The simultaneous determination of diazepam and its three metabolites in dog plasma by high-performance liquid chromatography with mass spectroscopy detection |
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| Authors: | Liu Z Short J Rose A Ren S Contel N Grossman S Unger S |
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| Institution: | Department of Drug Metabolism and Pharmacokinetics, The DuPont Pharmaceuticals Company, Stine-Haskell Research Center, PO Box 30, Newark, DE 19714, USA. peter.z.liu@dupontpharma.com |
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| Abstract: | A fast, sensitive and specific LC/MS/MS method for the simultaneous determination of diazepam and its three metabolites, oxazepam, temazepam and desmethyldiazepam, in dog plasma is described. The method consists of an automated 96-well solid phase extraction procedure and electrospray LC/MS/MS analysis. D(5)-Diazepam is used as the internal standard for all the compounds. Intra-day and inter-day assay coefficients of variations are less than 12.7%. The lower limit of quantitation (LLOQ) is 1 nM for each analyte, based on 0.1 ml aliquots of dog plasma. The analytical run time was 5 min. Linearity is observed over the range of 1--500 nM. This method has been used to support the discovery of pharmacokinetic studies. |
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