Simultaneous Gas Chromatographic Determination of Diazapam and its Major Metabolites in Human Plasma, Urine, and Saliva

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Abstract
A GLC analysis method was developed for the simultaneous determination of diazepam (I) [a mild tranquilizer, skeletal muscle relaxant, anticonvulsant and hypnotic], and its major metabolites N-desmethyldiazepam (II), oxazepam (III) and hydroxydiazepam (IV) to human plasma, urine and saliva. Medazepam (V) was used as the internal standard to control extraction efficiency and permit precise and accurate determination of I-IV. Extraction and GLC analysis of physiological fluid samples in triplicate required only 40 min using the developed method. Three human subjects were given either 20 or 5 mg of I via oral administration and serial specimens of blood, urine and saliva collected. All samples were analyzed using the developed assay method. The method could evidently be applied to pharmacokinetic studies of I where plasma, urine and/or saliva is to be monitored.