SIST EN 17503:2023
(Main)Soil, sludge, treated biowaste and waste - Determination of polycyclic aromatic hydrocarbons (PAH) by gas chromatography (GC) and high performance liquid chromatography (HPLC)
Soil, sludge, treated biowaste and waste - Determination of polycyclic aromatic hydrocarbons (PAH) by gas chromatography (GC) and high performance liquid chromatography (HPLC)
This European Standard specifies a method for quantitative determination of 16 polycyclic aromatic hydrocarbons (PAH) (see Table ) in soil, sludge, sediment, treated biowaste, and waste, using GC-MS and HPLC-UV-DAD/FLD covering a wide range of PAH contamination levels (see Table 2).
When using fluorescence detection, acenaphthylene cannot be measured.
Table —Target analytes of this European Standard
Target analyte CAS-RNa
Naphthalene 91–20–3
Acenaphthene 83–32–9
Acenaphthylene 208–96–8
Fluorene 86–73–7
Anthracene 120–12–7
Phenanthrene 85–01–8
Fluoranthene 206–44–0
Pyrene 129–00–0
Benz[a]anthracene 56–55–3
Chrysene 218–01–9
Benzo[b]fluoranthene 205–99–2
Benzo[k]fluoranthene 207–08–9
Benzo[a]pyrene 50–32–8
Indeno[1,2,3-cd]pyrene 193–39–5
Dibenz[a,h]anthracene 53–70–3
Benzo[ghi]perylene 191–24–2
a CAS-RN Chemical Abstracts Service Registry Number.
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used for the extraction of the sample and the clean-up of the extract
Under the conditions specified in this European Standard, lower limit of application from 10 μg/kg (expressed as dry matter) for soils, sludge and biowaste to 100 μg/kg (expressed as dry matter) for solid waste can be achieved. For some specific samples (e.g. bitumen) the limit of 100 μg/kg cannot be reached.
Sludge, waste and treated biowaste may differ in properties as well as in the expected contamination levels of PAH and presence of interfering substances. These differences make it impossible to describe one general procedure. This European Standard contains decision tables based on the properties of the sample and the extraction and clean-up procedure to be used.
The method may be applied to the analysis of other PAH not specified in the scope, provided suitability is proven by proper in-house validation experiments.
Boden, Schlamm, behandelter Bioabfall und Abfall - Bestimmung von polycyclischen aromatischen Kohlenwasserstoffen (PAK) mittels Gaschromatographie (GC) und Hochleistungs-Flüssigkeitschromatographie (HPLC)
Dieses Dokument legt verschiedene Verfahren für die quantitative Bestimmung von 16 polycyclischen aromatischen Kohlenwasserstoffen (PAK) (siehe Tabelle 2) in Boden, Schlamm, behandeltem Bioabfall und Abfall mittels GC MS oder HPLC UV DAD/FLD fest, wobei ein breites Spektrum an PAK Kontaminationsniveaus (siehe Tabelle 2) abgedeckt wird.
ANMERKUNG Das Verfahren kann auf Sedimente angewandt werden, sofern der Anwender die Validität nachweist.
Bei Anwendung der Fluoreszenzdetektion kann Acenaphthylen nicht bestimmt werden.
[Tabelle 2]
Die Nachweisgrenze hängt von den zu bestimmenden Substanzen, der verwendeten technischen Ausrüstung, der Qualität der für die Extraktion der Probe verwendeten Chemikalien und der Aufreinigung des Extrakts ab.
Unter den in diesem Dokument festgelegten Bedingungen kann eine untere Anwendungsgrenze zwischen 10 μg/kg (angegeben als Trockenmasse) für Böden, Schlamm und Bioabfall und 100 μg/kg (angegeben als Trockenmasse) für Feststoffabfall erreicht werden. Bei einigen bestimmten Proben (z. B. Bitumen) kann die Grenze von 100 μg/kg nicht erreicht werden.
Schlamm, Abfall und behandelter Bioabfall können sich in Bezug auf ihre Eigenschaften sowie auf die erwarteten PAK Kontaminationsniveaus und das Vorhandensein von störenden Substanzen unterscheiden. Aufgrund dieser Unterschiede ist es unmöglich, ein allgemeines Verfahren darzulegen. Dieses Dokument enthält Entscheidungstabellen, die auf den Eigenschaften der Probe und den anzuwendenden Extraktions und Aufreinigungsverfahren basieren.
Das Verfahren kann für die Analyse weiterer PAK angewendet werden, die im Anwendungsbereich nicht genannt sind, sofern die Eignung des Verfahrens durch hinreichende interne Validierungsuntersuchungen bewiesen wurde.
Die Probenahme ist nicht Bestandteil dieser Norm. In Abhängigkeit von den Materialien bedürfen die folgenden Normen der Berücksichtigung, z. B. EN 14899, ISO 5667 12 und EN ISO 5667 13.
Sols, boues, biodéchets traités et déchets - Dosage des hydrocarbures aromatiques polycycliques (HAP) par chromatographie en phase gazeuse et chromatographie liquide à haute performance
Le présent document spécifie différentes méthodes de dosage de 16 hydrocarbures aromatiques polycycliques (HAP) (voir Tableau 2) dans les sols, les boues, les biodéchets traités et les déchets faisant appel à des méthodes CG-SM ou HPLC-UV-DAD/FLD couvrant une large gamme de niveaux de contamination par les HAP (voir Tableau 2).
NOTE La méthode peut être appliquée aux sédiments sous réserve que l’utilisateur ait démontré la validité.
L’utilisation d’une méthode de détection fluorimétrique ne permet pas le mesurage de l’acénaphtylène.
[Tableau 2 - Analytes cible du présent document]
La limite de détection dépend des composés à analyser, de l’équipement utilisé, de la qualité des réactifs chimiques utilisés pour l’extraction de l’échantillon et la purification de l’extrait.
Dans les conditions spécifiées dans le présent document, la limite inférieure d’application allant de 10 μg/kg (exprimée par rapport à la matière sèche) pour les sols, boues et biodéchets jusqu’à 100 μg/kg (exprimée par rapport à la matière sèche) pour les déchets solides peut être atteinte. Pour certains échantillons particuliers (bitume, par exemple), la limite de 100 μg/kg ne peut pas être atteinte.
Les boues, les déchets et les biodéchets traités peuvent présenter des propriétés, des niveaux de contamination différents par les HAP attendus et des substances interférentes. En raison de ces différences, il est impossible de décrire un mode opératoire général. Le présent document contient des tables de décision basées sur les propriétés de l’échantillon ainsi que sur le mode opératoire d’extraction et de purification à utiliser.
La méthode peut être appliquée à l’analyse d’autres HAP non spécifiés dans le domaine d’application, à condition que son adéquation ait été prouvée par des expériences de validation interne.
L’échantillonnage n’est pas couvert par la présente norme. Selon le type de matériau, il est nécessaire de prendre en compte par exemple les normes suivantes : l’EN 14899, l’ISO 5667-12 et l’EN ISO 5667-13.
Tla, blato, obdelani biološki odpadki in odpadki - Določevanje policikličnih aromatskih ogljikovodikov (PAH) s plinsko kromatografijo (GC) in s tekočinsko kromatografijo visoke ločljivosti (HPLC)
Ta evropski standard določa metodo za kvantitativno določevanje 16 policikličnih aromatskih ogljikovodikov (PAH) (glej preglednico) v tleh, blatu, usedlinah, obdelanih bioloških odpadkih in odpadkih z metodama GC-MS in HPLC-UV-DAD/FLD, ki zajemata široko območje ravni onesnaženja PAH (glej preglednico 2).
S fluorescenčnim zaznavanjem ni mogoče izmeriti acenaftilena.
Preglednica – Ciljni analiti tega evropskega standarda
Ciljni analit CAS-RNa
Naftalen 91–20–3
Acenaften 83–32–9
Acenaftilen 208–96–8
Fluoren 86–73–7
Antracen 120–12–7
Fenantren 85–01–8
Fluoranten 206-44-0
Piren 129-00-0
Benz[a]antracen 56–55–3
Krizen 218–01–9
Benzo[b]fluoranten 205–99–2
Benzo[k]fluoranten 207–08–9
Benzo[a]piren 50–32–8
Indeno[1,2,3-cd]piren 193–39–5
Dibenzo[a,h]antracen 53–70–3
Benzo[ghi]perilen 191–24–2
a Registrska številka Chemical Abstracts Service CAS-RN.
Meja detekcije je odvisna od determinant, uporabljene opreme, kakovosti kemikalij, uporabljenih za ekstrakcijo vzorca in očiščenje izvlečka.
Pod pogoji, določenimi v tem evropskem standardu, je mogoče doseči spodnjo mejo uporabe od 10 μg/kg (izraženo kot suha snov) za tla, blato in biološke odpadke do 100 μg/kg (izraženo kot suha snov) za trdne odpadke. Za nekatere specifične vzorce (npr. bitumen) ni mogoče doseči meje 100 μg/kg.
Blato, odpadki in obdelani biološki odpadki se lahko razlikujejo glede lastnosti in tudi glede predvidenih stopenj onesnaženja s policikličnimi aromatskimi ogljikovodiki ter prisotnosti motečih snovi. Zaradi teh razlik ni možen opis enotnega splošnega postopka. Ta evropski standard vključuje tabele odločanja, ki temeljijo na lastnostih vzorca, ter postopek za ekstrakcijo in očiščenje, ki ga je treba uporabiti.
Metoda se lahko uporablja za analize drugih policikličnih aromatskih ogljikovodikov, ki ne spadajo na področje uporabe tega standarda, če je njena ustreznost dokazana s primernimi internimi poskusi za validacijo.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 17503:2023
01-marec-2023
Nadomešča:
SIST EN 15527:2009
SIST EN 16181:2018
SIST ISO 13877:1999
Tla, blato, obdelani biološki odpadki in odpadki - Določevanje policikličnih
aromatskih ogljikovodikov (PAH) s plinsko kromatografijo (GC) in s tekočinsko
kromatografijo visoke ločljivosti (HPLC)
Soil, sludge, treated biowaste and waste - Determination of polycyclic aromatic
hydrocarbons (PAH) by gas chromatography (GC) and high performance liquid
chromatography (HPLC)
Boden, Schlamm, behandelter Bioabfall und Abfall - Bestimmung von polycyclischen
aromatischen Kohlenwasserstoffen (PAK) mittels Gaschromatographie (GC) und
Hochleistungs-Flüssigkeitschromatographie (HPLC)
Sols, boues, biodéchets traités et déchets - Dosage des hydrocarbures aromatiques
polycycliques (HAP) par chromatographie en phase gazeuse et chromatographie liquide
à haute performance
Ta slovenski standard je istoveten z: EN 17503:2022
ICS:
13.030.01 Odpadki na splošno Wastes in general
13.080.10 Kemijske značilnosti tal Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST EN 17503:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 17503:2023
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SIST EN 17503:2023
EN 17503
EUROPEAN STANDARD
NORME EUROPÉENNE
February 2022
EUROPÄISCHE NORM
ICS 13.030.01; 13.080.10 Supersedes EN 15527:2008, EN 16181:2018
English Version
Soil, sludge, treated biowaste and waste - Determination of
polycyclic aromatic hydrocarbons (PAH) by gas
chromatography (GC) and high performance liquid
chromatography (HPLC)
Sols, boues, biodéchets traités et déchets - Dosage des Boden, Schlamm, behandelter Bioabfall und Abfall -
hydrocarbures aromatiques polycycliques (HAP) par Bestimmung von polycyclischen aromatischen
chromatographie en phase gazeuse et Kohlenwasserstoffen (PAK) mittels
chromatographie liquide à haute performance Gaschromatographie (GC) und Hochleistungs-
Flüssigkeitschromatographie (HPLC)
This European Standard was approved by CEN on 3 January 2022.
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
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17503:2022 E
worldwide for CEN national Members.
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EN 17503:2022 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 8
5 Interferences . 8
6 Safety remarks . 9
7 Reagents . 10
8 Apparatus . 14
9 Sample storage and preservation . 15
10 Procedure . 16
11 Performance characteristics. 29
12 Precision . 29
13 Test report . 30
Annex A (informative) Repeatability and reproducibility data . 31
A.1 Materials used in the interlaboratory comparison study . 31
A.2 Interlaboratory comparison results . 32
Annex B (informative) Examples of instrumental conditions and chromatograms . 39
B.1 Measurement of PAH with GC-MS . 39
B.2 Measurement of PAH with HPLC fluorescence . 45
B.3 Example for measurement conditions of PAH with GC-MS/MS . 51
Bibliography . 53
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EN 17503:2022 (E)
European foreword
This document (EN 17503:2022) has been prepared by Technical Committee CEN/TC 444
“Environmental characterization of solid matrices”, the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by August 2022, and conflicting national standards shall be
withdrawn at the latest by August 2022.
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 15527:2008 and EN 16181:2018.
Any feedback and questions on this document should be directed to the users’ national standards body.
A complete listing of these bodies can be found on the CEN website.
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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Introduction
Polycyclic aromatic hydrocarbons (PAH) are ubiquitous because they are released in appreciable
quantities every year into the environment through the combustion of organic matters such as coal, fuel
oils, petrol, wood, refuse and plant materials. Since some of these PAH compounds are carcinogenic or
mutagenic, their presence in the environment (air, water, soil, sediment and waste) is regularly
monitored and controlled. At present determination of PAH is carried out in these matrices in most of the
routine laboratories following the prescribed steps specified for sampling, pre-treatment, extraction,
clean-up by measurement of specific PAH by means of gas chromatography in combination with mass
spectrometric detection (GC-MS) or by high performance liquid chromatography (HPLC) in combination
with UV-DAD- or fluorescence detection (HPLC-UV-DAD/FLD). Both the GC-MS and the HPLC methods
are included in this horizontal standard.
This document was developed by the merging of EN 16181:2018, initially elaborated as a CEN Technical
Specification in the European project 'HORIZONTAL' and validated by CEN/TC 400 with the support of
BAM, with EN 15527, published by CEN/TC 292.
Considered the different matrices and possible interfering compounds, this document does not contain
one single possible way of working. Several choices are possible, in particular relating to clean-up.
Quantification with both GC-MS-detection and HPLC-DAD-UV/FLD is possible. Four different extraction
procedures are described and three clean-up procedures. The use of internal and injection standards is
described in order to have an internal check on choice of the extraction and clean-up procedure. The
method is as far as possible in agreement with the method described for polychlorinated biphenyls (PCB)
in EN 17322. It has been tested for ruggedness.
This document is applicable and validated for several types of matrices as indicated in Table 1 (see also
Annex A for the results of the validation).
Table 1 — Matrices for which this document is applicable and validated
Matrix Materials used for validation
Soil Sandy soil
Mix of soil from an industrial area in Brandenburg, Germany and
PAH-free German reference soil
Sludge Mix of municipal waste water treatment plant sludge from the
vicinity of Berlin, Germany
Biowaste Mix of compost from the vicinity of Berlin, Germany
Contaminated soil, building debris, waste wood, roofing tar,
Waste
shredder light fraction
WARNING — Persons using this document should be familiar with usual laboratory practice. This
document does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices and to ensure compliance
with any national regulatory conditions.
WARNING — It is absolutely essential that tests conducted according to this document be carried out by
suitably trained staff.
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1 Scope
This document specifies different methods for quantitative determination of 16 polycyclic aromatic
hydrocarbons (PAH) (see Table 2) in soil, sludge, treated biowaste, and waste, using GC-MS or HPLC-UV-
DAD/FLD covering a wide range of PAH contamination levels (see Table 2).
NOTE The method can be applied to sediments provided that validity is demonstrated by the user.
When using fluorescence detection, acenaphthylene cannot be measured.
Table 2 —Target analytes of this document
a
Target analyte
CAS-RN
Naphthalene 91–20–3
Acenaphthene 83–32–9
Acenaphthylene 208–96–8
Fluorene 86–73–7
Anthracene 120–12–7
Phenanthrene 85–01–8
Fluoranthene 206–44–0
Pyrene 129–00–0
Benz[a]anthracene 56–55–3
Chrysene 218–01–9
Benzo[b]fluoranthene 205–99–2
Benzo[k]fluoranthene 207–08–9
Benzo[a]pyrene 50–32–8
Indeno[1,2,3-cd]pyrene 193–39–5
Dibenz[a,h]anthracene 53–70–3
Benzo[ghi]perylene 191–24–2
a
CAS-RN Chemical Abstracts Service Registry Number.
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used
for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this document, the lower limit of application from 10 μg/kg (expressed
as dry matter) for soils, sludge and biowaste to 100 μg/kg (expressed as dry matter) for solid waste can
be achieved. For some specific samples (e.g. bitumen) the limit of 100 μg/kg cannot be reached.
Sludge, waste and treated biowaste can differ in properties as well as in the expected contamination
levels of PAH and presence of interfering substances. These differences make it impossible to describe
one general procedure. This document contains decision tables based on the properties of the sample and
the extraction and clean-up procedure to be used.
The method can be applied to the analysis of other PAH not specified in the scope, provided suitability is
proven by proper in-house validation experiments.
5
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Sampling is not part of this standard. In dependence of the materials, the following standards need to be
considered, e.g. EN 14899, ISO 5667-12 and EN ISO 5667-13.
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 15002, Characterization of waste — Preparation of test portions from the laboratory sample
EN 15934, Sludge, treated biowaste, soil and waste — Calculation of dry matter fraction after
determination of dry residue or water content
EN 16179, Sludge, treated biowaste and soil — Guidance for sample pretreatment
EN ISO 5667-15, Water quality — Sampling — Part 15: Guidance on the preservation and handling of
sludge and sediment samples (ISO 5667-15)
EN ISO 16720, Soil quality — Pretreatment of samples by freeze-drying for subsequent analysis (ISO 16720)
EN ISO 22892, Soil quality — Guidelines for the identification of target compounds by gas chromatography
and mass spectrometry (ISO 22892)
ISO 8466-1, Water quality — Calibration and evaluation of analytical methods — Part 1: Linear calibration
function
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 https://www.electropedia.org/
• ISO Online browsing platform: available at https://www.iso.org/obp
3.1
critical pair
pair of PAH that are separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic separation
meets minimum quality criteria
EXAMPLE Figure 1 shows an example of a chromatogram of a critical pair.
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∆ t
R 2 ×
YY +
ab
(1)
where:
R peak separation
Δt difference in retention times of the two peaks a and b in seconds (s)
Y peak width at the base of peak a in seconds (s)
a
Y peak width at the base of peak b in seconds (s)
b
Figure 1 — Example of a chromatogram of a critical pair
3.2
internal standard
compound added in a known amount to the sample from the beginning of the protocol and enabling
analytical coverage throughout the procedure, and that is used to correct for losses during sample
preparation and analysis by accounting for all-system matrix effects (recoveries, ionization effect,
variability of the detector response of the instrument for example)
Note 1 to entry: isotopically labelled mostly deuterated PAH or native PAH unlikely to be present in the sample
[SOURCE: EN ISO 21253-2:2019, 3.10]
3.3
injection standard
standard mixture added to a sample before injection into the GC-MS apparatus, to monitor variability of
instrument response and to calculate internal standard recovery
[SOURCE: ISO 28540:2011, 3.4]
7
=
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3.4
extraction standard
PAH that is unlikely to be present in samples added to the sample prior to extraction, used for checking
the extraction efficiency and not used for quantification purposes
3.5
sediment
solid material, both mineral and organic, deposited in the bottom of a water body
[SOURCE: ISO 5667-12:2017, 3.5]
4 Principle
Due to the multi matrix character of this document, different procedures for different steps (modules)
are allowed. Which modules should be used depends on the sample. A recommendation is given in this
document. Performance criteria are described and it is the responsibility of the laboratories applying this
document to show that these criteria are met. Using of spiking internal standards allows an overall check
on the efficiency of a specific combination of modules for a specific sample. But the use of internal
standards does not necessarily give the information regarding the extensive extraction efficiency of the
native PAH bonded to the matrix.
After pre-treatment, the sample is extracted with a suitable solvent.
The extract is concentrated by evaporation. If necessary, interfering compounds are removed by a clean-
up method suitable for the specific matrix, after the concentration step.
If a solvent exchange for HPLC analysis is necessary, the concentrated extract is taken up in an
appropriate less volatile water miscible polar solvent and the non-polar extract residue is removed under
a gentle flux of inert gas.
The extract is analysed by GC-MS using a capillary column with a stationary phase of low polarity or by
HPLC-UV-DAD/FLD with an appropriate reversed phase column.
PAH are identified and quantified with GC-MS by comparison of relative retention times and relative peak
heights (or peak areas) with respect to internal standards added, and with HPLC by using the
corresponding variables of the extraction standard solutions. 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 affect the
sample during the contact time. Avoid plastics and other organic materials during sampling, sample
storage or extraction. Keep the samples from direct sunlight and prolonged exposure to light.
During storage of the samples, losses of PAH may occur due to adsorption on the walls of the containers.
The extent of the losses depends on the storage time.
5.2 Interference with GC-MS
Substances that co-elute with the target PAH can interfere with the determination. These interferences
can lead to incompletely resolved signals and can, depending on their magnitude, affect accuracy and
precision of the analytical results. Peak overlap does not allow an interpretation of the result.
Asymmetrical peaks and peaks broader than the corresponding peaks of the reference substance suggest
interferences.
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Chromatographic separation between dibenz[a,h]anthracene and indeno[1,2,3-cd]pyrene are mostly
critical. Due to their molecular mass differences, quantification can be made by mass selective detection.
When incomplete resolution is encountered, peak integration shall be checked and, if necessary,
corrected.
Sufficient resolution (e.g. 0,8) between the peaks of benzo[b]fluoranthene and benzo[k]fluoranthene as
well as of benzo[a]pyrene and benzo[e]pyrene shall be set as quality criteria for the capillary column.
Benzo[b]fluoranthene and benzo[j]fluoranthene cannot be separated. It can happen that the signal of
triphenylene is not completely separated from the signals of benz[a]anthracene and chrysene. In this case
it shall be stated in the report.
5.3 Interference with the HPLC
Substances that show either fluorescence or quenching and co-elute with the PAHs to be determined can
interfere with the determination. These interferences can lead to incompletely resolved signals and can,
depending on their magnitude, affect accuracy and precision of the analytical results. Peak overlap does
not allow an interpretation of the result. Asymmetrical peaks and peaks being broader than the
corresponding peaks of the reference substance suggest interferences. This problem can arise for
naphthalene and phenanthrene depending on the selectivity of the phases used.
Incomplete removal of the solvents used for sample extraction and clean-up can lead to poor
reproducibility of the retention times and wider peaks or double peaks especially for the 2- and 3-ring
PAHs. Extracts shall be diluted sufficiently with acetonitrile for the HPLC-analysis, otherwise the
detection of naphthalene and 3-ring PAH can be interfered with by a broad toluene peak.
Usually the signal of perylene is incompletely resolved from the signal of benzo[b]fluoranthene, but by
choosing a selective wavelength, the perylene peak can be suppressed.
6 Safety remarks
Certain PAH are highly carcinogenic and shall be handled with extreme care. Avoid contact with solid
materials, solvent extracts and solutions of standard PAH. It is strongly advised that standard solutions
are prepared centrally in suitably equipped laboratories or are purchased from suppliers specialized in
their preparation.
PAH contamination of vessels may be detected by irradiation with 366 nm UV-light.
Solvent solutions containing PAH 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 should be followed with respect to all hazards associated with this method.
9
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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 Toluene, C H .
7 8
7.2.3 Petroleum ether, boiling range 40 °C to 60 °C.
Hexane-like solvents with a boiling range between 40 °C and 98 °C are allowed.
7.2.4 Sodium sulfate, Na SO .
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
O.
7.2.5 Distilled water or water of equivalent quality, H
2
7.2.6 Sodium chloride, NaCl.
7.2.7 Keeper substance (High boiling compound, e.g. octane, nonane).
7.3 Reagents for clean-up
7.3.1 Clean-up A using aluminium oxide
7.3.1.1 Aluminium oxide, Al O
2 3.
2
Basic or neutral, specific surface 200 m /g, activity Super I [9].
7.3.1.2 Deactivated aluminium oxide.
Deactivated with approximately 100 g/kg 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 of two weeks.
The activity depends on the water content. It can be necessary to adjust the water content.
NOTE Commercially available aluminium oxides with 100 g/kg mass fraction water can also be used.
7.3.2 Clean-up B using silica gel 60 for column chromatography
7.3.2.1 Silica gel 60, particle size 63 µm to 200 µm.
7.3.2.2 Silica gel 60, water content (mass fraction) w(H O) = 10 %.
2
Silica gel 60 (7.3.2.1), heated for at least 3 h at 450 °C, cooled down and stored in a desiccator containing
magnesium perchlorate or a suitable drying agent. Before use, heat for at least for 5 h at 130 °C in a drying
oven. Then allow cooling in a desiccator. Put the silica gel in a stopperad flask and add 10 % water (7.2.5)
10
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(mass fraction), shake for 5 min intensively until all lumps have disappeared and then for 2 h in a shaking
device (8.1.3). Store the deactivated silica gel in the absence of air, use it for a maximum of two weeks.
7.3.3 Clean-up C using gel permeation chromatography (GPC)
1)
7.3.3.1 Bio-Beads® S-X3 .
7.3.3.2 Ethyl acetate, C H O .
4 8 2
7.3.3.3 Cyclohexane, C H .
6 12
®
Preparation of GPC, for example: Put 50 g Bio-Beads S-X3 (7.3.3.1) into a 500 ml Erlenmeyer flask and
add 300 ml elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 1: 1 (volume
fraction) in order to allow the beads to swell. After swirling for a short time until no lumps are left,
maintain the flask closed for 24 h. Drain the slurry into the chromatography tube for GPC. After
approximately three days, push in the plungers of the column so that a filling level of
approximately 35 cm is obtained. To further compress the gel, pump approximately 2 l of elution mixture
−1
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.
7.4 Reagents for chromatographic analysis
7.4.1 GC-Analysis
7.4.1.1 Carrier gas for GC-MS.
Operating gases of high purity and in accordance with the manufacturer’s specifications.
7.4.2 HPLC-analysis
7.4.2.1 Acetonitrile (CH CN) or methanol (CH OH), HPLC purity grade.
3 3
7.4.2.2 Ultra-pure water, HPLC purity grade.
7.4.2.3 Helium, He, of suitable purity for degasification of solvents.
7.5 Standards
7.5.1 General
Choose the internal and/or extraction standards whose physical and chemical properties (such as
extraction behaviour, retention time) are similar to those of the compounds to be analysed. For GC-MS a
minimum of four deuterated internal standards shall be used as internal standards for evaluation of
results.
7.5.2 Calibration substances and internal standards
Table 3 contains native and deuterated PAH to be used for calibration. Verify the stability of the internal
standards regularly.
1)
Bio-Beads® is an example of a suitable product available commercially. This information is given for
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
Equivalent products can be used if they can be shown to lead to the same results.
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Table 3 — Native PAH and deuterated PAH
PAH reference substances Internal standard substances (deuterated PAHs)
Naphthalene (CAS-RN 91-20-3) Naphthalene-d8 (CAS-RN 1146-65-2)
Acenaphthene (CAS-RN 83-32-9) Acenaphthene-d10 (CAS-RN 15067-26-2)
Acenaphthylene (CAS-RN 208-96-8) Acenaphthylene-d8 (CAS-RN 93951-97-4)
Fluorene (CAS-RN 86-73-7) Fluorene-d10 (CAS-RN 81103-79-9)
Anthracene (CAS-RN 120-12-7) Anthracene-d10 (CAS-RN 1719-06-8)
Phenanthrene (CAS-RN 85-01-8) Phenanthrene-d10 (CAS-RN 1517-22-2)
Fluoranthene (CAS-RN 206-44-0) Fluoranthene-d10 (CAS-RN 93951-69-0)
Pyrene (CAS-RN 129-00-0) Pyrene-d10 (CAS-RN 1718- 52–1)
Benz[a]anthracene (CAS-RN 56-55-3) Benz[a]anthracene-d12 (CAS-RN 1718-53-2)
Chrysene (CAS-RN 218-01-9) Chrysene-d12 (CAS-RN 1719-03-5)
Benzo[b]fluoranthene (CAS-RN 205-99-2) Benzo[b]fluoranthene-d12 (CAS-RN 93951-98-5)
Benzo[k]fluoranthene (CAS-RN 207-08-9) Benzo[k]fluoranthene-d12 (CAS-RN 93952-01-3)
a
(CAS-RN 205-82-3)
Benzo[j]fluoranthene
Benzo[a]pyrene (CAS-RN 50-32-8) Benzo[a]pyrene-d12 (CAS-RN 63466-71-7)
a
Benzo[e]pyrene (CAS-RN 192-97-2)
Indeno[1,2,3-cd]pyrene (CAS-RN 193-39-5) Indeno[1,2,3-cd]pyrene-d12 (CAS-RN 203578-33-0)
Dibenz[a,h]anthracene (CAS-RN 53-70-3) Dibenz[a,h]anthracene -d14 (CAS-RN 13250-98-1)
Benzo[ghi]perylene (CAS-RN 191-24-2) Benzo[ghi]perylene-d12 (CAS-RN 93951-66-7)
a
Not part of 16 target analytes, but applicable for resolution check for the separation with benzo[a]pyrene for
GC measurements.
13
NOTE 1 C -labelled PAH standards can also be used as internal standards.
12
NOTE 2 Certified solutions of PAH, and single solid PAH substances with certified purity are available from a
limited number of suppliers.
When highly contaminated samples are analysed, an aliquot of the extract is often used for further clean-
up. This makes the costs of analyses caused by the use of deuterated standard very high. In these cases,
it is allowed to add the internal standard in two steps. Step 1 addition of unlabelled internal standards to
the sample. Step 2 addition of deuterated compounds to the aliquot of the extract used for clean-up.
For HPLC 6-methylchrysene, 1-methylnaphthalene or other alkylated PAH which are not present in the
sample and which are sufficient separated from the target PAH can be used as internal or extraction
standard.
7.5.3 Injection standard
If required for GC-MS a deuterated PAH such as 1-methylnaphthalene-d10, triphenylene-d12 and
perylene-d12 can be added to the final extract before GC-MS injection t
...
SLOVENSKI STANDARD
oSIST prEN 17503:2020
01-maj-2020
Trdni matriksi z vidika okolja - Določevanje policikličnih aromatskih
ogljikovodikov (PAH) s plinsko kromatografijo (GC) in s tekočinsko kromatografijo
visoke ločljivosti (HPLC)
Environmental solid matrices - Determination of polycyclic aromatic hydrocarbons (PAH)
by gas chromatography (GC) and high performance liquid chromatography (HPLC)
Umweltrelevante feste Matrizes - Bestimmung von polycyclischen aromatischen
Kohlenwasserstoffen (PAK) mittels Gaschromatograpie (GC) und Hochleistungs-
Flüssigkeitschromatographie (HPLC)
Matrices solides environnnementales - Determination des hydrocarbures aromatiques
polycycliques (HAP)par chromatographie en phase gazeuse (GC) et chromatographie
liquid a haute performance (HPLC)
Ta slovenski standard je istoveten z: prEN 17503
ICS:
13.030.01 Odpadki na splošno Wastes in general
13.080.10 Kemijske značilnosti tal Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
oSIST prEN 17503:2020 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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DRAFT
EUROPEAN STANDARD
prEN 17503
NORME EUROPÉENNE
EUROPÄISCHE NORM
May 2020
ICS 13.030.01; 13.080.10 Will supersede EN 15527:2008, EN 16181:2018
English Version
Environmental solid matrices - Determination of polycyclic
aromatic hydrocarbons (PAH) by gas chromatography
(GC) and high performance liquid chromatography (HPLC)
Matrices solides environnnementales ¿ Determination Umweltrelevante feste Matrizes ¿ Bestimmung von
des hydrocarbures aromatiques polycycliques polycyclischen aromatischen Kohlenwasserstoffen
(HAP)par chromatographie en phase gazeuse (GC) et (PAK) mittels Gaschromatograpie (GC) und
chromatographie liquid a haute performance (HPLC) Hochleistungs-Flüssigkeitschromatographie (HPLC)
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 444.
If this draft becomes a European Standard, 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.
This draft European Standard was established by CEN 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.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.
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. prEN 17503:2020 E
worldwide for CEN national Members.
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Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 8
5 Interferences . 8
6 Safety remarks . 9
7 Reagents . 9
8 Apparatus . 13
9 Sample storage and preservation . 15
10 Procedure . 16
11 Performance characteristics. 29
12 Precision . 29
13 Test report . 30
Annex A (informative) Repeatability and reproducibility data . 31
A.1 Materials used in the interlaboratory comparison study . 31
A.2 Interlaboratory comparison results . 31
Annex B (informative) Examples of instrumental conditions and chromatograms . 41
B.1 Measurement of PAH with GC-MS . 41
B.2 Measurement of PAH with HPLC fluorescence . 47
B.3 Example for measurement conditions of PAH with GC-MS/MS . 53
Bibliography . 55
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European foreword
This document (prEN 17503:2020) has been prepared by Technical Committee CEN/TC 444 “Test
methods for environmental characterization of solid matrices”, the secretariat of which is held by NEN.
This document is currently submitted to the CEN Enquiry.
This document will supersede EN 15527:2008 and EN 16181:2018.
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Introduction
Polycyclic aromatic hydrocarbons (PAH) are ubiquitous because they are released in appreciable
quantities every year into the environment through the combustion of organic matters such as coal, fuel
oils, petrol, wood, refuse and plant materials. Since some of these PAH compounds are carcinogenic or
mutagenic, their presence in the environment (air, water, soil, sediment and waste) is regularly
monitored and controlled. At present determination of PAH is carried out in these matrices in most of the
routine laboratories following the preceding steps for sampling, pre-treatment, extraction, clean-up by
measurement of specific PAH by means of gas chromatography in combination with mass spectrometric
detection (GC-MS) or by high performance liquid chromatography (HPLC) in combination with UV-DAD-
or fluorescence-detection (HPLC-UV-DAD/FLD). Both the GC-MS and the HPLC methods are included in
this horizontal standard.
This document was developed by merging of EN 16181:2018 [1], initially elaborated as a CEN Technical
Specification in the European project 'HORIZONTAL' and validated by CEN/TC400 with the support of
BAM, with EN 15527 [2], published by CEN/TC292.
Considered the different matrices and possible interfering compounds, this document does not contain
one single possible way of working. Several choices are possible, in particular relating to clean-up.
Quantification with both GC-MS-detection and HPLC-DAD-UV/FLD is possible. Four different extraction
procedures are described and three clean-up procedures. The use of internal and injection standards is
described in order to have an internal check on choice of the extraction and clean-up procedure. The
method is as far as possible in agreement with the method described for PCB (EN17322). It has been
tested for ruggedness.
This document is applicable and validated for several types of matrices as indicated in Table 1 (see also
Annex A for the results of the validation).
Table 1 — Matrices for which this document is applicable and validated
Matrix Materials used for validation
Soil Sandy soil
Mix of soil from an industrial area in Brandenburg, Germany and
PCB-free German reference soil
Sludge Mix of municipal waste water treatment plant sludge from the
vicinity of Berlin, Germany
Biowaste Mix of compost from the vicinity of Berlin, Germany
Contaminated soil, building debris, waste wood, roofing tar,
Waste
shredder light fraction
WARNING — Persons using this document should be familiar with usual laboratory practice. This
document does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices and to ensure compliance
with any national regulatory conditions.
WARNING — It is absolutely essential that tests conducted according to this document be carried out by
suitably trained staff.
4
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1 Scope
This document specifies different methods for quantitative determination of 16 polycyclic aromatic
hydrocarbons (PAH) (see Table 2) in soil, sludge, sediment, treated biowaste, and waste, using GC-MS or
HPLC-UV-DAD/FLD covering a wide range of PAH contamination levels (see Table 2).
When using fluorescence detection, acenaphthylene cannot be measured.
Table 2 —Target analytes of this document
Target analyte a
CAS-RN
Naphthalene 91–20–3
Acenaphthene 83–32–9
Acenaphthylene 208–96–8
Fluorene 86–73–7
Anthracene 120–12–7
Phenanthrene 85–01–8
Fluoranthene 206–44–0
Pyrene 129–00–0
Benz[a]anthracene 56–55–3
Chrysene 218–01–9
Benzo[b]fluoranthene 205–99–2
Benzo[k]fluoranthene 207–08–9
Benzo[a]pyrene 50–32–8
Indeno[1,2,3-cd]pyrene 193–39–5
Dibenz[a,h]anthracene 53–70–3
Benzo[ghi]perylene 191–24–2
a
CAS-RN Chemical Abstracts Service Registry Number.
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used
for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this document, lower limit of application from 10 μg/kg (expressed as
dry matter) for soils, sludge and biowaste to 100 μg/kg (expressed as dry matter) for solid waste can be
achieved. For some specific samples (e.g. bitumen) the limit of 100 μg/kg cannot be reached.
Sludge, waste and treated biowaste may differ in properties as well as in the expected contamination
levels of PAH and presence of interfering substances. These differences make it impossible to describe
one general procedure. This document contains decision tables based on the properties of the sample and
the extraction and clean-up procedure to be used.
The method can be applied to the analysis of other PAH not specified in the scope, provided suitability is
proven by proper in-house validation experiments.
5
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2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 5667-15, Water quality - Sampling - Part 15: Guidance on the preservation and handling of sludge
and sediment samples (ISO 5667-15)
EN ISO 16720, Soil quality - Pretreatment of samples by freeze-drying for subsequent analysis (ISO 16720)
EN ISO 22892, Soil quality - Guidelines for the identification of target compounds by gas chromatography
and mass spectrometry (ISO 22892)
ISO 8466-1, Water quality — Calibration and evaluation of analytical methods and estimation of
performance characteristics — Part 1: Statistical evaluation of the linear calibration function
ISO 18512, Soil quality — Guidance on long and short term storage of soil samples
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at http://www.iso.org/obp
3.1
critical pair
pair of PAH that shall be separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic
separation meets minimum quality criteria
6
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t
R2 (1)
YY
ab
Where:
R peak separation
Δt difference in retention times of the two peaks a and b in seconds (s)
Y peak width at the base of peak a in seconds (s)
a
Y peak width at the base of peak b in seconds (s)
b
Figure 1 — Example of a chromatogram of a critical pair
3.2
internal standard
isotopically labelled mostly deuterated PAH or native PAH unlikely to be present in the sample, added to
samples before extraction and used for quantification of the PAH content
3.3
injection standard
PAH that is unlikely to be present in samples, added to the sample extract before injection into the gas
chromatograph, to monitor variability of instrument response and the recovery of the internal standards
3.4
extraction standard
PAH that is unlikely to be present in samples added to the sample prior to extraction. Used for checking
the extraction efficiency and not used for quantification purposes
7
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3.5
sediment
solid material, both mineral and organic, deposited in the bottom of a water body
[SOURCE: ISO 5667-12:2017 definition 3.5]
4 Principle
Due to the multi matrix character of this document, different procedures for different steps (modules)
are allowed. Which modules should be used depends on the sample. A recommendation is given in this
European Standard. Performance criteria are described and it is the responsibility of the laboratories
applying this European Standard to show that these criteria are met. Using of spiking standards (internal
standards) allows an overall check on the efficiency of a specific combination of modules for a specific
sample. But it does not necessarily give the information regarding the extensive extraction efficiency of
the native PAH bonded to the matrix.
After pre-treatment, the sample is extracted with a suitable solvent.
The extract is concentrated by evaporation. If necessary, interfering compounds are removed by a clean-
up method suitable for the specific matrix, before the concentration step.
If a solvent exchange for HPLC analysis is necessary, the concentrated extract is taken up in an
appropriate less volatile water miscible polar solvent and the non-polar extract residue is removed.
The extract is analysed by GC-MS using a capillary column with a stationary phase of low polarity or by
HPLC-UV-DAD/FLD with an appropriate reversed phase column.
PAH are identified and quantified with GC-MS by comparison of relative retention times and relative peak
heights (or peak areas) with respect to internal standards added, and with HPLC by using the
corresponding variables of the extraction standard solutions. 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 affect the
sample during the contact time. Avoid plastics and other organic materials during sampling, sample
storage or extraction. Keep the samples from direct sunlight and prolonged exposure to light.
During storage of the samples, losses of PAH may occur due to adsorption on the walls of the containers.
The extent of the losses depends on the storage time.
5.2 Interference with GC-MS
Substances that co-elute with the target PAH may interfere with the determination. These interferences
may lead to incompletely resolved signals and may, depending on their magnitude, affect accuracy and
precision of the analytical results. Peak overlap does not allow an interpretation of the result.
Asymmetrical peaks and peaks broader than the corresponding peaks of the reference substance suggest
interferences.
Chromatographic separation between dibenz[a,h]anthracene and indeno[1,2,3-cd]pyrene are mostly
critical. Due to their molecular mass differences, quantification can be made by mass selective detection.
When incomplete resolution is encountered, peak integration shall be checked and, if necessary,
corrected.
8
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Sufficient resolution (e.g. 0,8) between the peaks of benzo[b]fluoranthene and benzo[k]fluoranthene as
well as of benzo[a]pyrene and benzo[e]pyrene shall be set as quality criteria for the capillary column.
Benzo[b]fluoranthene and benzo[j]fluoranthene cannot be separated. Triphenylene may not be
completely separated from benz[a]anthracene and chrysene. In this case it shall be stated in the report.
5.3 Interference with the HPLC
Substances that show either fluorescence or quenching and co-elute with the PAHs to be determined may
interfere with the determination. These interferences may lead to incompletely resolved signals and may,
depending on their magnitude, affect accuracy and precision of the analytical results. Peak overlap does
not allow an interpretation of the result. Asymmetrical peaks and peaks being broader than the
corresponding peaks of the reference substance suggest interferences. This problem may arise for
naphthalene and phenanthrene depending on the selectivity of the phases used.
Incomplete removal of the solvents used for sample extraction and clean-up may lead to poor
reproducibility of the retention times and wider peaks or double peaks especially for the 2- and 3-ring
PAHs. Extracts shall be diluted sufficiently with acetonitrile for the HPLC-analysis, otherwise the
detection of naphthalene and 3-ring PAH can be interfered with by a broad toluene peak.
Usually perylene is incompletely resolved from benzo[b]fluoranthene, but by choosing a selective
wavelength, the perylene peak can be suppressed.
6 Safety remarks
Certain PAH are highly carcinogenic and shall be handled with extreme care. Avoid contact with solid
materials, solvent extracts and solutions of standard PAH. Contact of solutions of standard with the body
should be prevented. It is strongly advised that standard solutions are prepared centrally in suitably
equipped laboratories or are purchased from suppliers specialized in their preparation.
PAH contamination of vessels may be detected by irradiation with 366 nm UV-light.
Solvent solutions containing PAH 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 should be followed with respect to all hazards associated with this method.
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 Toluene, C H .
7 8
7.2.3 Petroleum ether, boiling range 40 °C to 60 °C
Hexane-like solvents with a boiling range between 40 °C and 98 °C are allowed.
7.2.4 Sodium sulfate, Na SO .
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
9
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7.2.5 Distilled water or water of equivalent quality, H O.
2
7.2.6 Sodium chloride, NaCl
7.2.7 Keeper substance. High boiling compound, i.e. octane, nonane
7.3 Reagents for clean-up
7.3.1 Clean-up A using aluminium oxide
7.3.1.1 Aluminium oxide, Al O
2 3
2
Basic or neutral, specific surface 200 m /g, activity Super I [11].
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 of two weeks.
NOTE 1 The activity depends on the water content. It might be necessary to adjust the water content.
NOTE 2 Commercially available aluminium oxides with 10 % mass fraction water can also be used.
7.3.2 Clean-up B using silica gel 60 for column chromatography
7.3.2.1 Silica gel 60, particle size 63 µm to 200 µm
7.3.2.2 Silica gel 60, water content: mass fraction w(H O) = 10 %.
2
Silica gel 60 (7.3.2.1), heated for at least 3 h at 450 °C, cooled down and stored in a desiccator containing
magnesium perchlorate or a suitable drying agent. Before use, heat at least for 5 h at 130 °C in a drying
oven. Then allow cooling in a desiccator and add 10 % water (7.2.5) (mass fraction) in a flask. Shake for
5 min intensively until all lumps have disappeared and then for 2 h in a shaking device (8.1.3). Store the
deactivated silica gel in the absence of air, use it for a maximum of two weeks.
7.3.3 Clean-up C using gel permeation chromatography (GPC)
7.3.3.1 Bio-Beads® S-X3
®
NOTE Bio-Beads is an example of a suitable product available commercially. This information is given for
convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.
Equivalent products can be used if they can be shown to lead to the same results.
7.3.3.2 Ethyl acetate, C4H8O2
7.3.3.3 Cyclohexane, C H
6 12
®
Preparation of GPC, for example: Put 50 g Bio-Beads S-X3 (7.3.3.1) into a 500 ml Erlenmeyer flask and
add 300 ml elution mixture made up of cyclohexane (7.3.3.3) and ethyl acetate (7.3.3.2) 1:1 (volume
fraction) in order to allow the beads to swell. After swirling for a short time until no lumps are left,
maintain the flask closed for 24 h. Drain the slurry into the chromatography tube for GPC. After
approximately three days, push in the plungers of the column so that a filling level of
approximately 35 cm is obtained. To further compress the gel, pump approximately 2 l of elution mixture
−1
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.
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7.4 Reagents for chromatographic analysis
7.4.1 GC-Analysis
7.4.1.1 Carrier gas for GC-MS
Operating gases of high purity and in accordance with the manufacturer’s specifications.
7.4.2 HPLC-analysis
7.4.2.1 Acetonitrile (CH CN) or methanol (CH OH), HPLC purity grade
3 3
7.4.2.2 Ultra-pure water, HPLC purity grade.
7.4.2.3 Helium, He, of suitable purity for degasification of solvents
7.5 Standards
7.5.1 General
Choose the internal and/or extraction standards whose physical and chemical properties (such as
extraction behaviour, retention time) are similar to those of the compounds to be analysed. For GC-MS a
minimum of four deuterated internal standards shall be used as internal standards for evaluation of
results.
7.5.2 Calibration substances and internal standards
Table 3 contains native and deuterated PAH to be used for calibration. Verify the stability of the internal
standards regularly.
Table 3 — Native PAH and deuterated PAH
PAH reference substances Internal standard substances deuterated PAHs
Naphthalene (CAS-RN 91-20-3) Naphthalene-d8 (CAS-RN 1146-65-2)
Acenaphthene (CAS-RN 83-32-9) Acenaphthene-d10 (CAS-RN 15067-26-2)
Acenaphthylene (CAS-RN 208-96-8) Acenaphthylene-d8 (CAS-RN 93951-97-4)
Fluorene (CAS-RN 86-73-7) Fluorene-d10 (CAS-RN 81103-79-9)
Anthracene (CAS-RN 120-12-7) Anthracene-d10 (CAS-RN 1719-06-8)
Phenanthrene (CAS-RN 85-01-8) Phenanthrene-d10 (CAS-RN 1517-22-2)
Fluoranthene (CAS-RN 206-44-0) Fluoranthene-d10 (CAS-RN 93951-69-0)
Pyrene (CAS-RN 129-00-0) Pyrene-d10 (CAS-RN 1718- 52–1)
Benz[a]anthracene (CAS-RN 56-55-3) Benz[a]anthracene-d12 (CAS-RN 1718-53-2)
Chrysene (CAS-RN 218-01-9) Chrysene-d12 (CAS-RN 1719-03-5)
Benzo[b]fluoranthene (CAS-RN 205-99-2) Benzo[b]fluoranthene-d12 (CAS-RN 93951-98-5)
Benzo[k]fluoranthene (CAS-RN 207-08-9) Benzo[k]fluoranthene-d12 (CAS-RN 93952-01-3)
(CAS-RN-205–82–
a
Benzo[j]fluoranthene
3)
Benzo[a]pyrene (CAS-RN 50-32-8) Benzo[a]pyrene-d12 (CAS-RN 63466-71-7)
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PAH reference substances Internal standard substances deuterated PAHs
a
(CAS-RN 192-97-2)
Benzo[e]pyrene
Indeno[1,2,3-cd]pyrene (CAS-RN 193-39-5) Indeno[1,2,3-cd]pyrene-d12 (CAS-RN 203578-33-0)
Dibenz[a,h]anthracene (CAS-RN 53-70-3) Dibenz[a,h]anthracene -d14 (CAS-RN 13250-98-1)
Benzo[ghi]perylene (CAS-RN 191-24-2) Benzo[ghi]perylene-d12 (CAS-RN 93951-66-7)
a
Not part of 16 target analytes, but applicable for resolution check for the separation with benzo[a]pyrene for
GC measurements
13
NOTE 1 C12-labelled PAH standards can also be used as internal standards.
NOTE 2 Certified solutions of PAH, and single solid PAH substances with certified purity are available from a
limited number of suppliers.
When highly contaminated samples are analysed, an aliquot of the extract is often used for further clean-
up. This makes the costs of analyses caused by the use of deuterated standard very high. In these cases,
it is allowed to add the internal standard in two steps. Step 1 addition of unlabelled internal standards to
the sample. Step 2 addition of deuterated compounds to the aliquot of the extract used for clean-up.
For HPLC 6-methylchrysene, 1-Methylnaphthalene or other alkylated PAH which are not present in the
sample and which are sufficient separated from the target PAH can be used as internal or extraction
standard.
7.5.3 Injection standard
If required for GC-MS a deuterated PAH such as 1-methylnaphthalene-d10, triphenylene-d12 and
perylene-d12 can be added to the final extract before GC-MS injection to verify the recovery of the
deuterated internal standards.
7.6 Preparation of standard solutions
7.6.1 General
The procedure for the preparation of standard solutions for GC-MS and HPLC is the same only different
solvents are used.
Because of the dangerous nature of the substances to be used, commercially available - preferably
certified - standard solutions or mixed standard solutions are preferred. It is recommended to avoid skin
contact.
The working standard solutions shall be in the same solvent as the extract.
Store the primary and diluted standard solutions in a dark place at a temperature of (5 ± 3) °C. The
solutions are stable for at least one year, provided that evaporation of solvent is negligible.
Components present in mixed standard solutions should be separated by the chromatographic columns
used.
7.6.2 Preparation of calibration standard solutions for GC-MS
Prepare individual concentrated primary standard solutions of about 0,4 mg/ml in hexane-like solvent
(7.2.3) by weighing approximately 10 mg of each of the calibration standards (7.5.2 Table 3 left column)
to the nearest 0,1 mg and dissolving them in 25 ml of hexane-like solvent.
Combine small quantities (2 ml to 10 ml) of these individual primary standard solutions into a mixed
standard solution of PAH.
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Because of the dangerous nature of the substances to be used, commercially available - preferably
certified - standard solutions or mixed standard solutions are preferred. It is recommended to avoid skin
contact.
7.6.3 Preparation of internal standard solution for GC-MS
Prepare a concentrated primary internal
...
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