Persistant Organic Pollutants

Size: px

Start display at page:

Download "Persistant Organic Pollutants"

Jane Cummings
5 years ago
Views:

1 Persistant Organic Pollutants Fast screening and precise confirmation with GC-MSMS and GC-HRMS. Hans-Joachim Huebschmann GC-MS Technology Manager Dirk Krumwiede Product & Application Specialist for Advanced GC-MS Techniques Inge de Dobbeleer EU GC-MS Specialist Thermo Fisher Scientific, Bremen, Germany

2 Persistant Organic Pollutants POPs Stockholm Convention 4, signed by >1 UN member states Global agreement on the termination or restriction of the production of persistant organic pollutants (POPs) Reduce contamination. Dirty Dozen Worldwide Ban Are initially listed in the Stockholm convention as POPs Synthetically but also unintenionally produced halogenated chemicals Dioxins (PCDDs, PCDFs), Polychlorinated Byphenyls (PCBs), Toxaphens, Chlorinated Paraffins (PCs), Brominated Flame Retardants (BFRs), Organochlorine Pesticides (OCPs), addtional compounds added 9 Dioxin/Furans and DL-PCBs Still today of highest safety concern about dietary exposure Food scandals about contaminations are constantly in press Regulatory bodies such as the US Food and Drug Administration (FDA), the US Environmental Protection Agency (EPA) and the EU Commission authorities (EC) have produced detailed testing guidelines. 2

3 General structure of PCDD and PCDF Dioxins are a group of halogenated oxygen-containing aromatic hydrocarbons Polychlorinated Dibenzofurans (PCDFs) 135 possible congeners 1 toxic congeners (2,3,7,8 chloro substitution) Polychlorinated Dibenzo(p)dioxins (PCDDs) 75 possible congeners 7 toxic congeners (2,3,7,8 chloro substitution) 3

4 Dioxin like PCB s PCBs are a group of chlorinated aromatic hydrocarbons Polychlorinated Biphenyles (PCB) 9 possible congeners 4 non-ortho congeneres = coplanar (no chlorine substitution in position 2,2`,6,6`) 8 mono-ortho congeneres = 1 chlorine in position 2 or 2`or 6 or 6` 12 PCBs with dioxin-like properties; WHO-TEFs established in 1997 and revised 5 4

Dioxin Incidents Huge Economical Impact Dioxin and PCB incidents are expensive. Egg production enterprise with 5 million hens would cost in excess of $3 million.

The total costs of the Belgian food crisis are estimated up to $1 billion.

5 Dioxin Incidents Huge Economical Impact Dioxin and PCB incidents are expensive. Egg production enterprise with 5 million hens would cost in excess of $3 million. Broiler enterprise producing 3 million broilers per week would exceed $85 million. Times Beach, Missouri, road dust covered by oil spread. The cleanup cost with a total of $11 million. The total costs of the Belgian food crisis are estimated up to $1 billion. Sources to the Environment Combustion processes Landfill sites Improper waste managment Sewedge sludge Accumulation in Food Chain Regular control required In environment, feed, and food Repeated Dioxin cases from only recent years: 1998 Milk from citrus pulp pellet feed 1999 Belgian PCB/dioxin in eggs, poultry 1999 Clay and zeolithes for feed Choline chloride 2 Carbosan Copper 4 Potato pulp 5 Hydrochloric acid 7 Indian Guar Gum thickener 8 Irish Pork Meat 8 Italian Mozarella Cheese 11 German incident in cattle feed 5

6 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

7 In Brief: The Integrated POPs Analysis Workflow Integrated POPs Analysis Workflow Fast Screening analysis & Precise Confirmation analysis Combined use of high selective MS Technologies: Screening: Confirmation : TSQ Quantum XLS (Triple Quadropole MSMS) DFS (Magnetic Sector HRMS) 7

8 The POPs Analysis Workflow in the Lab Sample Preparation Screening Negative (TEQ < ML) Analysis Done GC-MS/MS Positive (TEQ >ML) Confirmation Negative Analysis Done GC-HRMS Positive Final TEQ Calculation Confirmation Report 8

9 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

10 Dioxins, Furans & DL-PCBs : common sample preparation workflow (1613) Sample Preparation 13 C Surrogates Matrix specific processing or more often here 3-7 days Extraction Clean-up & fractionation 2,3,7,8-[ 37 Cl 4 ]-TCDD Clean- up standard Concentration Instrumental Analysis 13 C Internal standard ( injection standard) 1

11 Extraction of dioxins Sample Preparation Objective to fully recover targets from matrix Disrupt matrix interactions Strongly bound so extraction brings a lot of co-extracted material Solid (semi-solid) state samples Acid digest Pressurized Liquid Extraction (PLE) ASE ( Dionex) Soxhlet Liquid samples Liquid/Liquid SPE 11

12 Multi-layer Silica open column clean-up Sample Preparation 4g 1g Anhydrous Na 2 SO 4 Remove residual water Silica layer as separator 8g 1g 4g 1g Acid Silica layer Removes bases Hydrolyses fats Hydrolyses other organics (except stable species) Basic Silica layer Removes acids 12

13 Separation capabilities Florisil Alumina (acidic or basic) Sample Preparation Dioxins/ dl PCBS Carbon 2-1% DCM in hexane 5% DCM in hexane 2-1% DCM in hexane 5% DCM in hexane Hexane (or DCM in hexane) Reverse Toluene PCBS dioxins PCBS dioxins Comprehensive fractionation difficult Practically & economically best effort with combinations 13 o-pcbs

14 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

15 Screening for POPs needs Screening Industrial Analysis - Food Industry Increasing Food Safetey requirements Ensure consistent Quality for raw materials and products High Brand Consciousness, prevent any damages Collaborate with external contract labs for Certification Government Control Labs Fast in case of accidents High number of samples need to be screened Requirements for Quality Control and at Incidents Manage high sample numbers by screening Provide higher efficiency and flexibility, faster results Run indicated samples on HRMS for confirmation analysis 15

16 Definitions for Screening and Confirmation Methods Screening Screening: High throughput analysis Low false negative rate (< 1%) Limited precision (<3 % RSD) Sensitivity 25% below of ML Confirmation: HRMS methods required High precision (<15 % RSD) Higher sensitivity to monitor lower background and action levels. Maximum Level (ML) TEQ 3 % RSD 15 % RSD Food Safety Screening Confirmation 16 Source: EU Directives 96/23/EC, 1883/6

Why Dioxins Screening with GC-MS/MS? Screening Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance Single Quad MS SIM Mode 19.61 RT: 23.76 23.55 24.11 25.

17 Why Dioxins Screening with GC-MS/MS? Screening Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance Single Quad MS SIM Mode RT: RT: TSQ Quantum XLS MRM Mode RT: RT: RT: Time (min) Matrix Matrix Matrix 17

18 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

19 Triple Quadrupole MS Principle of Operation Screening Quantitation of target compounds in matrix samples Select Fragment Detect Ion Source Detector Q1 selects the precursor ion TCDD ion: m/z Argon Collision Gas Q3 selects the product ion fragments to m/z

20 More Selectivity for More Sensitivity Screening Target POPs Signal 1. u Mass Resolution in Q1 determines selectivity Other technologies use a wide Q1 mass window More matrix gets the into the collision cell, and sacrifices selectivity. Matrix Background

21 Higher Mass Resolution for More Selectivity Dioxin Ratio Mass Dioxin Quan Mass Screening H-SRM Highly Selective SRM Mode.4 u Isobaric Matrix Background TSQ Quantum XLS Less matrix gets into the collision cell less noise. Increased mass resolution increases selectivity for higher S/N. 21

22 No Chemical Noise - Curved Collision Cell Screening Only Product Ions leave the coll. cell Exit lens assembly Entrance lens assembly, controls collision energy Ions and Neutrals Neutrals get pumped away Collision cell housing, Argon, ca. 4 mtorr Square quadrupole rods collision cell 22

23 Structure Specific Selectivity TCDD M Screening Structure Specific Loss of COCl 3 Cl O Cl Cl O Cl Product Ions m/z m/z SRM Precursor Ions m/z m/z (mainlib) 2,3,7,8-Tetrachlorodibenzo-p-dioxin

24 TSQ Quantum XLS Fish Extract with all PCBs Fish extract blank RT: Relative Abundance Screening Tri-CBs: m/z m/z 186. Tetra-CBs: m/z m/z 222. Penta-CBs: m/z m/z Hexa-CBs: m/z m/z Hepta-CBs: m/z m/z 358. Octa-CB: m/z m/z

25 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

moves on wheels Toroidal ESA for highest sensitivity Low power

26 Magnetic Sector HRMS: Thermo Scientific DFS Confirmation Benchtop-like operation (autotune) Most compact instrument in its class One unit, moves on wheels Toroidal ESA for highest sensitivity Low power consumption Unique Dual GC configuration Fit to purpose data evaluation software 26

27 Magnetic Sector HRMS Ion Optics of the DFS Confirmation Magnet TOROIDAL ESA ENS EXS Magnet field plus electric sector analyser = double focussing 27

28 Magnetic Sector HRMS: Resolution Setting Confirmation #668 R: 162 ST: 1. FX: 3.27E7 F: + p EI MS [ ] #624 R: 9857 ST: 1. CR: 2.24E6 F: + p EI MS [ ] Relative Intensity Relative Intensity m/z m/z R 1, R 1, DFS: Tune peak of FC43 calibrant, mz 414, screenshots at R 1, and R 1, 28

29 High Selectivity with High Resolution? Confirmation Mass peaks at Different Resolution Settings: Low Resolution TCDD Red: TCDD mass trace at R 1. Green: TCDD mass trace at R 1. High Resolution Blue: another chlorinated compound mass trace Yet another chlorinated compound Accurate masses are used for Target Compound Analysis 29

30 High Mass Resolution Selectivity Blood Sample Confirmation RT: SM: 5G R = 1, RT: 14. AA: tcdd DFS GC-HRMS Quantitation Mass m/z For the native TCDD RT: AA: 1262 Relative Abundan nce RT: AA: 2991 RT: AA: 1979 R = 5, RT: AA: 11 R = 1, RT: AA: 1564 RT: 13.9 AA: 1777 RT: 14.1 AA: 1834 RT: AA: RT: AA: 535 RT: AA: RT: AA: 13 RT: 14. AA: 1143 RT: AA: 154 RT: AA: RT: AA: 3823 RT: AA: Time (min) Chemical noise from matrix interferences shows up at lower mass resolution ONLY HRMS at R=1, selects the dioxin peak. 3

31 Application Example: Hexa DL-PCB Standard at very low concentration RT: RT: 21.1 AA: 551 RT: AA: fg RT: AA: 47 Isotope Dilution native - Quan mass 5 RT: 21.7 AA: 473 RT: AA: 491 RT: AA: 361 native - Ratio mass 5 RT: 21.7 AA: 2637 RT: AA: RT: AA: C labeled IS - Quan mass 5 RT: 21.7 AA: 2124 RT: AA: RT: AA: Time (min) 13 C labeled IS Ratio mass 31

32 Application Example PCDF and PCDD (Tetradioxins/furans in Fish) Confirmation RT: RT: AA: RT: AA: RT: RT: RT:.5 RT: AA: 397 AA: 4162 AA: AA: RT: RT: 19.2 RT: RT: AA: 7557 AA: 933 AA: 6899 AA: 1512 RT: AA: RT: AA: Relative Abundance RT: RT: RT:.5 RT: AA: AA: 5795 AA: 5231 AA: RT: RT: 19.2 RT: RT: AA: 7144 AA: AA: 8119 AA: 8345 RT: AA: RT: RT:.3 AA: 66 AA: 1312 RT: AA: RT: RT:.3 AA: 7927 AA: Time (min) N 3 m 3 3 M d 7 N 3 m/z= MS dk_diopcb 7 NL: 7.7E5 m/z= MS dk_diopcb 7 NL: 8.85E5 m/z= MS dk_diopcb 7 RT: RT:.18 RT: AA: AA: RT: AA: 2375 RT:.32 AA: RT: AA: 1757 RT: AA: 52 RT:.51 AA: 3528 native RT:.18 RT: AA: 572 AA: 4123 Relative Abundance RT: AA: 281 RT:.32 AA: RT: AA: RT: AA: 8922 RT:.51 RT: AA: 4946 AA: 1382 RT: AA: RT:.31 AA: labeled IS RT:.62 AA: 23 RT: AA: RT:.31 AA: 497 RT:.51 AA: Time (min) N 1 m 3 3 M d 7 N 1 m/z= MS dk_diopcb 7 NL: 1.29E6 m/z= MS dk_diopcb 7 NL: 1.55E6 m/z= MS dk_diopcb 7 TR-Dioxin (TR-5ms type) m column (.1 µm film) 32

33 DFS Sensitivity: TCDD Specification fg SN :1(4 σ) Confirmation Relative Abu undance S/N 239:1 (4σ) Highest sensitivity for: Quantifying at lowest levels High precision Reducing sample amounts Time (min) 33

34 Sensitivity Standard: Check & confirm instrument performance routinely Confirmation One standard with 6 different native TCDD isomers at different concentrations (2 fg/µl) RT: Relative Abundance Qualifier (ratio) mass trace 1478-TCDD 1379-TCDD 1378-TCDD 1368-TCDD 2378-TCDD 1234-TCDD Quantification mass trace 2 fg 5 fg 25 fg 1 fg 5 fg fg Time (min) 34

35 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

36 Screening + Confirmation using MS/MS + HRMS Dioxin Conference Presentation: Triple-Quadrupole GC-MS/MS Technique for PCDD/F and DL-PCB Determination in Milk Purpose: Develop a method to analyze PCDDs, PCDFs and DL-PCBs by a triple quadrupole GC-MS/MS system for a fast screening of dioxin- positive milk samples Buffalo Mozarella Collaboration with: ISS Rome, the National Health Institute, Italy, Nicola Iacovella, Anna Maria Ingelido, Elena De Felip, Gianfranco Brambilla Posters presented at Dioxin 9, Beijing, China ISCC, Riva, Italy Dioxin 1, San Antonio, TX, USA 36

37 Buffalo Milk Sample, TCDD Mass Traces GC-MS/MS (5 µl injection PTV solv. split ) GC-HRMS 1/1 concentration RT: Rela ative Abundance RT:.25 AA: 1532 RT:.28 AA: 1828 SM: 5G RT:.54 AA: Screening: GC-MS/MS RT:.82 AA: 4328 RT:. AA: RT:.69 AA: RT:.54 AA: RT:.68 AA: 2959 RT: 21.5 AA: RT: 21.5 AA: RT: 21.5 AA: RT: 21.5 AA: RT: AA: fast - yes / no RT: AA: ca. 15 fg RT: AA: RT: AA: Time (min) RT: AA: 934 RT: 21. AA: 5779 RT: 21. AA: 128 NL: 4.81E3 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 3.33E3 TIC F: + c EI SRM ms [ , ] MS BuffaloMilkA_Diox NL: 2.E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 2.9E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox RT: Relative Abundance Please note: Peak areas are not comparable between GC-MSMS and GC-HRMS Confirmation: GC-HRMS SM: 5G Higher - precision - sensitivity 1/1 conc.- ca. 15 fg RT: AA: RT: AA: RT: AA: 491 RT: AA: 561 RT: AA: RT: AA: RT: 18. AA: Time (min) NL: 1.54E2 m/z= MS 819_25956 A_16 NL: 1.82E2 m/z= MS 819_25956 A_16 NL: 7.87E4 m/z= MS 819_25956 A_16 NL: 9.67E4 m/z= MS 819_25956 A_16 37

38 Buffalo Milk Sample, PCDF GC-MS/MS (5 µl injection PTV solv. split ) GC-HRMS 1/1 concentration RT: Re elative Abundance RT: AA: RT: 24.1 AA: SM: 5G RT: AA: RT: AA: RT: AA: 578 RT: AA: Screening: GC-MS/MS RT: AA: RT: AA: RT: AA: RT: 25.9 AA: 2156 RT: AA: 2158 RT: 26.8 AA: RT: AA: RT: AA: NL: 2.56E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 4.34E4 TIC F: + c EI SRM ms [ , ] MS BuffaloMilkA_Diox NL: 1.22E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 9.84E5 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox RT: Relative Abundance Please note: Peak areas are not comparable between Time (min) GC-MSMS and GC-HRMS Confirmation: GC-HRMS SM: 5G RT:.79 AA: 117 RT:.79 AA: 441 1/1 conc. RT: AA: 458 RT: AA: 2569 RT:.78 AA: RT: AA: RT: 21.5 AA: 142 RT:.78 AA: 732 RT: AA: RT: AA: Time (min) NL: 1.11E3 m/z= MS 819_25956 A_16 NL: 6.16E2 m/z= MS 819_25956 A_16 NL: 2.78E4 m/z= MS 819_25956 A_16 NL: 1.78E4 m/z= MS 819_25956 A_16 38

39 Buffalo Milk Sample, HxCDD GC-MS/MS (5 µl injection PTV solv. split ) GC-HRMS 1/1 concentration RT: Re elative Abundance SM: 5G Screening: GC-MS/MS RT: AA: RT: AA: 5188 RT: 31.5 AA: 823 RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: 4914 RT: AA: Time (min) NL: 1.69E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 9.42E3 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 2.57E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 1.65E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox Please note: RT: Relative Abundance Confirmation: GC-HRMS SM: 5G RT: AA: 839 RT: AA: 2188 RT: MA: RT: AA: 628 RT: AA: 1638 RT: AA: RT: 26.7 AA: 631 RT: AA: RT: AA: 2427 RT: AA: 5935 Peak areas are not RT: AA: 74 comparable between Time (min) GC-MSMS and GC-HRMS NL: 4.85E2 m/z= MS 819_25956 A_16 NL: 4.19E2 m/z= MS 819_25956 A_16 NL: 6.E4 m/z= MS 819_25956 A_16 NL: 4.88E4 m/z= MS 819_25956 A_16 39

40 TSQ Quantum XLS Dioxin Screening Method TEQ LOQ of 1.5 pg/g ( lowest ML level) certifies the TSQ Quantum XLS for EU dioxin screening LOQ defined According to EU Directive 1883/6: Screening presicion < 3 % RSD 1 Var3 = *x Calibration runs at 32 fg injected amount, 21% RSD Courtesy N. Iacovella, G. Brambilla, ISS, Rome, Italy

41 TSQ Quantum XLS - TCDD in Buffalo Milk Samples Blank (GC-MS/MS) Buffalo Milk (GC-MS/MS) Buffalo Milk (GC-HRMS),17 pg/g fat 2,3,7,8-TCDD ṮCDD Labeled ISTD 2,3,7,8 41 Courtesy N. Iacovella, G. Brambilla, ISS, Rome, Italy

42 TSQ Quantum GC Intra-lab Trueness (routine) 42 Courtesy N. Iacovella, G. Brambilla, ISS, Rome, Italy

43 TSQ Quantum GC Precision (routine) Basis for RSD % evaluation 43 Courtesy N. Iacovella, G. Brambilla, ISS, Rome, Italy

44 TSQ Quantum XLS vs. GC-HRMS of Certified Material TSQ HRMS CERTIFIED LOQs 1. Results in [pg TEQ/g fat] TSQ 3,22 (5% RSD) HRMS 3.23 (2% RSD) ,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 1,2,3,7,8,9-HxCDF 2,3,4,6,7,8-HxCDF 1,2,3,4,6,7,8-HpCDF 1,2,3,4,7,8,9-HpCDF OCDF 44 Courtesy N. Iacovella, G. Brambilla, ISS, Rome, Italy

45 Split Extract Approach for POPs Workflow Analysis Sample Aliquote 1 Aliquote 2 simplified sample prep. aliquote 1 Raw extract Split into two aliquotes (5/5 or 75/25, ) full sample preparation store in fridge aliquote 2 Screening GC-MSMS Analysis Done compliant Negative (TEQ < ML) Positive (TEQ >ML) (dump aliquote 2) Confirmation GC-HRMS Negative (TEQ < ML) Analysis Done compliant Positive (TEQ >ML) Confirmation Report 45

46 Ongoing Project: Simplify Screening Sample Prep Buffalo milk, HxCDD: full sample cleanup - simplified & faster cleanup RT: SM: 5G RT: AA: RT: 31.5 AA: 823 Screening: GC-MS/MS Full sample clean-up RT: AA: NL: 1.69E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox RT: SM: 5G Screening: GC-MS/MS Simplified clean-up RT: AA: 7386 RT: 32.8 AA: 1381 NL: 1.38E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP Relative Abundance RT: AA: 5188 RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: RT: AA: 4914 RT: AA: NL: 9.42E3 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 2.57E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 1.65E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox lative Abundance Rel RT: AA: 3259 RT: AA: 384 RT: 32.8 AA: 62 RT: 32.6 AA: Same matrix sample RT: AA: RT: AA: 1947 RT: 32.7 AA: NL: 6.E3 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP NL: 1.71E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP NL: 1.9E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP Time (min) Time (min) 46

47 Ongoing Project: Simplify Screening Sample Prep Buffalo milk, HxCDF: full sample cleanup - simplified & faster cleanup RT: SM: 5G RT: 3.22 AA: RT: AA: RT: AA: Screening: GC-MS/MS Full sample clean-up NL: 2.76E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox RT: SM: 5G RT: 3.49 AA: RT: 3.3 AA: 11 Screening: GC-MS/MS Simplified clean-up RT: AA: 91 RT: AA: 291 RT: AA: 133 NL: 6.8E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP Relative Abundance RT: AA: 118 RT: 3.2 AA: 9386 RT: 3. AA: RT: AA: RT: AA: RT: AA: NL: 1.89E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox NL: 1.4E6 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox Relative Abundance RT: 3.9 AA: RT: 3.49 AA: RT: 3.28 AA: RT: AA: Same matrix sample RT: AA: RT: AA: RT: AA: RT: AA: 7873 NL: 3.79E4 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP NL: 7.98E5 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP RT: 3. AA: RT: AA: RT: AA: NL: 6.32E5 TIC F: + c EI SRM ms [ ] MS BuffaloMilkA_Diox RT: 3.28 AA: RT: AA: 2741 RT: AA: RT: 3.5 AA: 2959 NL: 4.85E5 TIC F: + c EI SRM ms [ ] MS BuffaloMilkD_NoPP Time (min) Time (min) 47

TargetQuan POPs Quantitation for TSQ and DFS Features for dioxin applications:

and upper boundary calculation User definable summation Of calculated amounts or

mass and up to two masses based on abundance EPA 1613 Rev.

48 TargetQuan POPs Quantitation for TSQ and DFS Features for dioxin applications: Toxicity equivalents (TEQ) according to WHO definition including lower, medium and upper boundary calculation User definable summation Of calculated amounts or TEQs for reporting of sum TEQ values Isotopic ratio confirmation One quantitation mass and up to two masses based on abundance EPA 1613 Rev.B compliant Allows quantification based on average response of selected compound Retention time correction 48

49 TargetQuan For TSQ Quantum XLS and DFS By Analysis One Compound Sequentially Selected Files By Compound One Analysis Sequentially Selected Entry 49

50 1. The Integrated POPs Analysis Workflow 2. Sample preparation 3. Screening and Confirmation Analysis Requirements 4. GC-MS/MS (Triple Quad MS) for Screening Analysis 5. GC-HRMS (Magnetic Sector MS) for Confirmation 6. How GC-MS/MS and GC-HRMS Work Together 7. Conclusions

The combination GC-MS/MS The Screening Solution Well known instrumentation in every trace analysis lab Very high sensitivity and selectivity for matrix samples Provides TEQ results for PCDD/Fs and

51 The combination GC-MS/MS The Screening Solution Well known instrumentation in every trace analysis lab Very high sensitivity and selectivity for matrix samples Provides TEQ results for PCDD/Fs and DL-PCBs Fast - with high throughput capability, automated runs GC-HRMS The Confirmation Solution Compliant with international regulations Highest sensitivity and precsion for low level analyses Highly productive routine method high throughput operation GC-MS/MS + GC-HRMS Work Seamlessly Together Use integrated sample prep workflow for screening and confirmation High precision data using labelled internal standards Common TargetQuan software platformtraining Aspect GC-MS standard operation for both instruments Extract then can be used in many cases on both instruments Efficient management of the laboratory sample flow Only indicated samples go to HRMS confirmation analysis 51

52 Complete Method Setup for DL-PCBs and PCDD/Fs For TSQ Quantum XLS Screening and DFS GC-HRMS Confirmation 52

53 Thank you for your attention Questions welcome! contact: 53

Prep TM Automated Extraction and Clean-up System for PCDDs, PCDFs and PCBs

Power-Prep Prep TM Automated Extraction and Clean-up System for PCDDs, PCDFs and PCBs Jef Focant and Hamid Shirkhan Dioxin Lab. BELGIUM FMS Inc. WWW.FMS-INC.Com USA Dioxins Analysis! Extraction (Soxhlet,