Pharmaceutical Service Center, Faculty of Pharmacy, Tanta University, Egypt.
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Dec 5;205:398-409. doi: 10.1016/j.saa.2018.07.018. Epub 2018 Jul 9.
Simple, rapid, sensitive, accurate, precise and earth-friendly spectrophotometric methods were developed for the simultaneous analysis of ledipasvir (LED) and sofosbuvir (SOF) without interference of both sunset yellow dye and copovidone excipients (the most probable interferents) in their combined dosage form. These proposed methods were based on measurement of LED in synthetic mixtures and combined dosage form by first derivative (D) spectrophotometry at 314 nm over the concentration range of 2-50 μg mL with coefficient of determination (R) > 0.9999, mean percentage recovery of 99.98 ± 0.62. On the other hand, SOF in synthetic mixtures and combined dosage form was determined by five methods. Method I is based on the use of D spectrophotometry at 274.2 nm (zero crossing point of LED). Method II involves the application of conventional dual wavelength method (DW) at the absolute difference between SOF zero order amplitudes at 261 nm (λ of SOF) and 364.7 nm. At these wavelengths, the absolute difference between LED zero order amplitudes was observed to equal zero. Method III depends on isosbestic point method (ISP) in which the total concentration of both drugs was measured at isosbestic point at 262.7 nm. Concentration of SOF could be obtained by subtraction of LED concentration. While, method IV depends on absorbance correction method (absorption factor method), which is based on determination of SOF concentration at 262.7 nm (λ) and LED at 333 nm (λ of LED). Finally, method V depends on absorbance ratio method (Q-analysis) in which 262.7 nm (λ) and 261 nm (λ of SOF) were selected to determine SOF concentration. The linearity range for all methods for SOF determination was 2-50 μg mL with coefficient of determination (R) > 0.9999. Methods I, II & III were also applied for determination of SOF concentration in single dosage form. Their mean percentage recoveries were 100.35 ± 1.85, 99.97 ± 0.54 and 100.03 ± 0.49, for the three methods respectively. The proposed methods were validated according to international conference of harmonization (ICH) requirements and statistically compared to published reference methods. The ANOVA test confirmed that there is no significant differences between the proposed methods, and can be used for routine analysis of LED and SOF in commercial tablets. These developed methods were applied to estimate the average content and uniformity of dosage unit for LED/SOF combined dosage form and SOF single dosage form according to British pharmacopeia (BP) requirements.
建立了同时分析雷迪帕韦（LED）和索磷布韦（SOF）的简单、快速、灵敏、准确、精密且环保的分光光度法，在其复方制剂中，不受日落黄染料和共聚维酮赋形剂（最可能的干扰物）的干扰。这些方法是基于在合成混合物和复方制剂中测量 LED，在 314nm 处进行一阶导数（D）分光光度法，浓度范围为 2-50μg/mL，相关系数（R）＞0.9999，平均回收率为 99.98％±0.62％。另一方面，通过五种方法测定了合成混合物和复方制剂中的 SOF。方法 I 基于在 274.2nm 处使用 D 分光光度法（LED 的零交点）。方法 II 涉及应用常规双波长法（DW），在 SOF 零阶幅度在 261nm（SOF 的λ）和 364.7nm 处的绝对差值。在这些波长下，观察到 LED 零阶幅度的绝对差值为零。方法 III 依赖等吸收点法（ISP），其中两种药物的总浓度在 262.7nm 等吸收点处进行测量。通过减去 LED 浓度即可获得 SOF 的浓度。而方法 IV 依赖于吸光度校正法（吸收因子法），它基于在 262.7nm（λ）处测定 SOF 浓度和在 333nm（LED 的λ）处测定 LED 浓度。最后，方法 V 依赖于吸光度比法（Q 分析），其中选择 262.7nm（λ）和 261nm（SOF 的λ）来确定 SOF 浓度。所有方法测定 SOF 的线性范围均为 2-50μg/mL，相关系数（R）＞0.9999。方法 I、II 和 III 也被应用于测定单剂量制剂中的 SOF 浓度。它们的平均回收率分别为 100.35％±1.85％、99.97％±0.54％和 100.03％±0.49％。根据国际协调会议（ICH）的要求验证了所提出的方法，并与已发表的参考方法进行了统计学比较。方差分析（ANOVA）检验证实，所提出的方法之间没有显著差异，可用于 LED 和 SOF 的商业片剂的常规分析。根据英国药典（BP）的要求，这些方法被应用于估计 LED/SOF 复方制剂和 SOF 单剂量制剂的平均含量和单位剂量均匀度。
