Development and validation of a stability indicating RP-HPLC method for determination of flucytosine and its process related impurities in injectable pharmaceuticals

Abstract

A simple, sensitive, reproducible and cost effective reversed-phase liquid chromatography (RPHPLC) method coupled with a photodiode array detector was developed and validated for determination of Flucytosine and its related substances in pharmaceutical dosage forms, especially for injectable solution. The separation was achieved from octadecylsilyl silica gel, C18 (4.6 mm x 250 mm, 5µ) column with a mobile phase consisting of HPLC grade Water and Methanol (95:5) at a flow rate of 1ml/min with UV detection at 260 nm at 30 ° C column temperature. Total run time was 10 min within which main compound and other known (Fluorouracil) and unknown impurities were separated. Stability indicating capability was established by force degradation experiments and separation of known degradation products. This chromatographic method was optimized using the samples generated from forced degradation studies and the impurity spiked solution. Good resolution between the peaks corresponds to process-related impurities and degradation products from the analyte were achieved. The method was validated for Accuracy, Repeatability, Reproducibility and Robustness, Linearity, LOQ, LOD were established for Flucytosine and its impurities in a single RPHPLC method. Therefore, this method can be used as a more convenient and efficient option for the analysis of Flucytosine assay and it’s related substances in injectable pharmaceutical dosage form to establish the quality of the drug product during routine analysis with consistent and reproducible results.