticides of 5 ng/g should normally be obtained. However, 10 ng/g applies for heptachlor aldrin, endrin, dieldrin, and endosulfan (α-, β- and sulphate). For the ndl-PCBs an LOQ of 0,5 to 1,0 ng/g should be obtained. The LOQs mentioned apply to the individual compounds (i.e. not the sum of two or more compounds). Individual laboratories are responsible for ensuring that the equipment that they used will achieve these LOQs. On customers' demand the standard may be applied to solely the analysis of PCBs or OCPs.

Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von OCP und PCB mittels GC-MS

Dieses Dokument legt ein gaschromatographisches massenspektrometisches Verfahren (GC-MS) für die Bestimmung von Organochlorpestiziden (OCP) und polychlorierten Biphenylen (PCB) in Futtermitteln und Öl fest.
Das Verfahren ist auf Futtermittel mit einem Massenanteil von weniger als 20 % und für öl /fetthaltige Proben anwendbar, die Rückstände von einem oder mehreren der folgenden OCP und PCB und einige von deren Isomeren und Abbauprodukten enthalten:
   Aldrin;
   Dieldrin;
   Chlordan, als die Summe von Chlordanisomeren und Oxychlordan;
   Dichlordiphenyltrichlorethan (DDT), als die Summe der Isomere von op’ DDT, pp’ DDT, pp’ TDE (pp’ DDD) und pp’ DDE;
   Endosulfan, als die Summe von α /β-Isomeren und Endosulfan-Sulfat;
   Endrin, als die Summe von Endrin und Delta-Keto-Endrin;
   Heptachlor, als die Summe von Heptachlor und Heptachlorepoxid;
   Hexachlorbenzol (HCB);
   Hexachlorcyclohexan-Isomere α-HCH (α BHC), β HCH (β BHC), γ HCH (γ-BHC oder Lindan);
   Photoheptachlor;
   cis- und trans-Nonachlor;
   nicht-dioxinähnliche PCB (ndl-PCB), als Summe von PCB 28, 52, 101, 138, 153 und 180.
Das Verfahren wurde vollständig durch einen Ringversuch für die in Tabelle 1 aufgeführten Substanzen und entsprechenden Bereiche (ng/g) validiert.
Das Verfahren wurde für Oxychlordan, Endrin-Keton, cis- und trans-Nonachlor und Photoheptachlor in allen Matrices nicht vollständig validiert.
Das Verfahren ist nicht anwendbar auf Chlorcamphen (Toxaphen), eine komplexe Mischung von polychlorierten Champhenen. Chlorocamphen hat ein sehr ausgeprägtes chromatographisches Profil und ist leicht erkennbar durch GC/ECD. Eine positive Identifizierung der Toxaphen-Isomere kann mit einer Massenspektrometrie mit negativen Ionen bei chemischer Ionisation (NCI-MS), einer Elektronenstoß-Tandem-Massenspektrometrie (EI-MS × MS) oder einer hochauflösenden Elektronenstoß-Massenspektro-metrie (EI-HRMS) durchgeführt werden, die nicht im Rahmen des Anwendungsbereichs dieses Verfahrens sind.
In der Regel sollte für die betreffenden Organochlorpestizide eine Bestimmungsgrenze (LOQ) von 5 ng/g erreicht werden. Jedoch gilt bei Heptachlor, Aldrin, Endrin, Dieldrin und Endosulfan (α , β  und Sulfat) eine Bestimmungsgrenze von 10 ng/g. Für die ndl-PCB sollte eine Bestimmungsgrenze von 0,5 ng/g bis 1,0 ng/g erreicht werden. Die erwähnten Bestimmungsgrenzen gelten für die einzelnen Verbindungen (d. h. nicht die Summe von zwei oder mehr Verbindungen). Die einzelnen Laboratorien sind dafür verantwortlich, dass die von ihnen verwendeten Geräte diese Bestimmungsgrenzen erreichen. Auf Kundenwunsch darf die Norm nur auf die Analyse von PCB oder OCP angewendet werden.

Aliments des animaux - Méthodes d’échantillonnage et d’analyse - Détermination des pesticides organochlorés (POC) et des polychlorobiphényles (PCB) par GC/MS

Le présent document décrit une méthode de chromatographie en phase gazeuse couplée à la spectrométrie de masse (GC-MS) pour la détermination des pesticides organochlorés (POC) et des polychlorobiphényles (PCB) dans les aliments pour animaux et les huiles.
La méthode est applicable aux aliments pour animaux constitués de moins de 20 % en masse et aux échantillons d’huile/de graisses contenant des résidus d’un ou de plusieurs des POC et PCB suivants et de certains de leurs isomères et produits de dégradation :
 aldrine ;
 dieldrine ;
 chlordane, correspondant { la somme des isomères de chlordane et d’oxychlordane ;
 dichlorodiphényltrichloréthane (DDT), correspondant { la somme des isomères de op’-DDT, pp’-DDT, pp’-TDE (pp’-DDD) et pp’-DDE ;
 endosulfan, correspondant à la somme des isomères α-/β et de l’endosulfan sulfate ;
 endrine, correspondant { la somme de l’endrine et du delta-keto-endrine ;
 heptachlore, correspondant { la somme de l’heptachlore et de l’heptachlore époxyde ;
 hexachlorobenzène (HCB) ;
 isomères des hexachlorocyclohexanes α-HCH (α-BHC), β-HCH (β-BHC), γ-HCH (γ-BHC ou lindane) ;
 photoheptachlore ;
 cis-nonachlore et trans-nonachlore ;
 PCB qui ne sont pas de type dioxine (NDL-PCB), correspondant à la somme des PCB 28, 52, 101, 138, 153 et 180.
La méthode a été intégralement validée par un essai interlaboratoires au regard des substances et des plages de concentrations correspondantes (ng/g) notées dans le Tableau 1.
La méthode n’a pas été intégralement validée pour l’oxychlordane, l’endrine-cétone, le cis-nonachlore, le trans-nonachlore et le photoheptachlore dans toutes les matrices.
La méthode ne s’applique pas au camphéchlore (toxaphène), mélange complexe de camphènes polychlorés. Le camphéchlore possède un profil chromatographique très particulier et est facilement identifiable par chromatographie en phase gazeuse avec détecteur à capture d’électrons (GC/ECD). L’identification positive des isomères du toxaphène peut être effectuée par spectrométrie de masse par ionisation chimique négative (NCI-MS), spectrométrie de masse en tandem par impact d’électrons (EI MS/MS) ou par spectrométrie de masse haute résolution par impact d’électrons (EI-HRMS), qui ne relèvent pas du domaine d’application de la présente méthode.
Il convient généralement d’obtenir une limite de quantification (LQ) des pesticides organochlorés mentionnés de 5 ng/g. Cependant, 10 ng/g s’applique à l’heptachlore, l’aldrine, l’endrine, le dieldrine et l’endosulfan (α-, β- et sulfate). Pour les NDL-PCB, il convient d’obtenir une LQ de 0,5 ng/g à 1,0 ng/g. Les LQ mentionnées s’appliquent aux composés individuels (c’est-à-dire non applicables à la somme de deux composés ou plus). Il incombe à chaque laboratoire de garantir que les appareils qu’ils utilisent atteignent ces LQ. Sur demande des clients, la norme peut être appliquée uniquement à l’analyse des PCB ou des POC.

Krma: metode vzorčenja in analize - Določevanje OCP in PCB z GC/MS

General Information

Status
Published
Public Enquiry End Date
24-Oct-2018
Publication Date
01-Apr-2020
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
31-Mar-2020
Due Date
05-Jun-2020
Completion Date
02-Apr-2020

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SLOVENSKI STANDARD
SIST EN 15741:2020
01-maj-2020
Nadomešča:
SIST EN 15741:2009
Krma: metode vzorčenja in analize - Določevanje OCP in PCB z GC/MS

Animal feeding stuffs: Methods of sampling and analysis - Determination of OCPs and

PCBs by GC/MS
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von OCP und
PCB mittels GC-MS

Aliments des animaux - Méthodes d’échantillonnage et d’analyse - Détermination des

pesticides organochlorés (POC) et des polychlorobiphényles (PCB) par GC/MS
Ta slovenski standard je istoveten z: EN 15741:2020
ICS:
65.120 Krmila Animal feeding stuffs
SIST EN 15741:2020 en,fr,de

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

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SIST EN 15741:2020
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SIST EN 15741:2020
EN 15741
EUROPEAN STANDARD
NORME EUROPÉENNE
March 2020
EUROPÄISCHE NORM
ICS 65.120 Supersedes EN 15741:2009
English Version
Animal feeding stuffs: Methods of sampling and analysis -
Determination of OCPs and PCBs by GC/MS

Aliments des animaux : Méthodes d'échantillonnage et Futtermittel: Probenahme- und

d'analyse - Détermination des pesticides organochlorés Untersuchungsverfahren - Bestimmung von OCP und

(POC) et des polychlorobiphényles (PCB) par GC/MS PCB mittels GC/MS
This European Standard was approved by CEN on 6 January 2020.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this

European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references

concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN

member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by

translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management

Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,

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

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

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

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 15741:2020 E

worldwide for CEN national Members.
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SIST EN 15741:2020
EN 15741:2020 (E)
Contents Page

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

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

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

2 Normative references .................................................................................................................................... 7

3 Terms and definitions ................................................................................................................................... 7

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

5 Reagents and materials ................................................................................................................................. 8

6 Apparatus ........................................................................................................................................................ 14

7 Sampling .......................................................................................................................................................... 18

8 Preparation of test sample ....................................................................................................................... 19

9 Procedure........................................................................................................................................................ 19

9.1 General ............................................................................................................................................................. 19

9.2 Test portions of animal feed stuff and oil samples .......................................................................... 19

9.3 Extraction of the feed test portion ......................................................................................................... 20

9.4 Clean-up ........................................................................................................................................................... 20

9.5 Gas chromatography ................................................................................................................................... 21

10 Calculation and expression of results ................................................................................................... 21

10.1 General ............................................................................................................................................................. 21

10.2 Calibration criteria ...................................................................................................................................... 22

10.3 Identification and confirmation.............................................................................................................. 22

10.4 Calculation ...................................................................................................................................................... 22

10.5 Recovery .......................................................................................................................................................... 24

11 Precision .......................................................................................................................................................... 25

11.1 Interlaboratory test ..................................................................................................................................... 25

11.2 Repeatability and precision within participating laboratories .................................................. 25

11.3 Reproducibility and precision between participating laboratories .......................................... 25

12 Test report ...................................................................................................................................................... 25

13 Important considerations to this method ........................................................................................... 25

13.1 Alternative extraction techniques ......................................................................................................... 25

13.2 Different amounts of silica ........................................................................................................................ 25

13.3 Different clean-up step ............................................................................................................................... 25

13.4 Different internal standard ...................................................................................................................... 25

13.5 Optimal GC-MS (MS) settings .................................................................................................................... 26

Annex A (informative) Description of PTV injection system (in case this is available) .................. 27

Annex B (informative) Collaborative trial........................................................................................................ 28

Bibilography ................................................................................................................................................................. 48

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SIST EN 15741:2020
EN 15741:2020 (E)
European foreword

This document (EN 15741:2020) has been prepared by Technical Committee CEN/TC 327 “Animal

feeding stuffs: Methods of sampling and analysis”, 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 September 2020, and conflicting national standards shall

be withdrawn at the latest by September 2020.

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 15741:2009.

In comparison with the previous edition, the following technical modification has been made:

Regarding non dioxin-like PCBs (ndl-PCBs), this document contains two approaches that can be followed.

Method 1 concerns the original extraction and clean-up methods of the previous edition of this standard,

but combined with more sensitive detection approaches. In method 2, the extraction and clean-up

methods have been modified in order to increase the test portion. The detection of method 2 concerns

the original detection method of the previous edition of this standard.

This document has been prepared under a standardization request given to CEN by the European

Commission and the European Free Trade Association.

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

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

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

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

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

Kingdom
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SIST EN 15741:2020
EN 15741:2020 (E)
Introduction

WARNING — The use of this document can involve hazardous materials, operations and equipment. This

standard does not purport to address all the safety problems associated with its use. It is the

responsibility of the user of this European Standard to establish appropriate safety and health practices

and determine the applicability of regulatory limitations prior to use.
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SIST EN 15741:2020
EN 15741:2020 (E)
1 Scope

This document specifies a gas chromatographic mass spectrometric (GC-MS) method for the

determination of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in compound

feeds and feed materials (oils and fats, fish meal).

The method is applicable to compound feeds consisting of less than 20 % water by mass and oil/fatty

samples containing residues of one or more of the following OCPs and PCBs and some of their isomers

and degradation products:
— aldrin;
— dieldrin;
— chlordane, as the sum of chlordane isomers and oxychlordane;

— dichlorodiphenyltrichloroethane (DDT), as the sum of isomers o,p'-DDT, p,p'-DDT, p,p'-TDE (p,p'-

DDD), and p,p'-DDE;
— endosulfan, as the sum of α-/β-isomers and endosulfan-sulphate;
— endrin, as the sum of endrin and delta-keto-endrin;
— heptachlor, as the sum of heptachlor and heptachlor epoxide;
— hexachlorobenzene (HCB);

— hexachlorocyclohexane isomers α-HCH (α-BHC), β-HCH (β-BHC), γ-HCH (γ-BHC or lindane);

— photo heptachlor;
— cis- and trans-nonachlor;

— non dioxin-like PCBs (ndl-PCBs), as the sum of PCB 28, 52, 101, 138, 153 and 180.

The method has been fully validated by a collaborative trial for the substances and corresponding ranges

(µg/kg) noted in Table 1.
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EN 15741:2020 (E)
Table 1 — Residue compound and range of (µg/kg) collaborative trial
Compound Range (µg/kg)
all ndl-PCBs 0,7 to 39
aldrin 10 to 34
dieldrin 12 to 97
endrin 13 to 88
a a
cis-chlordane 16 to 24
trans-chlordane 7 to 25
b b
p,p’-DDT 19 to 200
o,p’-DDT 8 to 87
c c
pp’-TDE 9 to 103
pp'-DDE 21 to 264
α-endosulfan 15 to 165
β-endosulfan 26 to 331
d d
endosulfan sulphate 29 to 61
heptachlor 15 to 365
heptachlor epoxide 15 to 382
e e
HCB 8 to 170
α-HCH 21 to 247
β-HCH 6 to 84
f f
γ-HCH 17 to 186
NOTE The following information is to be taken into consideration:

a) Cis-chlordane has not been fully validated for chicken feed, pig feed and fish oil.

b) p,p’-DDT has not been fully validated for pig feed and vegetable oil.
c) pp’-TDE has not been fully validated for pig feed and fish meal.

d) Endosulfan sulphate has not been fully validated for pig feed and vegetable oil.

e) HCB has not been fully validated for fish oil.
f) γ-HCH has not been fully validated for fish oil.

The method has not been fully validated for oxychlordane, endrin ketone, cis- and trans-nonachlor and

photo heptachlor in all matrices.

For those matrix-analyte combinations where the validation data were regarded insufficient, the results

obtained with this method can only be regarded as screening results, unless the laboratory performs an

in-house validation to show that satisfactory results can be obtained.

The method is not applicable to chlorocamphene (toxaphene), a complex mixture of polychlorinated

camphenes. Chlorocamphene has a very distinctive chromatographic profile and is easily recognizable by

GC/ECD. Positive identification of the toxaphene isomers can be performed by negative chemical

ionization mass spectrometry (NCI-MS), electron impact tandem mass spectrometry (EI MS × MS) or

electron impact high resolution mass spectrometry (EI-HRMS), which is not within the scope of this

method.
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SIST EN 15741:2020
EN 15741:2020 (E)

A limit of quantification (LOQ) for the mentioned organochlorine pesticides of 6 to 29 µg/kg should

normally be obtained (see Table 1). For the ndl-PCBs an LOQ of 0,5 to 1,0 µg/kg should be obtained. The

LOQs mentioned apply to the individual compounds (i.e. not the sum of two or more compounds).

Individual laboratories are responsible for ensuring that the equipment that they used will achieve these

LOQs. On customers' demand the standard may be applied to solely the analysis of PCBs or OCPs.

2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

EN ISO 6498, Animal feeding stuffs – Guidelines for sample preparation (ISO 6498)

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
limit of detection

smallest measured content, from which it is possible to deduce the presence of the analyte with

reasonable statistical certainty

Note 1 to entry: The limit of detection is numerically equal to 3 times the standard deviation of the mean of blank

determinations (n > 10).
3.2
limit of quantification

lowest content of the analyte which can be measured with reasonable statistical certainty

Note 1 to entry: If both accuracy and precision are constant over a concentration range around the limit of

detection, then the limit of quantification is numerically equal to 6 times the standard deviation of the mean of blank

determinations (n > 10).
3.3
feed additives
substances that are added to animal nutrition

Note 1 to entry: see Regulation (EC) No.1831/2003 of the European Parliament and of the Council on additives

for use in animal nutrition for more information.
4 Principle
4.1 General

In order to check for the presence of organochlorine pesticides (OCPs), a test portion of animal feeding

stuff is fortified with internal standard ( C -PCB mix) and extracted with ethyl acetate. The extract is

concentrated and subsequently purified by:

— gel permeation chromatography (GPC), with a mixture of cyclohexane/ethyl acetate as eluting

solvent;
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SIST EN 15741:2020
EN 15741:2020 (E)
— chromatography on partially deactivated silica gel.

The collected fraction containing the compounds of interest is concentrated and re-dissolved in a solution

containing another internal standard (PCB 209) as a reference standard. After concentration an aliquot

of the extract is injected into a GC-MS using a splitless injector. In case more sensitivity is necessary or

less volume reduction is wanted, injection of a larger volume by means of a Programmed Temperature

Vaporizer (PTV) injector is possible. An example is described in Annex B.
For ndl-PCBs, two approaches can be followed.
4.2 Ndl-PCBs; method 1

A test portion of animal feeding stuff is fortified with internal standard ( C -PCB mix), and is extracted

with ethyl acetate. The extract is concentrated and subsequently purified by:

— gel permeation chromatography, with a mixture of cyclohexane /ethyl acetate as eluting solvent;

— chromatography on partially deactivated silica gel.

The collected fraction containing the compounds of interest is concentrated and re-dissolved in a solution

containing another internal standard (PCB 209) as a reference standard. After concentration an aliquot

of the extract is injected into a GC-MS-MS or GC-HRMS using a splitless injector. As an alternative,

injection by means of a PTV injector is possible. An example is described in Annex B.

4.3 Ndl-PCBs; method 2

A test portion of animal feeding stuff is fortified with internal standard ( C -PCB mix), and is extracted

with ethyl acetate. The extract is concentrated and subsequently purified by:
— concentrated sulfuric acid;

— gel permeation chromatography (GPC), with a mixture of cyclohexane/ethyl acetate as eluting

solvent;
— chromatography on partially deactivated silica gel.

The collected fraction containing the compounds of interest is concentrated and re-dissolved in a solution

containing another internal standard (PCB 209) as a reference standard. After concentration an aliquot

of the extract is injected into a GC-MS, using a splitless injector. As an alternative, injection by means of a

PTV injector is possible. An example is described in Annex B.
5 Reagents and materials
5.1 General

Use only reagents of recognized analytical grade and with a purity suitable for OC and PCB residue

analysis. Check the purity of the reagents by performing a blank test under the same conditions as used

in the method. The chromatogram should not show any interfering impurity at the retention time of

compounds of interest.
5.2 Chemicals
5.2.1 Ethyl acetate
5.2.2 Cyclohexane
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SIST EN 15741:2020
EN 15741:2020 (E)
5.2.3 Ethyl acetate/Cyclohexane = 1+1 parts by volume

Mix 500 ml of ethyl acetate (5.2.1) with 500 ml of cyclohexane (5.2.2) and mix thoroughly. Store at room

temperature in a tightly closed glass bottle.
5.2.4 Hexane
5.2.5 Decane
5.2.6 Hexane/Decane = 95+5 part by volume

Mix 950 ml of hexane (5.2.4) with 50 ml of decane (5.2.5) and mix thoroughly. Store at room temperature

in a tightly closed glass bottle.
5.2.7 Iso-octane
5.2.8 Toluene
5.2.9 Silica gel, deactivated with 3,5 % water

Heat silica gel 60 (63µm to 200µm = 70 mesh to 230 mesh), at 130 °C for at least 5 h, allow to cool in a

desiccator and store in a tightly stopped container in the desiccator. Add 3,5 ml water dropwise from a

burette, with a continuous swirling, to 96,5 g dried silica gel in a 300 ml Erlenmeyer flask with a ground

joint. Immediately stopper the flask with a ground stopper and shake vigorously for 5 min until all lumps

have disappeared. Next, shake for 2 h on a mechanical shaker, then store in a tightly stoppered container.

Deactivated silica gel is tenable during approximately 2 weeks if carefully stored.

5.2.10 Hexane/toluene = 3+7 parts by volume

Mix 30 ml of n-hexane (5.2.4) with 70 ml of toluene (5.2.8) and mix thoroughly. Store at room

temperature in a tightly closed glass bottle.
5.2.11 Concentrated H SO
2 4
5.2.12 Internal standard (PCB 209)
5.2.12.1 PCB 209 stock solution 1, 100 µg/ml

Weigh 10 mg (±0,01 mg) of PCB 209 (5.2.12) in a brown medicine glass bottle of 100 ml and add iso-

octane (5.2.7) to achieve a concentration of 100 μg/ml. Store the solution in a refrigerator at 4°C (±3°C).

The solution is tenable under these conditions during at least 5 years if the weight of the solution is

carefully controlled. Alternatively, use a commercially available standard solution of 100 µg/ml.

5.2.12.2 PCB 209 stock solution 2, 10,0 µg/ml

Dilute 10,0 ml of PCB 209 Stock solution 1 (5.2.12.1) to 100,0 ml with hexane (5.2.4). Store the solution

in a refrigerator at 4°C (±3°C). The solution is tenable under these conditions during at least 5 years if the

weight of the solution is carefully controlled.
5.2.12.3 PCB 209 working solution, concentration 1 000 ng/ml

Dilute 10 ml of PCB 209 Stock solution 2 (5.2.12.2) to 100,0 ml with hexane (5.2.4). Store the solution in

a refrigerator at 4 °C (±3 °C). The solution is tenable under these conditions during at least 5 years if the

weight of the solution is carefully controlled.
5.3 Internal standards ( C mass labelled PCBs)
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SIST EN 15741:2020
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5.3.1 Internal standards ( C mass labelled PCBs), 1 000 ng/ml
13 13
C PCB 28 (2,4,4’ trichlorobiphenyl, C ); CAS Number: 208263-76-7;
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13 13
C PCB 52 (2,2’,5,5’ tetrachlorobiphenyl, C ); CAS Number: 208263-80-3;
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13 13
C PCB 101 (2,2’,4,5,5’ pentachlorobiphenyl, C ); CAS Number: 104130-39-4;
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13 13
C PCB 138 (2,2’,3’,4,4’,5 hexachlorobiphenyl, C ); CAS Number: 208263-66-5;
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13 13
C PCB 153 (2,2’,4,4’,5,5’ hexachlorobiphenyl, C ); CAS Number: 185376-58-3;
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13 13
C PCB 180 (2,2’,3,4,4’,5,5’ heptachlorobiphenyl, C ); CAS Number: not available.
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Alternatively, use a certified mixture at a concentration of 1 000 ng/ml.
5.3.2 Internal standards ( C mass labelled PCBs), 100 ng/ml

Dilute 1,0 ml of internal standards ( C mass labelled PCBs) (5.3.1) to 10,0 ml with hexane (5.2.4). Store

the solution in a refrigerator at 4 °C (±3 °C). The solution is tenable under these conditions during at least

5 years if the weight is carefully controlled.
5.4 PCB congeners stock standard solution
5.4.1 PCB congeners stock standard solution, 10 µg/ml
PCB 28 (2,4,4’ trichlorobiphenyl); CAS Number: 7012-37-5;
PCB 52 (2,2’,5,5’ tetrachlorobiphenyl); CAS Number: 35693-99-3;
PCB 101 (2,2’,4,5,5’ pentachlorobiphenyl); CAS Number: 37680-73-2;
PCB 138 (2,2’,3’,4,4’,5 hexachlorobiphenyl); CAS Number: 35065-28-2;
PCB 153 (2,2’,4,4’,5,5’ hexachlorobiphenyl); CAS Number: 35065-27-1;
PCB 180 (2,2’,3,4,4’,5,5’ heptachlorobiphenyl); CAS Number: 35065-29-3.
Alternatively, use a certified mixture at a concentration of 10 µg/ml.
5.4.2 PCB congeners working standard solution, 2,0 µg/ml

Dilute 2,0 ml of PCB congeners stock standard solution (5.4.1) to 10,0 ml with hexane (5.2.4). Store the

solution in a refrigerator at 4 °C (±3 °C). The solution is tenable under these conditions during at least

5 years if the weight is carefully controlled.
5.4.3 PCB congeners work standard solution, 0,2 µg/ml

Dilute 1,0 ml of PCB congeners stock standard solution (5.4.2) to 10,0 ml with hexane (5.2.4). Store the

solution in a refrigerator at 4 °C (±3 °C). The solution is tenable under these conditions during at least

5 years if the weight is carefully controlled.
5.5 OC-pesticide reference standards, as follows
Each with a purity of not less than 99 %:
Aldrin:
(1R,4S,4aS,5S,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5,8-
dimethanonaphthalene; CAS Number: 309-00-2.
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SIST EN 15741:2020
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Dieldrin:

(1R,4S,4aS,5R,6R,7S,8S,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,7,8,8a-octahydro-6,7epoxy-1,4:5,8-

dimethanonaphthalene; CAS Number: 60-57-1.
Chlordane, α isomer:

1,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-ethano-1H-indene; α isomer; CAS Number: 5103-

71-9.
Chlordane, β isomer:

1,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-ethano-1H-indene; β isomer; CAS Number: 5103-

74-2.
Oxychlordane :

4,7-Methanoindan, 1,2,4,5,6,7,8,8-octachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-, exo, endo-; CAS Number:

27304-13-8.
o,p'-DDT :

1,1,1-trichloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)ethane; CAS Number: 789-02-6;

p,p'-DDT :
1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane; CAS Number: 50-29-3.
pp'-TDE :
(pp'-DDD) 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane; CAS Number: 72-54-8.
pp'-DDE :
1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene; CAS Number: 72-55-9.
Endosulfan, α stereoisomer:

6,9-Methano-2,4,3-benzodioxathiepin, 6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-, 3-oxide,

(3α,5aβ, 6α,9α,9aβ); CAS Number: 959-98-8.
Endosulfan, β stereoisomer :

6,9-Methano-2,4,3-benzodioxathiepin, 6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-, 3-oxide,

(3α,5aα,6β,9β,9aα); CAS Number: 33213-65-9.
Endosulfan-sulphate;

6,9-Methano-2,4,3-benzodioxathiepin, 6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-, 3,3-dioxide;

CAS Number: 1031-07-8.
Endrin

(1R,4S,4aS,5S,6S,7R,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,7,8,8a-octahydro-6,7-epoxy-1,4:5,8-dd;

CAS Number: 72-20-8.
Delta-keto endrin

2,5,7-Metheno-3H-cyclopenta[a]pentalen-3-one,3b,4,5,6,6,6a-hexachlorodecahydro-, 2α,

3aβ,3bβ,4β,5β,6aβ,7α,7aβ,8R*; CAS Number 53494-70-5.
Heptachlor

1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene; CAS Number: 76-44-8.

β-Heptachlor epoxide

1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene(exo); CAS Number: 1024-57-3.

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SIST EN 15741:2020
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HCB
hexachlorobenzene; CAS Number: 118-74-1.
α-HCH (α-BHC)
α-1,2,3,4,5,6-hexachlorocyclohexane; CAS Number: 319-84-6.
β-HCH (β-BHC)
β-1,2,3,4,5,6-hexachlorocyclohexane; CAS Number: 319-85-7.
γ-HCH (γ-BHC; lindane)
γ-1,2,3,4,5,6-hexachlorocyclohexane; CAS Number: 58-89-9.
Nonachlor, cis;
CAS Number 5103-73-1.
Nonachlor, trans;
CAS Number 39765-80-5.
Photo heptachlor;
CAS Number 33442-83-0.

Alternatively, use a certified mixture at a precise or certified concentration (approximately 10 µg/ml).

5.5.1 Pesticide stock solution 1, 100 µg/ml

Weigh 10 mg (±0,01 mg) of each individual pesticide (5.5) in separate brown medicine glass bottles of

100 ml and add iso-octane (5.2.7) to achieve a concentration of 100 μg/ml. Store the solutions in a

refrigerator at 4 °C (±3 °C). The solutions are tenable under these conditions during at least 5 years if the

weight is carefully controlled. Alternatively, use a commercially available standard solution with a precise

or certified concentration of approximately 100 µg/ml.

NOTE Dissolve ß-HCH in 10 ml toluene (5.2.8) to achieve complete solvability and dilute further with iso-

octane (5.2.7) to achieve a concentration of 100 μg/ml
5.5.2 Pesticide stock solution 2, 2,5 µg/ml

Pipet 2,5 ml of each individual pesticide stock solutions 1 (5.5.1) into a 100,0 ml graduated flask and

dilute with hexane (5.2.4) to 100,0 ml. Store the solution in a refrigerator at 4 °C (±3 °C). The solution is

tenable under these conditions during at least 5 years if the weight is carefully controlled.

5.5.3 Pesticide working solution, 0,5 µg/ml

Pipet 2,0 ml of pesticide stock solution 2 (5.5.2) into a 10,0 ml graduated flask and dilute with hexane

(5.2.4) to 10,0 ml. Store the solution in a refrigerator at 4 °C (±3 °C). The solution is tenable under these

conditions during at least 5 years if the weight is carefully controlled.
5.6 Calibration solutions

Prepare calibration mixtures according to Table 2 in a final volume of 1,0 ml of an alkane solvent (5.2.6

or 5.2.7) and store them at 4 °C (±3 °C).
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SIST EN 15741:2020
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Table 2 — Calibration mixtures
C -PCB PCB 209
PCB 0,2 μg/ml OC 0,5 μg/ml OC 5,0 μg/ml
Level 1 000 ng/ml Solvent
mix 100 ng/ml
(5.4.3) (5.5.3) (5.5.2)
(5.2.12.3)
(5.3.2)
μl ng/ml μl ng/ml μl ng/ml μl ng/ml µl ng/ml µl
1 0 0 0 0 0 n.a. 100 10 50 50 850
2 5 1 20 10 0 n.a. 100 10 50 50 825
3 10 2 50 25 0 n.a. 100 10 50 50 790
4 25 5 250 125 0 n.a. 100 10 50 50 575
5 100 20 0 n.a. 100 500 100 10 50 50 650
6 250 50 0 n.a. 250 1 250 100 10 50 50 350
5.7 Glass vial, 100 ml, including PTFE-lined screwcaps
5.8 Glass wool
Heated at 160 °C – 200 °C during at least 24 h.
5.9 Sodium sulp
...

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