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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