Twinning Bulgaria Italia

Training 3.3
Simitli 1717-19 February 2009

Analytical Methods for PAH, PBDE,

C10-13
C10
13 Chloroalkanes and
alkylphenols
Stefano POLESELLO
Istituto di Ricerca sulle AcqueAcque-CNR
Sezione di Brugherio
polesello@irsa.
l
ll @i
cnr.it
it

ALKYLPHENOLS
Status
Priority Substance

Standard

Principle

Nonylphenols3)

ISO 188571:2005

GC/MS

Octylphenol (4-(1,1,3,3)Tetramethylbutylphenol

ISO 188571:2005

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).

inland
waters

other
waters

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
18857-1:2005
188571 200 )
1:2005)
With derivatisation with MSTFA (ISO
(ISO 1885718857-2:draft
2:draft))

Extraction
z Water

LLE with toluene ((ISO

ISO 1885718857-1:2005; ASTM D7065D7065-06
06))
Continuous LLE (ASTM
(ASTM WK1521)
WK1521
SPE with C18, SDVB ((ISO
ISO 1885718857-2: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 1885718857-1:2005
z Compounds:

4n
4n--NP, 44-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:

GCGC-MS in SIM (m/z=107/135)

z Quantification:

ESTD e ISTD: 13C6-ring

ringg-labelled 44-nnonylphenol m/z 113

ISO 1885718857-1,
1 2005
2005, Water quality determination of selected alkylphenols
Part 1: Method for nonnon-filtered samples using liquid extraction and gas
chromatography with mass selective detection.

ISO 1885718857-1:2005

GC with derivatizazion:
derivatizazion:
ISO 1885718857-2: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:
z Detection:

MSTFA Trifluoro
Trifluoro--N-methyl
methyly -N(trimethylsilyl)acetamide

selected ion monitoring

z Quantification:

ESTD and ISTD: 13C6-ring

ring--labelled

ISO 1885718857-2:
2:draft
draft (ESTD+ISTD
ESTD+ISTD))

ISO 1885718857-2: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)
g )
surface ((0.05 to 0.2 g/L)
wastewater (0.1 to 25 g/L)
calibration solution

Intercalibration trial

matrix

variables

NAP
%

xref

REC
%

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

26

13.3

66.8

75.0

89.0

17.9

26.8

22

26.7

82.8

90.0

92.0

15.6

18.8

30

1400 1400 100.1

100 1 0.45
0 45

32 0
32.0

30

6.3

2020 1800 112.2 0.64

31.5

2 surface

4-tert-OP
4 NP
4-NP

3 waste

4-tert-OP
4-NP

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 with electron impact mass spectrometric detection at low resolution ((GC-LRMS))

Gas chromatography with electron impact mass spectrog resolution (GC-HRMS)

(
)
metric detection at high

Gas chromatography with electron capture negative

ionisation mass spectrometric
p
detection (GC-ECNI-MS)
(
)

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

CEN-ISO Method for sediment

GC-ECNI-MS
Experimental Conditions:
Matrix:
Compounds:

Sediment, suspended particulate matter (SPM)

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
BDE-47:
BDE-99:
BDE-100:
BDE-153:
BDE-154:
BDE-209:

RSD
22/24
/
36/45
20/20
32/37
35/45
48/78

in % Assessment
acceptable
further improvement
p
necessary
y
acceptable
further improvement necessary
further improvement necessary
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
Principle

inland
waters

other
waters

Fluoranthene

ISO 17993:
2002

HPLC/Fluo

Naphthalene

ISO 17993:
2002

HPLC/Fluo

EN ISO
15680:2003

Purge/Trap +
Therm Desorp.
Therm.
Desorp

Benzo(a)pyrene

ISO 17993:
2002

HPLC/Fluo

Benzo(b)fluoranthene4)

ISO 17993:
2002

HPLC/Fluo

Benzo(k)fluoranthene4)

ISO 17993:
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 liquidliquid 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

z

z
z
z

the Po river;
S
Simultaneous
bulk sampling by 8
teams with different
methodologies
(also from IWP
IWP-Sofia-BG);
Sofia BG);
Use of bulk sampling and short
term integrating methods (large
volume approaches);
Homogeneity tests during
exercise;
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

C18

C18

Randall
extraction

HLB 60

DCM

Envichrom
P

Soxhlet

SPM

DCM/EA

n.a.

DCM

SPM

SPE cart. / HLB 200

solvent
SPE elution

EA

MeOH
MTBE

Analytical
method

LC-MS (tq)

LC-MS (tq)

LC-fluor

LC-fluor

GC-MS

GC-MS

GC-MS

GC-MS (it)

H2O-ACN

H2O-MeOH

H2O-MeOH

n.a.

n.a.

n.a.

n.a.

C18

Phenylhexyl

Phenylhexyl

DB-5

DB-5

DB-1

ZB-5

LC solvents H2O-ACN

+ Acetone

Column

C18

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

MSTFA

Ions or
wavelength
monitored

219 > 133

227 > 112
205 > 106
205 > 133

219 > 133

219 > 106
225 > 112
205 > 133

230 nm exc. 230 nm exc. 149(for NP) 135

302 nm em
302 nm em
135(for
OP)
121
107

135
107
57
220
77

207 > 179

221 > 179
235 > 179
278 > 179
292 > 179

IST

4n-NP d8

4n-NP d6

no

Atrazine d5

4-Bromophenol

no

no

no

4n-NP d8

4n-NP
d8

Alkylphenols

standard solution
300

LC-FL
LC
FL
250

NP
n-OP

LC-MS

GC-MS

GC-MS

200

n
ng/mL

GC-MS
150

100

LC-MS
50

CMA00

CMA01

CMA02

CMA03

CMA04

CMA05

CMA06

RSD 5 labs 23-29%

CMA07

Alkylphenols
River Seveso extract
1200

GC-MS

NP
tert-OP

1000

LC-MS

ng//mL

800

GC-MS

600

GC-MS

LC-FL

400

LC MS
LC-MS
200

CMA00

CMA01

CMA02

CMA03

CMA04

CMA05

CMA06

RSD 5 labs % 29 (NP)

CMA07

Alkylphenols

river Po
120

GC-MS

100

LC-FL

LC-MS

ng/L

80

60

GC MS
GC-MS

LC-MS
40

< 22

GC-MS

GC-MS

20

5.1

< 10

< 10

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

22.00
20.00

CMA00

CMA01

CMA02

CMA03

CMA04

CMA05

CMA06

CMA07

89

18.00
16.00

value below
LOQ

12.00
10.00
8.00
6.00
4.00
2.00

cd
)p
yr
en
e

In
de
no
(1
,2
,3
-

er
yl
en
e
Be
nz
o(
g,
h,
i )p

ne
flu
or
an
th
e

Be
nz
o(
b+
j+
k)

Be
nz
o(
a)
py
re
ne

Fl
uo
ra
nt
he
ne

0.00
An
th
ra
ce
ne

[ng/L]

14.00

Po river: PAH sum whole water results

(SPM = 45 mg/L)
LLE

135

40

ng//L

30 AVERAGE OF

20

5
REFERENCE
METHODS
SPM+DISS

10

SPE
WITH
FILTRATION

DISS

CENTRIFUGE

SPEEDISK

SPM

WITHOUT
FILTRATION

LLE
< 10

FILTR + LLE
SPM+DISS

SPE
WITHOUT
FILTRATION

< 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

[ng/L
L]

0.5
0.4

CMA00
CMA03

0.3

CMA04

0.2

CMA05
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
SPM+DISS

[ng/L
L]

0.8
0.6

AVERAGE OF
5
REFERENCE
METHODS

0.4

SPM+DISS

SPEEDISK
WITHOUT
FILTRATION

CENTRIFUGE

0.2

SPM

FILTR + LLE
SPM+DISS

CMA04

CMA05

0
CMA00

CMA03

CMA06

Total PBDEs

Results for river Po

water expressed as total 3,5
3
concentrations (ng/L) 2,5
Grey bars are < LOD

2
1,5
1
0,5

160
140
120
100
80
60
40
20
0

Conclusion:
0
Further work is needed to develop suitable and
Total
PAHs for PS monitoring in natural surface
robust
methods
waters and harmonise labs at EC level
Total alkylphenols
120
100
80
60
40

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

20
0
CMA00

CMA01

CMA02

CMA03

CMA04

Participant

CMA05

CMA06

CMA07

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