RP-hplc method validation for quantitative analysis of pemetrexed disodium hemipentahydrate

Abstract

A simple, sensitive, and reliable method has been developed for quantitative analysis of an Anti-neoplastic drug Pemetrexed Disodium Hemipentahydrate (PDH) using reversed phase high performance liquid chromatography (RP-HPLC). This method is developed and validated as per the International Council on Harmonisation (ICH) guidelinesQ2(R1). The effective chromatographic separations were achieved on Zorbax SB C8, (4.6 x 150mm) 3.5μ column with isocratic elution programme. The mobile phase is as homogeneous mixture of 0.17% glacial acetate acid with 5.3 pH and acetonitrile in the ratio of 89:11. The delivered flow rate of mobile phase is 2.0 mL/min using UV/PDA detector. The column oven temperature is 30°C and the injection volume is 20.0μL. This method is linear with correlation coefficient being 0.99992 and %Y intercept of 0.16. The repeatability and intermediate precision are evaluated and %RSD for these assays is 0.09 and 0.28, respectively, with overall % RSD of 0.22. Robustness studies do not shows significant change for the system suitability criteria like tailing factor, theoretical plates and %RSD. The values of these criteria are well within acceptance limit. The degradation of Pemetrexed under various stress condition is studied and all known as well as degradants impurities are well separated from the PDH peak. This RP-HPLC method is precise, accurate, and rapid; it also qualifies all the criteria of linearity, stability as well as robustness.