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Assay of the major (4-hydroxylated) metabolites of diphenylhydantoin in human urine


A modified gas chromatographic procedure for the determination of unconjugated and conjugated 4-hydroxydiphenylhydantoin (4-OH-DPH) in urine has been developed.

Unconjugated 4-OH-DPH is determined after selective extraction with toluene — ether (1:1). For the assay of conjugated 4-OH-DPH, the urine is pre-extracted withisoamylalcohol before acid hydrolysis to avoid interference by the dihydrodiol metabolite of DPH. The sensitivity of the method is 0.1 µg per ml. The method has been used to determine the urinary metabolites in two adult volunteers, during steady state plasma concentrations of DPH and in the elimination phase.

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To the memory of Balzar Alexanderson

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Karlén, B., Garle, M., Rane, A. et al. Assay of the major (4-hydroxylated) metabolites of diphenylhydantoin in human urine. Eur J Clin Pharmacol 8, 359–363 (1975).

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Key words

  • Diphenylhydantoin metabolites
  • human urine
  • gas chromatography
  • 4-hydroxydiphenylhydantoin