Development and Validation of an Isocratic, Sensitive and Facile RP-HPLC Method for Rapid Analysis of 5-Fluorouracil and Stability Studies Under Various Stress Conditions

Abstract

The objective of the present investigation was to develop and validate a RP-HPLC method for rapid analysis of a cytotoxic drug 5-fluorouracil in bulk drug, marketed injection, water-in-oil (w/o) nanoemulsion and double water-in-oil-in-water (w/o/w) nanoemulsion. The chromatographic identification was achieved on Lichrosphere 250 mm × 4.0 mm RP C₈ column having a 5 μm packing as a stationary phase using methanol:water (50:50 % v/v) as a mobile phase, at a flow rate of 1 mL/min with UV detection at 254 nm. The proposed method was validated for linearity, accuracy, precision, robustness, sensitivity and specificity. The utility of the method was verified by assay of drug in various pharmaceutical formulations. The proposed method was found to be precise, accurate, robust, sensitive and specific. The amount of 5-fluorouracil in marketed injection, developed water-in-oil nanoemulsion and double water-in-oil-in-water was found to be more than 99 %. The proposed method successfully resolved drug peak in the presence of its degradation products which established stability-indicating property of the proposed method. Overall, these results indicated that the proposed method can be successfully employed for routine analysis of 5-fluorouracil in bulk drug and various pharmaceutical dosage forms.