Pharmaceutical Chemistry Department, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
Bioavailability Center, National Organization for Drug Control and Research (NODCAR), Giza P.O. Box 29, Egypt.
Molecules. 2020 Oct 10;25(20):4611. doi: 10.3390/molecules25204611.
A simple, rapid, sensitive, and precise reversed-phase liquid chromatographic method was developed and validated for the simultaneous determination of four direct-acting antivirals, sofosbuvir (SF), ledipasvir (LD), declatasvir (DC), and simeprevir (SM), in their respective pharmaceutical formulations. Effective chromatographic separation was achieved on an Agilent Eclipse plus C8 column (250 mm × 4.6 mm, 5 µm) at 40 °C with gradient elution using a mobile phase composed of acetonitrile:phosphate buffer (pH 6.5). The quantification of SF and DC was based on peak area measurements at 260 nm, while the quantification of LD and SM was achieved at 330 nm. The linearity was acceptable from 1.0 to 20.0 μg/mL for the studied drugs, with correlation coefficients >0.999. The analytical performance of the newly proposed HPLC procedure was thoroughly validated according to ICH guidelines in terms of linearity, precision (RSD%, 0.39-1.57), accuracy (98.05-101.90%), specificity, limit of detection (LOD) (0.022-0.039 μg/mL), limit of quantification (LOQ) (0.067-0.118 μg/mL), and robustness. The validated HPLC method was successfully used to analyze the abovementioned drugs in their pure and dosage forms without interference from common excipients present in commercial formulations.
建立并验证了一种用于同时测定四种直接作用抗病毒药物（索磷布韦、来迪派韦、达拉他韦和西美瑞韦）的简单、快速、灵敏和精确的反相高效液相色谱法。在 40°C 下，采用乙腈-磷酸盐缓冲液（pH 6.5）作为流动相，在 Agilent Eclipse plus C8 柱（250mm×4.6mm，5μm）上实现了有效的色谱分离。SF 和 DC 的定量基于 260nm 处的峰面积测量，而 LD 和 SM 的定量则在 330nm 处进行。在所研究的药物浓度为 1.0 至 20.0μg/mL 范围内，线性良好，相关系数＞0.999。根据 ICH 指南，新提出的 HPLC 方法在线性、精密度（RSD%，0.39-1.57）、准确度（98.05-101.90%）、专属性、检测限（LOD）（0.022-0.039μg/mL）、定量限（LOQ）（0.067-0.118μg/mL）和稳健性方面进行了全面验证。该验证的 HPLC 方法成功地用于分析纯品和制剂中的上述药物，且无来自商业制剂中常见赋形剂的干扰。
