A NEW RELATED SUBSTANCES METHOD DEVELOPMENT AND VALIDATION OF TWO ANTI-CANCER DRUGS BY USING EFFECTIVE LIQUID CHROMATOGRAPHIC METHOD

Objective: The present study was aimed at developing and successively validating novel, simple, responsive and stable RP-HPLC method for the measurement of active pharmaceutical ingredients of mitomycin and fluorouracil and their related substances. Methods: Using the impurity-spiked solution, the chromatographic approach was optimized. The chromatographic method used Luna C18 column of dimensions 150x4.6 mn, 3.5 μm, using gradient elution with a mobile phase of acetonitrile and 0.1 percent orthophosphoric acid. A flow rate of 1 ml/min and a detector wavelength of 260 nm using the PDA detector was provided in the instrumental settings. Recovery, specificity, linearity, accuracy, robustness, ruggedness was determined as a part of method validation and the results were found to be within the acceptable range. Results: According to the ICH guidelines, the developed approach was validated. The calibration charts plotted were linear with a regression coefficient of R2>0.999. Conclusion: The method developed was found to be applicable to routine analysis and to be used for the measurement of active pharmaceutical ingredients (i. e, Mitomycin and Fluorouracil and its related impurities). Since there is no HPLC method reported in the literature for the estimation of Mitomycin, Fluorouracil and its related impurities; there is a need to develop quantitative methods under different conditions to achieve improvement in specificity, selectivity etc