A rapid solid-phase extraction and HPLC/DAD procedure for the simultaneous determination and quantification of different benzodiazepines in serum, blood and post-mortem blood

Abstract

A rapid and quantitative method for the determination of benzodiazepines using high-performance liquid chromatography (HPLC) with diode-array detection (DAD) is reported. The drugs were extracted from serum, blood or post-mortem blood using C18 extraction columns. Brotizolam was used as internal standard. Experiments with spiked serum/blood samples resulted in recoveries between 75% and 94% for all investigated benzodiazepines. Excellent linearity was obtained over the concentration range 5-1500 ng benzodiazepine/ml. The limit of detection was approximately 2 ng/ml. The detection of low therapeutic serum levels of highly potent benzodiazepines is also possible.

A preliminary study on the stability of benzodiazepines in blood and plasma stored at 4° C

“…Besides high pressure liquid chromatography [20], gas chromatography with electron capture detection seemed to be most appropriate regarding sensitivity, but it failed to detect some important metabolites and to identify major degradation products, for example arising from 7-nitro-1,4benzodiazepines. Moreover, it seemed important to confirm particular peaks by mass spectrometry to avoid unreliably high concentrations or false positive results arising from potential matrix interferences as demonstrated by lorazepam.…”

First Experience with the REMEDi HS Urine Benzodiazepine Assay

“…The hydrolyzed samples were extracted according to a previously published method (18). The hydrolyzed samples were extracted according to a previously published method (18).…”

“…These methods cannot determine the exact identity of the benzodiazepine present in the sample and show varying degrees of cross-reactivity. For specific identification confirmatory techniques are used, such as highperformance liquid chromatography (HPLC) followed by UV detection (12)(13)(14)(15)(16)(17)(18)(19), or gas chromatography mostly coupled with mass spectrometry (GC/MS) (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). Another commonly used screening technique is thin layer chromatography (TLC) in conjunction with acid hydrolysis, which generates various benzophenone derivatives for benzodiazepines and their glucuronides (11).…”

Liquid chromatography–electrospray ionisation mass spectrometry for the determination of nine selected benzodiazepines in human plasma and oral fluid

“…Finally, some references related to the determination of BZP in biological samples by HPLC-DAD [29][30][31][32] and capillary electrophoresis [33][34][35] must be included in order to complete a full revision of the analytical methodology.…”