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  • 1 Forensic Science Laboratory Madivala Bangalore 560 068 India
  • | 2 University of Mysore Department of Chemistry Manasagangothri Mysore 570 006 India
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A high-performance thin-layer chromatographic method combined with densitometric measurement and UV scanning was used to optimize a new mobile phase consisting of hexane and ethyl acetate (7:3 v/v) for the estimation of diazepam in spiked human blood. Efficacy and ease of extraction with three solvents, acetonitrile, diethyl ether, and chloroform, each at three different pH conditions were studied. Validation parameters for diazepam were found to be linear in the range of 0.5–20 μg/spot with r = 0.998, limit of detection (LOD), 0.2 μg, limit of quantification (LOQ), 0.5 μg. Usefulness of the new mobile phase for the separation of six common benzodiazepines, namely, nitrazepam, clonazepam, lorazepam, chlordiazepoxide, alprazolam, and clozapine from diazepam, was proved through their retention factor and UV pattern.

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