BIO-ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS DETERMINATION OF LEDIPASVIR AND SOFOSBUVIR DRUGS IN HUMAN PLASMA BY RP-HPLC METHOD
Objective: A novel, sensitive and accurate high-performance liquid chromatography with ultraviolet/visible light detection (HPLC-UV/VIS) method for the quantification of ledipasvir and Sofosbuvir in plasma was developed and validated.
Methods: The analytes were extracted by liquid-liquid extraction method and chromatograph using a mobile phase consisting of acetonitrile and buffer solution, Methanol and Acetonitrile in the ratio of 200:600:200 (v/v) using Oyster BDS RP-C18 column. The flow rate 1.0 ml/min and UV detection at 238 nm were employed. The retention time for Ledipasvir and Sofosbuvir was 4.61 and 9.09 min respectively. Linearity for ledipasvir and Sofosbuvir was found to be in the range of 250-2000 ng/ml for both drugs respectively. Intra-and inter-day precision was less than 2% coefficient of variation.
Results: The method was validated as per the USFDA guidelines and the results were within the acceptance criteria for selectivity, sensitivity, linearity, precision, accuracy, recovery stability of the solution, the stability of solution in plasma and dilution integrity.
Conclusion: Majority of the HPLC method should be useful for monitoring human plasma drug concentrations, and pharmacokinetic studies in patients diagnosed with the Ledipasvir and Sofosbuvir formulations.
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