Method for the determination of 5,5-diphenylbarbituric acid and its separation from 1,3-dimethoxymethyl-5,5-diphenylbarbituric acid in plasma by high-performance liquid chromatography

David L. Thacker, Darrell R. Abernethy, Arthur Raines, David A. Flockhart

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A method is described for the measurement of 5,5-diphenylbarbituric acid in plasma using high-performance liquid chromatography with UV detection. Briefly, the compounds are separated on a C18 reversed-phase column using a mobile phase of 50 mM sodium acetate (pH 4.5) and methanol. The flow-rate is 1.0 ml/min and 25 μl are injected and detected at 215 nm. The method is specific and sensitive in the range of concentrations tested, with a limit of quantification of 0.25 μg/ml. The calibration curves are linear for concentrations between 0.25 and 10 μg/ml. Intra-day and inter-day coefficients of variation are less than 8.5 and 10.5%, respectively, over the linear range. Intra-day and inter-day bias are less than 7.0 and 8.0%, respectively. A pharmacokinetic study conducted in male Beagle dogs administered 10 mg/kg of 1,3-dimethoxy-methyl-5,5-cliphenyibarbituric acid or 8 mg/kg of 5,5-diphenylbarbituric acid intravenously demonstrates the utility of this method.

Original languageEnglish (US)
Pages (from-to)149-155
Number of pages7
JournalJournal of Chromatography B: Biomedical Applications
Issue number1-2
StatePublished - Jun 12 1998



  • 1,3-Dimethoxymethyl-5,5-diphenylbarbituric acid
  • 5,5-Diphenylbarbituric acid

ASJC Scopus subject areas

  • Chemistry(all)

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