The present work is aimed at development and validation of RP HPLC method which is simple, specific, precise, and accurate for estimation of Sofosbuvir and its process-related impurity in bulk and pharmaceutical dosage forms. Extensive literature survey revealed no method for estimation of the above said. The chromatographic separation was achieved on Agilent Eclipse XDB-C18, 4.6 × 250 mm, 5 μm with mobile phase composed of 0.1% trifluoroacetic acid in 1000 ml of water:acetonitrile (50:50) using an isocratic mode of elution. Detection was made using UV detector at 260.0 nm and LC solution software for analysis of data. The developed method was validated according to ICH guidelines. The linearity of calibration curve for Sofosbuvir in concentration range of 160-480 μg/ml was good. The curve was linear for its process related impurity (Phosphoryl) in concentration range of 10-30 μg/ml. There exists a good correlation between peak area and analyte concentration. Retention time for Sofosbuvir was found to be 3.674 min and its impurity was 5.704 min. Relative standard deviation values for Sofosbuvir is 1.741 and its process related impurity is 0.043. LOD for Sofosbuvir and its impurity was found to be 0.01% (0.04 μg) and 0.03% (0.12 μg) respectively. LOQ for Sofosbuvir and its impurity was found to be 0.50% (0.125 μg) and 1.50% (0.375 μg) respectively. All the results reveal that the proposed method was found to be highly sensitive, simple, precise, accurate, robust, and fast. Large number of samples can be analyzed in shorter time due to shorter retention times, so it can be successfully applied for routine analysis of Sofosbuvir and related phosphoryl impurity in bulk and pharmaceutical dosage forms.

中文翻译：

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开发和验证用于估计散装和药物剂型中 Sofosbuvir 和相关杂质的 RP HPLC 方法

目前的工作旨在开发和验证 RP HPLC 方法，该方法简单、特异性、精确和准确，用于估计 Sofosbuvir 及其在散装和药物剂型中与工艺相关的杂质。广泛的文献调查发现没有方法可以估计上述情况。色谱分离是在 Agilent Eclipse XDB-C18，4.6 × 250 mm，5 μm 上实现的，流动相由 0.1% 三氟乙酸在 1000 ml 水:乙腈 (50:50) 中组成，使用等度洗脱模式。使用 260.0 nm 的 UV 检测器和用于数据分析的 LC 溶液软件进行检测。根据 ICH 指南对开发的方法进行了验证。Sofosbuvir 在 160-480 μg/ml 浓度范围内的校准曲线线性良好。浓度范围为 10-30 μg/ml 的过程相关杂质（磷酰基）的曲线呈线性。峰面积与分析物浓度之间存在良好的相关性。发现 Sofosbuvir 的保留时间为 3.674 分钟，其杂质为 5.704 分钟。Sofosbuvir 的相对标准偏差值为 1.741，其工艺相关杂质为 0.043。发现 Sofosbuvir 及其杂质的 LOD 分别为 0.01% (0.04 μg) 和 0.03% (0.12 μg)。发现 Sofosbuvir 及其杂质的 LOQ 分别为 0.50% (0.125 μg) 和 1.50% (0.375 μg)。所有结果表明，所提出的方法具有高度灵敏、简单、精确、准确、稳健和快速的特点。由于更短的保留时间，可以在更短的时间内分析大量样品，