“Bio-Analytical Method Development and Validation for The Estimation of Capecitabine in Plasma by Using RP-HPLC Method”

Abstract

The present study focuses on the development and validation of High-Performance Liquid Chromatography (HPLC) and UV spectrophotometric methods for the quantitative estimation of Capecitabine, an oral chemotherapeutic agent, in pharmaceutical formulations and human plasma. The HPLC method was optimized using a mobile phase consisting of Methanol and 0.05% Orthophosphoric Acid (65:35 v/v) with a flow rate of 0.8 mL/min and detection at 284 nm. The chromatographic separation was achieved on an Agilent C18 column (250 × 4.6 mm, 5 µm particle size), yielding a sharp and symmetric peak at a retention time of 4.347 minutes. Validation of the developed method, as per ICH Q2(R1) guidelines, demonstrated excellent linearity in the range of 10–50 µg/mL (r2=0.999r^2 = 0.999r2=0.999). Recovery studies confirmed high accuracy, with recovery rates between 97.7% and 98.5%. Precision, expressed as %RSD, was below 2% for both intra-day and inter-day analyses, indicating reproducibility. The robustness of the method was confirmed by minor deliberate variations in chromatographic conditions, which showed negligible impact on results. In addition, the UV spectrophotometric method, based on Capecitabine's λmax at 284 nm, provided a complementary tool for rapid preliminary screening. The developed methods were successfully applied to analyze marketed formulations, achieving a % label claim of 98.23%, and were suitable for pharmacokinetic studies in human plasma. These validated methods offer a reliable, reproducible, and efficient approach for the routine quality control of Capecitabine in pharmaceutical and clinical settings, contributing to better therapeutic monitoring and compliance with regulatory requirements.