Analysis of Pesticides and Contaminants using HS-SPME-GC/MS and DRS Detection. Blair Surridge, Tyler Dyer, Rene Bilideau, Paloma Bertolin Presoti
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1 Analysis of Pesticides and Contaminants using HS-SPME-GC/MS and DRS Detection Blair Surridge, Tyler Dyer, Rene Bilideau, Paloma Bertolin Presoti
2 Applied Research at Camosun College Provides funding to support collaborations with local companies in commercialization, technology transfer, or technology adaptation type projects
3 Project Goals Develop a HS-SPME-GC/MS method for the analysis of a large number of pesticides and contaminants in fruits and vegetables Evaluate the use of DRS as a detection and quantification method for pesticides and contaminants.
4 Outline Selection of Pesticides and Contaminants Development of a HS-SPME Method AOAC QuEChERS Method 27.1 SPME vs QuEChERS Method Performance Characteristics Analysis of Fruits/Vegetables Samples
5 Selection of Pesticides Compound selection was based on frequency of detections 378 samples samples
6 Selection of Pesticides A page-by-page review was carried out
7 Compounds Selected For Method Validation 73 compounds prepared into 4 solutions were purchased for use in the project Fluoranthene 2,4,4'-Trichlorobiphenyl (PCB 28) 2,2',4,5,5'-Pentachlorobiphenyl (PCB 11) 2,2',5,5'-Tetrachlorobiphenyl (PCB 52) 2,2',4,5,5'-Hexachlorobiphenyl (PCB 153) 2,2',3,4,4',5,5'-Heptachlorobiphenyl (PCB 18) Phenanthrene Pyrene bis(2-ethylhexyl)phthalate 2,4'-DDE 2,4'-DDD 2,4'-DDT a-chlordane g-chlordane Chlorothaloni Chlorpyrifos Diazinon Dimethoate Ethion Fenthion Imazalil Malathion Methyl parathion Mirex Phosalone Phosmet Prochloraz Trichlorfon Azoxystrobin Bifenthrin Buprofezin Captan Carbaryl Carbofuran Clofentezine Cypermethrin Difenoconazole Diphenylamine Fludioxonil Metalaxyl Myclobutanil Permethrin Pirimicarb Pyraclostrobin Pyrifenox Pyrimethanil Simazine Thiabendazole Triadimenol Atrazine Trifloxystrobin Aldrin a-bhc b-bhc g-bhc d-bhc p,p'-ddd (4,4') p,p'-dde (4,4') p,p'-ddt (4,4') Dieldrin Endosulfan I Endosulfan II Endosulfan sulfate Endrin Endrin aldehyde Endrin ketone Heptachlor Heptachlor epoxide (isomer B) Methoxychlor
8 GC/MS Conditions GC : Agilent 789 with CTC CombiPal including SPME/Headspace Column: ZB-5MS, 3m x.25mm x 25µm (Phenomenex) Injection volume: 1µL Inlet Conditions: Pulsed Split less at 27 o C, constant pressure Retention Time Locking: Methyl chlorpyrifos, 12.7 min at 22.5 psi Mass Spec : Agilent 5975C 35 Column Oven Temperature Program Ionization mode: EI (+) ion Source Temp: 25 o C Acquisition: SIM (5 time segments) mass scan: 5-4 m/z Temperature (oc) Time (mins)
9 Abundance Example Chromatogram TIC from SIM acquisition 2 TIC: 8dec14_1.D\DATASIM.MS
10 Sample Homogenization Use of an Osterizer food processor
11 Headspace SPME Method sample + aqueous extracting solvent SPME extraction on to fiber (extraction temperature and time) Desorption in GC inlet Analysis by GC/MS (SIM & open scan acquisition)
12 Effect of Solution ph 12 1 Response NaH2PO4 ph 8 NaCl ph 7 NaH2PO4 ph4 All solvent solutions contained 1%MeOH Incubation temperature 1 C, Incubation time using a 15ng/g spiked organic grape sample
13 Effect of Extraction Temperatures Diphenylamine Phosmet C + 6 C + 8 C + 1 C + 12 C + 14 C + 4 C + 6 C + 8 C + 1 C + 12 C + 14 C + Phenanthrene Optimal Temperature: 11 o C Diazinon C + 6 C + 8 C + 1 C + 12 C + 14 C C + 6 C + 8 C + 1 C + 12 C + 14 C +
14 Effect of Extraction Time 25 Diphenylamine 35 3 Cyprodinil Response min 45min 6min Incubation Time (min) Phenanthrene min 45min 6min Diazinon min 45min 6min 1min 45min 6min
15 Extraction Times (cont d) Atrazine Bifenthrin min 45min 6min 1min 45min 6min Optimal Time: 2 min Myclobutanil Diazinon min 45min 6min min 45min 6m
16 Effect of Fiber Type 45 4 Response Atrazine Phosalone Phosmet Diazinon Methyl Parathion PCB 11 Cyprodinil Diphenylamine Fiber Incubation temperature 11 C, Incubation time using a 15ng/g spiked organic grape sample
17 Effect of Fiber Type %Difference of DMS/DVB vs the other fibers % Difference PDMS/DVB vs Polyacrylate PDMS/DVB vs 3um PDMS Comparisons PDMS/DVB vs DVB/CAR/PDMS PDMS/DVB vs 1um PDMS
18 Developed Headspace SPME Method 1. gram of sample + 3mL of 1% MeOH in NaCl sat d water SPME for utes with a 65µm PDMS/DVB fiber at 11 o C Desorption in GC inlet at 27 o C for 2 minutes Analysis by GC/MS (SIM & open scan acquisition)
19 SPME Calibration Data Carried out in Grape Matrix External standard No sample manipulation steps! Incubation temperature: 11 C Incubation time: SPME Fiber: 65 um PDMS/DVB Level Concentration (ng/ml) Response CS CS CS CS CS CS CS CS
20 SPME Calibration In Matrix No significant difference observed Response Diphenylamine Concentration (ng/g) Grape Tomato No Matrix Raddish Phenanthrene Grape Tomato No Matrix Raddish Concentration (ng/g)
21 QuEChERS Method 27.1 Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) Method AOAC 27.1 Use of dspe material containing(c18 and PSA)
22 SPME vs QuEChERS Level Ciprodinil Concentration (ng/g) Response CS CS CS CS CS CS CS CS ~1x Level Ciprodinil Concentration (ng/g) Response CS CS CS CS CS CS CS CS
23 Detection Limits (in Strawberries) Selected Analytes RT (min) Ion (m/z) SPME LOD (ng/g) QuEChERS LOD (ng/g) % Recovery QuEChERS % Recovery SPME Organophosphates Chlorpyrifos Ethion Fenthion Malathion NA Organochlorines Aldrin a-bhc b-bhc a-chlordane g-chlordane Chlorothalonil ,4 -DDT Dieldrin Organonitrogens Buprofezin Cyprodinil Na 13 Diphenylamine Triazine Permethrin NA Contaminants Anthracene Phenanthene PCB PCB
24 Summary of DRS Detection Agilent s G1672AA (Pesticide and endocrine disruptor RTL and mass spectral library, 926 entries) NIST 8 mass spec library Extraction method used 1ppm DRS 1ppm SIM 5ppm DRS 5ppm SIM 1ppm DRS 1ppm SIM.5ppm DRS.5ppm SIM.5ppm DRS.5ppm SIM QuEChERS Organic Grape Matrix 64/68 68/68 56/68 68/68 37/68 68/68 28/68 59/68 7/68 28/68 QuEChERS No Matrix 68/68 68/68 56/68 68/68 42/68 68/68 25/68 59/68 7/68 28/68 SPME N/A N/A N/A N/A 53/68 68/68 52/68 68/68 39/68 61/68 QuEChERS Strawberry Matrix 63/68 68/68 N/A 68/68 43/68 68/68 24/68 58/68 6/68 28/68 QuEChERS Potato Matrix 64/68 68/68 N/A 68/68 4/68 68/68 23/68 54/68 6/68 22/68
25 MRL Data Select Compounds Grape MRL values (ppm) QuEChERS DRS detection SPME DRS Detection Organophosphates Chlorpyrifos.1 No Yes Malathion 8. Yes Yes Phosalone 5. Yes Yes Diazinon.75 Yes Yes Phosmet 1 Yes Yes α-chlordane 1. No Yes Organochlorines Heptachlor.2 No Yes α-bhc 1. Yes Yes 2,4 -DDT 1. Yes Yes Heptachlor epoxide.5 No Yes Aldrin.2 No Yes Organonitrogens Thiabendazole 1 Yes Yes Cyprodinil 2 Yes Yes Buprofezin 1. Yes Yes Diphenylamine 5. Yes Yes Trifloxystrobin 2. Yes Yes Pyrimethanil 5. Yes Yes
26 Results from Survey A 13 local samples (domestic, import, and unknown)were tested using both the standard AOAC extraction protocol and the developed HS-SPME methods Below is a list of detection information from the results Sample Type Compound Name Chem station amount (ppm) DRS amount (ppm) QuEChERS SIM detection? SPME SIM detection? DRS detect (Q or S)? Match Quality Green Grapes Cyprodinil Yes Yes Both 98 2 MRL Value (ppm) Buprofezin Yes Yes Both 81 1 California Strawberry Cyprodinil Yes Yes Both 85 2 Quinoxyfen.8 Yes Yes Both 86.9 BC strawberry Cyprodinil Yes Yes Both 97 2 Fludioxonil.7.72 Yes Yes Both 84 3 Boscalid (Nicobifen).59 Yes Yes Both Potato Chlorpropham.79 Yes Yes Both Bisphenol A.34 Yes Yes Both 9 N/A Basil Azoxystrobin Yes Not enough sample Q Metalaxyl Yes Not enough sample Q
27 Results from Survey More results Sample Type Compound Name Chem station amount (ppm) DRS amount (ppm) QuEChERS SIM detection? SPME SIM detection? DRS detect (Q or S)? Spartan Apple Pyrimethanil.12 - Yes Yes no Paprika Ethoxyquin N/A 9.8 Yes Yes Both Chili Powder Ethoxyquin N/A Yes Yes Both Garlic Powder Anthracene.7.6 Yes Yes Both Cherry Tomato Buprofezin.7.5 Yes Yes Both Piperonyl butoxide 5.14 Yes Yes Both Pyriproxyfen.26 Yes No Red Pepper Chlorpyrifos Yes Yes Both Tea Bifenthrin Raspberry Iprodione.7 Yes Yes Both
28 E.G.Chromatogram - Cyprodinil Measured concentration was 2.1 ppm from grape sample
29 Hot off the press!
30 Conclusions A simple and highly automated screening method capable of measuring a diverse set of pesticides and contaminants was developed and evaluated More work is needed regarding.. quantitation and the use of internal standards.. Use of triphenylphosphate Confirmation is required for the detected pesticides above the MRL level
31 Acknowledgments Special thanks to the following organizations Special thanks to the students Paloma Bertolin Tyler Dyer Brenna Stanford SCIENCE WITHOUT BORDERS BRAZILIAN SCHOLARSHIP PROGRAM
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