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 microg/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 microL sample size and a 20 microL injection volume. Within-day and between-day coefficients of variation were below 4 and 10%, respectively.
Simultaneous determination of 5'-deoxy-5-fluorouridine, 5-fluorouracil and its main metabolites in body fluids by HPLC
GUERRIERI, Antonio;
1992-01-01
Abstract
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 microg/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 microL sample size and a 20 microL injection volume. Within-day and between-day coefficients of variation were below 4 and 10%, respectively.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
