1

THE SUPELCO “DIOXIN PREP SYSTEM - FLORISIL VERSION” : A MULTI-LAYER SILICA GEL COLUMN

CONNECTED IN SERIES TO AN ACTIVATED FLORISIL MICRO-COLUMN FOR RAPID DETERMINATIONS OF PCDD/Fs AND PCBs IN ENVIRONMENTAL SAMPLES

Eric Aries1, David R. Anderson1, Nicholas Ordsmith2, Lisa Fitzpatrick3 and Fiona Barclay3

1CORUS R, D and T, Swinden Technology Centre, Rotherham, UK S60 3AR2HALL ANALYTICAL LABORATORIES, Manchester, UK M23 9YJ3SIGMA ALDRICH COMPANY Ltd., Poole, Dorset, UK BH12 4QH

POPs Workshop_30March2006

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Overview

• Brief review on standard analytical procedures for PCDD/Fs and PCB analyses

• Collaboration between Corus / Hall Analytical / Supelco on the Dioxin Prep Systems

• Supelco Dioxin Prep Systems

Dual Carbon Column VersionFlorisil VersionValidation dataSample throughput

• Acknowledgements

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Standard analytical procedures for PCB and PCDD/F analyses

Method US EPA 1613 B : PCDD/Fs in solids, waste waters and sedimentsMethod US EPA 23 : PCDD/Fs in stack emissionsMethod EN1948 : European method for PCDD/Fs and PCBs in stack emissionsMethod 1668A : PCBs in water, soil sediment and tissue

ISOTOPE DILUTION : 13C12 PCDD/F and PCB internal standardsHigh Resolution Gas Chromatography / High Resolution Mass Spectrometry

HRGC / HRMS

SEVERAL DAYS OF SAMPLE PREPARATIONExtraction (Soxhlet, Dionex ASE)Solvent concentrationSuccessive clean-up steps (Silica, Florisil, Alumina, Activated Carbon)Data processing

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Supelco Dioxin Prep SystemCollaboration Corus / Hall Analytical / Supelco

2002 : Development and validation of the Dioxin Prep System – Dual Carbon Column Version in Japan for fast determinations of PCDD/Fs.(Japan Quality Assurance Organisation and Kawajyu Techno Service)

2003 : Evaluation of the Dioxin Prep System – Dual Carbon Column Version by Corus and Hall Analytical prior to its UK commercial launch.

2004 : A collaboration agreement is signed byCorus, Hall and Supelco to develop and validate an enhanced version of the Dioxin Prep System.

2005 : Development and validation of the Supelco Dioxin Prep System - Florisil Version byCorus and Hall Analytical Laboratories.

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Testing LaboratoriesTrace Organic Analysis Laboratory (Corus R, D and T)Hall Analytical Laboratories

Hall Analytical LaboratoriesManchester, UKContract laboratoryUKAS (ISO 17025) accreditationfor PCDD/F and PCB analyses2 HRGC – HRMS instruments

Corus R, D & TSwinden Technology Centre, Rotherham, UKResearches into the emissions and the formation of PCDD/Fs and PCBs from UK steel plants (iron ore sintering, EAF steel-making)UKAS (ISO 17025) accreditation for PCDD/F and PCB analysesMicromass Autospec Ultima HRGC - HRMS

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Corus and Hall Analytical standard procedures for determination of PCDD/Fs and PCBs inenvironmental samples

EXTRACTION (ASE 200)

MULTI-LAYERED SILICA CHROMATOGRAPHY

FLORISIL CHROMATOGRAPHY PCDD/FsHRGC / HRMS

US EPA 23 or 1613BEN-1948

EMISSION OR DUST SAMPLES

Addition of 13C12 PCBs& 2,3,7,8-PCDD/Fs

PCBsSaturated

hydrocarbons

Saturatedhydrocarbons

PCBs HRGC / HRMSMethod

US EPA 1668 A

BASIC ALUMINA CHROMATOGRAPHY

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Sample throughput at Trace Organic Analysis Laboratory (Corus) using standard analytical methods

1 fully trained technician / 8 samples

Extraction and concentration (ASE 200) = 1 to 2 days

Multi-layer silica clean-up = 1 day

Florisil step = 1 day

PCDD/FsAnalysis overnight

Data processing = 1 day

Basic Alumina step = 1 day

PCBsAnalysis overnight

Data processing = 1 day

PCDD/Fs : 4 to 5 working daysPCBs : 5 to 6 working daysAdditional Tasks : Packing and activation of columns

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SUPELCO “DIOXIN PREP SYSTEMS”

The dual-carbon column versionDescription Analytical methodsValidation data

The Florisil versionDescription Analytical methodsValidation data

Sample throughput using Supelco Dioxin Prep system - Florisil Version

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Supelco Dioxin Prep System - Dual Carbon Column Version

Vacuummanifold

Dual carbon reversible tube

Multi-layer Silica gel column

Carboxen 1016(75 m2 / g)

Carboxen 1000(1200 m2 / g)

Silica gellayer

Silica gellayer

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Dual - carbon column fitted to an empty silica column

Back elution of dual-carbon column

PCDD/F FRACTION

HRGC / HRMS

Toluene100 mL

Analytical procedureDioxin Prep System – Dual Carbon Column Version

Addition of 13C12 2,3,7,8-PCDD/Fsand WHO-12 PCBs

EXTRACTION (ASE 200)

WASTE 200 ml n-hexane

ENVIRONMENTAL SAMPLES

CLEAN - UP

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Analysis of a waste sinter dust (QC material) for PCDD/Fs using the Dioxin Prep System- Dual Carbon Column Version

0

100

200

300

400

2,3,

7,8-TCDD

1,2,3,

7,8-PeC

DD

1

,2,3,4

,7,8-H

xCDD

1

,2,3,6

,7,8-H

xCDD

1

,2,3,7

,8,9-H

xCDD

1,2

,3,4,6

,7,8-H

pCDD

OCDD

2,3,

7,8-TCDF

1,2,3,

7,8-PeC

DF

2,3,4,

7,8-PeC

DF

1

,2,3,4

,7,8-H

xCDF

1

,2,3,6

,7,8-H

xCDF

2

,3,4,6

,7,8-H

xCDF

1

,2,3,7

,8,9-H

xCDF

1,2

,3,4,6

,7,8-H

pCDF

1,2

,3,4,7

,8,9-H

pCDF

OCDF

Total I-T

EQ (ng/kg

)

ng / kg

Corus data using standard US EPA 23 analytical method (N = 59)Corus data using Dioxin Prep system (N = 2)Hall Analytical data using Dioxin Prep system (N = 3)

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Supelco Dioxin Prep System - Florisil VersionSchematic

250 mL solvent reservoir(28449-U)

24/24mm PP Connector(21216-U)

Multi-layered silica tube(28397-U)

Florisil micro-column

6.35/10 mm Reducing union(28398-U)

6.35/10 mm Reducing union(28398-U)

Vacuum Manifold(28403-U)

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Supelco Dioxin Prep System - Florisil VersionMulti-layer silica and activated Florisil

Glass filter

AgNO3(10%) 3g

Multi-layerSilica

Silica gel 0.9 g

KOH(2%) 3gH2SO4(44%) 4.5g

H2SO4(22%) 6g

MULTI - LAYER SILICA

Designed to meet requirements of Japanese Industrial StandardMethods K- 0311 / K - 0312

ACTIVATED FLORISIL

Empty Florisil tubes and connectors.

Flame sealed ampoules containing 1g activated Florisil (60/100 mesh)

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Analytical procedureDioxin Prep System - Florisil Version

WASTE + PCBs25 ml Hexane/DCM (98 / 2, v/v)

WASTE(Mainly saturated

hydrocarbons)

ALUMINA CLEAN - UP PCBs HRGC / HRMS

PCDD/Fs ELUTION50 mL DCM

PCDD/Fs

ENVIRONMENTAL SAMPLES

Addition of 13C12 labelled internal standards

EXTRACTION

WASTE + PCBs 175 ml n-hexane

Multi-layer silica

Florisil micro-column

CLEAN-UP

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Determination of PCDD/Fs in DX-3 certified reference sediment using the Dioxin Prep System – Florisil Version

0

1000

2000

3000

4000

5000

6000

2,3,7

,8-TC

DD1,2

,3,7,

8-Pe

CDD

1,2,3

,4,7,

8-Hx

CDD

1,2,3

,6,7,

8-Hx

CDD

1,2,3

,7,8,

9-Hx

CDD

1,2,3

,4,6,

7,8-

HpCD

DOC

DD2,

3,7,8

-TCD

F1,

2,3,7

,8-P

eCDF

2,3,

4,7,8

-PeC

DF

1,2,

3,4,7

,8-Hx

CDF

1,2,

3,6,7

,8-Hx

CDF

2,3,

4,6,7

,8-Hx

CDF

1,2,

3,7,8

,9-Hx

CDF

1,2,3

,4,6,

7,8-H

pCDF

1,2,3

,4,7,

8,9-H

pCDF

OCDF

TeCD

DsPe

CDDs

HxCD

DsHp

CDDs

TeCD

FsPe

CDFs

HxCD

FsHp

CDFs

ng / kg

Certified values (ng / kg) DX-3 Replicate 1 (ng / kg)DX-3 Replicate 2 (ng / kg) DX-3 Replicate 3 (ng / kg)DX-3 Replicate 4 (ng / kg)

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Determination of WHO-12 PCBs in DX-3 certified reference sediment using the Dioxin Prep System – Florisil Version

0.00

5.00

10.00

15.00

20.00

25.00

PeCB

105

PeCB

114

PeCB

118

PeCB

123

HxCB

156

HxCB

157

HxCB

167

HpCB

189

TeCB

77

TeCB

81

PeCB

126

HxCB

169

ng / g

Certified values (ng / g)DX-3 Replicate 1 (ng / g)DX-3 Replicate 2 (ng / g)DX-3 Replicate 3 (ng / g)DX-3 Replicate 4 (ng / g)

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Determination of ‘non dioxin-like’ PCBs in DX-3 certified reference sediment using the Dioxin Prep System - Florisil Version

0

2

4

6

8

10

12

14

16

18

20ng / g

TrCB

28

TePC

B 52

PeCB

101

HxCB

153

HxCB

138

HpCB

180

HxCB

149

OcCB

194

OcCB

202

OcCB

205

NoCB

206

NoCB

208

DeCB

209

Information values (ng/g)DX-3 concentrations (ng/g) [Replicate 1] DX-3 concentrations (ng/g) [Replicate 2] DX-3 concentrations (ng/g) [Replicate 3] DX-3 concentrations (ng/g) [Replicate 4]

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Typical recoveries of 13C12 labelled PCDD/Fs and PCBs using the Dioxin Prep System - FlorisilVersion

0.0

20.0

40.0

60.0

80.0

100.0

120.0

13C-

2,3,7,

8-TC

DD

13C-

1,2,3,

7,8-P

eCDD

13C-

1,2,3,

6,7,8-

HxCD

D

13C-

1,2,3,

4,6,7,

8-Hp

CDD

13C-

OCDD

13C-

2,3,7,

8-TC

DF

13C-

1,2,3,

7,8-P

eCDF

13C-

1,2,3,

6,7,8-

HxCD

F

13C-

1,2,3,

4,6,7,

8-Hp

CDF

13C-

PeCB

105

13C-

PeCB

114

13C-

PeCB

118

13C-

PeCB

123

13C-

PeCB

156

13C-

PeCB

157

13C-

PeCB

167

13C-

PeCB

189

13C-

PeCB

7713

C-Pe

CB 81

13C-

PeCB

126

13C-

PeCB

169

%

DX-3 Replicate 1 (%) DX-3 Replicate 2 (%)DX-3 Replicate 3 (%) DX-3 Replicate 4 (%)

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Fish tissue (WMF-01, Wellington Laboratories) and sandy soil (CRM 529, Supelco) – Validation data using the Dioxin Prep System - Florisil Version

0

2

4

6

8

10

12

14

16

18

20

2,3,

7,8-

TCD

D

1,2,

3,7,

8-Pe

CD

D

1,2,

3,6,

7,8-

HxC

DD

2,3,

4,7,

8-Pe

CD

F

ng / kg

0

20000

40000

60000

80000

100000

120000

140000

160000

180000

PeCB

105

PeCB

114

PeCB

118

PeCB

123

HxCB

156

HxCB

157

HxCB

167

HpCB

189

TeCB

77

TeCB

81

PeCB

126

HxCB

169

ng / kg

Certified values (ng / kg)WMF-01 Replicate 1 (ng / kg)WMF-01 Replicate 2 (ng / kg)WMF-01 Replicate 3 (ng / kg)WMF-01 Replicate 4 (ng / kg)

0

1000

2000

3000

4000

5000

6000

7000

2,3,

7,8-

TCDD

1,2,

3,7,

8-Pe

CDD

1,2,

3,4,

7,8-

HxCD

D1,

2,3,

6,7,

8-Hx

CDD

1,2,

3,7,

8,9-

HxCD

D

2,3,

7,8-

TCDF

1,2,

3,7,

8-Pe

CDF

2,3,

4,7,

8-Pe

CDF

1,2,

3,4,

7,8-

HxCD

F1,

2,3,

6,7,

8-Hx

CDF

2,3,

4,6,

7,8-

HxCD

F1,

2,3,

7,8,

9-Hx

CDF

ng / kg Certified values (ng / kg)CRM529 Replicate 1 (ng / kg)CRM529 Replicate 2 (ng / kg)CRM529 Replicate 3 (ng / kg)CRM529 Replicate 4 (ng / kg)

WMF-01 Fish Tissue - PCDD/F data WMF-01 Fish Tissue - PCB data

Soil CRM 529 - PCDD/F data

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Sample throughput at Trace Organic Analysis Laboratory (Corus) using the Supelco Dioxin Prep System – Florisil Version

1 fully trained technician / 9 samples

Extraction and concentration (ASE 200) = 1 to 2 days

Multi-layer silica clean-up + Florisil = 1 day

Basic Alumina step = 1 day PCDD/FsAnalysis overnight

Data processing = 1 dayPCBsAnalysis overnight

Data processing = 1 day

PCDD/Fs : 3 to 4 working days PCBs : 4 to 5 working days No additional tasks (column packing and activation)

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Acknowledgements• Shelley Richardson

Corus R, D & T

• Laurent Ragava, Florian Mure

University of Aix-Marseille III, France

• Rasika Fernando

University of Sheffield

• Sue and Katie Hall Analytical Laboratories