A new ultra-high-performance liquid chromatography method for the simultaneous quantification of sofosbuvir, daclatasvir and ledipasvir was developed. Two combinations of these direct-acting antivirals are used in hepatitis C virus infection therapy and show high efficacy and safety. Fractional factorial design was used for screening the most influential factors on separation and time analysis. These significant factors were optimized using a central composite design. The optimum resolution was carried out by using a Waters XBridge C18 column (150 mm, 4.6 mm ID, 5 μm) at a temperature of 35°C ± 2°C and acetonitrile/sodium perchlorate buffer (10 mM, pH = 3.2) (40: 60 v/v) as mobile phase at a flow rate of 1.5 mL min-1. UV detection was set at λ = 210 nm. A short chromatographic separation time was achieved. The developed method was validated according to the accuracy profile approach and was found specific, precise, faithful and accurate. The detection limits were between 0.07 and 0.13 μg mL-1. Hence, this novel method can be employed for the routine quality control analysis and in dissolution profile studies of generics containing these products.

中文翻译：

---

化学高效辅助的超高效液相色谱方法的开发，用于以药物剂型同时定量Sofosbuvir，Daclatasvir和Ledipasvir。

开发了一种新的超高效液相色谱方法，用于同时定量索非布韦，达卡他韦和ledipasvir。这些直接作用抗病毒药的两种组合用于丙型肝炎病毒感染治疗，并显示出高疗效和安全性。分数阶乘设计用于筛选分离和时间分析中最有影响力的因素。这些重要因素使用中央复合设计进行了优化。最佳分离度是通过使用Waters XBridge C18色谱柱（150 mm，4.6 mm ID，5μm）在35°C±2°C的温度和乙腈/高氯酸钠缓冲液（10 mM，pH = 3.2）下进行的（ 40:60 v / v）作为流动相，流速为1.5 mL min-1。紫外线检测设定为λ= 210 nm。色谱分离时间短。所开发的方法已根据准确度分布图方法进行了验证，并被发现具有特定性，精确性，忠实性和准确性。检出限在0.07和0.13μgmL-1之间。因此，这种新方法可用于常规质量控制分析和含有这些产品的仿制药的溶出度研究。