Rapid Determination of 5 Antiviral Compounds in Indian Generic for Anti-COVID-19 by HPLC

OBJECTIVE  To develop an HPLC method for rapid determination of 5 antiviral compounds(molnupiravir, oseltamivir, daclatasvir, nirmatrelvir, and ritonavir) in Indian generic for anti-COVID-19.

METHODS  The samples were ultrasonically extracted with methanol, and separated on an Agilent Poroshell 120 EC-C₁₈ column(50 mm×4.6 mm, 2.7 μm) at 35 ℃ by gradient elution with formic acid aqueous solutionpH(3.0±0.1)-acetonitrile as mobile phase at a flow rate of 1.5 mL·min⁻¹ , the injection volume was 10 μL. The detection wavelength for diode array detector was set at 210 nm. Quantification analysis was performed by the external method.

RESULTS The calibration curves for the 5 compounds showed good linearity in a certain range of mass concentration with their correlation coefficients ≥ 0.999 7. The limits of detection and limits of quantitation for 5 compounds were 24−25 μg·g⁻¹ and 80−84 μg·g⁻¹. The average recoveries of 5 compounds at three spiked levels(low, middle and high) ranged from 96.6%−104.3%, with RSD of 0.9%−1.9%.

CONCLUSION  The method is rapid and accurate, and can be used for the rapid screening of 5 antiviral components in Indian generic for anti-COVID-19, indicating its extensive application prospect.