Forensic Toxicology An ultrafast RapidFireMSMS method was developed for simultaneous qualitative analysis of common barbiturates in human urine The need for greater analytical capacity and throughp ID: 475128
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High-Throughput Triple Quadrupole Forensic Toxicology An ultrafast RapidFire/MS/MS method was developed for simultaneous qualitative analysis of common barbiturates in human urine. The need for greater analytical capacity and throughput for the analysis of barbiturates in forensic toxicology laboratories has placed demands on the traditional analytical technologies of HPLC and LC/MS/MS. We developed a method utilizing the Agilent RapidFire/MS/MS system to analyze butalbital, phenobarbital, secobarbital, amobarbital, and pentobarbital in urine with much faster sample cycle times and similar analytical results compared to LC/MS/MS assays. Phenobarbital OO Butalbital HOOOH2CH3CH3C OOO O H pentobarbital. these drugs reliably and accurately. analytical technologies. We strived study, we developed a method to conditions Triple Quadrupole Mass Spectrometer using Agilent MassHunter Triple experiments (Table1). Buffer A (pump 1)5 mM Ammonium acetate in LC/MS grade water; 1.5 mL/min Buffer B (pump 2)5 % LC/MS grade acetonitrile in water; 1.25 mL/min Buffer C (pump 3)50 % LC/MS grade methanol: 50 % HPLC grade isopropanol; Injection volume10 LSPE cartridgeAgilent RapidFire cartridge F (reversed-phase phenyl, G9208A)RF State 1sip sensorRF State 23,000 msRF State 32,000 msRF State 47,000 msRF State 51,000 ms Triple quadrupole conditions Gas temperature325 °C ow13 L/minNebulizer25 psi ow11 L/minSheath gas temperature400 °CNozzle voltage2,000 VCapillary voltage2,500 VESI modeNegative Table 1. RapidFire/MS/MS Conditions. AnalyteQ1 Q3 Dwell Fragmentor CE CAV Butalbital-d5228.142.1509552Phenobarbital 231.142.15095162Amobarbital and pentobarbital (isobars)225.142.15091162Butalbital223.142.15095122Secobarbital237.1 42.150100202 3Results and DiscussionSamples were prepared by spiking barbiturates into drug-free human cient mode of analysis. Data analysis were performed by Agilent MassHunter c. procedure. Ten microliters of each urine Thermal Microplate Sealer, mixed for 55657585 95105115125135145155165175 within the measured range (Table 2). 2 = 0.9995 02,0004,0006,0008,00010,000 2 = 0.9995 02,0004,0006,0008,00010,000 Amo/Pentobarbital 02,0004,0006,0008,00010,000 30 35 2 = 0.9981 02,0004,0006,0008,00010,000 16 Table 2. Interday and Intraday accuracy and precision (n=6) for RapidFire/MS/MS analysis of (ng/mL)Interday % AccuracyInterday % Precision Intraday % Accuracy Intraday % Precision 50100.45.0101.62.3250 99.05.0 97.94.51,000 99.42.6101.83.15,000 99.95.8101.15.110,000101.42.3102.65.7 (ng/mL)Interday % Accuracy Interday % Precision Intraday % Accuracy Intraday % Precision 50100.06.3101.11.1250 99.14.8 97.76.41,000 96.92.7 99.43.95,000101.14.4 99.94.010,000102.62.8101.61.5 (ng/mL)Interday % Accuracy Interday % Precision Intraday % Accuracy Intraday % Precision 50 99.66.0101.01.1250102.14.5 96.96.11,000 96.74.4102.02.25,000 99.34.5100.23.010,000102.34.4101.91.4 Pentobarbital Interday % Accuracy Interday % Precision Intraday % AccuracyIntraday % Precision 50100.06.2101.61.5250 99.45.9 98.97.61,000 95.74.1 99.04.65,000101.24.2101.61.310,000103.42.3101.91.5 5Carryover was assessed by analyzing the AUC of the blank calculated as % cant carryover (0%) was determined for all barbiturates cant matrix effect was observed ()This method consisting of dilute and shoot sample preparation followed by cient mode of qualitatively measuring barbiturates in signi cant carryover was observed for any analyte. 10.50×104-ESI MRM Frag = 100.0 V CID@20.0 (237.1 & 42.1) sequence1-ESI MRM Frag = 95.0 V CID@16.0 (231.1 & 42.1) sequence1-ESI MRM Frag = 91.0 V CID@16.0 (225.1 & 42.1) sequence1-ESI MRM Frag = 95.0 V CID@12.0 (223.1 & 42.1) sequence1 6 82 40×103 210×104 1030507090110130150170190210230250270 Acquisition time ( sec ) CountsCountsCountsCounts SecobarbitalPhenobarbital Butalbital shoot methodology. There is also the per hour. The analytical results were 1. June Feng, Lanqing Wang, Ingrid Dai, Tia Harmon, and John T. Bernert. Metabolites in Urine by LC/MS/MS. Journal of Analytical Toxicologywww.agilent.com/chemForensic Use Only. © Agilent Technologies, Inc., 2014