In the present study two different RSLC columns, Acclaim RSLC 120 C18, 5.0 µm, 4.6 × 150 mm (column A) and Acclaim RSLC 120 C18, 2.2 µm, 2.1 × 100 mm (Column B) were utilized for the analysis of velpatasvir (VPS) in presence of sofosbuvir (SFV), where due to the encountered fluorescent properties of VPS fluorescent detection at 405 nm after excitation at 340 nm (Method 1) was used for its detection where the non-fluorescent SFV did not interfere. The same columns were further utilized for the simultaneous determination of SFV and VPS either in bulk form or in their combined tablet, where UV- spectrophotometric detection at 260 nm was selected for the simultaneous analysis of both drugs (Method 2). A mobile phase consisting of NaH2PO4, pH 2.5 (with phosphoric acid) and acetonitrile in a ratio of 60:40 v/v was used for both methods. The mobile phase was pumped at a flow rate of 1.0 mL/min when using column, A and 0.5 mL/min when using column B. The methods showed good linearity over the concentration ranges of 1.0–5.0 and 2.5–10.0 ng/mL for VPS when utilizing Method 1 A and B respectively. Where the linearity concentration range was from 30.0–150.0 to 120–600.0 ng/mL for VPS and SFV respectively when applying Method 2. Both methods 1 and 2 were performed by utilizing the two analytical columns. The different chromatographic parameters as retention time, resolution, number of theoretical plates (N), capacity factor, tailing factor and selectivity were carefully optimized. The results show that comparing the performance of the two utilized columns revealed that shorter column (2.1 mm × 100 mm) with small particle packing was superior to the longer column (4.6 × 150 mm) for the analysis of the studied drugs allowing a reduction of the analysis time by 70% without any detrimental effect on performance. This prompts the decrease of the investigation costs by saving money on organic solvents and expanding the overall number of analyses per day.

中文翻译：

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两种新颖的 UPLC 方法利用两种不同的分析柱和不同的检测方法来同时分析维帕他韦和索磷布韦：应用于其联合配制的片剂

在本研究中，使用两种不同的 RSLC 色谱柱 Acclaim RSLC 120 C18，5.0 µm，4.6 × 150 mm（A 柱）和 Acclaim RSLC 120 C18，2.2 µm，2.1 × 100 mm（B 柱）来分析维帕他韦（ VPS) 存在索磷布韦 (SFV) 时，由于在 340 nm 激发后 VPS 荧光检测在 405 nm 处遇到荧光特性（方法 1），因此使用非荧光 SFV 不干扰的检测。相同的色谱柱进一步用于同时测定散装形式或组合片剂中的 SFV 和 VPS，其中选择 260 nm 的紫外分光光度检测来同时分析两种药物（方法 2）。两种方法均使用由 NaH2PO4、pH 2.5（含磷酸）和乙腈（v/v 比例为 60:40）组成的流动相。使用色谱柱 A 时，流动相的泵送流速为 1.0 mL/min；使用色谱柱 B 时，泵送流动相的流速为 0.5 mL/min。该方法在 1.0–5.0 和 2.5–10.0 ng/mL 的浓度范围内表现出良好的线性。分别使用方法 1 A 和 B 时的 VPS。应用方法 2 时，VPS 和 SFV 的线性浓度范围分别为 30.0–150.0 至 120–600.0 ng/mL。方法 1 和方法 2 均通过使用两个分析柱进行。不同的色谱参数如保留时间、分辨率、理论塔板数 (N)、容量因子、拖尾因子和选择性都经过仔细优化。结果表明，比较两种所用色谱柱的性能表明，对于所研究药物的分析，具有小颗粒填充的较短色谱柱 (2.1 mm × 100 mm) 优于较长色谱柱 (4.6 × 150 mm)，从而可以减少分析时间缩短了 70%，且对性能没有任何不利影响。这可以节省有机溶剂的费用并扩大每天的分析总数，从而降低调查成本。