Development Of Validated Bioanalytical Method For Estimation Of Remdesivir In Human Plasma By Rp-Hplc

Abstract

Remdesivir, formerly GS-5734, has recently become the first antiviral drug approved by the U.S. Food and Drug Administration (FDA) to treat COVID-19, the disease caused by SARS-CoV-2. Therapeutic dosing and pharmacokinetic studies require a simple, sensitive, and selective, precise validated method to quantify drug concentrations in clinical samples. Therefore, we developed a rapid and sensitive RP-HPLC method for the estimation of remdesivir in human plasma. Sample processing involved a simple protein precipitation extraction technique by acetonitrile as the extraction solvent. Methods: Separation is carried out on Sunfire-C18 plasma 5 µm (4.6 × 250 mm) column as a stationary phase, Mobile Phase: methanol + 0.1% OPA (orthophosphoric acid) in ratio (80: 20), at a flow rate of 0.7 ml/min.  The sample was detected at a wavelength of 247 nm and the sample size was 20μl. The objective of method validation, according to ICH principles, is to show that an analytical technique is appropriate for the intended use. The standard calibration plot was found linear over a range of 5 to 25 µg/ml and the correlation coefficient was found to be (R2 =0.997). The % RSD value of intraday and interday precision is 0.14 and 0.63 respectively. The LOD and LOQ were found to be 0.0.0654991 and 0.1984822 respectively for Remdesivir. The retention time was observed at 5.2 min for remdesivir. The developed method was eventually applied for quantification of the marketed formulation satisfactory result was obtained. Conclusions: It can be concluded that the developed bioanalytical method is capable of quantifying Remdesivir from spiked human plasma samples. The method meets the requirements of the USFDA Guidelines and can be applied to Bioavailability/ Bioequivalence studies of Remdesivir.