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James W. King, Leslie J. King, Solid-Phase Extraction and On-Disc Derivatization of the Major Benzodiazepines in Urine Using Enzyme Hydrolysis and Toxi-Lab VC MP3 Column, Journal of Analytical Toxicology, Volume 20, Issue 4, July-August 1996, Pages 262–265, https://doi.org/10.1093/jat/20.4.262
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Abstract
Because of the increase in use of the newer benzodiazepines, we explored the opportunity to develop a gas chromatographic-mass spectrometric (GC-MS) method that encompasses most of the widely prescribed benzodiazepines in use today. The benzodiazepines included in our study are nordiazepam, oxazepam, temazepam, Iorazepam, α-hydroxyalprazolam, α-hydroxytriazolam, desalkylflurazepam, and 2-hydroxyethylflurazepam. Using 1.0 mL of urine as the matrix, we added the enzyme Glusulase and incubated the specimens for 2 h to obtain the free drugs. The hydrolyzed samples were then loaded onto a Toxi-Lab Spec VC MP3 column containing a 15-mg disc. On-disc derivatization was accomplished by adding N-methyl-N-(t-butyldimethylsilyl) trifluroacetamide (MTBSTFA) with 1% TBDMSCI to the disc. The derivatives were then placed in a GC vial and analyzed by GC-MS in the selected ion monitoring mode. These results were then compared to confirmed positives by the traditional acid hydrolysis GC-MS method.