Specific and Sensitive High Performance Liquid Chromatographic Determination of Alprazolam or Triazolam

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Abstract
A specific and highly sensitive liquid chromatographic procedure was developed for the rapid determination of intact [psychomimetic drugs] alprazolam [8-chloro-6-phenyl-1-methyl-4H-s-triazolo[4,3-a][1,4]benzodiazepine, Xanax] or triazolam [8-chloro-6-(o-chlorophenyl)-1-methyl-4H-s-triazolo[4,3-a][1,4]benzodiazepine, Halcion] in dog serum using the structurally similar triazolobenzodiazepines as mutual internal standards. The procedure consists of extracting 1 ml of alkali buffered serum with toluene, evaporating an aliquot of the toluene to dryness and quantitating the redissolved residue by HPLC [high performance liquid chromatography] using UV detection (221 nm). Samples were chromatographed on a microparticulate reverse phase column using a mobile phase composed of acetonitrile:isopropanol:water (94:5:1) and a flow rate of 0.75 ml/min. Metabolites of alprazolam and triazolam did not interfere in the assay. The lower limit of detection was .apprx. 1 ng/ml of serum extracted. The utility of the analytical methodology for the determination of alprazolam or triazolam in pharmacokinetic studies in the dog was demonstrated.