目的: 建立HPLC-MS法检测奈玛特韦原料药3个非对映异构体。方法: 采用InfinityLab Poroshell SB-C18（150 mm×3.0 mm,2.7 μm）色谱柱,2根串联使用,以缓冲液（0.1%甲酸溶液）（A）-乙腈（B）为流动相,以水-乙腈-甲醇（61∶19.5∶19.5）为稀释液,进行梯度洗脱,柱温80 ℃,进样量2 μL,样品温度5 ℃,以电喷雾离子（ESI+）作为离子源,SIM模式扫描。结果: 奈玛特韦与3个非对映异构体能够完全分离（分离度>1.5）,供试品溶液在3 d内稳定性良好;非对映异构体1的定量限为0.04%,非对映异构体2的定量限为0.04%,非对映异构体3的定量限为0.05%;非对映异构体3线性相关系数>0.99,范围为杂质定量限浓度~150%指标浓度;非对映异构体3平均回收率（n=9）为97.2%,RSD为3.1%;重复性和中间精密度符合规定。经检测,3批奈玛特韦原料药24个月长期稳定性3个非对映异构体结果均符合质量标准。结论: 该方法简便快速,灵敏度高,专属性强,可用于奈玛特韦原料药3个非对映异构体的测定。
Objective: To establish an HPLC-MS method for determination of three diastereoisomers in nirmatrelvir API. Methods: The analytical column was an InfinityLab Poroshell SB-C18 (150 mm×3.0 mm,2.7 μm), the two columns connected in series in series. The mobile phase was buffer (0.1% formic acid) (A) -acetonitrile (B), the diluent was water-acetonitrile-methanol (61 ∶ 19.5 ∶ 19.5). The whole run carried out by gradient elution. The column temperature was 80 ℃,the injection volume was 2 μL, and the sample temperature was 5 ℃. Results: Nirmatrelvir was separated completely from three diastereoisomers (the resolution>1.5). The test solution was stable for at least 3 d. The LOQ of diastereoisomer 1 was 0.04%, the LOQ of diastereoisomer 2 was 0.04%, the LOQ of diastereoisomer 3 was 0.05%. The linear correlation coefficient of diastereoisomer 3 was >0.99. The linear range was LOQ-150% of specification. The average recovery (n=9) of diastereoisomer 3 was 97.2%, RSD = 3.1%. The repeatability and intermediate precision completely met the requirements. The three diastereoisomers contents in three batches of nirmatrelvir API 24 months long-term stability test were all completely met the requirements, respectively. Conclusion: This method is simple, rapid, sensitive and specific to be used for the determination of three diastereoisomers in nirmatrelvir API.
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