DEVELOPMENT AND VALIDATION OF RAPID AND SENSITIVE HPLC METHOD FOR THE DETERMINATION OF 5-FLUOROURACIL IN HUMAN SERUM

Summary This study describes a simple and fast high-performance liquid chromatography method for the determination of 5-Fluorouracil in serum. Samples were collected from adult cancer patients receiving high dose 5-Fluorouracil at Mahathma Gandhi Memorial hospital (Warangal, AP.India) at various time intervals after the end of each infusion. Serum was deproteinized with trichloroacetic acid and the supernatant was injected into a 250×4.6 mm octadecylsilane column. Mobile phase was methanol: water (10:90) with a flow rate of 1ml/min. Ultraviolet detection was done at 313 nm and at ambient temperature. Para aminoacetophenone was used as internal standard. 5-Fu and internal standard [thymine] retention times were 4.6 and 9.5 minutes, respectively. Results showed that reproducibility (precision) of method within a day was 98 to 99.8 percent and between days was 95.6 to 99.7 percent. The recovery of the method was between 88.6 and 99.7 percent. The quantitation limit of the method for 5-Fu was 10µM. This method is suitable for quantitation of 5Fu after infusion of high doses of this drug and has good accuracy, precision and quantitation limit.