Rapid and ecofriendly ultra performance liquid chromatographic analytical methodology for the simultaneous determination of four drugs included in COVID-19 treatment protocol

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The global spread of COVID-19 disease has increased the demand for new therapeutic approaches. The clinical studies revealed that remdesivir and molnupiravir have been effective against COVID-19. Remdesivir (REM) is the first FDA-approved antiviral drug for COVID-19, while molnupiravir is the first oral antiviral drug for SARS-CoV-2 infections. In addition to antivirals, anticoagulants such as apixaban (APX) and antibiotics such as levofloxacin (LVF) are also included in the treatment protocols. This study suggests and validates a rapid and selective method for the determination of the four drugs involved in the COVID-19 treatment protocol. The simultaneous separation and determination of REM, MLP, LVF, and APX in human plasma were achieved using UPLC with a fluorophenyl column and a mobile phase composed of acetonitrile and water acidified with formic acid (pH = 3) in the ratio of 45: 55, v/v, at a flow rate of 0.1 mL/min. The PDA detector was adjusted at 254 nm. The method could simultaneously determine the four drugs in a single run in less than 6 min. The method exhibited linearity over the concentration ranges of 1.0–15.0 µg/mL, 0.1–5.0 µg/mL, 0.1–2.0 µg /mL, and 0.1–5.0 µg/mL, for LVF, MLP, APX and REM, respectively. The proposed method was validated following the International Council on Harmonization Q2(R1) guidelines and the findings were compared statistically to previously reported methods. The recovery rates of the drugs from spiked human plasma were satisfactory. The greenness of the suggested method was evaluated using three metrics, namely, Analytical Eco-scale, Green Analytical Procedure Index (GAPI) metric, and the Analytical GREEnness measure approach (AGREE).