Rapid Detection of Pesticides in Fruit Juice without Sample Preparation - Waters Corporation Food Safety
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This presentation compares wo methods for the detection of low-level pesticide residues in fruit juice. One involves the use of QuEChERS sample preparation and the other a 'dliute and shoot' approach. Sample preparation is utilised to remove the matrix effects associated with mass spectrometry (MS), using a 'dilute and shoot' approach requires the use of highly sensitive MS detection. It can be seen from the results shown that the 'dilute and shoot' approach can be used in many cases.
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Rapid Detection of Pesticides in Fruit Juice without Sample Preparation - Waters Corporation Food Safety
- 1. ©2015 Waters Corporation 1 Rapid Detection of Pesticides in Fruit Juice using tandem quadrupole LC/MS/MS without the need for Sample Preparation
- 2. ©2015 Waters Corporation 2 Presentation Overview Overview of pesticide residue analysis Experimental – System Solution Results – Multi-Residue Story o Sample preparation comparison • Both scenarios working well… • One method is better than the other… o Robustness and repeatability data Conclusion
- 3. ©2015 Waters Corporation 3 Food Contaminant Testing What are the challenges? Changing Legislation Demanding Performance Multitude of Food Types Variety of Compounds
- 4. ©2015 Waters Corporation 4 Ultra Trace Analysis An Analogy Whole product Representative sample Ultra trace detail
- 5. ©2015 Waters Corporation 5 Ultra trace analysis What are the challenges? Challenging matrix/analyte combinations Low LOQs • Low target levels Amount of sample • Mass/volume possible to extract Matrix effects • Complex matrices Reproducibility • Trace levels can affect statistics
- 6. ©2015 Waters Corporation 6 What impact does this have on the laboratory? Demand for Flexible methods – Many analytes – Many matrices – Simple set-up / modification Method performance – High sensitivity – Quantitative accuracy Data management – Automated processing – Reporting – Archive and back-up
- 7. ©2015 Waters Corporation 7 Pesticide Residue Analysis Recent trend for pesticide residue analysis is to run multi- analyte methods Many labs now running QuEChERS (or variations of…) Experimental Questions – Is it possible to use dilute-and-shoot for pesticide residue analysis? – How does QuEChERS compare with dilute-and-shoot? – Are we able to reduce to sample prep time even further for the detection of pesticides in food and beverage products? – Can we meet the regulatory detection limits required? – Can we produce repeatable data with a dilute-and-shoot approach for pesticide residues in fruit juices?
- 8. ©2015 Waters Corporation 8 • Automatically sets up instrument, • Optimises MRMs and performs system suitability Setup & Optimize Complete Analytical System • Generates optimised methods, time scheduled MRMs Generate / Select Methods to Run • Methods generated automatically by QuanPedia, • Clear flagging when data is out of regulatory tolerance • Reduced risk of errors Review Data & Prepare Reports • Real time QC, Intelligent ‘on the fly’ decision making Check QC & Calibration Standards • Long term system monitoring, Lab management tool Track System & Method Performance / Trends QUAN Workflow
- 9. ©2015 Waters Corporation 9 Experimental Information Multi-Residue Method
- 10. ©2015 Waters Corporation 10 Pesticide Residues Targeted Screening Workflow
- 11. ©2015 Waters Corporation 11 Sample Preparation Two methods employed Measure out 10 mL of fruit juice (spiked at 10ng/mL)+ 15mL of 1% acetic acid in Acetonitrile into DisQuE tube 1 Centrifuge and transfer 1 mL of the extract into DisQuE tube 2 Centrifuge Dilute 10 times with water (~ 15x dilution) Dilute and Shoot Measure out 10 mL of fruit juice (spiked at 10ng/mL) Dilute 10 and 100 times with water Filter and Inject
- 12. ©2015 Waters Corporation 12 UPLC system ACQUITY UPLC System Column: ACQUITY UPLC BEH C18 2.1 x 100 mm, 1.7 m Column temp 40 C Mobile phase A Water + 0.1% Formic acid Mobile phase B Methanol + 0.1% Formic acid Flow Rate 0.45 mL/min Inject. volume 10 L Gradient ACQUITY UPLC & Xevo TQ-S Conditions Min % A %B 0.00 95 5 0.25 95 5 7.75 0 100 8.50 0 100 8.51 95 5 10.00 95 5
- 13. ©2015 Waters Corporation 13 ACQUITY UPLC & Xevo TQ-S Conditions MS System XEVO TQ-S Ionization mode ESI Positive Capillary voltage 3.0 kV Cone voltage 30.0 V Source temp 150 C Desolvation temp 500 C Desolvation gas 1000 L/hr Cone gas 50 L/hr Routine targeted quantification with information rich qualitative data Specifically designed for sensitivity AND robustness
- 14. ©2015 Waters Corporation 14 • Automatically sets up instrument, • Optimises MRMs and performs system suitability IntelliStart • Generates optimised methods, time scheduled MRMs Quanpedia • Methods generated automatically by QuanPedia, • Clear flagging when data is out of regulatory tolerance • Reduced risk of errors TargetLynx • Real time QC, Intelligent ‘on the fly’ decision making QCMonitor • Long term system monitoring, Lab management tool TrendPlot Waters QUAN Workflow
- 15. ©2015 Waters Corporation 15 Quanpedia A central data base for quantitative LC/MS methods Based on compound name A tool to aid MS method creation – Automatically creates data acquisition methods – Automatically creates data processing methods
- 16. ©2015 Waters Corporation 16 Quanpedia Quanpedia – manage compounds and methods
- 17. ©2015 Waters Corporation 17 Quanpedia
- 18. ©2015 Waters Corporation 18 Quanpedia improves data quality Manual MS method 1 MRM per compound 6.6 % RSD 8 data points across peak Quanpedia MS method 1 MRM per compound 2.4 % RSD 12 data points across peak Quinoxyfen @ 1ppb Manual average = 23 % RSD Quanpedia average = 8.6 % RSD 80 pesticides spiked, acquired using 402 compound MS method
- 19. ©2015 Waters Corporation 19 90 Multi-Analytes Screened Acaricide Monocrotophos Herbicide Atraton Herbicide Aziprotryne Acaricide/Insecticide Methamidophos Herbicide Prometryn Herbicide Propaquizafop Acaricide/Insecticide Dimethoate Herbicide MCPA Herbicide Isoproturon Acaricide/Insecticide Dichlorvos Herbicide Isoxaben Herbicide Terbuthylazine Acaricide/Insecticide Methiocarb Herbicide Asulam Herbicide Tebutam Acaricide/Insecticide Parathion Herbicide Dimethachlor Herbicide Thidiazuron Acaricide/Insecticide EPN Herbicide Picloram Herbicide Diuron acaricides/insecticides Phosmet Herbicide Isocarbamide Herbicide Siduron Breakdown product 2,6-Dichlorbenzamid Herbicide Tebuthiuron Herbicide Hexazinone Fungicide Oxadixyl Herbicide Diphenamid Herbicide Propanil Fungicide Hexaconazole Herbicide Quizalofop-ethyl Herbicide Clodinafop-propargyl Fungicide Carbendazim Herbicide Cyanazine Herbicide Dimethenamid Fungicide Difenoconazole Herbicide Prometon herbicides Haloxyfop Fungicide Pyracarbolid Herbicide Naptalam herbicides Propisochlor Fungicide Propiconazole Herbicide Napropamide herbicides Butachlor Fungicide Thiabendazole Herbicide Rimsulfuron herbicides 2,4-DB Fungicide Flutriafol Herbicide Propyzamide herbicides MCPB Fungicide Imazalil Herbicide Atrazine Insecticide Cyromazine Fungicide Boscalid Herbicide Terbutryn Insecticide Dicrotophos Fungicide Fenamidone Herbicide Desmetryn Insecticide Thiodicarb Fungicide Tebuconazole Herbicide Ametryn Insecticide Fenpropathrin Fungicide Benomyl Herbicide Simazine Insecticide Methomyl Fungicide Azoxystrobin Herbicide Metazachlor Insecticide Acephate Fungicide/Herbicide Ethoxyquin Herbicide Alachlor Insecticide Imidacloprid Fungicide Tridemorph Herbicide Mecoprop Insecticide Trichlorfon Fungicide Octhilinone (2-Octyl-4-isothiazoline-3-one) Herbicide Propachlor Insecticide Phoxim Insecticide Phorate Insecticide Propoxur insecticides Tetramethrin Insecticide Pymetrozine Insecticide Diflubenzuron insecticides Fenitrothion Insecticide Pirimicarb Insecticide Buprofezin insecticides Allethrin Insecticide Diazinon Insecticide Deltamethrin insecticides Rotenone Insecticide Carbofuran Insecticide/Acaricide Azinphos-methyl Metabolite Aldicarb sulfone Insecticide Parathion-methyl Insecticide/Acaricide Aldicarb Metabolite Atrazine-desethyl Insecticide Carbaryl Insecticide/Nematicide Oxamyl Metabolite Aldicarb sulfoxide Insecticide Fenthion Insecticide/Nematicide Phosphamidon Metabolite Atrazine-desisopropyl Insecticide Acetamiprid Insecticide/Nematicide Dichlofenthion Metabolite Carbofuran-3-hydroxy Insecticide Sulfotep Insecticide Phenothrin
- 20. ©2015 Waters Corporation 20 Results and Discussion
- 21. ©2015 Waters Corporation 21 Developing a Suitable Method Analysis of food samples can be challenging – need to develop a sample preparation that can: o Reduce matrix effects (e.g. peak distortion, ion suppression…) o Extract relevant compounds o Provide a robust, repeatable method Method development to investigate different extraction techniques can be time intensive Useful to have tools that support this process and help improve lab efficiency LC-MS/MS Analysis Modify analytical method LC-MS/MS Analysis Modify sample preparation
- 22. ©2015 Waters Corporation 22 Spiked Orange Juice without RADAR
- 23. ©2015 Waters Corporation 23 On of the ways to determine sample prep efficiency… What is RADAR? – Rapid electronics allow instrument to switch between MRM & full scan – Added information can be gained from full scan data in routine analyses • Intelligent decision making • Improves method development • Monitor matrix components • Detect un-targeted adulterants How do I set it up on the instrument? - Parallel MRM with full scan data acquisition
- 24. ©2015 Waters Corporation 24 Apple juice Grape juice Orange juice Q Q Q 100X 100X 100X 10X 10X 10X Juice Samples – Matrix Contribution QuEChERS versus Dilute-and-Shoot
- 25. ©2015 Waters Corporation 25 Sample prep comparison When both options work… 0.7 ng/mL 1 ng/mL 0.1 ng/mL Atraton @ 10 ng/mL 10x 10x 100X 100X100X 10x Quechers QuechersQuechers
- 26. ©2015 Waters Corporation 26 Sample prep comparison When both options work… 0.7 ng/mL 1 ng/mL 0.1 ng/mL Atraton @ 10 ng/mL 10x 10x 100X 100X100X 10x Quechers QuechersQuechers
- 27. ©2015 Waters Corporation 27 Sample prep comparison When QuEChERS works… Rotenone @ 10 ng/mL
- 28. ©2015 Waters Corporation 28 Sample prep comparison When Dilute & Shoot Doesn’t work… Rotenone @ 10 ng/mL Sample Diluted 10x
- 29. ©2015 Waters Corporation 29 Q - diluted Q - diluted Q - diluted Sample prep comparison When Dilute-and-Shoot works… Q 10X Q 10X Q 10X Poor peak shape Shift in RT
- 30. ©2015 Waters Corporation 30 • Automatically sets up instrument, • Optimises MRMs and performs system suitability IntelliStart • Generates optimised methods, time scheduled MRMs Quanpedia • Methods generated automatically by QuanPedia, • Clear flagging when data is out of regulatory tolerance • Reduced risk of errors TargetLynx • Real time QC, Intelligent ‘on the fly’ decision making QCMonitor • Long term system monitoring, Lab management tool TrendPlot Waters QUAN Workflow
- 31. ©2015 Waters Corporation 31 TargetLynx Reporting Designed for food safety customers
- 32. ©2015 Waters Corporation 32 Linearity and Repeatability Compound name: Atraton Correlation coefficient: r = 0.997944, r^2 = 0.995892 Calibration curve: 22116.2 * x+ 949.701 Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None ng/mL 0 10 20 30 40 50 60 70 80 90 100 Response 0 500000 1000000 1500000
- 33. ©2015 Waters Corporation 33 Dilute-and-Shoot Calibration Curve Compoundname:Atraton Correlationcoefficient:r =0.998102,r^2=0.996207 Calibrationcurve:0.0161973* x+0.00307921 Responsetype:Internal Std( Ref16),Area* ( ISConc./ISArea) Curvetype:Linear,Origin:Exclude,Weighting:1/x,Axis trans:None ng/mL 0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 Response 0.00 1.00 2.00 3.00 4.00 5.00
- 34. ©2015 Waters Corporation 34 • Automatically sets up instrument, • Optimises MRMs and performs system suitability IntelliStart • Generates optimised methods, time scheduled MRMs Quanpedia • Methods generated automatically by QuanPedia, • Clear flagging when data is out of regulatory tolerance • Reduced risk of errors TargetLynx • Real time QC, Intelligent ‘on the fly’ decision making QCMonitor • Long term system monitoring, Lab management tool TrendPlot Waters QUAN Workflow
- 35. ©2015 Waters Corporation 35 Compound Repeatability & Robustness Dilute-and-Shoot Approach
- 36. ©2015 Waters Corporation 36 Pressure Repeatability Dilute-and-Shoot Approach 1st injection 100th injection 155th injection
- 37. ©2015 Waters Corporation 37 Conclusions Variety of different fruit juice samples have been analysed Direct comparison of QuEChERS and dilute-and-shoot for pesticide residue screening Analysis was performed using a targeted multi-analyte screening solution: o Setting up the method – IntelliStart, Quanpedia o Sample preparation- DisQUE and ‘dilute-and-shoot’ o Reliable chromatography - ACQUITY UPLC & ACQUITY BEH C18 columns o Robust and Repeatable results - Xevo TQ-S o Matrix monitoring tools - RADAR o Review of quantitative data –TargetLynx, TrendPlot