Development and utilization of a combined LC–UV and LC–MS/MS method for the simultaneous analysis of tegafur and 5-fluorouracil in human plasma to support a phase I clinical study of oral UFT®/leucovorin

Tegafur is a 5-fluorouracil (5-FU) prodrug widely used outside the United States to treat colorectal cancer as well as cancers of the head and neck. The resulting plasma concentrations of tegafur are much higher than those of 5-FU; thus, analytical methods are needed that are sensitive enough to detect low plasma concentrations of 5-FU and robust enough to simultaneously analyze tegafur. Previous LC–MS/MS methods have either failed to demonstrate the ability to simultaneously measure low 5-FU and high tegafur plasma levels, or failed to be applicable in clinical studies. Our goal was to develop a method capable of measuring low concentrations of 5-FU (8–200 ng/ml) and high concentrations of tegafur (800–20,000 ng/ml) in human plasma and to subsequently evaluate the utility of the method in patient samples collected during a phase I clinical study where oral doses of either 200 mg or 300 mg UFT®/LV (uracil and tegafur in a 4:1 molar ratio plus leucovorin) were administered. A combined LC–MS/MS and LC–UV method was developed utilizing negative ion atmospheric pressure ionization (API). The method provides an accuracy and precision of <10% and <6%, respectively, for both analytes. Material recoveries from the liquid–liquid extraction technique were 97–110% and 86–91% for tegafur and 5-FU, respectively. Utilization of this method to determine tegafur and 5-FU plasma concentrations followed by noncompartmental pharmacokinetic analyses successfully estimated pharmacokinetic parameters (CMAX, tMAX and AUC0–10 h) in the clinical study patients. Overall, this method is ideal for the simultaneous bioanalysis of low levels of 5-FU and relatively higher levels of its prodrug, tegafur, in human plasma for clinical pharmacokinetic analysis.

► Published analytic methods for tegafur are suboptimal for pharmacokinetic studies.
► An analytic method was developed for the anticancer drug tegafur and its metabolite.
► Accuracy, precision and recovery of the analytes from plasma were acceptable.
► The method was used to successfully determine human pharmacokinetic parameters.