Advanced Tools to Screen for Targeted and Nontargeted Contaminants in Food Samples André Schreiber AB SCIEX, Concord, ntario (Canada)
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 2 2011 AB SCIEX
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 3 2011 AB SCIEX
High Resolution Accurate Mass LC-MS/MS Highest Sensitivity in MS and MS/MS mode High Resolution ~ 30 000 at all speeds EasyMass Accuracy ~ 1 ppm RMS SmartSpeed Acquisition 10 ms accumulation time Linear Dynamic Range ~ 4 orders of magnitude 4 2011 AB SCIEX
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 5 2011 AB SCIEX
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 6 2011 AB SCIEX
Multi-Pesticide Screening and Quantitation with Identification based on MS/MS Library Searching 7 2011 AB SCIEX
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 (100-1000 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 8 2011 AB SCIEX
MS Resolution of TripleTF 5600 System Resolution 26946 @ 189 Da Resolution 27130 @ 192 Da FWHH = 7 mda Δt ~ 0.3 nsec Resolution 31775 @ 404 Da Resolution 34297 @ 732 Da 9 2011 AB SCIEX
Increased Selectivity through Resolution 6.8 μg/kg Imidacloprid in kale sample (5x dilution) XIC of 256.0596 ± 256mDa (1000ppm) S/N ~ 4 ± 25.6mDa (100ppm) S/N ~ 10 ± 2.56mDa (10ppm) S/N ~ 100 10 2011 AB SCIEX
MS/MS Resolution of TripleTF 5600 System 48 μg/kg Flonicamid in mustard greens sample (5x dilution) XIC of 230.0536 ± 2.3mDa Resolution 22900 @ 79 Da Resolution 29400 @ 230 Da 11 2011 AB SCIEX
XIC Manager Table and Confidence Settings 12 2011 AB SCIEX
Targeted Screening for ~300 Pesticides 5 ng/ml Standard 13 2011 AB SCIEX
Pesticides in Kohlrabi Sample Metalaxyl Methoxyfenozide Imidacloprid Spirotetramat Pyraclostrobin Cyprodinil TF-MS TF-MS/MS 14 2011 AB SCIEX
Pesticides in Chinese Broccoli Sample Metalaxyl Imidacloprid Spirotetramat Cyprodinil TF-MS TF-MS/MS 15 2011 AB SCIEX
Quantitation of Imidacloprid 23 μg/kg in Chinese Broccoli Sample 10 μg/kg MRL after dilution 16 2011 AB SCIEX
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 17 2011 AB SCIEX
Humulone and Cohumulone in Different Hops XIC of 363.2166 Da and 349.20010 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) 18 2011 AB SCIEX
MS Identification of Humulone Formula Finder Tool Resolution = 28700 Accuracy = 0.8 ppm 19 2011 AB SCIEX
MS/MS Identification of Humulone Fragment Interpretation Tool Resolution 20400 @ 57 Da Resolution 27800 @ 363 Da 20 2011 AB SCIEX
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 21 2011 AB SCIEX
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 = 3.2 10 μl injection AB SCIEX TripleTF 5600 LC/MS/MS system Negative polarity Electrospray Ionization (ESI) TF-MS survey (100-1000 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 22 2011 AB SCIEX
PCA Scores Plot Broccoli seeds Brussels sprouts Broccoli Cabbage Kale 23 2011 AB SCIEX
PCA Loadings Plot after PCVG Broccoli seeds PCVG 4 Brussels sprouts Broccoli Cabbage Kale 24 2011 AB SCIEX
PCA Profile Plot Broccoli seeds PCVG 4 Brussels sprouts Broccoli Cabbage Kale Broccoli marker 420.0463 at 14.1 min 25 2011 AB SCIEX
Empirical Formula Calculation C 12 H 23 N 9 S 3 26 2011 AB SCIEX
Search for C 12 H 23 N 9 S 3 in the Internet 27 2011 AB SCIEX
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) 28 2011 AB SCIEX
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. 2011 AB SCIEX. 29 2011 AB SCIEX
Questions and Answers 30 2011 AB SCIEX