Simultaneous determination of 5′-deoxy-5-fluorouridine, 5-fluorouracil and its main metabolites in body fluids by HPLC

F. Palmisano, F. Berardi, M. De Lena, A. Guerrieri, V. Lorusso, P. G. Zambonin

Research output: Contribution to journalArticle


A reversed phase high performance liquid chromatographic method has been developed for the simultaneous determination of the pro-drug 5′-deoxy-5-fluorouridine (5′-dFUR), its metabolite 5-fluorouracil (5-FU) and some fluorinated pyrimidines involved in the metabolic activation process of 5-FU. The method has been used to monitor the bioavailability of 5′-dFUR and 5-FU in patients undergoing anticancer chemotherapy. Serum sample treatment involves addition of internal standard (5-bromouracil), protein precipitation with saturated ammonium sulphate solution and liquid-liquid extraction with ethyl acetate-isopropanol (90 : 10 v/v). Urine is simply diluted with mobile phase and injected. The average recovery from serum (at 0.5 μg/mL level) was 95.0%±1.4 for 5′-dFUR and 86.3%±3.5 for 5-FU. A linear response extending over four decades of concentration was observed. Detection limits in the low ng/ mL range were obtained using a 250 μL sample size and a 20 μL injection volume. Within-day and between-day coefficients of variation were below 4 and 10%, respectively.

Original languageEnglish
Pages (from-to)413-417
Number of pages5
Issue number9-10
Publication statusPublished - May 1992



  • Chemotherapy
  • Column liquid chromatography
  • Fluoropyrimidine drugs
  • Pharmacokinetics of doxifluridine

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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