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Analytical Methods for PAH, PBDE, C10-13 Chloroalkanes andC10 13 Chloroalkanes and

alkylphenols

Stefano POLESELLOStefano POLESELLOStefano POLESELLOStefano POLESELLOIstituto di Ricerca sulle AcqueIstituto di Ricerca sulle Acque--CNRCNRSezione di BrugherioSezione di Brugherio

l ll @i itl ll @i itpolesello@irsa. cnr.itpolesello@irsa. cnr.it

ALKYLPHENOLS

Status

ALKYLPHENOLS

Priority Substance Standard Principle

inland otherinland waters

other waters

Nonylphenols3) ISO 18857-1:2005 GC/MS C C

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

ISO 18857-1:2005 GC/MS A D

LOQ = 0.1 µg/L for NP and 0.0033 µg/L for OPµg/L for NP and 0.0033 µg/L for OP

Blank interferences for NPBlank interferences for NP

At my knowledge, the only CRM available for NP is No validation for whole water

WWTP sludge (www.eurofins.dk).

Alkylphenols: analytical methodsAlkylphenols: analytical methodsAlkylphenols: analytical methodsAlkylphenols: analytical methods

Liquid chromatographyLiquid chromatography–– ESI (ESI (--) (LC) (LC--MS/MS) (MS/MS) (ASTM WK15211ASTM WK15211)–– Fluorimeter (Fluorimeter (IRSA method, ItalyIRSA method, Italy))

Gas chromatography with MS detection (Gas chromatography with MS detection (ASTM ASTM WK1521WK1521; ASTM D7065 – 06)

Wi h d i i i (Wi h d i i i (ISO 188ISO 188 1 2001 200 ))–– Without derivatisation (Without derivatisation (ISO 18857ISO 18857--1:20051:2005))–– With derivatisation with MSTFA (With derivatisation with MSTFA (ISO 18857ISO 18857--2:draft2:draft))

ExtractionExtractionExtractionExtractionWaterWater–– LLE with toluene (LLE with toluene (ISO 18857ISO 18857--1:2005; ASTM D70651:2005; ASTM D7065--0606))–– Continuous LLE (Continuous LLE (ASTM WK1521ASTM WK1521)–– SPE with C18, SDVB (SPE with C18, SDVB (ISO 18857ISO 18857--2:draft ; ASTM 2:draft ; ASTM

WK15211WK15211)

Solid samples (SPM, sediment)Solid samples (SPM, sediment)–– Soxhlet or ASE extraction withSoxhlet or ASE extraction with

MEOH (MEOH (IRSA method, Italy)IRSA method, Italy)Acetone/esanoAcetone/esano

LCLC::–– single peak for each isomeric mixtures of a single single peak for each isomeric mixtures of a single

compound…compound…–– …but often broad peaks…but often broad peaks–– Possibility to determine ethoxylatesPossibility to determine ethoxylates–– Columns: preferred polar embedded or reverse Columns: preferred polar embedded or reverse

phase such as pphase such as phenylhexylhenylhexyl

GCGC–– Different peaks for about every isomers…Different peaks for about every isomers…–– ...but integration must be the sum of a group of ...but integration must be the sum of a group of

peakspeaks–– Variability of NP without derivatisation (Adsorption?)Variability of NP without derivatisation (Adsorption?)

GC without derivatisation:GC without derivatisation:ISO 18857ISO 18857--1:20051:2005

Compounds:Compounds: 4n4n--NP, 4NP, 4--NP NP (branched)(branched), 4, 4--terttert--OPOPRange : Range : 0.005 to 0.2 µg/L; drinking, surface 0.005 to 0.2 µg/L; drinking, surface

d t td t tand waste watersand waste watersm/zm/z 135 + 107135 + 107Extraction: Extraction: LLE with tolueneLLE with tolueneClaenClaen--up: up: silica columnsilica columnDetection: Detection: GCGC--MS in SIM (m/z=107/135)MS in SIM (m/z=107/135)Quantification:Quantification: ESTD e ISTD: ESTD e ISTD: 1313CC66--ringring--labelled 4labelled 4--nn--66 gg

nonylphenol m/z 113nonylphenol m/z 113

ISO 18857ISO 18857 1 2005 Water quality1 2005 Water quality determination of selected alkylphenolsdetermination of selected alkylphenolsISO 18857ISO 18857--1, 2005, Water quality 1, 2005, Water quality –– determination of selected alkylphenols determination of selected alkylphenols PartPart 1: Method for non1: Method for non--filtered samples using liquid extraction and gas filtered samples using liquid extraction and gas chromatography with mass selective detection. chromatography with mass selective detection.

ISO 18857ISO 18857 1:20051:2005ISO 18857ISO 18857--1:20051:2005

GC GC withwith derivatizazionderivatizazion::ISO 18857ISO 18857--2:2:draftdraft

Analytes:Analytes: NP, OP, OP(1,2)EO, NP(1,2)EO and BPANP, OP, OP(1,2)EO, NP(1,2)EO and BPARange:Range: 0 005 to 0 2 µg/L (OP and OP(1 2)EO)0 005 to 0 2 µg/L (OP and OP(1 2)EO)Range: Range: 0.005 to 0.2 µg/L (OP and OP(1,2)EO)0.005 to 0.2 µg/L (OP and OP(1,2)EO)

0.030 to 0.2 µg/L (NP and NP(1,2)EO)0.030 to 0.2 µg/L (NP and NP(1,2)EO)0 05 to 0 2 µg/L (BPA)0 05 to 0 2 µg/L (BPA)0.05 to 0.2 µg/L (BPA)0.05 to 0.2 µg/L (BPA)(drinking water, ground and surface water) (drinking water, ground and surface water)

Range :Range : 0.1 to 50 µg/L (waste water)0.1 to 50 µg/L (waste water)gg µg ( )µg ( )Extraction: Extraction: SPE (styreneSPE (styrene--divinylbenzene) without filtrationdivinylbenzene) without filtrationDerivatizazion: Derivatizazion: MSTFA TrifluoroMSTFA Trifluoro--NN--methylmethyl--NN--yy

(trimethylsilyl)acetamide(trimethylsilyl)acetamideDetection: Detection: selected ion monitoringselected ion monitoringQuantification:Quantification: ESTD and ISTD: ESTD and ISTD: 1313CC66--ringring--labelledlabelled

ISO 18857ISO 18857 2:2:draftdraft ((ESTD+ISTDESTD+ISTD))ISO 18857ISO 18857--2:2:draftdraft ((ESTD+ISTDESTD+ISTD))

ISO 18857ISO 18857--2:2:draftdraft((spikedspiked water water extract+ISTDextract+ISTD))

Intercalibration ISO 18857-1:2005

Bavarian Water Management Agency Munich, Germany according Bavarian Water Management Agency Munich, Germany according to ISO 5725to ISO 5725

•17 labs from 6 countries• 3 water samples (duplicate)• 3 water samples (duplicate)

• surface – (0.005 to 0.05 µg/L)• surface – (0.05 to 0.2 µg/L)( µg )• wastewater (0.1 to 25 µg/L)

• calibration solution

Intercalibration trial

matrix variables N NAP% X xref

REC% SR CVR% ref % R R

1 surface 4-tert-OP 26 7.1 16.6 19.0 87.3 4.16 25.14-NP 22 15.4 28.7 23.0 124.9 16.4 57.2

2 surface 4-tert-OP 26 13.3 66.8 75.0 89.0 17.9 26.8

4 NP4-NP 22 26.7 82.8 90.0 92.0 15.6 18.8

3 waste 4-tert-OP 30 0 1400 1400 100 1 0 45 32 03 waste 30 0 1400 1400 100.1 0.45 32.0

4-NP 30 6.3 2020 1800 112.2 0.64 31.5

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

PBDEPBDE

Standardised method does not exist for water: – Pentabromodiphenylether (PDBE) p y ( )– Method ISO: only for sediment (SPM?)– Under development (CEN method)– Under development (CEN method)

LOQ = 0.066 ng/L

POLYBROMINATED DIPHENYLETHERS (PBDEs)

Technical mixtures

Penta-BDEMixture 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-BDEOcta-BDEMixture of hepta-, octa- and nonabromodiphenylethers;BDE-183 can be used as indicator compound

Deca-BDEDecabromodiphenylether makes up more than 97% 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

No standardised method for the determination of PBDE in No standardised method for the determination of PBDE in water available! (CEN Method under development for whole water)

ISO 22032:2006 Determination of selected polybrominated in sediment: diphenylethers (PBDE) in sediment and

l d sewage sludge – Method using extraction and gas chromatography/mass

spectrometry

EQS = Σ0.5 ng/L; Other Surface Waters =0.2 ng/LLOQ = 0 066 ng/LLOQ = 0.066 ng/L

Methods for PBDE analysis in water are available:

EPA Method 527 employs solid-phase extraction with analysis by gas chromatography/mass spectrometry – MDL is fairly high (0.39 μg/l)

EPA Method 1614 2007 applies HRGC/HRMS for the EPA Method 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 The authors enriched 100 and 2500 L of water, respectively, on XAD resin. SPME has been proposed to extract selected BDE congeners from water samples by Polo et al 2004 et al. 2004.

PBDEsAnalytical Methods

Gas chromatography with electron capture detection (GC-ECD)

Gas chromatography with atomic emission detection (GC-AED)

Gas chromatography with electron impact mass spectro-metric detection at low resolution (GC-LRMS)

g p y ( )

( )

Gas chromatography with electron impact mass spectro-metric detection at high resolution (GC-HRMS)g ( )

Gas chromatography with electron capture negative ionisation mass spectrometric detection (GC-ECNI-MS)p ( )

CEN-ISO Method for sedimentGC-ECNI-MS

Experimental Conditions:Experimental Conditions:

Matrix: Sediment, suspended particulate matter (SPM) Compounds: BDE 47 BDE 99 BDE 100 BDE 153Compounds: BDE-47, BDE-99, BDE-100, BDE-153,

BDE-154, (BDE-183, BDE-209)Limit of determination: 0.01 to 0.05 µg/kg (test portion 10 g)Extraction: Soxhlet extraction with toluene orExtraction: Soxhlet extraction with toluene or

hexane/acetoneClean-up: Multi-layer column chromatography on silica gelDetection: GC-ECNI-MS in the selected ion monitoringDetection: GC ECNI MS in the selected ion monitoring

mode (m/z=79/81 for all congeners except BDE-209)

Quantification: ISTD: BDE-77, BDE-181 not occurringQuantification: ISTD: BDE 77, BDE 181 not occurring in technical mixtures; 13C12-labelled BDE-209

PBDEPBDEInterlaboratory studies on ISO Method

First interlaboratory study on PBDEs in 2000J de Boer and W P Cofino Chemosphere 46 (2002) 625-633J. de Boer and W. P. Cofino Chemosphere 46 (2002), 625 633

Results for the two sediment samples:

Congener RSD in % AssessmentBDE-47: 22/24 acceptableBDE-99: 36/45 further improvement necessary/ p yBDE-100: 20/20 acceptableBDE-153: 32/37 further improvement necessaryBDE-154: 35/45 further improvement necessaryBDE-209: 48/78 not under control

PAHPAH

Polycyclic aromatic hydrocarbons

AA-EQS:

Polycyclic aromatic hydrocarbons

Anthracene 100 ng/LFluoranthene 100 ng/LFluoranthene 100 ng/LBenzo(a)pyrene 50 ng/LBenzo(b)fluoranthene+Benzo(k)fluoranthene Σ30 ng/LBenzo(g,h,i)perylene+ Indeno(1,2,3-cd)pyrene Σ2 ng/L

PAH: LOQ = from 0 6 ng/L to 16 ng/Lng/L to 16 ng/LPAH: LOQ = from 0. 6 ng/L to 16 ng/Lng/L to 16 ng/L

StatusPriority Substance Standard

Principle

Status

inland waters

other waters

Fluoranthene ISO 17993: 2002 HPLC/Fluo A A

ISO 17993:Naphthalene ISO 17993: 2002 HPLC/Fluo A A

EN ISO 15680:2003

Purge/Trap + Therm Desorp A A15680:2003 Therm. Desorp.

Benzo(a)pyrene ISO 17993: 2002 HPLC/Fluo C C

Benzo(b)fluoranthene4) ISO 17993: 2002 HPLC/Fluo C C

Benzo(k)fluoranthene4) ISO 17993: C CBenzo(k)fluoranthene4)2002 HPLC/Fluo C C

Available Standard Method

EN ISO 17993: 2003Determination of 15 polycyclic aromatic hydrocarbons (PAH) p y y y ( )in water by HPLC with fluorescence detection after liquid-liquid extractionMatrix: Drinking, ground, waste and surface waterMatrix: Drinking, ground, waste and surface water

Method Descriptionhigh performance liquid chromatography (HPLC) with 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 A l ti l M th dOther Analytical Methods

EPA 8270c claims detection limit of 10 μg/L EPA 8270c claims detection limit of 10 μg/L which is obviously too high.

Other analytical methods based on this standard using modern GC/MS equipment however may tt i th i d l LOQ attain the required low LOQs.

R kRemarks

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 and Indeno[1,2,3-cd]pyrene.[g, , ]p y [ , , ]py

In addition, not enough validation data are available di th l i f f t l i regarding the analysis of surface water samples in

particular samples containing substantial amounts of SPM.

CEN Mandate for the development of analytical methods in progress!

Chemical Monitoring ActivityOn-Site WorkshopOn Site Workshop

Comparison of analytical approaches for WFD chemical monitoring: a field experimentg p

a JRC IES RWER Initiative

1st CMA On-Site Technical 1st CMA On-Site Technical Workshop

10-11 October 2006,in Ferrara Italyin Ferrara, Italy

River Po - Pontelagoscuro

2nd CMA On-Site Technical WorkshopWorkshop

17-18 September 2008in Budapest, Hungary

River Danube - Budapest

Practical sampling exercise on the Po river;SSimultaneous bulk sampling by 8 teams with different methodologies(also from IWP Sofia BG);(also from IWP-Sofia-BG);Use of bulk sampling and short–term integrating methods (large volume approaches);volume approaches);Homogeneity tests during exercise;Acquisition of accompanyingAcquisition of accompanying data, as SPM, pH, temp.

• CMA on‐site organized by JRC IES (Ispra) and the Province of ( ) f h i l i fFerrara (Oct. 2006) for the implementation of WFD.

• 7 laboratories from 7 different countries

• Analytes: PBDEs, PAHs, nonyl and octylphenols

• 3 samples

– Standard 

– extract water from the river :– Corresponding to 14.6 L of Po River waterconcerning PBDEs and PAHs

d f f– Correspondig to 1 L of water from Seveso River concerning alchilfenoli

S li d l i f P Ri– Sampling and analysis of Po River

AlkylphenolsCMA01 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. /solvent

HLB 200 C18 C18 Randallextraction

HLB 60 DCM EnvichromP

Soxhlet

SPE elution EA MeOH +MTBE

Acetone SPM DCM/EA n.a. DCM SPM

Analytical LC-MS (tq) LC-MS (tq) LC-fluor LC-fluor GC-MS GC-MS GC-MS GC-MS (it)methodLC 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 2.1×150mm 2.1 × 100 mm 4.6×250 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 orwavelengthmonitored

219 > 133227 > 112205 > 106205 > 133

219 > 133219 > 106225 > 112205 > 133

230 nm exc.302 nm em

230 nm exc.302 nm em

149(for NP)135(forOP)121107

135 1351075722077

207 > 179221 > 179235 > 179278 > 179292 > 179107 77 292 > 179

IST 4n-NP d8 4n-NP d6 no no 4n-NP d8 4n-NPd8

Atrazine d5 4-Bromophenol

Alkylphenolsstandard solutionstandard solution

300

NPLC-FL

200

250

NP

n-OPLC-MSGC-MS

GC-MS

LC FL

150

200

ng/m

L

GC-MS

100

n

LC-MS

0

50

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

RSD 5 labs 23-29%

AlkylphenolsRiver Seveso extractRiver Seveso extract

1200

1000 NPtert-OP

LC-MS

GC-MS

600

800

/mL

GC-MS GC-MSLC-FL

400

600

ng/

LC MS200

LC-MS

0

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

RSD 5 labs % 29 (NP)

AlkylphenolsAlkylphenolsriver Poriver Po

120 GC-MS

80

100

LC-MS LC-FL

60

80

ng/L

GC MS40

< 22

LC-MS

GC-MS

GC-MS

GC-MS

0

20 < 10< 105.1

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

CMA on site field intercomparison:CMA on-site field intercomparison:PAH in river water

Methods used:CMA00: SPE- LC/MS/MS/ /CMA01: LLE-GC/MSCMA02: SPE-GC/MSCMA02: SPE GC/MSCMA03: LLE-GC/MSCMA04: ASE-GC/MS (only SPM)CMA04: ASE GC/MS (only SPM)CMA05: LLE-GC/MSCMA06: SPE-GC/MSCMA06: SPE GC/MSCMA07: SPE-GC/MS

CMA on-site field intercomparison:

CMA00 CMA01 CMA02 CMA03

PAH in river water

20.00

22.00

CMA00 CMA01 CMA02 CMA03CMA04 CMA05 CMA06 CMA07

89

14.00

16.00

18.00 value below LOQ

8.00

10.00

12.00

[ng/

L]

2.00

4.00

6.00

0.00

Anthrac

ene

Fluoran

thene

zo(a)

pyren

e

)fluora

nthene

h,i)pery

lene

2 3-cd)p

yrene

A Fl

Benz

Benzo

(b+j+k

)fl

Benzo

(g,h

Inden

o(1,2,

3

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

LLE40

135

30SPEEDISK

WITHOUT /L

SPECENTRIFUGE

SPM

AVERAGE OF 5

REFERENCE

20 SPEWITH

FILTRATION FILTR + LLE SPM+DISS

FILTRATION

ng/

LLEWITHOUT

FILTRATION

< 10 < 10

METHODS

SPM+DISS

10 DISSSPM+DISS

0CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

Grey bars are the blank value determined LOQs of the labs.

CMA on site field intercomparison:CMA on-site field intercomparison:PBDE in river water

Methods used:CMA00: SPE- GC/MS/MS/ /CMA03: LLE-GC/MSCMA04: ASE-GC/MS (only SPM)CMA04: ASE GC/MS (only SPM)CMA05: LLE-GC/MSCMA06: SPE-GC/MSCMA06: SPE GC/MS

CMA on-site field intercomparison:CMA on site field intercomparison:

PBDE in river water

0.5

0.3

0.4

L]

CMA00CMA03CMA04

0.2

[ng/

L CMA05CMA06

0

0.1

0BDE-28 BDE-47 BDE-99 BDE-100 BDE-153 BDE-154

Po river: PBDE sum whole water results(SPM 45 /L)(SPM = 45 mg/L)

1 FILTR + LLE

0.8

1 SPM+DISS

0.6

L]

SPEEDISK

WITHOUT FILTRATION

AVERAGE OF 5

REFERENCE

0.4[ng/

L REFERENCE METHODS

SPM+DISSFILTR + LLECENTRIFUGE

0.2

FILTR + LLE SPM+DISS

CENTRIFUGE

SPM

0CMA00 CMA03 CMA04 CMA05 CMA06

Total PBDEsResults for river Po

2,5

3

3,5water expressed as total concentrations (ng/L)

1

1,5

2Grey bars are < LOD

0

0,5

Total PAHs

Conclusion:Further work is needed to develop suitable and

robust methods for PS monitoring in natural surface

120140160

Total PAHs

Total alkylphenols

robust methods for PS monitoring in natural surface waters and harmonise labs at EC level

6080

100120

80

100

120

02040

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07 20

40

60

0

20

CMA00 CMA01 CMA02 CMA03 CMA04 CMA05 CMA06 CMA07

Participant

POLYCHLOROALKANES C to C Challenges 1POLYCHLOROALKANES C10 to C13 – Challenges 1

Complex mixture of many thousands of positional isomers6304 positional isomers assuming no more than one bound chlorine atom on any carbon atomatom on any carbon atom

Incomplete chromatographic separationgas chromatograms of short chain PCAs show broad humps covering

Lack of specificity when using GC ECNI MS at standard ion

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

Lack of specificity when using GC-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

t ti l i t f f PCB t h d hl d l t d

Lack of appropriate calibration standards

potential interferences from PCBs, toxaphenes, and chlordane related compounds

Lack of appropriate calibration standardsDepending on the degree of chlorination of the calibration standard results can differ from by a factor of ten

POLYCHOROALKANES C10 to C13 – Methods10 13

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-HRMS)

ionisation mass spectrometric detection at low resolution (GC-ECNI-MS)

POLYCHLOROALKANES C10 to C13I t l b t t dInterlaboratory study

First interlaboratory study on PCAs in 1998G. 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 concentrationIn case of the two PCA solutions deviations from of up to 74 and 310 %, respectively, were observedFish sample1 (only three results) RSD = 27%Fish sample 2 RSD = 47%

Conclusion:Conclusion:

Analysis of PCAs is not under control

POLYCHLOROALKANES C10 to C13 – StandardisationPOLYCHLOROALKANES C10 to C13 Standardisation

Analysis of short-chain PCAs is just on the research level at the momenton the research level at the moment

EC gives a mandate to CEN for developing a suitable method