A validated thin‐layer chromatography (TLC) method combined with fluorescence detection mode was developed for the selective determination of a recently approved anti‐hepatitis C virus (HCV) drug (velpatasvir). The separation was performed on silica gel 60 F₂₅₄ plates using ethylacetate:methanol:triethylamine (48:1.5:1.0, v/v/v) as a mobile phase. Plates were scanned in the fluorescence mode after excitation at 335 nm. This method provided an excellent separation of velpatasvir from sofosbuvir with R_F values of 0.22 and 0.46 for velpatasvir and sofosbuvir, respectively, after scanning the developed plates in the ultraviolet detection mode at 335 nm. The calibration curve was linear over the range 4–40 ng/band with a correlation coefficient of 0.9994. The developed procedure was validated according to ICH guidelines with a detection limit of 1.30 ng/band and quantitation limit of 3.95 ng/band. The suggested method could selectively determine velpatasvir with high sensitivity in a synthetic tablet powder containing a co‐formulated anti‐HCV drug (sofosbuvir) without any interference from excipients or sofosbuvir. In addition, the method was successfully applied for determination of velpatasvir in spiked human plasma with adequate % recovery.

中文翻译：

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薄层色谱/荧光检测方法用于灵敏和选择性地确定丙型肝炎病毒抗病毒药物（velpatasvir）：应用于人体血浆。

开发了一种经过验证的薄层色谱（TLC）方法与荧光检测模式相结合，用于选择性测定最近批准的抗丙型肝炎病毒（HCV）药物（velpatasvir）。分离在硅胶60 F ₂₅₄板上进行，使用乙酸乙酯：甲醇：三乙胺（48：1.5：1.0，v / v / v）作为流动相。在335nm激发后，以荧光模式扫描板。该方法可通过R _F很好地将维帕他韦与索非布韦分开在335 nm紫外检测模式下扫描显影后的板后，velpatasvir和sofosbuvir的C值分别为0.22和0.46。校准曲线在4-40 ng / band范围内呈线性，相关系数为0.9994。根据ICH指南验证了开发的程序，检测限为1.30 ng / band，定量限为3.95 ng / band。所建议的方法可以选择性地测定含有共同配制的抗HCV药物（sofosbuvir）的合成片剂粉末中的维帕他韦，而不会受到赋形剂或sofosbuvir的干扰。另外，成功地适用于在加标人血浆具有足够判定velpatasvir的方法％回收率。