HPLC ANALYSIS AND VALIDATION OF 5-FLUOROURACIL AND ITS METABOLITES INRAT PLASMA

An isocratic, reverse phase HPLC method, for the simultaneous determin ation of 5-fluorouracil (5-Fu), 5-fluoro-5,6-dihydrouracil (FuDH), and 5-fluorouridine (Furd), in rat plasma has been developed. The method utilizes a Spherisorb(R) column and a mobile phase composition of 100 mu M phosphate buffer at pH 3.0. The extraction procedure from the pla sma samples has been optimized to give maximum recovery of the analyte s. The method has been validated as per USP guidelines in terms of lin earity, accuracy and precision. The limit of quantitation for 5-Fu, Fu DH, and Furd was found to be 0.2 mu g/mL, 0.3 mu g/mL and 0.8 mu g/mL respectively.