Abstract
We hereby present a fast, high-throughput, and clinical LC-MS/MS assay for the simultaneous analysis of benzodiazepines in human urine. The assay is used as both a confirmations and semi-quantitative assay for the abovementioned drugs of abuse following immunoassay urine drug screens. Urine levels are reported in the range of 25 ng/mL–500 ng/mL for each of the 22 analytes, based on a six-level calibration and using a subset (10) of stable isotopically labeled analogues as internal standards. The urine sample is clarified, diluted ten times in internal standard reagent, and thereafter injected into the LC-MS/MS instrument. Reversed-phase liquid chromatography is used to separate the mixture, and the TSQ Endura triple-quadrupole (QqQ) MS instrument performs detection via positive-mode electrospray ionization multiple reaction monitoring.
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References
Morrow AL, Paul SM (1988) Benzodiazepine enhancement of γ-aminobutyric acid-mediated chloride ion flux in rat brain synaptoneurosomes. J Neurochem 50(1):302–306. https://doi.org/10.1111/j.1471-4159.1988.tb13264.x
Fraser AD, Meatherall R (1996) Comparative evaluation of five immunoassays for the analysis of alprazolam and triazolam metabolites in urine: effect of lowering the screening and GC-MS cut-off values. J Anal Toxicol 20(4):217–223. https://doi.org/10.1093/jat/20.4.217
ElSohly MA, Gul W, Murphy TP, Avula B, Khan IA (2007) LC-(TOF) MS analysis of benzodiazepines in urine from alleged victims of drug-facilitated sexual assault. J Anal Toxicol 31(8):505–514. https://doi.org/10.1093/jat/31.8.505
Marin SJ, Coles R, Merrell M, McMillin GA (2008) Quantitation of benzodiazepines in urine, serum, plasma, and meconium by LC-MS-MS. J Anal Toxicol 32(7):491–498. https://doi.org/10.1093/jat/32.7.491
Salomone A, Gerace E, Brizio P, Gennaro MC, Vincenti M (2011) A fast liquid chromatography-tandem mass spectrometry method for determining benzodiazepines and analogues in urine. Validation and application to real cases of forensic interest. J Pharm Biomed Anal 56(3):582–591. https://doi.org/10.1016/j.jpba.2011.06.007
Rossi B, Freni F, Vignali C, Stramesi C, Collo G, Carelli C, Moretti M, Galatone D, Morini L (2021) Comparison of two immunoassay screening methods and a LC-MS/MS in detecting traditional and designer benzodiazepines in urine. Molecules 27(1). https://doi.org/10.3390/molecules27010112
Bertol E, Vaiano F, Borsotti M, Quercioli M, Mari F (2013) Comparison of immunoassay screening tests and LC-MS-MS for urine detection of benzodiazepines and their metabolites: results of a national proficiency test. J Anal Toxicol 37(9):659–664. https://doi.org/10.1093/jat/bkt063
Bertol E, Vaiano F, Furlanetto S, Mari F (2013) Cross-reactivities and structure-reactivity relationships of six benzodiazepines to EMIT((R)) immunoassay. J Pharm Biomed Anal 84:168–172. https://doi.org/10.1016/j.jpba.2013.05.026
Wang J, Yang Z, Lechago J (2013) Rapid and simultaneous determination of multiple classes of abused drugs and metabolites in human urine by a robust LC-MS/MS method - application to urine drug testing in pain clinics. Biomed Chromatogr 27(11):1463–1480. https://doi.org/10.1002/bmc.2945
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Gqamana, P.P., Zhang, Y.V. (2024). High-Throughput Quantitative LC-MS/MS Analysis of Benzodiazepines in Human Urine. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Drug Analysis. Methods in Molecular Biology, vol 2737. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3541-4_10
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DOI: https://doi.org/10.1007/978-1-0716-3541-4_10
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