Twinning Bulgaria Italia

Training 3.3
Simitli 17-
17-19 February 2009

Analytical Methods for PAH, PBDE,

C10-13
C10 13 Chloroalkanes and
alkylphenols

Stefano POLESELLO
Istituto di Ricerca sulle Acque-
Acque-CNR
Sezione di Brugherio
polesello@irsa.
l ll @i cnr.it
it
 ALKYLPHENOLS
Status

Priority Substance Standard Principle

inland other
waters waters
ISO 18857-
Nonylphenols3) C C
1:2005 GC/MS
Octylphenol (4-(1,1,3,3)- ISO 18857-
A D
Tetramethylbutylphenol 1:2005 GC/MS

LOQ = 0.1 g/L for NP and 0.0033 g/L for OP

Blank interferences for NP
No validation for whole water
At my knowledge, the only CRM available for NP is
WWTP sludge (www.eurofins.dk).
 Alkylphenols: analytical methods
z Liquid chromatography
ESI (-
(-) (LC
(LC--MS/MS) ((ASTM
ASTM WK15211)
WK15211
Fluorimeter (IRSA
(IRSA method, Italy)
Italy)

z Gas chromatography with MS detection (ASTM

(ASTM
WK1521 ASTM D7065 06)
WK1521;
Without
Wi h d
derivatisation
i i i (ISO (ISO 188 1 200 )
18857-1:2005
18857- 1:2005)
With derivatisation with MSTFA (ISO
(ISO 18857- 2:draft))
18857-2:draft
 Extraction
z Water
LLE with toluene ((ISO
ISO 18857-
18857-1:2005; ASTM D7065- 06))
D7065-06
Continuous LLE (ASTM
(ASTM WK1521)
WK1521
SPE with C18, SDVB ((ISOISO 18857-
18857-2:draft ; ASTM
WK15211)
WK15211

z Solid samples (SPM, sediment)

Soxhlet or ASE extraction with
z MEOH (IRSA
(IRSA method, Italy)
z Acetone/esano
z LC:
single peak for each isomeric mixtures of a single
compound
but often broad peaks
Possibility to determine ethoxylates
Columns: preferred polar embedded or reverse
phase such as phenylhexyl
phenylhexyl

z GC
Different peaks for about every isomers
...but integration must be the sum of a group of
peaks
Variability of NP without derivatisation (Adsorption?)
 GC without derivatisation:
ISO 18857-
18857-1:2005
z Compounds: 4n
4n--NP, 4-
4-NP (branched)
(branched),, 4
4--tert
tert--OP
z Range : 0.005 to 0.2 g/L; drinking, surface
and
d waste
t waters
t
z m/z 135 + 107
z Extraction: LLE with toluene
z Claen
Claen--up: silica column
z Detection: GC-
GC-MS in SIM (m/z=107/135)
z Quantification: ESTD e ISTD: 13C6-ring
ring-
g-labelled 4-
4-n-
nonylphenol m/z 113

ISO 18857-
18857-1,
1 2005
2005, Water quality determination of selected alkylphenols
Part 1: Method for non-
non-filtered samples using liquid extraction and gas
chromatography with mass selective detection.
ISO 18857-
18857-1:2005
 GC with derivatizazion:
derivatizazion:
ISO 18857-
18857-2:draft
2:draft
z Analytes: NP, OP, OP(1,2)EO, NP(1,2)EO and BPA
z Range: 0.005
0 005 to 0
0.2
2 g/L (OP and OP(1
OP(1,2)EO)
2)EO)
0.030 to 0.2 g/L (NP and NP(1,2)EO)
0 05 to 0
0.05 0.2
2 g/L (BPA)
(drinking water, ground and surface water)
z Range
g : 0.1 to 50 g
g/L ((waste water))
z Extraction: SPE (styrene
(styrene--divinylbenzene) without filtration
z Derivatizazion: MSTFA Trifluoro
Trifluoro--N-methyl
methyl-
y -N-
(trimethylsilyl)acetamide
z Detection: selected ion monitoring
z Quantification: ESTD and ISTD: 13C6-ring
ring--labelled
ISO 18857-
18857-2:
2:draft
draft (ESTD+ISTD
ESTD+ISTD))
 ISO 18857-
18857-2:draft
2:draft
(spiked water extract+ISTD
extract+ISTD))
 Intercalibration
ISO 18857-1:2005
Bavarian Water Management Agency Munich, Germany according
to ISO 5725

17 labs from 6 countries

3 water samples (duplicate)
surface (0.005 to 0.05 g/L)
surface ((0.05 to 0.2 g/L)
g )
wastewater (0.1 to 25 g/L)
calibration solution
 Intercalibration trial

NAP REC
matrix variables N X xref SR CVR
% %
1 surface 4-tert-OP 26 7.1 16.6 19.0 87.3 4.16 25.1
4-NP 22 15.4 28.7 23.0 124.9 16.4 57.2
4-tert-OP
2 surface 26 13.3 66.8 75.0 89.0 17.9 26.8

4 NP
4-NP
22 26.7 82.8 90.0 92.0 15.6 18.8

4-tert-OP
3 waste 30 0 1400 1400 100.1
100 1 0.45
0 45 32 0
32.0

4-NP
30 6.3 2020 1800 112.2 0.64 31.5

Octylphenol: 19
19-1400
1400 ng/L ; reproducibility: 19 e 57 %
Nonylphenol: 23 -1800 ng/L; reproducibility: 25 e 32 %
 PBDE

z Standardised method does not exist for water:

Pentabromodiphenylether
p y ((PDBE))
Method ISO: only for sediment (SPM?)
Under development (CEN method)

LOQ = 0.066 ng/L

POLYBROMINATED DIPHENYLETHERS
(PBDEs)
Technical mixtures

) Penta-BDE
Mixture of tetra-, penta-, and hexabromodiphenylethers;
BDE-47,, BDE-99,, BDE-100,, BDE-153 and BDE-154
constitute more than 80% of the technical product

) Octa-BDE
Mixture of hepta-, octa- and nonabromodiphenylethers;
BDE-183 can be used as indicator compound

) Deca-BDE
Decabromodiphenylether makes up more than 97%
 PBDE: sum of BDE-28, BDE-47, BDE-99, BDE-100,
BDE-153, BDE-154
CAS Number: 32534-81-9 Log KOW: 6.57

z No standardised method for the determination of PBDE in

water available! (CEN Method under development for whole
water)

z ISO 22032:2006 Determination of selected polybrominated

in sediment: diphenylethers (PBDE) in sediment and
sewage sludge
l d
Method using extraction and gas chromatography/mass
spectrometry

EQS = 0.5 ng/L; Other Surface Waters =0.2 ng/L

LOQ = 0.066
0 066 ng/L
 Methods for PBDE analysis in water are
available:

z EPA Method 527 employs solid-phase extraction with

analysis by gas chromatography/mass spectrometry
MDL is fairly high (0.39 g/l)

z EPA Method 1614

1614, 2007 applies HRGC/HRMS for the
analysis of PBDE in water, soil, sediment and tissue.
MDL for BDE 99 is 0.04 ng/l.
Large volume methods

There are a few reports on extremely low levels of PBDEs in

surface water samples.
The authors enriched 100 and 2500 L of water,
water
respectively, on XAD resin. SPME has been proposed to
extract selected BDE congeners from water samples by Polo
et al.
al 2004.
2004
 PBDEs
Analytical Methods

) Gas chromatography with electron capture detection (GC-ECD)

) Gas chromatography
g p y with atomic emission detection (GC-AED)
( )

) Gas chromatography with electron impact mass spectro-

metric detection at low resolution ((GC-LRMS))

) Gas chromatography with electron impact mass spectro-

g resolution (GC-HRMS)
metric detection at high ( )

) Gas chromatography with electron capture negative

ionisation mass spectrometric
p detection (GC-ECNI-MS)
( )
 CEN-ISO Method for sediment
GC-ECNI-MS
Experimental Conditions:

Matrix: Sediment, suspended particulate matter (SPM)

Compounds: BDE 47 BDE-99,
BDE-47, BDE 99 BDE-100,
BDE 100 BDE-153,
BDE 153
BDE-154, (BDE-183, BDE-209)
Limit of determination: 0.01 to 0.05 g/kg (test portion 10 g)
Extraction: Soxhlet extraction with toluene or
hexane/acetone
Clean-up: Multi-layer column chromatography on silica gel
Detection: GC-ECNI-MS
GC ECNI MS in the selected ion monitoring
mode (m/z=79/81 for all congeners except
BDE-209)
Quantification: ISTD: BDE-77,
BDE 77, BDE-181
BDE 181 not occurring
in technical mixtures; 13C12-labelled BDE-209
 PBDE
Interlaboratory studies on ISO Method

First interlaboratory study on PBDEs in 2000

J de Boer and W.
J. W P.P Cofino Chemosphere 46 (2002),
(2002) 625
625-633
633

Results for the two sediment samples:

Congener RSD in % Assessment

BDE-47: 22/24 acceptable
BDE-99: 36/45
/ further improvement
p necessary
y
BDE-100: 20/20 acceptable
BDE-153: 32/37 further improvement necessary
BDE-154: 35/45 further improvement necessary
BDE-209: 48/78 not under control
 PAH
Polycyclic aromatic hydrocarbons
AA-EQS:
z Anthracene 100 ng/L
z Fluoranthene 100 ng/L
z Benzo(a)pyrene 50 ng/L
z Benzo(b)fluoranthene+Benzo(k)fluoranthene
30 ng/L
z Benzo(g,h,i)perylene+ Indeno(1,2,3-cd)pyrene
2 ng/L
PAH: LOQ = from 0
0. 6 ng/L to 16 ng/L

Status
Priority Substance Standard
inland other
Principle waters waters
ISO 17993:
Fluoranthene A A
2002 HPLC/Fluo
ISO 17993:
Naphthalene A A
2002 HPLC/Fluo
EN ISO Purge/Trap +
A A
15680:2003 Therm Desorp.
Therm. Desorp
ISO 17993:
Benzo(a)pyrene C C
2002 HPLC/Fluo
ISO 17993:
Benzo(b)fluoranthene4) C C
2002 HPLC/Fluo
ISO 17993:
Benzo(k)fluoranthene4) C C
2002 HPLC/Fluo
Available Standard Method

EN ISO 17993: 2003

Determination of 15 p
polycyclic
y y aromatic hydrocarbons
y (PAH)
( )
in water by HPLC with fluorescence detection after liquid-
liquid extraction
Matrix: Drinking, ground, waste and surface water

Method Description
high performance liquid chromatography (HPLC) with
fluorescence detection for the determination of 15 selected
polycyclic aromatic.

Limit of Quantification (LOQ):

Drinking and ground water: > 5 ng/L
Surface water: > 10 ng/L
Oth Analytical
Other A l ti l Methods
M th d

EPA 8270c claims detection limit of 10 g/L

which is obviously too high.

Other analytical methods based on this standard

using modern GC/MS equipment however may
attain
tt i the
th required
i d low
l LOQs.
LOQ
 R
Remarks
k

z The methods do not attain low enough LOQs and

uncertainties for compliance checking with the AA-EQS for
the sum of Benzo[b]fluoroanthene and
Benzo[k]fluoroanthene as well as the sum of
Benzo[g,h,i]perylene
[g, , ]p y and Indeno[1,2,3-cd]pyrene.
[ , , ]py

z In addition, not enough validation data are available

regarding
di the
th analysis
l i off surface
f water
t samples
l in
i
particular samples containing substantial amounts of SPM.

z CEN Mandate for the development of analytical

methods in progress!
 Chemical Monitoring Activity
On-Site
On Site Workshop
Comparison of analytical approaches for WFD chemical
monitoring:
g a field experiment
p
a JRC IES RWER Initiative
1st CMA On-Site Technical
Workshop
10-11 October 2006,
in Ferrara
Ferrara, Italy

River Po - Pontelagoscuro

2nd CMA On-Site Technical

Workshop
17-18 September 2008
in Budapest, Hungary

River Danube - Budapest

z Practical sampling exercise on
the Po river;
z S
Simultaneous bulk sampling by 8
teams with different
methodologies
(also from IWP
IWP-Sofia-BG);
Sofia BG);
z Use of bulk sampling and short
term integrating methods (large
volume approaches);
z Homogeneity tests during
exercise;
z Acquisition of accompanying
data, as SPM, pH, temp.
 CMAonsiteorganizedbyJRCIES(Ispra)andtheProvinceof
Ferrara(Oct.2006)fortheimplementationofWFD.
( )f h i l i f
7laboratoriesfrom7differentcountries
Analytes:PBDEs,PAHs,nonylandoctylphenols
3samples
Standard
extract water from the river :
Correspondingto14.6LofPoRiverwater
concerningPBDEsandPAHs
Correspondigto1LofwaterfromSevesoRiver
d f f
concerningalchilfenoli
SamplingandanalysisofPoRiver
S li d l i f P Ri
 Alkylphenols
CMA01 CMA03 CMA 06 CMA02 CMA05 CMA07 CMA04

Volume [L] 0.4 0.5 1.0 11.82 0.2 2.3 1.0 4150

Filtration no no yes yes yes no no n.a.

Extraction SPE SPE SPE Filtration SPE LLE SPE Centrifuge

SPE cart. / HLB 200 C18 C18 Randall HLB 60 DCM Envichrom Soxhlet
solvent extraction P
SPE elution EA MeOH + Acetone SPM DCM/EA n.a. DCM SPM
MTBE

Analytical LC-MS (tq) LC-MS (tq) LC-fluor LC-fluor GC-MS GC-MS GC-MS GC-MS (it)
method
LC solvents H2O-ACN H2O-ACN H2O-MeOH H2O-MeOH n.a. n.a. n.a. n.a.

Column C18 C18 Phenylhexyl Phenylhexyl DB-5 DB-5 DB-1 ZB-5

Length 2.1150mm 2.1 100 mm 4.6250 mm 4.6 x 250 mm 30 m 30 m 60 m 30 m

Deriv. n.a. n.a. no no no no no MSTFA

Ions or 219 > 133 219 > 133 230 nm exc. 230 nm exc. 149(for NP) 135 135 207 > 179
wavelength 227 > 112 219 > 106 302 nm em 302 nm em 135(for 107 221 > 179
monitored 205 > 106 225 > 112 OP) 57 235 > 179
205 > 133 205 > 133 121 220 278 > 179
107 77 292 > 179

IST 4n-NP d8 4n-NP d6 no no 4n-NP d8 4n-NP Atrazine d5 4-Bromophenol

d8
 Alkylphenols
standard solution
300

LC-FL
LC FL NP
250
LC-MS n-OP
GC-MS
GC-MS
200
GC-MS
ng/mL

150
n

100

LC-MS
50

0
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

RSD 5 labs 23-29%

 Alkylphenols
River Seveso extract
1200

GC-MS NP
1000
tert-OP
LC-MS
800

LC-FL
ng//mL

GC-MS GC-MS
600

400

LC MS
LC-MS
200

0
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

RSD 5 labs % 29 (NP)

 Alkylphenols
river Po
120
GC-MS

100

LC-MS LC-FL
80
ng/L

60

GC MS
GC-MS
LC-MS
40
GC-MS GC-MS
< 22
20
5.1 < 10 < 10

0
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07
 CMA on
on-site
site field intercomparison:
PAH in river water
Methods used:
/ /
CMA00: SPE- LC/MS/MS
CMA01: LLE-GC/MS
CMA02: SPE
SPE-GC/MS
GC/MS
CMA03: LLE-GC/MS
CMA04: ASE
ASE-GC/MS
GC/MS (only SPM)
CMA05: LLE-GC/MS
CMA06: SPE
SPE-GC/MS
GC/MS
CMA07: SPE-GC/MS
 CMA on-site field intercomparison:
PAH in river water

CMA00 CMA01 CMA02 CMA03

22.00 CMA04 CMA05 CMA06 CMA07
20.00 89
18.00
16.00 value below
LOQ
14.00
[ng/L]

12.00
10.00
8.00
6.00
4.00
2.00
0.00
ne

ne

e
ne
ne

en
en
he

e
re
ce

th

yr
yl
py
nt
ra

er
an

)p
ra

a)
th

i )p

cd
or
uo

o(
An

flu

h,

-
,3
nz
Fl

g,
k)

,2
Be

o(
j+

(1
nz
b+

no
Be
o(

de
nz

In
Be
 Po river: PAH sum whole water results
(SPM = 45 mg/L)
LLE
40

135
30 AVERAGE OF
CENTRIFUGE SPEEDISK
5
SPE
REFERENCE SPM WITHOUT
ng//L

METHODS FILTRATION WITHOUT

20 SPE
LLE FILTRATION
SPM+DISS WITH
FILTRATION < 10 FILTR + LLE < 10
SPM+DISS
DISS
10

0
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

Grey bars are the blank value determined LOQs of the labs.
 CMA on
on-site
site field intercomparison:
PBDE in river water
Methods used:
/ /
CMA00: SPE- GC/MS/MS
CMA03: LLE-GC/MS
CMA04: ASE
ASE-GC/MS
GC/MS (only SPM)
CMA05: LLE-GC/MS
CMA06: SPE
SPE-GC/MS
GC/MS
 CMA on
on-site
site field intercomparison:
PBDE in river water

0.5

0.4 CMA00
CMA03
0.3 CMA04
L]
[ng/L

CMA05
0.2 CMA06

0.1

0
BDE-28 BDE-47 BDE-99 BDE-100 BDE-153 BDE-154
 Po river: PBDE sum whole water results
(SPM = 45 mg/L)
/L)
FILTR + LLE
1
SPM+DISS

0.8
SPEEDISK
AVERAGE OF
5 WITHOUT
0.6 REFERENCE FILTRATION
L]
[ng/L

METHODS
0.4 SPM+DISS
CENTRIFUGE FILTR + LLE
SPM SPM+DISS
0.2

0
CMA00 CMA03 CMA04 CMA05 CMA06
 Results for river Po Total PBDEs
water expressed as total 3,5
3
concentrations (ng/L) 2,5
Grey bars are < LOD 2
1,5
1
0,5
Conclusion:
0
Further work is needed to develop suitable and
Total
robust PAHs for PS monitoring in natural surface
methods
160 waters and harmonise labs at EC level
140
120
Total alkylphenols
120
100
80 100
60 80
40
60
20
0 40
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07
20

0
CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07
Participant
 POLYCHLOROALKANES C10 to C13 Challenges 1

) Complex mixture of many thousands of positional isomers

6304 positional isomers assuming no more than one bound chlorine
atom on any carbon atom

) Incomplete chromatographic separation

gas chromatograms of short-chain
short chain PCAs show broad humps covering
10 to 15 minutes

) Lack of specificity when using GC

GC-ECNI-MS
ECNI MS at standard ion
source operating conditions
Monitoring of less charcteristic m/z = 70-73 ions (Cl2-; HCl2-)
 POLYCHLOROALKANES C10 to C13 Challenges 2

) Lack of specificity when using GC-ECNI-MS detection at

nominal mass
potential
t ti l interferences
i t f ffrom PCB
PCBs, ttoxaphenes,
h and
d chlordane
hl d related
l t d
compounds

) Lack of appropriate calibration standards

Depending on the degree of chlorination of the calibration standard
results can differ from by a factor of ten
 POLYCHOROALKANES C10 to C13 Methods

) Dehydrochlorination of the PCAs and subsequent

determination of the resulting aliphatic hydrocarbons

) Gas chromatography with electron impact

ionisation mass spectrometric detection (GC-MS)
) Gas chromatography with electron capture negative
ionisation mass spectrometric detection at high
resolution (GC-ECNI-HRMS)
) Gas chromatography with electron capture negative
ionisation mass spectrometric detection at low
resolution (GC-ECNI-MS)
 POLYCHLOROALKANES C10 to C13
I t l b
Interlaboratory
t study
t d

First interlaboratory study on PCAs in 1998

G. T. Tomy et al. Analytical Chemistry 71 (1999), 446-451

Results:
) Seven laboratories
) Two PCA solutions, two fish extracts, PCA-60 commercial
formulation of stated concentration
) In case of the two PCA solutions deviations from of up to 74
and 310 %, respectively, were observed
) Fish sample1 (only three results) RSD = 27%
) Fish sample 2 RSD = 47%
Conclusion:
) Analysis of PCAs is not under control
 POLYCHLOROALKANES C10 to C13 Standardisation

) Analysis of short-chain PCAs is just

on the research level at the moment

) EC gives a mandate to CEN for

developing a suitable method