Department of Analytical Chemistry, Faculty of Pharmacy, Minia University, Minia, Egypt.
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut, Egypt.
Luminescence. 2020 Nov;35(7):1048-1055. doi: 10.1002/bio.3816. Epub 2020 Apr 29.
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 F 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 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.
建立了一种经验证的薄层层析（TLC）方法，结合荧光检测模式，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 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 激发下，以荧光模式扫描平板。该方法提供了 velpatasvir 与 sofosbuvir 的极好分离，在 335nm 下以紫外检测模式扫描开发平板后，velpatasvir 和 sofosbuvir 的 R 值分别为 0.22 和 0.46。校准曲线在 4-40ng/带范围内呈线性，相关系数为 0.9994。根据 ICH 指南对所开发的程序进行了验证，检测限为 1.30ng/带，定量限为 3.95ng/带。该方法可以选择性地测定含有共配方抗 HCV 药物（sofosbuvir）的合成片剂粉末中的 velpatasvir，具有高灵敏度，无赋形剂或 sofosbuvir 的任何干扰。此外，该方法成功应用于加标人血浆中 velpatasvir 的测定，回收率足够。
