UHPLC-MS/MS: A SELECTIVE BIOANALYTICAL TECHNIQUE TO DETECT AND QUANTIFY REPURPOSED COVID-19 PROTOCOL DRUGS IN HUMAN PLASMA

Document Type : Original Article

Authors

1 Pharmaceutical Chemistry department, Faculty of Pharmacy, Cairo University, Cairo, Egypt

2 Pharmaceutical Chemistry department, Faculty of Pharmacy, Cairo University, Cairo, Egypt Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al Baha University, Saudi Arabia

3 Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University Cairo, Egypt

Abstract

The SARS-CoV-2 caused COVID-19 disease first appeared during late 2019 and announced in March, 2020 as a global pandemic by the World Health Organization. COVID-19 was deemed to be no longer a public health emergency of worldwide concern in May 2023. Still, the virus is a potentially dangerous health risk and is still spreading throughout communities. In the current study a sensitive and rapid developed method was validated for the quantitative estimation of the drugs namely (apixaban, moxifloxacin-HCl, remdesivir and paracetamol) used in the management of COVID-19 in spiked human plasma simultaneously. The extraction solvent was diethyl ether: dichloromethane (70:30) used to extract the cited drugs and azithromycin (IS) from human plasma (liquid-liquid extraction). ultra-high performance liquid chromatography with tandem mass detection was employed for separation and quantification. In an Agilent Poroshell 120 EC-C18 (4.6 x 50 mm, 2.7 µm) column, a mobile phase consisting of methanol: water with 0.1% formic acid (93:7 v/v) was pumped in isocratic elution at a flow rate of 0.3 mL/min. with a total run time of 3 minutes. Multiple reaction monitoring mode (MRM) transitions were measured at the positive ionization mode for the four drugs and IS. The new approach was bio-analytically validated in accordance with the European Medicines Agency (EMA) requirements. The validated method is precise, selective, accurate and can be used in many applications including bioequivalence studies, therapeutic drug monitoring and routine drug analysis.

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