高效液相色谱-串联质谱法同时测定药品中奈玛特韦等5种小分子抗病毒药物

Simultaneous Determination of Five Small Molecule Antivirals Including Nematovir in Pharmaceuticals by High Performance Liquid Chromatography- Tandem Mass Spectrometry

建立了一种同时检测奈玛特韦、利托那韦、莫诺拉韦、奥司他韦和达卡他韦5种小分子抗病毒药物的高效液相色谱-质谱分析方法。样品经甲醇溶解超声提取后,经CNWShell C18 Core-Shell色谱柱(100×2.1 mm,2.6μm)分离,以0.1%甲酸水溶液和甲醇为流动相,梯度洗脱,电喷雾正离子模式下,同时采用多反应监测(MRM)和二级全扫(MS^(2))模式扫描分析。5种抗病毒药物在一定浓度范围内线性良好;检出限(S/N=3)为0.15~0.5μg/g;定量限(S/N=10)为0.5~1.2μg/g;加标平均回收率为90.0%~97.4%,精密度(n=6)均小于5%。采用本方法对10份样品进行测定,发现7份样品不含标示的有效成分奈玛特韦,有效成分利托那韦含量远低于标示含量,且检出奥司他韦或达卡他韦等非法添加化学成分。所建立方法灵敏、快速、可靠,可用于药品制剂中5种抗病毒药物的定性及定量分析。

A HPLC-MS/MS method was established for the simultaneous detection of five small molecule antivirals,namely nirmatrelvir,ritonavir,molnupiravir,oseltamivir and daclatasvir.Samples were extracted by methanol with ultrasound extraction,and separated on a CNWShell C18 Core-Shell column(100×2.1 mm,2.6 μm) by gradient elution using 0.1% formic acid solution and methanol as the mobile phase.The electrospray ion source was operated in the positive ion mode and both multiple reaction monitoring and product ion scanning were used in this method.There were good linearities for five small molecule antivirals in their linear ranges.The limits of detection(LODs) were 0.15-0.5 μg/g and the limits of quantitation were 0.5-1.2 μg/g.Average recoveries of three spiked levels ranged from 90% to 97.4%(n=6) with the relative standard deviations(RSDs) less than 5%.The method was used to test 10 samples and 7 samples were found not to contain the active ingredient nirmatrelvir as labelled.Active ritonavir was found to be much lower than labelled,and illegally added chemicals such as oseltamivir or daclatasvir were detected.The method was proved to be sensitive,rapid and accurate for the qualitative and quantitative analysis of 5 small-molecule antivirals.