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Abstract

DEVELOPMENT AND VALIDATION OF LIQUID CHROMATOGRAPHY, MASS SPECTROMETER METHODS FOR DETERMINATION OF REMDESIVIR AND ITS METABOLITE GS- 441524 IN PLASMA AND THEIR APPLICATION IN COVID-19 RELATED CLINICAL STUDIES

J. N. Suresh Kumar*, M. Keerthana, N. Varsha, P. Asha, T. Shanthi Kumari, P. Vijay Babu and B. Satya Prasad

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Abstract

A simple method based on liquid chromatography coupled to triple quandrupole mass spectroscopy detection using 50μL of plasma was developed and fully validated for quantification GS-41524. of remdesivir and its active metabolites GS-Remdesivir is a Phosphoramidate prodrug designed to have activity against a broad spectrum of viruses. RDV is rapidly distributed in to cells and tissues and simultaneously metabolised in to GS-441524 in plasma. LC-MS methods were validated for determination of 2analytes in human plasma that involved two key aspects to guarantee their precision, accuracy and robustness. Chromatographic separation was achieved on an Acquity UPLC HSS T3 column [2.1×50mm, 1.8μm] with a run time of 3.4min. The calibration ranges were 4-4000, 2-2000, respectively for RDV, GS- 441524. Precision evaluated at 2.5,400 and 4000μg for Remdesivir and 12.5, 2000μgml for GS-441524 were lower than 14.7% and accuracy was in the 89.6-110.2% range. The validated method was successfully applied on covid-19 related clinical studies. A simple precipitation was carried out using 75μL of methanol containing the internal standard remdesivir.

Keywords: Covid-19, GS-441524, Remdesivir, LC-MS, Plasma.


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