SIST EN 15549:2008

SLOVENSKI STANDARD
01-julij-2008
.DNRYRVW]UDND6WDQGDUGQDPHWRGD]DGRORþHYDQMHNRQFHQWUDFLMHEHQ]R>D@SLUHQD
%>D@3Y]XQDQMHP]UDNX
Air quality - Standard method for the measurement of the concentration of benzo[a]
pyrene in ambient air
Luftbeschaffenheit - Messverfahren zur Bestimmung der Konzentration von Benzo[a]
pyren in Luft
Méthode de mesurage de la concentration du benzo [a] pyrene dans l'air ambiant
Ta slovenski standard je istoveten z: EN 15549:2008
ICS:
13.040.20 Kakovost okoljskega zraka Ambient atmospheres
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN 15549
NORME EUROPÉENNE
EUROPÄISCHE NORM
March 2008
ICS 13.040.20
English Version
Air quality - Standard method for the measurement of the
concentration of benzo[a]pyrene in ambient air
Qualité de l'air - Méthode normalisée pour le mesurage de Luftbeschaffenheit - Messverfahren zur Bestimmung der
la concentration de benzo[a]pyrène dans l'air ambiant Konzentration von Benzo[a]pyren in Luft
This European Standard was approved by CEN on 2 February 2008.
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 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 Management Centre has the same status as the
official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, 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 and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36  B-1050 Brussels
© 2008 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 15549:2008: 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 Symbols and abbreviations.7
5 Principle of the method.8
6 Requirements.9
7 Reagents and gases.11
8 Apparatus .12
9 Sampling.13
10 Sample preparation .14
11 Analysis .15
12 Quantification.17
13 Quality control .21
14 Determination of measurement uncertainty .23
15 Interferences .25
16 Reporting of results.26
Annex A (informative) Sampling systems .27
Annex B (informative) Extraction methods (examples of experimental conditions) .30
Annex C (informative) Example for clean-up procedure.32
Annex D (informative) Parameters for analysis (examples) .33
Annex E (informative) Assessment of performance indicators and uncertainty contributions .35
Annex F (informative) Results of laboratory and field tests .44
Bibliography.50

Foreword
This document (EN 15549:2008) has been prepared by Technical Committee CEN/TC 264 “Air quality”, the
secretariat of which is held by DIN.
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 2008, and conflicting national standards shall be
withdrawn at the latest by September 2008.
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.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association, and supports essential requirements of EU Directive(s).
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, 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 and the United Kingdom.
Introduction
The European Directive 2004/107/EC prescribes the reference methodology for the measurement of
benzo[a]pyrene (BaP) in ambient air and states that the method shall be a method based on manual PM10
sampling systems described in EN 12341 or equivalent.
Hence, this European Standard describes a method in which the sampling systems fulfil this requirement.
However, in the course of drafting this European Standard it became clear that in certain circumstances, in the
presence of oxidants such as ozone, BaP may be degraded. In specific situations this may result in losses of
BaP of > 50 %. It has been shown that the degradation due to ozone can be substantially reduced by
including an ozone denuder in the sampling system.
To date only a limited number of experiments has been performed in order to evaluate the particular
conditions under which the denuder systems can be efficiently used. Consequently, the application of ozone
denuders lacks sufficient validation to be a normative part of this European Standard.
In order to have a complete picture of the performance of ozone denuder systems further information is
required on:
• efficiency under variable atmospheric conditions,
• regeneration time after exposure to high humidity,
• maximum capacity for ozone,
• maximum sample volume and maximum sampling period,
• stability of catalyst,
• maximum period of use,
• particle losses.
Examples of sampling using an ozone denuder are given in Annex A.
The experimental evidence collected so far is presented in Annex F.
It is recommended that further work is undertaken to provide data of BaP comparisons with and without ozone
denuders.
1 Scope
This document specifies a measurement method for the determination of particulate benzo[a]pyrene (BaP) in
ambient air, which can be used in the framework of the Council Directive 96/62/EC [1] and the Directive
2004/107/EC [2]. This document specifies performance characteristics and performance criteria for the
measurement method when it is used as a reference method. The performance characteristics of the
measurement method are based on a sampling period of 24 h.
This document describes a measurement method which comprises sampling of BaP as part of the PM10
particles, sample extraction and analysis by high performance liquid chromatography (HPLC) with
fluorescence detector (FLD) or by gas chromatography with mass spectrometric detection (GC/MS). The
method is applicable for the measurement of BaP in the concentration range from approx. 0,04 ng/m to
approximately 20 ng/m .
The lower limit of the applicable range depends on the noise level of the detector and the variability of the
laboratory filter blank.
NOTE If the BaP concentration exceeds the calibration range the extract can be diluted.
2 Normative references
The following referenced documents are indispensable for the application 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 12341:1998, Air quality – Determination of the PM 10 fraction of suspended particulate matter – Reference
method and field test procedure to demonstrate reference equivalence of measurement methods
ENV 13005:1999, Guide to the expression of uncertainty in measurement
ISO 8258, Shewhart control charts (including ISO 8258:1993 Technical Corrigendum 1)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
calibration solution
solution used for calibration of the analytical instrument, containing the analyte of interest at a suitable
concentration, prepared by dilution of the stock standard solution
3.2
certified reference material (CRM)
reference material one or more of whose property values are certified by a technically valid procedure,
accompanied by or traceable to a certificate or other documentation that is issued by a certifying body
3.3
external standard solution
solution of the analyte of known concentration
3.4
field blank filter
filter that is taken through the same procedure as a sample, except that no air is drawn through it
3.5
internal standard solution
solution of a known substance of known concentration, added to the sample before chromatographic analysis
3.6
laboratory blank filter
unused filter that does not leave the laboratory and is taken through the same analytical procedure as the
sample
3.7
PM10
target specification for sampling the thoracic particles
[EN 12341:1998]
3.8
reagent blank solution
solution that contains all the reagents used during analysis of the sample, but without the sample and filter
matrix
[EN 14902:2005] [27]
3.9
stock standard solution
solution used for preparing calibration solutions, containing the analyte of interest at a concentration traceable
to national or international standards
3.10
surrogate standard solution
solution of a known substance and of known concentration, used to spike filters before extraction in order to
check the recovery efficiency
3.11
target value
concentration in the ambient air fixed with the aim of avoiding, preventing or reducing harmful effects on
human health and the environment, as a whole, to be attained where possible over a given period
NOTE This definition originates from [2]. The current value for BaP is 1 ng/m for the total content in the PM10
fraction averaged over a calendar year.
3.12
uncertainty (of measurement)
parameter associated with the result of a measurement, that characterises the dispersion of the values that
could reasonably be attributed to the measurand
[ENV 13005:1999]
4 Symbols and abbreviations
4.1 Symbols
a is the slope of linear calibration function;
A is the peak area or peak height of BaP or of its characteristic ion in the chromatogram of the
C
calibration solution;
A is the peak area or peak height of BaP or of its characteristic ion in the chromatogram of the sample
E
extract;
A is the peak area or peak height of the internal standard or of its characteristic ion in the chromatogram
IS
of the calibration solution;
A is the peak area or peak height of the internal standard or of its characteristic ion in the chromatogram
ISE
of the sample extract;
b is the intercept of the linear calibration function;
C is the concentration of BaP in ambient air in ng/m³;
D is the detection limit, expressed in ng/m³;
C
D is the absolute detection limit in the sample in ng;
M
f is the response factor of BaP;
m is the mean of laboratory filter blanks in ng;
m is the mass of BaP in the calibration solution in ng;
C
m is the certified mass in the CRM in ng;
CRM
m is the mass of BaP in the sample extract in ng;
E
m is the mass of BaP on the filter sample in ng;
F
m is the individual filter blank in ng;
i
m is the mass of the internal standard in the calibration solution in ng;
IS
m is the mass of the internal standard in the sample extract in ng;
ISE
m is the mass of BaP calculated from the regression equation at the level of the calibration standard in
reg
ng;
m is the mass of the surrogate standard in the sample extract in ng;
SSE
m is the mass of the surrogate standard added to the filter in ng;
SSF
m/z is the mass-to-charge ratio;
n is the number of analysed filters;
R is the recovery efficiency of BaP in %;
R is the peak resolution;
s
S is the standard deviation of laboratory filter blanks in ng;
lfb
s(m ) is the standard deviation of the replicate measurement results of the mass determined in ng;
E
t is the sampling time in h;
t is the Student factor for n measurements and a 95 % confidence interval;
n-1;0,95
t is the retention time for peak 1 in min;
R1
t is the retention time for peak 2 in min;
R2
VE is the volume of the extract in ml;
V is the volume of air sampled in m ;
V is the nominal daily sampling volume in m³;
n
w is the peak width of peak 1 in min;
w is the peak width of peak 2 in min;
X is the measured mass fraction of BaP in mg/kg;
a
X is the certified mass fraction of BaP in mg/kg.
ca
4.2 Abbreviations
BaP Benzo[a]pyrene
CRM Certified reference material
DAD Diode array detector
FLD Fluorescence detector
GC Gas chromatography
HPLC High performance liquid chromatography
MS Mass spectrometry
5 Principle of the method
The method is divided into two main parts: first the sampling in the field and second the analysis in the
laboratory. During sampling, particles are collected on a filter by sampling a measured volume of air by means
of a sampler equivalent to one of those described in EN 12341.
The sampling time is 24 h. The filter is transported to the laboratory. BaP is extracted using an organic
solvent. If necessary, the extract is cleaned up. The resulting solution is analysed by HPLC/FLD or GC/MS.
6 Requirements
6.1 Siting requirements
Specific siting requirements depend on the objectives of the measurements. For measuring in compliance with
Directive 2004/107/EC then the instructions for siting samplers given in [2] shall be followed.
6.2 Sampling requirements
The sampling system shall be equivalent to EN 12341 (see [2]).
NOTE 1 Equivalence can be demonstrated by performing a side-by-side comparison of the system with a PM10
reference sampler. Guidance for the performance of such comparisons is given in [3].
NOTE 2 In the presence of ozone BaP may degrade. In specific situations this may lead to losses of BaP of > 50 %.
Whenever these effects are expected to be significant, the PM10 sampler may be equipped with an ozone denuder (see
Annex A). The experimental evidence collected so far is presented in Annex F. However, the application of ozone
denuders lacks sufficient validation to be a normative part of this European Standard.
NOTE 3 Examples of sampling systems with and without denuder are presented in Annex A.
6.3 Analysis
6.3.1 Recovery efficiency
Using the external or internal standard method for quantification check the recovery efficiency periodically by
spiking laboratory blank filters with a known amount of BaP and process them as usual. The recovery
efficiency shall be between 80 % and 120 %, otherwise the surrogate standard method shall be used. The
recovery efficiency shall be checked with a frequency to ensure that 95 % probability for a correct
measurement is maintained (see 12.1.1).
Using the surrogate standard method (see 12.1.3) this initial recovery check is not necessary. The surrogate
recovery for field samples shall not be less than 50 %, otherwise the sample shall be discarded.
NOTE 1 If the surrogate recovery is constantly less than 70 %, this indicates problems with the sample preparation
procedure. These problems should be eliminated.
Check the recovery efficiency of the method for BaP in certified reference material (e. g. NIST 1649a) using
equation (1).
X
a
R = × 100
(1)
X
ca
where
R is the recovery efficiency of BaP in %;
X is the measured mass fraction of BaP in mg/kg;
a
X is the certified mass fraction of BaP in mg/kg.
ca
The recovery efficiency shall be between 80 % and 120 %.
NOTE 2 A certified reference material containing the same matrix as ambient PM10 particles collected on filters is not
available at the moment. Interferences occurring to field samples, e. g. chemical reactions of BaP during extraction, can
be identified, for example, by
• Repeating the extraction step with a different method and comparing the results;
• Comparing the ratio of BaP to at least one more stable and high-boiling PAH like benzo[e]pyrene or
benzo[k]fluoranthene: an indication for problems occurring during the sample preparation procedure is that deviations
(lower ratios) with respect to previous measurements at the same location and in the same season are observed.
Changing the sample preparation procedure (different extraction solvent, different purification procedure) can verify
the problem.
6.3.2 Detection limit
6.3.2.1 General
The detection limit shall be less than 0,04 ng/m³.
6.3.2.2 Determination based on laboratory filter blanks
Determine the detection limit from the standard deviation of at least ten laboratory filter blanks using equation
(2).
n
(m − m )
∑ i
i=1
S =
(2)
lfb
n−1
where
S is the standard deviation of laboratory filter blanks in ng;
lfb
m is the mean of laboratory filter blanks in ng;
m is the individual filter blank in ng;
i
n is the number of analysed filters.
The minimal detectable mass of BaP is calculated using equation (3).
D = t × S
(3)
M n−1;0,95 lfb
where
D is the minimal detectable mass of BaP in ng;
M
t is the Student factor for n measurements and a 95 % confidence interval;
n-1;0,95
S is the standard deviation of laboratory filter blanks in ng.
lfb
6.3.2.3 Determination based on the signal-to-noise ratio
Perform a chromatographic analysis without injecting any solution. Keep the chromatographic parameters as
used for the calibration and the detection of BaP. Calculate the detection limit as three times the average of
the height of the noise at the retention time of BaP ± 10 times the peak width at half peak height at the lowest
calibration level.
6.3.2.4 Calculation of the detection limit
The detection limit, expressed in ng/m³, is calculated introducing the nominal daily sampling volume according
to equation (4).
D
M
D = (4)
C
V
n
where
D is the detection limit, expressed in ng/m³;
C
D is the minimal detectable mass of BaP in ng;
M
V is the nominal daily sampling volume in m³.
n
For the nominal daily sampling volume data shall be used which are usually obtained during sampling.
NOTE The volume is, for example, approximately 1 630 m³ for the sampler type, which has been used in the field test
(see Annex F.2), or approximately 64 m³ for cuts of 50 mm diameter of the filter samples, collected with this sampler type.
6.4 Oven temperature for HPLC
The temperature of the HPLC column oven shall kept constant to at least ± 1 °C.
7 Reagents and gases
7.1 Solvents
High purity solvents (see 13.1), e.g. toluene, cyclohexane, dichloromethane, acetonitrile and water.
7.2 Gases
Helium (purity 99,999 %) used as carrier gas for GC/MS and for degasification of solvents of the HPLC
method.
7.3 External standard
BaP, e.g. a dilution of the stock standard solution (7.6).
7.4 Internal standard
 Methylated or halogenated PAH, e.g. 6-methylchrysene (for HPLC/FLD);
 deuterated or carbon-13-labelled PAH, e. g. perylene-d12 (for GC/MS).
Make sure that the standards contain less than 1 % (relative) of the native BaP.
7.5 Surrogate standard
 Methylated or halogenated PAH, e.g. 7-methylbenzo[a]pyrene (for HPLC/FLD);
 deuterated or carbon-13-labelled 5-ring PAH, e. g. BaP -d12 (for GC/MS).
Make sure that the standards contain less than 1 % (relative) of the native BaP.
7.6 Stock standard solution
Dilution of a solution of BaP with a concentration traceable to internationally accepted standards, e.g. NIST
1647e.
7.7 Certified reference material
Containing a certified concentration of BaP, in a matrix similar to PM 10 particles, e.g. NIST 1649a.
8 Apparatus
8.1 Sampling equipment
8.1.1 PM10 sampler
The sampling system shall be equivalent to EN 12341 (see [2]).
8.1.2 Greasing agent, if required, suitable for greasing the sampler impaction plate (see manufacturer's
instructions).
8.1.3 Quartz fibre or glass fibre filters, of a diameter suitable for use with the samplers (8.1.1), with a
separation efficiency of at least 99,5 % at an aerodynamic diameter of 0,3 µm. This criterion has also to be
met after pre-treatment of filters according to NOTE 1 in 13.6.
The purity of the filters shall be checked according to 13.7.
NOTE It is recommended that filter manufacturers determine the filter separation efficiency according to standard
methods such as EN 13274-7 [4] or EN 1822-1 [5].
8.1.4 Flow meter, with a measurement uncertainty that is sufficient to enable the volumetric flow rate of
the samplers (8.1.1) to be measured within ± 5 %. The calibration of the flow meter shall be traceable to
internationally accepted standards.
8.2 Sample preparation/extraction
The following apparatus is required:
• round-bottomed flask with reflux condenser; or
• Soxhlet assembly; or
• microwave digestion system; or
• accelerated solvent extraction apparatus; or
• sonication bath.
For examples or details of the procedure see Annex B.
8.3 Laboratory apparatus
8.3.1 HPLC/FLD apparatus
Liquid chromatograph fitted with injection system, a reverse phase column suitable for PAH analysis, a
temperature controlled oven, a pump system and a FLD (see also D.1). Furthermore a system for solvent
degassing (internal or external) is required.
NOTE If the BaP concentration in the extract is high enough a DAD can be used (see 15.2).
8.3.2 GC/MS apparatus
Gas chromatograph with split/splitless injector or on column injector, capillary column suitable for PAH
analysis, and a mass selective detector (see also D.2).
9 Sampling
9.1 Preparation of the equipment before sampling
Consult the manufacturer's instruction manual to determine the minimum voltage and power requirements of
the sampler and ensure that an adequate power supply is available at the sampling site.
Clean the sampler inlet, suction pipe, and all other parts of the sampler, such as filter a change mechanism
and filter cassettes, which may come in contact with the filter before use according to the manufacturer's
specifications. Similarly, inspect greased parts like impaction plates before use, clean them if necessary and
grease them again.
A leak test and flow rate calibration shall be carried out before deploying the sampler in the field in order to
identify problems with the sampler in an early stage.
9.2 Handling of filters
Handle small filters with blunt tweezers, so as to avoid contamination and damage. For large filters this
procedure might not be practicable. In this case handle them carefully using gloves made of an appropriate
material, touching only the outside edges of the filters.
9.3 Preparation of filters
Filters that have visibly been contaminated, e.g. during packing and/or transport shall be rejected.
Inspect each filter before use for pin holes and other imperfections, such as chaffing, loose material,
dislocation and non-uniformity. For example, use a magnifying lens with a light or check in front of an area
light. Reject any filter if its integrity is suspect.
Assign each filter a unique identifier and place it in a labelled, sealed container for storage and transportation
to the field.
NOTE The container should be made of appropriate material (e. g. glass, PTFE).
If the filter has to be marked for identification purposes, do not mark it in an area that will be analysed.
Establish a filter log (i.e. a chain of custody book/record) to document the use of each filter. Record the
number of filters in the filter log. If the sampler to be used is a sequential sampler that operates continuously
for a programmed period, load the required number of filters into a labelled filter cartridge and seal it ready for
transportation to the field. It shall be recorded which filter was located into which position in the cartridge.
Handle laboratory blank filters in the same way as real samples, but do not transport them and do not draw air
through them. Each batch of filters shall be checked before use by analysing one filter (see 13.6).
Prepare field blank filters and process them as far as possible as real samples. Transport them to the
sampling site, mount them into the sampler (without switching on the pump), dismount them, return them to
the laboratory and process them in the same way as real samples. At least one of every twenty filters shall be
analysed as field filter blank. If the field filter blank significantly exceeds the average laboratory filter blank, the
sources of contamination shall be investigated and eliminated. If the results of real samples are significantly
affected by the field filter blank, the samples shall be reanalysed, if possible.
9.4 Sample collection and storage
Set up the sampler in the field according to the manufacturer’s instructions, ensuring that the siting
requirements (see 6.1) are met. Then carry out a leak test and check the flow rate of the sampler using the
calibrated flow meter before use and at least every three months, following the manufacturer's instructions. If
the flow rate deviates from its nominal value by more than the deviation given by the manufacturer, calibrate
the sampler by adjusting the flow rate as necessary.
Take field filter blanks periodically at each site (at least once for every twenty filters used for sampling, see
9.3).
Load either an unexposed filter (for single filter devices) or a cartridge of unexposed filters (for sequential
samplers) into the sampler at the start of the sampling period. Program the sampler following the
manufacturer's instructions, start the timer and record the start time.
The sampling time is 24 h.
Collect the filter from the sampler at the end of the sampling period, replace it in its uniquely marked transport
container and seal the container for transportation to the laboratory (for single filter devices). For sequential
samplers, collect the used filters and prepare them for transportation to the laboratory.
If filters are folded for storage (for easier transportation), then it will be necessary to analyse the whole filter as
folding can affect the distribution of particles on the filter surface. In this case the marked part of the filter (see
9.3) shall be cut out before extraction.
The filters shall be extracted not later than two months after sampling. They shall be stored in the dark in a
closed vessel at a temperature below 20 °C.
NOTE Individual samples taken over a period of up to one month can be combined and analysed as a composite
sample [2]. Filter cuts of identical size of single days are extracted together. If the BaP content of these composite extracts
is divided by the sum of the air volumes sampled with the filter cuts the result is the mean value for that period. The
minimum time coverage of the sampling period has to be 33 %.
Record all details of each sample in the filter log, including stop time, flow rate, air sample volume in cubic
metre, any mechanical or electrical failures during the sampling period and any other data that could be
important for the evaluation of the sampling.
Clean and grease the inlet impaction plate, if applicable, according to the manufacturer’s instructions. Perform
intensive cleaning of the PM10 sampling head according to the manufacturer's instructions at least once every
6 months.
10 Sample preparation
10.1 Cleaning of the laboratory apparatus
Cleaning of extraction devices and labware shall be carried out in a way that blank values are avoided or
lower than a mass corresponding to a final concentration of 0,04 ng/m after usual work-up.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml the blank value
should be lower than 4 ng.
10.2 Extraction
The following techniques have been shown to meet the requirements of 6.3.1:
• extraction by reflux;
• Soxhlet extraction;
• accelerated solvent extraction;
• ultrasonic extraction;
• microwave extraction.
The methods are described in Annex B.
Any extraction method results in a solution of BaP and other substances in an organic solvent. For GC/MS the
extract can directly be analysed, if it is reduced to a known volume and if no further purification is necessary.
For HPLC/FLD the extract shall be carefully evaporated to dryness and shall be dissolved in a known volume
of acetonitrile. If necessary, the extract shall be cleaned before reducing it in volume to at least 10 % of the
final volume (see, for example, Annex C). If the solutions are not analysed immediately, they shall be stored in
the dark to avoid compound degradation and at a temperature less than 6 °C to avoid the evaporation of the
solvent. The maximum permissible storage period is one month.
11 Analysis
11.1 HPLC/FLD analysis
11.1.1 Principle of the method [6]
The organic extract containing BaP is filtered, if necessary purified by column chromatography (e.g. Annex C),
reduced in volume and dissolved in acetonitrile. An aliquot of the solution is injected into the HPLC/FLD
apparatus. BaP is identified by its retention time. The peak area and/or peak height is used as a measure of
its concentration in the sample.
NOTE In practice, extracts from PM10 particles may be carefully evaporated to dryness without reducing the
recovery of BaP.
The response and sensitivity of the FLD is constant over a long period, so that either the internal or the
external standard method can be used for quantification. If the clean-up procedure or a complex sample
matrix affects the recovery efficiency, the surrogate standard method can be used to correct losses during
sample preparation.
11.1.2 Reagents
11.1.2.1 Calibration solutions
Prepare calibration solutions of BaP from the stock solution at a minimum of five concentration levels by
adding appropriate volumes of the BaP stock solution to a volumetric flask and making up with acetonitrile.
The concentrations shall cover a working range corresponding to the expected range of concentrations found
in real samples. The lowest concentration shall be near, but above, the detection limit. The concentration of
these solutions shall be at a level corresponding to the target value ± 20 %.
NOTE 1 For the external standard method (see 12.1.1) these solutions are used for calibration purposes, for the
internal and surrogate standard method (see 12.1.2, 12.1.3) these solutions are used for the check of the lack of fit.
NOTE 2 For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.1.2.2 External standard solution
Use a calibration solution (11.1.2.1) with the concentration nearest to the one which is equivalent to the BaP
concentration at the target value ± 20 %.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.1.2.3 Internal standard solution
Dilute the internal standard (7.4) in acetonitrile. Add, for example, 10 µl of this solution to the sample extracts
(after clean-up, if any) before analysis. The concentration of the solution shall be equivalent to the BaP
concentration at the target value ± 20 %.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.1.2.4 Surrogate standard solution
Dilute the surrogate standard (7.5) in acetonitrile. Add, for example, 10 µl of this solution to the filter before
extraction. The concentration of the solution shall be equivalent to the BaP concentration at the target value
± 20 %.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.1.3 Proposed parameters for HPLC/FLD operation
See example in D.1.
11.1.4 Calibration
When using the external standard method for quantification, inject the calibration solutions (see 11.1.2.1;
aliquots of 20 µl, for example) directly into the HPLC system and plot the peak area or the peak height versus
the concentration. Calculate the calibration function using linear regression. The lack of fit of the calibration
function shall fulfil the requirements in 14.2.1, Table 1 (see E.4.4.2.3 for calculation).
11.1.5 Detection and measurement
Remove the sample extracts from cold storage and allow them to warm up to room temperature. Inject an
aliquot of the sample (for example 20 µl) into the HPLC system, identify BaP by its retention time. Dilute an
aliquot of the extract and reanalyse it, if the concentration of BaP in the extract is above the upper limit of the
working range. For quantification use either the external, or the internal or the surrogate standard technique.
11.2 GC/MS analysis
11.2.1 Principle of the method [7]
The organic extract containing BaP may be purified by column chromatography (see, for example, Annex C),
if necessary. Then the extract is concentrated. An aliquot of the solution is injected into the GC/MS system.
After separation in a capillary column BaP is detected by a mass spectrometric detector. The substance is
identified by its retention time and by the m/z values of specific ions; the peak area or the peak height is a
measure of its concentration in the sample.
For GC-MS analysis a combination of the internal standard method and the surrogate standard method is
required for quantification.
11.2.2 Reagents
11.2.2.1 Solutions for checking the lack of fit
Prepare calibration solutions of BaP from the stock solution at a minimum of five concentration levels by
adding appropriate volumes of the BaP stock solution to a volumetric flask and making up with the appropriate
solvent (see B.6). The lowest concentration shall be near, but above, the detection limit. The other
concentrations shall cover a working range corresponding to the expected range of concentrations found in
real samples. The concentration of one solution shall be at a level corresponding to the target value ± 20 %.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.2.2.2 Internal standard solution
Dilute the internal standard (7.4) in the appropriate solvent (see B.6). Add, for example, 10 µl of this solution
to the sample extracts (after clean-up, if any) before analysis. The concentration of the solution shall be
equivalent to the BaP concentration at the target value ± 20 %.
11.2.2.3 Surrogate standard solution
Dilute the surrogate standard (7.5) in the appropriate solvent (see B.6). Add, for example, 10 µl of this solution
to the filter before extraction. The concentration of the solution shall be equivalent to the BaP concentration at
the target value ± 20 %.
NOTE For example, with a sampling volume of 100 m and a final volume of the extract of 1 ml and the target value
of 1 ng/m³ the concentration should be 100 ng/ml.
11.2.3 Proposed parameters for GC/MS operation
See example in D.2.
11.2.4 Lack of fit
In order to determine the linear working range of the detector for BaP inject the solutions for checking the lack
of fit (see 11.2.2.1; aliquots of 2 µl, for example) directly into the GC/MS system and plot the peak area or
peak height versus the concentration. Check the lack of fit of the function (requirement see 14.2.1, Table 1;
determination see E.4.4.2.3).
11.2.5 Detection and measurement
Remove the sample extracts from cold storage and allow them to warm up to room temperature. Inject an
aliquot of the sample (for example, 2 µl) into the GC/MS system. Identify BaP by its retention time and its
molecular ion (m/z = 252) and an appropriate qualifier, see e.g. Annex D.2. Use the integrated abundance
(peak area) of the target ion (m/z =252) for quantification. Quantification shall be carried out by a combination
of the internal standard and the surrogate standard technique (see 12.1.2 and 12.1.3). Dilute an aliquot of the
extract and reanalyse it if the concentration of BaP in the extract is above the upper limit of the working range.
12 Quantification
12.1 HPLC/FLD analysis
12.1.1 External standard method
Calculate the mass of BaP in the extract according to equation (5), using the calibration function:
A − b
E
m = × V (5)
E E
a
where
m is the mass of BaP in the extract in ng;
E
A is the peak area or peak height of BaP in the extract;
E
b is the intercept of the linear calibration function;
a is the slope of linear calibration function in ml/ng;
VE is the volume of the extract in ml.
Use the external standard solution (11.1.2.2) to verify the proper operation of the HPLC/FLD system (at least
10 % of the samples).
Verify the recovery efficiency (see 6.3.1), if necessary including the clean-up step, with laboratory blank filters
(at least 5 % of the samples), spiked with the external standard solution (see 11.1.2.2).
12.1.2 Internal standard method
Prepare at least five solutions with BaP concentrations, which cover of the whole working range, and a
constant concentration of the internal standard (7.4), which shall be equivalent to the BaP concentration at the
target value ± 20 %.
NOTE The calibration solutions (see 11.1.2.1) may be used for this purpose by adding constant volumes of the
internal standard solution (see 11.1.2.3).
Inject these solutions and calculate the response factor f of BaP from the peak areas of both BaP and the
internal standard and the corresponding masses of these substances according to equation (6).
A × m
IS c
f =
(6)
A × m
c IS
where
f is the response factor of BaP;
A is the peak area or peak height of the internal standard in the chromatogram of the calibration

IS
solution;
A is the peak area or peak height of BaP in the chromatogram of the calibration solution;
c
m is the mass of BaP in the calibration solution in ng;
c
m is the mass of the internal standard in the calibration solution in ng.
IS
The average values of the response factors can be used for further analysis.
Add the internal standard solution (see 11.1.2.3) to the sample before injection.
The mass of BaP in the sample extracts is calculated according to equation (7):
f × A × m
E ISE
m =
(7)
E
A
ISE
where
f is the response factor of BaP;
A is the peak area or peak height of BaP in the chromatogram of the sample extract;
E
A is the peak area or peak height of the internal standard in the chromatogram of the sample extract;
ISE
m is the mass of the internal standard in the sample extract in ng;
ISE
m is the mass of BaP in the sample extract in ng.
E
Verify the recovery efficiency (see 6.3.1), if it is used, including the clean-up step, with laboratory blank filters
(at least 5 % of the samples), spiked with the external standard solution (see 11.1.2.2).
12.1.3 Surrogate standard method
Correct the mass of BaP for the recovery efficiency as follows: Calculate the concentration of the surrogate
standard either using a calibration function in analogy to 12.1.1 and equation (5) for the external calibration
method, or using response factors in analogy to 12.1.2 and equations (6) and (7) for the internal standard
method.
Add for example 10 µl of the surrogate standard solution (11.2.2.3) to the filter before the extraction step.
Evaporate the solvent and calculate the mass of BaP in the filter according to equation (8):
m × m
SSF E
m = (8)
F
m
SSE
where
m is the mass of BaP on the filter sample in ng.
F
m is the mass of the surrogate standard added to the filter in ng;
SSF
m is the mass of BaP in the extract in ng, calculated according to equation (5) or equation (7);
E
m is the mass of the surrogate standard in the sample extract in ng;
SSE
12.2 GC/MS Analysis
Prepare at least five solutions with BaP concentrations, which cover the whole working range, and a constant
concentration of the internal standard (7.4), which shall be the target value ± 20 %.
NOTE The solutions (see 11.2.2.1) may be used for this purpose by adding constant volumes of the internal standard
solution (see 11.2.2.2).
Inject these solutions and calculate the response factor f of BaP from the peak areas/peak heights of both
BaP and the internal standard and the corresponding masses of these substances according to equation (9).
...