Quantitation of Benzodiazepines in Urine, Serum, Plasma, and Meconium by LC-MS-MS

Abstract

A single method for confirmation and quantitation of a panel of commonly prescribed benzodiazepines and metabolites, alpha-hydroxyalprazolam, alpha-hydroxyethylflurazepam, alpha-hydroxytriazolam, alprazolam, desalkylflurazepam, diazepam, lorazepam, midazolam, nordiazepam, oxazepam, temazepam, clonazepam, and 7-aminoclonazepam, was developed for three specimen types, urine, serum/plasma, and meconium. Quantitation was by liquid chromatography tandem-mass spectrometry (LC-MS-MS) using a Waters Alliance-Quattro Micro system. The instrument was operated in multiple reaction monitoring mode with an electrospray ionization source in positive ionization mode. The method was evaluated for recovery, imprecision, linearity, analytical measurement range, specificity, and carryover. Average recovery and imprecision (within-run, between-run, and total % CV) were within +/- 15% of the target concentrations for urine (10 to 5000 ng/mL) and serum/plasma (10 to 2500 ng/mL) and within +/- 20% for meconium (10 to 5000 ng/g). In all, 205 patient specimens were analyzed, and the results compared to a previous in-house gas chromatography-MS method or LC-MS-MS results from an outside laboratory. Oxazepam glucuronide was evaluated as a hydrolysis control for the urine and meconium specimens.

Evaluation of the Short Term Photostability of Alprazolam Drug

“…In general, the benzodiazepines are broadly recommended drugs that are used as anxiolytics, sedative hypnotics, anticonvulsants or muscle relaxants [1][2] [3]. Aside from their therapeutic uses, benzodiazepines are frequently abused by drug addicts, so these drugs are often involved in both clinical and forensic cases.…”

“…Aside from their therapeutic uses, benzodiazepines are frequently abused by drug addicts, so these drugs are often involved in both clinical and forensic cases. Alprazolam {8-chloro-1-methyl-6-phenyl-4H-s-triazol-o(4,3-a)(1,4)benzodiazepine} ( Fig.1) belongs to benzodiazepines and is used for treatment of panic disorders, anxiety, depression and sleeping disorders [1]. Alprazolam is one of the most prescribed benzodiazepines.…”

“…It is rapidly absorbed and has a bioavailability of around 80-100% [4][2]. This molecule can be determined by a number of different methods such as: UV-VIS [5], TLC [6], GC-MS [7], LC-EI-MS [8][9], LC-MS-MS [1], HPLC-TOF/MS [10] in different materials such as: pharmaceutical formulations [5], human plasma [8][9], oral fluids [10], hair [7], blood [11], urine [2], etc. The stability of pharmaceutical molecules is a matter of great worry as it influences the safety and efficacy of the drug product [12].…”

Separation and Quantification of 1,4-benzodiazepines: HPLC versus CZE

“…3,4 Many studies on these drugs can be found in the literature. [5][6][7][8] Diverse analytical methods for determination of benzodiazepines in various matrices have been published, 9,10 e.g. potentiometry, 11 immunoassays, 12 spectrophotometry 13 and also separation techniques that represent a clear majority of the reported methods because they allow determination of individual compounds in often rather complicated mixtures.…”

DEVELOPMENT AND VALIDATION OF A GC-NPD/micro-ECD METHOD USING DUAL COLUMN FOR THE DETERMINATION OF BENZODIAZEPINE IN HUMAN WHOLE BLOOD AND PLASMA

“…Several analytical methods for the detection of multiple benzodiazepine drugs in human blood and urine samples have been reported using GC/MS, 7-10 LC/MS, 11,12 and LC/MS/ MS. [13][14][15][16][17][18] Some of these drugs and their metabolites possess polar groups in their molecules that require derivatization prior to GC/MS analysis.…”