Zaman Bakht, Hassan Waseem, Khan Adnan, Mushtaq Ayesha, Ali Nisar, Bilal Muhammad, Ahmed Dina A
Institute of Chemical Sciences, University of Peshawar, Peshawar 25120, Pakistan.
Department of Pharmacy, Quaid-i-Azam University, Islamabad 45320, Pakistan.
Antibiotics (Basel). 2022 Jul 5;11(7):897. doi: 10.3390/antibiotics11070897.
Analytical methods for the drug substance and degradation products (DPs) are validated by performing forced degradation studies. Forced degradation studies of Velpatasvir (VEL) drug substance and Velpatasvir copovidone solid dispersion (VEL-CSD) were performed under the stressed alkaline, acidic, oxidative and thermal conditions according to ICH guidelines ICH Q1A (R2). VEL is labile to degrade in stressed alkaline, acidic, and oxidative conditions. It is also photolabile and degraded during photostability studies as described by ICH Q1B, and showed no degradation on exposure to extreme temperature when protected from light. A sensitive stability indicating HPLC-UV method was developed and validated for the separation of VEL and eight DPs. The DPs of VEL are separated using gradient elution of mobile phase containing 0.05% Trifluoroacetic acid (TFA) and methanol over symmetry analytical column C18 (250 mm × 4.6 mm, 5 µm) with a flow rate of 0.8 mL min. Simultaneous detection of all DPs and VEL was performed on UV detector at 305 nm. The performance parameters like precision, specificity and linearity of the method were validated using reference standards as prescribed by ICHQ2 (R1). Limits of quantification and limits of detection were determined from calibration curve using the expression 10δ/slope and 3δ/slope respectively. The proposed method is stability-indicating and effectively applied to the analysis of process impurities and DPs in VEL drug substance and VEL-CSD.
通过进行强制降解研究来验证原料药和降解产物（DPs）的分析方法。根据国际协调会议（ICH）指南ICH Q1A（R2），在碱性、酸性、氧化和热应激条件下对维帕他韦（VEL）原料药和维帕他韦共聚维酮固体分散体（VEL-CSD）进行了强制降解研究。VEL在碱性、酸性和氧化应激条件下易降解。如ICH Q1B所述，它在光稳定性研究中也对光敏感且会降解，并且在避光条件下暴露于极端温度时未显示降解。开发并验证了一种灵敏的稳定性指示高效液相色谱-紫外（HPLC-UV）方法，用于分离VEL和8种降解产物。使用含有0.05%三氟乙酸（TFA）和甲醇的流动相在Symmetry分析柱C18（250 mm×4.6 mm，5 µm）上进行梯度洗脱，流速为0.8 mL/min，以分离VEL的降解产物。在305 nm波长下用紫外检测器同时检测所有降解产物和VEL。按照ICH Q2（R1）的规定，使用参考标准对该方法的精密度、特异性和线性等性能参数进行了验证。定量限和检测限分别根据校准曲线用表达式10δ/斜率和3δ/斜率确定。所提出的方法是稳定性指示的，并有效地应用于VEL原料药和VEL-CSD中工艺杂质和降解产物的分析。
