X500R QTOF - SCIEX

1 X500R QTOFA COMPENDIUM OF RELATED RESEARCH APPLICATIONS Food TestingPesticide Analysis in Food Antibiotic Analysis in Food Mycotoxin Analysis in FoodPesticide and Potency Testing for the Cannabis IndustryUsing the X500R qtof System and SCIEX OS Software to Identify and Quantify Food ResiduesUsing the X500R qtof System and SCIEX OS Software to Quickly Identify Unknowns in Food SamplesWater AnalysisPesticide Analysis in Water PPCP Analysis in Water Illicit Drugs Analysis in Water Forensic AnalysisForensic Identification and Quantitation Workflows Delivered on a Revolutionary Designed qtof and SCIEX OS SoftwareForensic Drug Screening Analysis - Urine Forensic Drug Screening Analysis - BloodContents X500R qtof FOOD TESTINGp 1 Pesticide analysis in food Elevate your food testing with the X500R qtof System Method details and access to HR-MS/MS libraries to detect, quantify, and confirm pesticides in food extracts using HPLC coupled with the X500R qtof system, powered by SCIEX OS Suggested sample preparation conditions based on the QuEChERS method (QuEChERS European standard method 15662).

2 Column Phenomenex Kinetex Biphenyl, 50 x mm, um Mobile Phase A 5 mM ammonium formate in water Mobile Phase B 5 mM ammonium formate in methanol Flow rate mL/min Column temperature 40 C Injection volume 2 uL Gradient profile Time (min) % B 0 10 10 30 60 80 95 95 10 10 Step 1 10g homogenized sample (hydrate with water if needed).Step 2 Add 10mL acetonitrile and internal standardStep 3 Extract with vigorous shaking for 1 minuteStep 4 Add QuEChERS salt mix (Phenomenex roQ KS0-8909),shake vigorously for 1 5 Centrifuge for 10 minutes at 9000 6 Transfer 1mL supernatent to dSPE tube (PhenomenexroQ KS0-8916,8913,8914, or 8915 depending on 7 Shake vigorously for 30 secondsStep 8 Dilute 100 L of extract to 1mL in water for MS analysisp 2 Review your results with utmost efficiency using SCIEX OS for simultaneous quantitation and MS/MS library confirmation. Download a free XIC compound list detailing a full list of pesticide compounds including molecular formula and accurate mass.)

3 Download a free trial of the pesticide high resolution MS/MS library, containing 557 compounds. Learn more at Suggested IDA (Information Dependent Acquisition) conditions for routine food contaminant testing as displayed in SCIEX OS. AB SCIEX is doing business as SCIEX . 2015 AB SCIEX . For Research Use Only. 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. Document number: RUO-MKT-07-2861-A p 1 Antibiotic analysis in food Elevate your food testing with the X500R qtof System Method details and access to HR-MS/MS libraries to detect, quantify, and confirm antibiotic vet drugs in tissue extracts using HPLC coupled with the X500R qtof system, powered by SCIEX OS Sample prep protocol adopted from: Mastovska & Lightfield, J. Chrom. A., 2008, 1202, 118-123 Column Phenomenex Gemini 3 m C18 110 column, 50 x Mobile Phase A formic acid in water Mobile Phase B formic acid in methanol Flow rate mL/min Column temperature 40 C Injection volume 10 uL Gradient profile Time (min) % B 0 2 2 80 99 99 11 2 15 2 Step 1 1g Tissue homogenized with 10mL extraction solvent(1:5) Water AcetonitrileStep 2 Vortex, shake vigorously for 5 minutes, centrifuge(5000rpm, 5 minutes)Step 3 Decant supernatent to 15mL tube & add 500mg 4 Vortex, shake 30 sec, centrifuge (5000rpm, 1 minute).

4 Step 5 Collect 5mL aliquot of extract and reduce in volume to< 6 Bring to final volume of 1mL w/water and filter for analysisp 2 Review your results with utmost efficiency using SCIEX OS for simultaneous quantitation and MS/MS library confirmation. Download a free XIC compound list detailing a full list of antibiotic compounds including molecular formula and accurate mass. Download a free trial of the antibiotic high resolution MS/MS library, containing 244 compounds. Learn more at Suggested IDA (Information Dependent Acquisition) conditions for routine food contaminant testing as displayed in SCIEX OS. AB SCIEX is doing business as SCIEX . 2015 AB SCIEX . For Research Use Only. 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. Document number: RUO-MKT-07-2858A p 1 Mycotoxin analysis in food Elevate your food testing with the X500R qtof System Method details and access to HR-MS/MS libraries to detect, quantify, and confirm mycotoxins in food extracts using HPLC coupled with the X500R qtof system, powered by SCIEX OS Software.

5 Sample prep protocol based : Sulyok M, Krska R, Schumacher R (2010) Food Chem 119:408-416 SCIEX OS can deliver faster method set-up Column Phenomenex Gemini C18, 150 x mm, 5 m Mobile Phase A 5 mM ammonium acetate + 1% acetic acid in waterMobile Phase B 5 mM ammonium acetate + 1% acetic acid in methanolFlow rate mL/min Column temperature 25 C Injection volume 5 uL Gradient profile Time (min) % B 0 0 2 0 14 100 18 10 0 0 Step 1 Add 4mL extraction solvent (79/20/1. V/V/V)Acetonitrile/Water/Acetic Acid per 1g 2 Extract on a rotary shaker for 90 minutesStep 3 4 Dilute 350 L of extract with 350 L 20/79/1 (v/v/v)Acetonitrile/Water/Acetice 5 Filter into vial for 2 Review your results with utmost efficiency using SCIEX OS for simultaneous quantitation and MS/MS library confirmation. Download a free XIC compound list detailing a full list of mycotoxin compounds include molecular formula and accurate mass.

6 Download a free trial of the mycotoxin high resolution MS/MS library, containing 288 compounds. Learn more at Suggested IDA (Information Dependent Acquisition) conditions for routine food contaminant testing as displayed in SCIEX OS. AB SCIEX is doing business as SCIEX . 2015 AB SCIEX . For Research Use Only. 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. Document number: RUO-MKT-07-2860-A p 1 Pesticide and Potency Testing for the Cannabis Industry Elevate your confidence in cannabis testing with the X500R qtof System Method details and access to HR-MS/MS libraries to detect, quantify, and confirm pesticides, mycotoxins, cannabinoids and terpenes in plant edible samples using HPLC coupled with the X500R qtof system, powered by SCIEX OS Software. SCIEX OS can deliver faster method set-up Column Restek Raptor ARC-18 50 x , m Mobile Phase A formic acid, 5mM ammonium formate in water Mobile Phase B formic acid, 5mM ammonium formate in acetonitrile Flow rate mL/min Column temperature 40 C Injection volume 5 L Gradient profile Time (min) % B 0 30 30 4 95 5 95 30 Step 1 Weight sampleStep 2 Sonicate in 9mL methanol for 5 minutesStep 3 Bring to 10mL final volumeStep 4 Centrifuge and draw off 1mL for 2 Review your results with utmost efficiency using SCIEX OS for simultaneous quantitation and MS/MS library confirmation.

7 Download a free XIC compound list detailing a full list of applicable compounds including molecular formula and accurate mass. Download a free trial of our All in One high resolution MS/MS library, containing 2135 compounds. Learn more at IDA (Information Dependent Acquisition) conditions for routine food contaminant testing as displayed in SCIEX OS. AB SCIEX is doing business as SCIEX . 2015 AB SCIEX . For Research Use Only. 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. Document number: RUO-MKT-07-2859 p 1 Perfect Balance to Elevate your Lab s PerformanceUsing the X500R qtof System and SCIEX OS Software to Identify and Quantify Food Residues Andr Schreiber1, April Quinn-Paquet2, and Jianru Stahl-Zeng31 SCIEX Concord, Ontario (Canada), 2 SCIEX Framingham, Massachusetts (USA), 3 SCIEX Darmstadt (Germany) OverviewHere we present results using a new method to identify andquantify pesticide residues in food using the SCIEX X500 RQTOF system.

8 Samples were extracted using a QuEChERSmethod and analyzed by LC-HR-MS/MS. Limits of quantitation of10 g/kg were achieved for every compound after 10x dilution ofthe extract to minimize possible matrix compounds were automatically identified by matchingretention time, accurate mass and isotope pattern of themolecular ion and MS/MS library searching using SCIEX OSsoftware. In the same data processing step, compounds werequantified and unknown samples were flagged when a user-defined reporting level was advancements in LC-MS/MS technology, including hybridsystems like quadrupole-quadrupole Time-of-Flight ( qtof ), nowprovide the ability to perform targeted and non-targetedscreening in food samples on a routine SCIEX X500R qtof system is a robust, high performancehigh resolution MS/MS system designed for routine useproviding: Sensitivity to easily detect compounds at maximum residuelevels Resolving power to remove interference from complex foodmatrices Linearity to quantify over up to 3 orders of magnitude Mass accuracy to identify compounds following regulatoryguidelines Confident identification using MS/MS spectra and ion ratios Industry leading robustness of Turbo V source and CurtainGas interfaceFull scan chromatograms are very rich in information and easilycontain thousands of ions from any residue present in thesample, including the food matrix itself.

9 Powerful software isneeded to explore the high resolution MS/MS spectra generatedto get answers and results from these complex SCIEX OS software is a single platform for MS control, dataprocessing, and reporting and provides: Simple software workflows that deliver reliable results Simultaneous identification and quantitation Quick data review and reporting utilizing customizableflagging and filtering of resultsExperimentalStandardsA standard mix of 200 pesticides was used to prepare serialdilutions for quantitative preparationEU proficiency test samples and food samples from a localsupermarket were extracted using a QuEChERS procedurefollowing guideline EN 15662/2007. Sample extracts were diluted10x to minimize possible matrix SeparationLC separation was performed using a SCIEX ExionLC ACsystem with a Phenomenex Kinetex Biphenyl (50 x )column and a fast gradient of water and methanol with 5 mMammonium formate buffer at a flow rate of mL/min (see Table1 for the gradient profile).

10 The injection volume was 5 2 Table 1. Gradient conditions used for the separation of pesticidesStep Time (min) A (%)B (%)0 DetectionThe SCIEX X500R qtof system with Turbo V source andElectrospray Ionization (ESI) was calibration was achieved using the integrated calibrantdelivery system (CDS) with the TwinSprayer probe (dual ESIneedle).High resolution data were acquired using an IDA methodconsisting of a TOF-MS survey (100-1000 Da for 100 msec) andup to 20 dependent MS/MS scans (50-1000 Da for 35 msec).MS/MS fragmentation was achieved using CE of 35 V with acollision energy spread (CES) of 15 background subtraction (DBS) was activated for bestMS/MS coverage, and no inclusion list was used to also allowretrospective unknown identification without the need for asecond injection to acquire MS/MS Acquisition and ProcessingAll data were acquired and processed using SCIEX OS softwareversion , which showcases a thoughtfully designed userinterface that is fast to learn and delivers improved and DiscussionX500R Performance CharacteristicsResolution > 20,000 (at full width half height) and mass accuracy<5 ppm are often sufficient to separate the analytes of interestfrom interfering matrices and, thus, are identified as the setrequirements for compound identification in various , 2 The X500R qtof system utilizes N-optics design to maximizeresolution while maintaining benchtop design and a minimizedfootprint.