Development and validation of a novel bioanalytical method for the simultaneous determination of glecaprevir and pibrentasvir in human plasma using reversed-phase high-performance liquid chromatography

Abstract

Background and objective For the simultaneous determination of glecaprevir (GPR) and pibrentasvir (PTR) in human plasma, a novel, accurate, and selective reversed-phase high-performance liquid chromatography method was developed and validated. Materials and methods Owing to structural resemblance, bictegravir was selected as an internal standard. Anticoagulant used was K2-EDTA. The GPR-PTR was the first of its kind approved drug by FDA for the treatment of chronic hepatitis C. Precipitation technique with acetonitrile was employed for the extraction of analyte from human plasma. Kromasil C18 column (5 μ, 150×4.6 mm) with an isocratic mobile phase of 0.1% orthophosphoric acid buffer pH 4.3, adjusted with dilute hydrochloric acid: acetonitrile in the ratio of 70 : 30 v/v, was used for the resolution. At a flow rate of 1 ml/min, the mobile phase was pumped. Using a photodiode array detector, effluents were monitored at 250 nm. Results Over concentration ranges of 5–200 μg/ml and 6.650–266.000 μg/ml, the method was found to be linear for GPR and PTR, respectively, in human plasma, with the precision and accuracy ranging from 0.76 to 9.05% and 90.55 to 98.98% for GPR respectively, whereas for PTR ranged from 0.74 to 9.52% and 91.56 to 105.61%, respectively. Conclusion The stability of the analyte was evaluated in plasma under different stress conditions.