SIST-TS CEN/TS 16189:2012

SLOVENSKI STANDARD
01-junij-2012
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Sludge, treated biowaste and soil - Determination of linear alkylbenzene sulfonates
(LAS) by high-performance liquid chromatography (HPLC) with fluorescence detection
(FLD) or mass selective detection (MS)
Schlamm, behandelter Bioabfall und Boden - Bestimmung von Linearen
Alkylbenzolsulfonaten (LAS) mittels Hochleistungs-Flüssigkeitschromatographie (HPLC)
mit Fluoroszenzdetektion (FLD) oder massenselektiver Detektion (MS)
Boues, bio-déchets traités et sols - Détermination des alkylbenzènesulfonates linéaires
(LAS) par chromatographie liquide à haute performance (CLHP) avec détection par
fluorescence (FLD) ou détection sélective de masse (SM)
Ta slovenski standard je istoveten z: CEN/TS 16189:2012
ICS:
13.030.20 7HNRþLRGSDGNL%ODWR Liquid wastes. Sludge
13.080.10 .HPLMVNH]QDþLOQRVWLWDO Chemical characteristics of
soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

TECHNICAL SPECIFICATION
CEN/TS 16189
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
February 2012
ICS 13.030.01
English Version
Sludge, treated biowaste and soil - Determination of linear
alkylbenzene sulfonates (LAS) by high-performance liquid
chromatography (HPLC) with fluorescence detection (FLD) or
mass selective detection (MS)
Boues, biodéchets traités et sols - Détermination des
Schlamm, behandelter Bioabfall und Boden - Bestimmung
alkylbenzènesulfonates linéaires (LAS) par von linearen Alkylbenzolsulfonaten (LAS) mittels
chromatographie liquide à haute performance (CLHP) avec Hochleistungs-Flüssigkeitschromatographie (HPLC) mit
détection par fluorescence (FLD) ou détection sélective de Fluoreszenzdetektion (FLD) oder massenselektiver
masse (SM) Detektion (MS)
This Technical Specification (CEN/TS) was approved by CEN on 24 April 2011 for provisional application.

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

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

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.

EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16189:2012: E
worldwide for CEN national Members.

Contents Page
Foreword .3
Introduction .4
1 Scope .5
2 Normative references .5
3 Terms and definitions .5
4 Principle .5
5 Interferences .6
5.1 Interferences from sampling .6
5.2 Interferences by HPLC-FLD and HPLC-MS .6
6 Reagents .6
7 Apparatus .9
8 Sample storage and sample pretreatment . 10
8.1 Sample storage . 10
8.2 Sample pretreatment . 10
9 Procedure . 10
9.1 Extraction . 10
9.1.1 Extraction of dried sludge . 10
9.1.2 Extraction of dried soil, sediment and treated biowaste . 10
9.2 Concentration (optional) . 11
9.3 Clean-up (optional) . 11
9.4 Blank test . 11
9.5 HPLC analysis . 11
9.5.1 General . 11
9.5.2 Fluorescence detection . 12
9.5.3 Mass selective detection . 12
9.6 Calibration . 12
9.6.1 General . 12
9.6.2 Initial calibration . 12
9.6.3 Verification of calibration . 13
9.7 Analysis of samples and identification . 13
10 Calculation and expression of results . 13
10.1 General . 13
10.2 Calibration . 14
10.3 Calculation . 14
11 Precision . 15
12 Test report . 15
Annex A (informative) Repeatability and reproducibility data . 16
A.1 Materials used in the interlaboratory comparison study . 16
A.2 Interlaboratory comparison results . 17
Annex B (informative) Examples of chromatographic conditions and chromatograms . 18
B.1 Isomeric separation of LAS . 18
B.2 Homologue separation of LAS . 19
Annex C (informative) Examples of clean-up procedures . 20
C.1 Clean-up procedure based on graphitised carbon black (GCB) columns . 20
C.2 Clean-up procedure based on strong anion exchange (SAX) columns . 20
Bibliography . 21

Foreword
This document (CEN/TS 16189:2012) has been prepared by Technical Committee CEN/TC 400 “Project
Committee - Horizontal standards in the fields of sludge, biowaste and soil”, the secretariat of which is held by
DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
The preparation of this document by CEN is based on a mandate by the European Commission
(Mandate M/330), which assigned the development of standards on sampling and analytical methods for
hygienic and biological parameters as well as inorganic and organic determinants, aiming to make these
standards applicable to sludge, treated biowaste and soil as far as this is technically feasible.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy,
Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia,
Spain, Sweden, Switzerland, Turkey and the United Kingdom.

Introduction
The anionic surfactant LAS (Linear Alkylbenzene Sulfonate) is found in the environment due to the use of LAS
in detergents. For more than 30 years LAS has been the largest single surfactant used in detergents, and the
use continues on a high level.
Although LAS is readily biodegradable during wastewater treatment, considerable amounts may still be found
in sludge of municipal origin. By the use of sludge for soil improvement LAS may end up in the agricultural
soil, where a rapid biodegradation takes place.
The method describes the determination of LAS in sludge, soil, treated biowaste and neighbouring fields. LAS
is the sodium salt of alkylbenzene sulfonic acids, and it consists of a mixture of the homologues C -LAS,
C -LAS, C -LAS, C -LAS and C -LAS. LAS is determined as the sum of the homologues.
11 12 13 14
This Technical Specification 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 Technical Specification is applicable and validated
Matrix Materials used for validation
Sludge Municipal sewage sludge
Biowaste Fresh compost
Soil Sludge amended soil
WARNING — Persons using this Technical Specification should be familiar with usual laboratory
practice. This Technical Specification does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this Technical
Specification be carried out by suitably trained staff.

1 Scope
This Technical Specification specifies a method for the determination of linear alkylbenzene sulfonate (LAS) in
sludge, treated biowaste and soil using high-performance liquid chromatography (HPLC) with a fluorescence
detector (FLD) or a mass selective detector (MS).
This Technical Specification specifies the determination of the sum of LAS. Under the conditions specified in
this Technical Specification, typically a limit of detection of 20 mg/kg (expressed as dry matter) for sludge and
of 0,2 mg/kg to 0,5 mg/kg for soil and treated biowaste may be achieved.
Lower limits of detection may be achieved by concentrating the extract by solvent evaporation.
NOTE The single LAS homologues C to C can be determined by this Technical Specification.
10 14
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
EN 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 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
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
analyte
mixture of homologues (i. e. C -LAS, C -LAS, C -LAS, C -LAS and C -LAS) where each homologue
10 11 12 13 14
consists of a mixture of four to six isomers depending on the length of the alkyl group
Note 1 to entry: The dominant homologues in detergents and environmental samples are C -LAS and C -LAS. C to
11 12 10
C refers to the chain length of the linear alkyl group.
4 Principle
After pretreatment, the test sample is extracted by shaking with methanol. If necessary, interfering compounds
are removed from the extract by a clean-up on a suitable column.
The extract is analysed by high performance liquid chromatography (HPLC) on a C - or C -column and
8 18
detection by fluorescence (FLD) or mass spectrometry (MS).
The identification is based on the retention times of the homologues and of the isomers of each homologue.
Another identification point is the pattern/fingerprint of the homologues, and the isomer fingerprint of each
homologue if a C -column is used for HPLC. By use of MS detection the relative intensities of two diagnostic
ions may also be used for the identification (optional).
The quantification is based on an internal standard procedure. The internal standard (C -LAS) is taken
through the whole analytical procedure.
Depending on the type of matrices from which LAS is extracted, different analytical pathways can be applied.
An overview of the analytical procedure for the matrix of interest is shown in Table 2.
5 Interferences
5.1 Interferences from sampling
Use sampling containers of materials (preferably glass or steel) that do not significantly affect the sample
during the contact through sampling and storage. Plastic containers may be used if it has been proven that
they do not significantly affect the sample.
5.2 Interferences by HPLC-FLD and HPLC-MS
The chromatographic analysis can be done on a C - or a C -reverse phase column, and the choice of
8 18
column determines the separation obtained. On the C -column (with methanol in mobile phase) the LAS
homologues are separated, however, there is no separation of the isomers. On the C -column (with
acetonitrile in mobile phase) the homologues are separated and there is a partial separation of the isomers of
each homologue. This is illustrated by the chromatograms in Annex B.
The selectivity of the fluorescence as well as the mass selective detector is high; however, interference from
co-eluting substances may occur. It is essential that the interfering peaks are not included in the calculations.
A peak is excluded if the retention time differs from the LAS standard mixture. Interfering peaks can best be
detected when a C -column is used for the LC analysis, due to the partial separation of the isomers. The
C -column is mandatory when fluorescence is used, due to the higher selectivity obtained. The interfering
peaks can usually be detected by comparing the fingerprints of the sample with the fingerprints of the LAS
standard mixture, although the isomer and homologue distribution in the environmental samples may differ
from the distribution in the standard mixture.
The highest selectivity is obtained by the use of a C -column and the MS detector. However, for most
applications the separation on a C -column is sufficient, when MS is used. When all isomers are eluted in one
peak, the integrations are less complicated, resulting in a higher precision and a lower limit of detection.
Table 2 — Choice of analytical procedure
Matrix FLD MS
C-column C-column C-column C -column
8 18 8 18
Sludge No Yes Yes Yes
a
Soil No (Yes) Yes Yes
a
Treated biowaste No (Yes) Yes Yes

a
For FLD the limit of detection will generally be inadequate for this type of matrix.

6 Reagents
6.1 General
Use only reagents of recognized analytical grade, unless otherwise specified.
The purity of the reagents used shall be checked by running a blank determination as described in 9.4.
6.2 Methanol, CH OH; HPLC-grade.
6.3 Acetonitrile, C H N; HPLC-grade.
2 3
- +
6.4 Ammonium acetate, c(CH COO NH ) = 0,01 mol/l.
3 4
6.5 Mobile phases for HPLC
6.5.1 For isomeric separation on C -column
 Mobile phase A: 0,01 mol/l ammonium acetate (6.4);
 Mobile phase B: Acetonitrile (6.3).
6.5.2 For homologue separation on C -column
 Mobile phase A: 0,01 mol/l ammonium acetate (6.4);
 Mobile phase B: Methanol (6.2).
6.6 Reagents for clean-up procedures
6.6.1 Clean-up procedure based on strong anion exchange (SAX)
6.6.1.1 SAX column
6.6.1.2 Acetic acid, CH COOH
6.6.1.3 Hydrochloric acid, HCl
6.6.1.4 Methanol, CH OH
6.6.2 Clean-up procedure based on graphitised carbon black (GCB)
6.6.2.1 GCB column
6.6.2.2 Hydrochloric acid, HCl
1)
6.6.2.3 Tetramethylammonium hydroxide, C H NO (CAS-RN 10424-65-4 ); pentahydrate.
4 13
6.6.2.4 Formic acid, HCOOH
6.6.2.5 Dichloromethane, CH Cl
2 2
6.6.2.6 Methanol, CH OH
6.7 Nitrogen, N , for solvent evaporation of sufficient purity.
6.8 Standards for calibration
6.8.1 General
The standards shall be stored in a freezer at a temperature of (–18 ± 3) °C.

1)
CAS-RN Chemical Abstracts Service Registry Number.
6.8.2 C -LAS, sodium linear undecylbenzene sulfonate, C H SO Na; 99 %.
11 17 27 3
6.8.3 C -LAS, sodium linear dodecylbenzene sulfonate, C H SO Na; 99 % (CAS-RN 2211-98-5).
12 18 29 3
6.8.4 C -LAS, sodium linear tridecylbenzene sulfonate, C H SO Na; 99 %.
13 19 31 3
6.8.5 C -C -LAS mixture of homologues and isomers, highest possible purity (CAS-RN 69669-44-9,
10 14
CAS-RN 25155-30-0).
6.9 Internal standard, C -LAS
Octylbenzene sulfonic acid, sodium salt C H SO Na (CAS-RN 6149-03-7).
14 21 3
The internal standard shall be stored in the freezer at a temperature of (–18 ± 3) °C.
6.10 Internal standard solution
Prepare the internal standard solution of the internal standard (6.9) by dilution to about 1 000 mg/l in methanol
(6.2).
It is essential that the same internal standard solution is used for calibration standard solutions and for
samples, blank tests and internal quality control samples.
Store the internal standard solution in a dark place at a temperature of (4 ± 3) °C. The solution is stable for at
least two years.
6.11 Stock solutions
Prepare individual stock solutions of 1 000 mg/l to 5
...