Application of green solvents for developing green analytical methodologies has grown dramatically in the past few years. The “more hazardous reagents” are replaced by more “environment-friendly solvents” without affecting method performance. In the present study, two simple and accurate chromatographic methods were developed and validated for determination of the new antiviral combination sofosbuvir (SBR) and ledipasvir (LPV). The first adopted method is high-performance thin-layer chromatography coupled to densitometric determination where silica gel 60 F254 plates were used as the stationary phase. Whereas, the running mobile phase used was toluene: ethanol: ammonia (4:1:0.2, v/v/v). Reversed-phase high-performance liquid chromatography with ultraviolet detection was the second method developed. The column used was Inertsil C18 column (150 × 4.6 mm, 5 μm) and the mobile phase was 20 mM potassium dihydrogen orthophosphate (adjusted to pH = 3 using acetic acid): ethanol (60:40, v/v) with a flow rate of 1.0 mL/min. The detection wavelength for both methods was 265 nm. The validation of both methods was done according to ICH guidelines where both methods were found to be accurate, reproducible, and selective. The linearity range for HPTLC and RP-HPLC methods were 0.8–25.6 and 0.4–12.8 μg/band and 6.0–100.0 and 4.0–80.0 μg/mL for sofosbuvir and ledipasvir, respectively. Comparison of the developed methods was done with reported HPLC method where no significant difference was found.

中文翻译：

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Sofosbuvir/Ledipasvir 联合剂型的同时色谱分析：在绿色分析化学中的应用

在过去几年中，绿色溶剂在开发绿色分析方法方面的应用急剧增长。在不影响方法性能的情况下，将“更危险的试剂”替换为更“环保的溶剂”。在本研究中，开发并验证了两种简单而准确的色谱方法，用于测定新的抗病毒组合索非布韦 (SBR) 和雷迪帕韦 (LPV)。第一种采用的方法是高效薄层色谱与光密度测定相结合，其中使用硅胶 60 F254 板作为固定相。而使用的流动相是甲苯：乙醇：氨（4:1:0.2，v/v/v）。带有紫外检测的反相高效液相色谱是第二种开发的方法。所用色谱柱为 Inertsil C18 色谱柱（150 × 4. 6 mm, 5 μm)，流动相为 20 mM 正磷酸二氢钾（使用乙酸调节至 pH = 3）：乙醇（60:40，v/v），流速为 1.0 mL/min。两种方法的检测波长均为 265 nm。根据 ICH 指南对这两种方法进行了验证，发现这两种方法都是准确、可重复和具有选择性的。HPTLC 和 RP-HPLC 方法的线性范围分别为 0.8–25.6 和 0.4–12.8 μg/条带，sofosbuvir 和 ledipasvir 的线性范围分别为 6.0–100.0 和 4.0–80.0 μg/mL。已开发方法与报告的 HPLC 方法进行了比较，未发现显着差异。两种方法的检测波长均为 265 nm。根据 ICH 指南对这两种方法进行了验证，发现这两种方法都是准确、可重复和具有选择性的。HPTLC 和 RP-HPLC 方法的线性范围分别为 0.8-25.6 和 0.4-12.8 μg/条带，sofosbuvir 和 ledipasvir 的线性范围分别为 6.0-100.0 和 4.0-80.0 μg/mL。已开发方法与报告的 HPLC 方法进行了比较，未发现显着差异。两种方法的检测波长均为 265 nm。根据 ICH 指南对这两种方法进行了验证，发现这两种方法都是准确、可重复和具有选择性的。HPTLC 和 RP-HPLC 方法的线性范围分别为 0.8–25.6 和 0.4–12.8 μg/条带，sofosbuvir 和 ledipasvir 的线性范围分别为 6.0–100.0 和 4.0–80.0 μg/mL。已开发方法与报告的 HPLC 方法进行了比较，未发现显着差异。