高效液相色谱法快速测定印仿新冠药中5种抗病毒成分

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

关键词:  快速筛查  仿制药  抗病毒  新冠用药  奈玛特韦  莫诺拉韦

Key words:  rapid screening  generic  antiviral  anti-COVID-19  nirmatrelvir  molnupiravir

摘要:

目的? 建立了同时快速测定印仿药中5种抗病毒成分(莫诺拉韦、奥司他韦、达卡他韦、奈玛特韦、利托那韦)的高效液相色谱分析方法,以便稽查人员及时发现可疑样品,守护消费者用药安全。方法? 样品经含甲醇超声提取,用Agilent Poroshell 120 EC-C18(50 mm×4.6 mm,2.7 μm)分离,以甲酸水溶液(pH 3.0)-乙腈为流动相进行梯度洗脱,流速1.5 mL·min-1,柱温35 ℃,二极管阵列检测器检测,检测波长为210 nm,外标法定量。结果? 5种抗病毒成分在一定质量浓度范围内线性良好,相关系数不低于0.9997。5种成分的检出限(LOD)为24~25 μg·g-1,定量下限(LOQ)为80~84 μg·g-1。5种局麻成分在低、中、高3个加标水平的平均回收率为96.1%~104.3%,相对标准偏差(RSD)为0.4%~1.5%。结论 该方法快速、准确,适用于印仿新冠药中5种成分抗病毒的快速筛查,具有广阔的应用前景。

Abstract:

ABSTRACT: OBJECTIVE A HPLC was developed for the rapid determination 5 antiviral compounds(molnupiravir，oseltamivir，daclatasvir，nirmatrelvir，and ritonavir)in indian generic for anti-COVID-19，so that inspectors can find the suspicious samples as soon as possible to protect the safety of consumers. METHODS The samples were ultrasonically extracted with methanol，and separated on an Agilent Poroshell 120 EC-C18 (50 mm×4.6 mm, 2.7 μm) at 35 ℃ by gradient elution with formic acid aqueous solution (pH 3.0)-acetonitrile as mobile phase at a flow rate of 1.5mL/min. The detection wavelength for diode array detector was set at 210 nm. Quantification analysis was performed by the extemal method. RESULTS The calibration curves for the 5 compounds showed good linearity in a certain range of mass concentration with their correlation coefficients not less than 0.9997. The limits of detection(LOD) and limits of quantitation(LOQ) for 5 compounds were 24~25 μg·g-1 and 80~84 μg·g-1. The average recoveries at three spiked levels ranged from 96.1%~104.3%，with relative standard deviations (RSD) of 0.4%~1.5%. CONCLUSION The method was rapid and accurate，and could be used for the rapid screening of antiviral components in indian generic for anti-COVID-19，indicating its extensive application prospect.