Validated Analytical Method for the Estimation of Gemcitabine from its Pharmaceutical Formulation by RP-HPLC

A simple, sensitive and rapid high performance liquid chromatographic method has been developed for the determination of Gemcitabine. Chromatographic separation was achieved on an Inertsil ODS-3V column (250mm x 4.6mm; 5μm) with a mobile phase consisting of water: acetonitrile (90: 10; pH 3.5 adjusted with orthophosphoric acid) with a flow rate of 1.0ml/min (UV detection at 270nm). Linearity was observed over the concentration range of 1–120μg/ml with a regression equation y=19773x+ 86063 and having a regression value (R2)= 0.998. The limit of detection (LOD) and limit of quantification (LOQ) values found to be 10ng/ml and 20ng/ml, respectively. The percentage recovery of the marketed formulation was found to be 100.08%. Validation revealed that the preferred method was specific, accurate, precise, reliable, robust, reproducible and suitable for the quantitative analysis.