Advanced Tools to Screen for Targeted and Nontargeted Contaminants in Food Samples. André Schreiber AB SCIEX, Concord, Ontario (Canada)
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1 Advanced Tools to Screen for Targeted and Nontargeted Contaminants in Food Samples André Schreiber AB SCIEX, Concord, ntario (Canada)
2 verview High resolution and accurate mass LC-MS/MS in food analysis AB SCIEX TripleTF 5600 system Applications Multi-pesticide screening and quantitation with identification based on MS/MS library searching Identification of bitter compounds in hops Identification of authenticity markers in broccoli AB SCIEX
3 Traditional, Current and Future of Food Analysis Traditional Single compound or group specific methods Extensive cleanup and extraction Instrumental analysis and data processing Current Multi-target screening using generic cleanup and chromatography Targeted Instrumental analysis Data processing Future Non-target screening using generic cleanup and chromatography Instrumental analysis using full scan techniques Targeted and retrospective non-targeted data processing Unknown screening AB SCIEX
4 High Resolution Accurate Mass LC-MS/MS Highest Sensitivity in MS and MS/MS mode High Resolution ~ at all speeds EasyMass Accuracy ~ 1 ppm RMS SmartSpeed Acquisition 10 ms accumulation time Linear Dynamic Range ~ 4 orders of magnitude AB SCIEX
5 The AB SCIEX TripleTF 5600 System I QJet Ion Guide Q0 High Pressure Cell High Frequency Q1 Ion compression optics Accelerator TF Analyzer 40 GHz Multichannel TDC Detector 30kHz Accelerator LINAC collision cell 15 kv Acceleration voltage Two-stage reflectron TurboV source AB SCIEX
6 EasyMass Accuracy Calibrant Delivery System (CDS) DuoSpray Source second sprayer dedicated for delivery of CDS solution Fully automated pumping system for delivery of up to four calibration solutions Automated calibration of MS and MS/MS at the beginning of the batch and in between samples AB SCIEX
7 Multi-Pesticide Screening and Quantitation with Identification based on MS/MS Library Searching AB SCIEX
8 Experimental QuEChERS extraction and 5x dilution Different brassica samples (broccoli, kale, cabbage ) UHPLC with Shimadzu UFLC XR system Restek Ultra Aqueous C18 (100x2.1 mm) 3 μm Water/methanol + 10 mm ammonium formate 10 μl injection AB SCIEX TripleTF 5600 LC/MS/MS system Positive polarity Electrospray Ionization (ESI) TF-MS survey ( Da) IDA: 10 dependent TF-MS/MS at CE=35 V with CES=15 V Data processing using PeakView with the XIC Manager and MultiQuant software AB SCIEX
9 MS Resolution of TripleTF 5600 System Resolution 189 Da Resolution 192 Da FWHH = 7 mda Δt ~ 0.3 nsec Resolution 404 Da Resolution 732 Da AB SCIEX
10 Increased Selectivity through Resolution 6.8 μg/kg Imidacloprid in kale sample (5x dilution) XIC of ± 256mDa (1000ppm) S/N ~ 4 ± 25.6mDa (100ppm) S/N ~ 10 ± 2.56mDa (10ppm) S/N ~ AB SCIEX
11 MS/MS Resolution of TripleTF 5600 System 48 μg/kg Flonicamid in mustard greens sample (5x dilution) XIC of ± 2.3mDa Resolution 79 Da Resolution 230 Da AB SCIEX
12 XIC Manager Table and Confidence Settings AB SCIEX
13 Targeted Screening for ~300 Pesticides 5 ng/ml Standard AB SCIEX
14 Pesticides in Kohlrabi Sample Metalaxyl Methoxyfenozide Imidacloprid Spirotetramat Pyraclostrobin Cyprodinil TF-MS TF-MS/MS AB SCIEX
15 Pesticides in Chinese Broccoli Sample Metalaxyl Imidacloprid Spirotetramat Cyprodinil TF-MS TF-MS/MS AB SCIEX
16 Quantitation of Imidacloprid 23 μg/kg in Chinese Broccoli Sample 10 μg/kg MRL after dilution AB SCIEX
17 Identification of Bitter Compounds in Hops H H 3 C H 3 C H 3 C H 3 C H H brewing H 3 C H H + H 3 C H H Humulone H 3 C cis-isohumulone H 3 C trans-isohumulone AB SCIEX
18 Humulone and Cohumulone in Different Hops XIC of Da and Da ±5 mda H H Nugget (herbal spicy) H 3 C H 3 C H H H 3 C H 3 C H H Galena (heavy bitter) Cascade (citrus aroma) Willamette (fruity, floral) AB SCIEX
19 MS Identification of Humulone Formula Finder Tool Resolution = Accuracy = 0.8 ppm AB SCIEX
20 MS/MS Identification of Humulone Fragment Interpretation Tool Resolution 57 Da Resolution 363 Da AB SCIEX
21 Identification of Authenticity Markers in Broccoli H 3 C S H H H H S N S H enzyme C H 3 S N C S Glucoraphanin Sulforaphane AB SCIEX
22 Experimental Liquid-liquid extraction and 5x dilution Different broccoli and samples (air dried, freeze dried, seeds) UHPLC with Shimadzu UFLC XR system Phenomenex Hydro-RP (150x4.6 mm) 4 μm Water/acetonitrile + 25 mm ammonium formate, ph = μl injection AB SCIEX TripleTF 5600 LC/MS/MS system Negative polarity Electrospray Ionization (ESI) TF-MS survey ( Da) IDA: 10 dependent TF-MS/MS at CE=35 V with CES=15 V Data processing using Principal Components Analysis (PCA) in MarkerView, Formula Finder, and Fragment Pane in PeakView software AB SCIEX
23 PCA Scores Plot Broccoli seeds Brussels sprouts Broccoli Cabbage Kale AB SCIEX
24 PCA Loadings Plot after PCVG Broccoli seeds PCVG 4 Brussels sprouts Broccoli Cabbage Kale AB SCIEX
25 PCA Profile Plot Broccoli seeds PCVG 4 Brussels sprouts Broccoli Cabbage Kale Broccoli marker at 14.1 min AB SCIEX
26 Empirical Formula Calculation C 12 H 23 N 9 S AB SCIEX
27 Search for C 12 H 23 N 9 S 3 in the Internet AB SCIEX
28 Summary High resolution and accurate mass LC-MS/MS AB SCIEX TripleTF 5600 system Targeted screening and quantitation Sensitivity, selectivity (resolution and mass accuracy), speed, and linear dynamic range Identification of target analytes MS/MS library searching Identification of unknown analytes Empirical formula calculation (accurate mass MS and MS/MS, isotopes, cluster information) Search for possible structures MS/MS interpretation (Confirmation using reference material and orthogonal technique) AB SCIEX
29 Acknowledgements/Trademarks/Licensing The authors would like to thank Jon Wong and Kai Zhang (FDA, Collage Park, MD) for providing pesticide standards and samples for analysis Sandra Stanley (FutureCeuticals, Momence, IL) for authenticity samples For Research Use nly. Not for use in diagnostic procedures. The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners. AB SCIEX is being used under license AB SCIEX AB SCIEX
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