Sigma-Aldrich (Poole,Dorset,UK) LGC Standards(Teddington,Surrey,UK) 1 mg/ml Sigma-Aldrich Greyhound Chromatography(Brikenhead,UK) / Waters ACQUITY UPL

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1 ACQUITY UPLC I-Class/Xevo TQD Mark Roberts and Michelle Wood Waters Corporation, MS Technologies Centre, Manchester, UK ACQUITY UPLC I-Class/Xevo TQD 2 2 MRM 178 / 950 MS ACQUITY UPLC/ACQUITY TQD MRM 100 LC/MS LC/MS 2 ACQUITY UPLC I-Class/Xevo TQD MRM 1-3 ACQUITY UPLC I-Class/Xevo TQD ACQUITY UPLC I-Class System Xevo TQD RADAR Technology PIC Scannning ACQUITY UPLC HSS C18 Waters UPLC /MS/MS 1

2 Sigma-Aldrich (Poole,Dorset,UK) LGC Standards(Teddington,Surrey,UK) 1 mg/ml Sigma-Aldrich Greyhound Chromatography(Brikenhead,UK) / Waters ACQUITY UPLC I-Class/Xevo TQD ACQUITY UPLC / TQD UPLC MS ACQUITY UPLC I-Class Xevo TQD A B ACQUITY UPLC HSS C x 150 mm 1.8 µm µl MRM 10 µl 5 mm ph mm ph µl/min 5 mm ph % B 13% 95% 15 ESI ESI 3.0 kv L/h 20 L/h MassLynx Multiple Reaction Monitoring MRM Targetlynx TM ChromaLynx TM ACQUITY TQD 1,2 ACQUITY UPLC 1,2 ACQUITY UPLC I-Class/Xevo TQD 2

3 FTN 5 ACQUITY UPLC I-Class 1 UPLC 100 µl RADAR PIC Xevo TQD 6 1 RADAR MRM Product Ion Confirmation scanning (PICs) MRM 1 ACQUITY UPLC I-Class/Xevo TQD MRM MS 178 MRM / ACQUITY UPLC MRM ACQUITY UPLC I-Class ACQUITY UPLC I-Class/Xevo TQD 3

4 +20 V +35 V +50 V +65 V +80 V +95 V -20 V ChromaLynx 950 ACQUITY TQD Xevo TQD MRM SSM 2 ACQUITY UPLC I-Class/Xevo TQD SSM TargetLynx SSM 10 MRM 0.3 ACQUITY UPLC I-Class ACQUITY UPLC I-Class/Xevo TQD 4

5 Total Ion Chromatogram Library Spectrum Acquired Spectrum Candidate Listing 3 SSM ChromaLynx 10 MRM ACQUITY UPLC I-Class methadone EDDP cocaine benzoylecgonine 4 ACQUITY UPLC I-Class/Xevo TQD MRM TargetLynx ACQUITY UPLC I-Class/Xevo TQD ACQUITY UPLC I-Class ACQUITY UPLC I-Class/Xevo TQD 5

6 ACQUITY UPLC/TQD (original configuration) ACQUITY UPLC I-Class/Xevo TQD (new configuration) RT RT RT Methadone EDDP Paracetamol Cocaine Benzoylecgonine Nicotine Caffeine Temazepam Oxazepam Theophylline Nordiazepam MRM 4 methadone TargetLynx ACQUITY UPLC I-Class/Xevo TQD 6

7 MRM 950 ChromaLynx 5 ACQUITY UPLC I-Class/Xevo TQD mirtazapine mirtazapine N-desmethyl ChromaLynx mirtazapine N-desmethyl ACQUITY TQD ACQUITY TQD Xevo TQD xylometazoline MRM 5 mirtazapine mirtazapine N-desmethyl ChromaLyn ACQUITY UPLC I-Class/Xevo TQD 7

8 PICs hydromorphone morphine Xevo TQD MRM Product Ion Confirmation scanning(pics) 6 MRM hydromorphone hydromorphone hydromorphone morphine MRM Hydromorphone morphine MRM 0.2 PIC PICs morphine hydromorphone TargetLynx hydromorphone morphine B A C 6 TargetLynx hydromorphone (A) 200 ng/ml hydromorphone (B) morphine (C) ACQUITY UPLC I-Class/Xevo TQD 8

9 RADAR RADAR MRM MRM RADAR MRM MRM RADAR MRM mirtazapine N-desmethyl desmethyl citalopram xylometazoline V RADAR 7 RADAR MRM MRM mirtazapine N-desmethyl 3.79 desmethyl citalopram 6.47 xylometazoline 7.42 A B 8 RADAR mirtazapine N-desmethyl 3.79 (A) xylometazoline 7.42 (B) 30 V ACQUITY UPLC I-Class/Xevo TQD 9

10 UPLC/MS ACQUITY UPLC I-Class MS RADAR PICs Xevo TQD 1. Roberts M, Lee R, Wood M. Targeted MRM Screening for Toxicants in Biological Samples by UPLC/MS/MS. Waters Application Note EN. 2. Lee R, Roberts M, Paccou A, Wood M..Development of a New UPLC/MS Method for Systematic Toxicological Analysis. Waters Application Note EN. 3. Humbert L, Grisel F, Richeval C, Lhermitte M. Screening of Xenobiotics by Ultra-Performance Liquid Chromatography Mass Spectrometry Using In-Source Fragmentation at Increasing Cone Voltages: Library Constitution and an Evaluation of Spectral Stability. Journal of Analytical Toxicology. 2010; Vol Rosano TG, Wood M, Swift TA. Postmortem Drug Screening by Non-Targeted and Targeted UltraPerformance Liquid Chromatography-Mass Spectrometry Technology. Journal of Analytical Toxicology. 2011; Vol ACQUITY UPLC I-Class System Brochure. Waters Brochure EN. 6. Xevo TQD Brochure. Waters Brochure EN TEL FAX F TEL FAX Waters ACQUITY UPLC ACQUITY UPLC PIC Xevo MassLynx Waters Corporation RADAR TargetLynx ChromaLynx The Science of What s Possible Waters Corporation 2013 Waters Corporation. Printed in Japan JA PDF

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MS Xevo G2 Q-Tof ESI+ 1 2 / pm-4 00 am 2 00 am # am # am #3 / pm am am #4 0.8 kv 20 V MS E ev MS E MASSLYNX UPLC-Tof-MS E John Archer, 1 Paul Dargan, 1 David Wood, 1 Nayan Mistry, 2 and Michelle Wood 2 1 Guy s and St. Thomas NHS Foundation Trust, Clinical Toxicology, London, UK 2 Waters Corporation, MS Technologies

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1 WSV SIR m/z Analyte RT (Min) SIR m/z Cone voltage (V) Ascorbic Acid (C).91 177 2 Thiamine (B1) 1.1 265 5 Nicotinic Acid (B3) 1.27 124 15 Pyridoxal ( ACQUITY UPLC H-Class ACQUITY QDa Mark E. Benvenuti, Dimple Shah, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA MS ACQUITY QDa UPLC HPLC UV ACQUITY QDa LC EC 1925/26 CFR 21 HPLC 1 LC-MS ACQUITY

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