Developing and validation a method for identifying a chosen bulk medication 5-flourouracil and its formulation.

Abstract

This study presents the development and validation of a stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative analysis of 5-Fluorouracil (5-FU), a widely used chemotherapeutic agent, in bulk drug and pharmaceutical formulations. The method utilized a C18 column (250 × 4.6 mm, 5 µm) with a mobile phase of 50 mM potassium dihydrogen phosphate buffer (pH 5.0) at a flow rate of 1.2 mL/min and UV detection at 254 nm. Validation was performed as per ICH Q2(R1) guidelines, assessing specificity, linearity, accuracy, precision, robustness, limit of detection (LOD), and limit of quantification (LOQ). The method demonstrated excellent linearity (R² = 0.999) over a concentration range of 5–25 µg/mL, with a regression equation of AUC = 69.15x + 2.276. Accuracy was confirmed with recoveries of 98.82–99.45% across 80%, 100%, and 120% levels, and precision studies showed %RSD values below 0.13%. The LOD and LOQ were 0.265 µg/mL and 0.832 µg/mL, respectively. Forced degradation studies under acidic, alkaline, oxidative, and thermal conditions confirmed the method’s stability-indicating capability, with degradation ranging from 4.10% (thermal) to 17.15% (alkaline). Analysis of marketed tablet formulations yielded a drug content of 99.74% of the labeled claim. The method is simple, precise, robust, and cost-effective, making it suitable for routine quality control, stability testing, and pharmacokinetic studies of 5-FU.