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 Aries 1, David R. Anderson 1, Nicholas Ordsmith 2, Lisa Fitzpatrick 3 and Fiona Barclay 3 1 CORUS R, D and T, Swinden Technology Centre, Rotherham, UK S60 3AR 2 HALL ANALYTICAL LABORATORIES, Manchester, UK M23 9YJ 3 SIGMA ALDRICH COMPANY Ltd., Poole, Dorset, UK BH12 4QH 1 POPs Workshop_30March2006
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 Version Florisil Version Validation data Sample throughput Acknowledgements 2
Standard analytical procedures for PCB and PCDD/F analyses Method US EPA 1613 B : PCDD/Fs in solids, waste waters and sediments Method US EPA 23 : PCDD/Fs in stack emissions Method EN1948 : European method for PCDD/Fs and PCBs in stack emissions Method 1668A : PCBs in water, soil sediment and tissue ISOTOPE DILUTION : 13 C 12 PCDD/F and PCB internal standards High Resolution Gas Chromatography / High Resolution Mass Spectrometry HRGC / HRMS SEVERAL DAYS OF SAMPLE PREPARATION Extraction (Soxhlet, Dionex ASE) Solvent concentration Successive clean-up steps (Silica, Florisil, Alumina, Activated Carbon) Data processing 3
Supelco Dioxin Prep System Collaboration 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 by Corus, 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 by Corus and Hall Analytical Laboratories. 4
Testing Laboratories Trace Organic Analysis Laboratory (Corus R, D and T) Hall Analytical Laboratories Hall Analytical Laboratories Manchester, UK Contract laboratory UKAS (ISO 17025) accreditation for PCDD/F and PCB analyses 2 HRGC HRMS instruments Corus R, D & T Swinden Technology Centre, Rotherham, UK Researches 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 analyses Micromass Autospec Ultima HRGC - HRMS 5
Corus and Hall Analytical standard procedures for determination of PCDD/Fs and PCBs in environmental samples EMISSION OR DUST SAMPLES Addition of 13 C 12 PCBs & 2,3,7,8-PCDD/Fs EXTRACTION (ASE 200) MULTI-LAYERED SILICA CHROMATOGRAPHY PCBs Saturated hydrocarbons FLORISIL CHROMATOGRAPHY PCDD/Fs HRGC / HRMS US EPA 23 or 1613B EN-1948 BASIC ALUMINA CHROMATOGRAPHY Saturated hydrocarbons PCBs HRGC / HRMS Method 6 US EPA 1668 A
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 Basic Alumina step = 1 day PCBs Analysis overnight Data processing = 1 day PCDD/Fs Analysis overnight Data processing = 1 day PCDD/Fs : 4 to 5 working days PCBs : 5 to 6 working days Additional Tasks : Packing and activation of columns 7
SUPELCO DIOXIN PREP SYSTEMS The dual-carbon column version Description Analytical methods Validation data The Florisil version Description Analytical methods Validation data Sample throughput using Supelco Dioxin Prep system - Florisil Version 8
Supelco Dioxin Prep System - Dual Carbon Column Version Carboxen 1016 (75 m 2 / g) Carboxen 1000 (1200 m 2 / g) Multi-layer Silica gel column Dual carbon reversible tube Silica gel layer Silica gel layer Vacuum manifold 9
Analytical procedure Dioxin Prep System Dual Carbon Column Version ENVIRONMENTAL SAMPLES EXTRACTION (ASE 200) Addition of 13 C 12 2,3,7,8-PCDD/Fs and WHO-12 PCBs Toluene 100 ml CLEAN - UP Dual - carbon column fitted to an empty silica column Back elution of dual-carbon column WASTE 200 ml n-hexane PCDD/F FRACTION HRGC / HRMS 10
Analysis of a waste sinter dust (QC material) for PCDD/Fs using the Dioxin Prep System- Dual Carbon Column Version 400 300 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) ng / kg 200 100 0 11 2,3,7,8-TCDD 1,2,3,7,8-PeCDD 1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDD 1,2,3,7,8,9-HxCDD 1,2,3,4,6,7,8-HpCDD OCDD 2,3,7,8-TCDF 1,2,3,7,8-PeCDF 2,3,4,7,8-PeCDF 1,2,3,4,7,8-HxCDF 1,2,3,6,7,8-HxCDF 2,3,4,6,7,8-HxCDF 1,2,3,7,8,9-HxCDF 1,2,3,4,6,7,8-HpCDF 1,2,3,4,7,8,9-HpCDF OCDF Total I-TEQ (ng/kg)
Supelco Dioxin Prep System - Florisil Version Schematic 250 ml solvent reservoir (28449-U) 24/24mm PP Connector (21216-U) Multi-layered silica tube (28397-U) 6.35/10 mm Reducing union (28398-U) Florisil micro-column 6.35/10 mm Reducing union (28398-U) Vacuum Manifold (28403-U) 12
Supelco Dioxin Prep System - Florisil Version Multi-layer silica and activated Florisil MULTI - LAYER SILICA Designed to meet requirements of Japanese Industrial Standard Methods K- 0311 / K - 0312 Silica gel 0.9 g Glass filter Multi-layer Silica AgNO 3 (10%) 3g H 2 SO 4 (22%) 6g H 2 SO 4 (44%) 4.5g KOH(2%) 3g ACTIVATED FLORISIL Empty Florisil tubes and connectors. Flame sealed ampoules containing 1g activated Florisil (60/100 mesh) 13
Analytical procedure Dioxin Prep System - Florisil Version ENVIRONMENTAL SAMPLES EXTRACTION Addition of 13 C 12 labelled internal standards PCDD/Fs ELUTION 50 ml DCM CLEAN-UP Multi-layer silica Florisil micro-column WASTE + PCBs 175 ml n-hexane WASTE + PCBs 25 ml Hexane/DCM (98 / 2, v/v) PCDD/Fs WASTE (Mainly saturated hydrocarbons) ALUMINA CLEAN - UP PCBs HRGC / HRMS 14
PeCDFs HxCDFs HpCDFs Determination of PCDD/Fs in DX-3 certified reference sediment using the Dioxin Prep System Florisil Version ng / kg 6000 5000 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) 4000 3000 2000 1000 0 15 2,3,7,8-TCDD 1,2,3,7,8-PeCDD 1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDD 1,2,3,7,8,9-HxCDD 1,2,3,4,6,7,8-HpCDD OCDD 2,3,7,8-TCDF 1,2,3,7,8-PeCDF 2,3,4,7,8-PeCDF 1,2,3,4,7,8-HxCDF 1,2,3,6,7,8-HxCDF 2,3,4,6,7,8-HxCDF 1,2,3,7,8,9-HxCDF 1,2,3,4,6,7,8-HpCDF 1,2,3,4,7,8,9-HpCDF OCDF TeCDDs PeCDDs HxCDDs HpCDDs TeCDFs
Determination of WHO-12 PCBs in DX-3 certified reference sediment using the Dioxin Prep System Florisil Version ng / g 25.00 20.00 15.00 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) 10.00 5.00 0.00 PeCB 105 PeCB 114 PeCB 118 PeCB 123 HxCB 156 HxCB 157 HxCB 167 HpCB 189 TeCB 77 TeCB 81 PeCB 126 HxCB 169 16
Determination of non dioxin-like PCBs in DX-3 certified reference sediment using the Dioxin Prep System - Florisil Version ng / g 20 18 16 14 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] 12 10 8 6 4 2 0 TrCB 28 TePCB 52 PeCB 101 HxCB 153 HxCB 138 HpCB 180 HxCB 149 OcCB 194 OcCB 202 OcCB 205 NoCB 206 NoCB 208 DeCB 209 17
13C-PeCB 81 13C-PeCB 126 13C-PeCB 169 Typical recoveries of 13 C 12 labelled PCDD/Fs and PCBs using the Dioxin Prep System - Florisil Version 120.0 100.0 80.0 60.0 40.0 20.0 0.0 18 13C-PeCB 77 % DX-3 Replicate 1 (%) DX-3 Replicate 2 (%) DX-3 Replicate 3 (%) DX-3 Replicate 4 (%) 13C-2,3,7,8-TCDD 13C-1,2,3,7,8-PeCDD 13C-1,2,3,6,7,8-HxCDD 13C-1,2,3,4,6,7,8-HpCDD 13C-OCDD 13C-2,3,7,8-TCDF 13C-1,2,3,7,8-PeCDF 13C-1,2,3,6,7,8-HxCDF 13C-1,2,3,4,6,7,8-HpCDF 13C-PeCB 105 13C-PeCB 114 13C-PeCB 118 13C-PeCB 123 13C-PeCB 156 13C-PeCB 157 13C-PeCB 167 13C-PeCB 189
Fish tissue (WMF-01, Wellington Laboratories) and sandy soil (CRM 529, Supelco) Validation data using the Dioxin Prep System - Florisil Version 20 18 16 14 ng / kg ng / kg 180000 160000 140000 120000 12 10 8 6 4 100000 80000 60000 40000 2 20000 0 0 19 2,3,7,8-TCDD 1,2,3,7,8- PeCDD 1,2,3,6,7,8- HxCDD 2,3,4,7,8- PeCDF PeCB 105 PeCB 114 PeCB 118 PeCB 123 HxCB 156 HxCB 157 HxCB 167 HpCB 189 TeCB 77 TeCB 81 PeCB 126 HxCB 169 WMF-01 Fish Tissue - PCDD/F data WMF-01 Fish Tissue - PCB data 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) 7000 6000 5000 Soil CRM 529 - PCDD/F data 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) 4000 3000 2000 1000 0 2,3,7,8-TCDD 1,2,3,7,8-PeCDD 1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDD 1,2,3,7,8,9-HxCDD 2,3,7,8-TCDF 1,2,3,7,8-PeCDF 2,3,4,7,8-PeCDF 1,2,3,4,7,8-HxCDF 1,2,3,6,7,8-HxCDF 2,3,4,6,7,8-HxCDF 1,2,3,7,8,9-HxCDF
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 PCBs Analysis overnight Data processing = 1 day PCDD/Fs Analysis 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) 20
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 21