Abstract Drug substances undergo degradation during manufacturing process and storage of finished pharmaceutical products which often results in the generation of degradation products. When process-related substances and degradation products are present as impurities, the therapeutic effect of the drug may alter or cause adverse effects. To address these issues, a mass compatible selective reverse phase UPLC method was developed and validated for separation of Velpatasvir (VPR) and some of its impurities. VPR was subjected to hydrolytic, oxidative, photolytic and thermal stress conditions as prescribed by ICH. Four process-related substances and eight stress degradation products (DPs) were separated on an Acquity Phenyl-Hexyl (100 × 2.1 mm x 1.8 µm) column using 0.1% formic acid and methanol as eluent with gradient program. The same method was applied to LC coupled tandem mass spectrometer for separation and structure elucidation of impurities. Out of eight degradation impurities; two degradation products (V1 and V2) were obtained in acid hydrolysis, another two DPs (V3 and V4) formed in alkaline hydrolysis. Four DPs (V5, V6, V7 and V8) were obtained in oxidative stress. V7 and V8 were obtained as major degradation products and their chemical structures were further confirmed by NMR experiments. The performance of this method was validated for parameters by following ICH Q2 (R1) guideline. This method can be utilized in the identification of impurities in bulk drug manufacturing and dosage form design studies.

中文翻译：

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UPLC 在线液相色谱-四极杆-飞行时间-串联质谱法分离 Velpatasvir 的强制降解和工艺相关杂质并进行结构解析

摘要 原料药在药品成品的生产过程和储存过程中会发生降解，这往往会导致产生降解产物。当工艺相关物质和降解产物作为杂质存在时，药物的治疗效果可能会改变或引起不良反应。为了解决这些问题，我们开发并验证了质量兼容的选择性反相 UPLC 方法，用于分离 Velpatasvir (VPR) 及其某些杂质。VPR 经受 ICH 规定的水解、氧化、光解和热应力条件。四种工艺相关物质和八种应力降解产物 (DP) 在 Acquity Phenyl-Hexyl (100 × 2.1 mm x 1.8 µm) 色谱柱上使用 0.1% 甲酸和甲醇作为洗脱液，采用梯度程序进行分离。将相同的方法应用于 LC 耦合串联质谱仪，用于杂质的分离和结构解析。八种降解杂质中；在酸水解中获得两种降解产物（V1 和 V2），在碱水解中形成另外两种 DP（V3 和 V4）。在氧化应激中获得了四个 DP（V5、V6、V7 和 V8）。V7 和 V8 作为主要降解产物获得，它们的化学结构通过核磁共振实验进一步证实。按照 ICH Q2 (R1) 指南验证了该方法的参数性能。该方法可用于原料药生产和剂型设计研究中的杂质鉴定。在酸水解中获得两种降解产物（V1 和 V2），在碱水解中形成另外两种 DP（V3 和 V4）。在氧化应激中获得了四个 DP（V5、V6、V7 和 V8）。V7 和 V8 作为主要降解产物获得，它们的化学结构通过核磁共振实验进一步证实。按照 ICH Q2 (R1) 指南验证了该方法的参数性能。该方法可用于原料药生产和剂型设计研究中的杂质鉴定。在酸水解中获得两种降解产物（V1 和 V2），在碱水解中形成另外两种 DP（V3 和 V4）。在氧化应激中获得了四个 DP（V5、V6、V7 和 V8）。V7 和 V8 作为主要降解产物获得，它们的化学结构通过核磁共振实验进一步证实。按照 ICH Q2 (R1) 指南验证了该方法的参数性能。该方法可用于原料药生产和剂型设计研究中的杂质鉴定。按照 ICH Q2 (R1) 指南验证了该方法的参数性能。该方法可用于原料药生产和剂型设计研究中的杂质鉴定。按照 ICH Q2 (R1) 指南验证了该方法的参数性能。该方法可用于原料药生产和剂型设计研究中的杂质鉴定。