Toxicological determination of benzodiazepines in serum: methods and concentrations associated with high-dose intravenous therapy with diazepam

Clinical Toxicology
R C KellyI Sunshine


This paper describes a rapid and simple procedure for the determination of benzodiazepines in biological samples. Five common benzodiazepines (diazepam, oxazepam, clorazepate, flurazepam, and chlordiazepoxide) and/or their major metabolites are extracted from a buffered serum sample at pH 9.2 by a mixture of toluene, hexane, and isoamyl alcohol. The phases are separated by centrifugation, and a small aliquot of the organic layer is injected into a gas chromatograph equipped with an electron-capture detector. The drugs are identified and quantitated by comparison with standards simultaneously processed similarly. A sample can be analyzed in about 30 min. The technique is illustrated by its application to sequential samples from a patient being treated for delirium tremens with large intravenous doses of diazepam. Serum diazepam, N-desmethyldiazepam, and oxazepam concentrations several times higher than usually encountered therapeutically were observed.


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