Abstract

NEW METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF LEDIPASVIR IN BULK DRUG FORM BY USING REVERSE PHASE HPLC TECHNIQUE

J. Devilal*, B. Durgaprasad, Narottam Pal and A. Srinivasa Rao

ABSTRACT

The present work is a method development and validation for the determination of Ledipasvir by using reverse phase High performance liquid chromatography (HPLC). The procedure involved use of isocratic elution where the stationary phase was a BDS column (250 mm, 4.6 mm, 5 μm), mobile phase 0.05% triflouroacetic acid in methanol and 0.05% triflouroacetic acid in acetonitrile (55:45). pH of the chromatographic system was maintained at 3.0, flow rate 1 ml/minute, eluent was monitored by PDA detector wavelength at 270 nm. Retention time was found to be 2.749 minutes, regression analysis shown the value of correlation coefficient 0.999. Value for limit of detection (LOD) was 1.064μgm/ml and limit of quantification (LOQ) was 3.224 μgm/ml. Linearity range was designed 15μg/ml to 300μg/ml for Ledipasvir. Accuracy study revealed percentage recovery 99.81%-100.10% and precision result in terms of standard deviation and percentage relative standard deviation was 1986.515 and 0.08511%. Test for intermediate results also were well within the limit for Ledipasvir. Robustness study results proved the suitability of the method under different chromatographic condition. The developed method was validated as per ICH guideline and was found to be an ideal one for regular analysis in the laboratory.

Keywords: HPLC, isocratic elution, simultaneous, Ledipasvir.