Abou-Omar Mona N, Annadi Abdelaziz M, El Zahar Noha M, Youssef Ahmed O, Amin Mohammed A, Attia Mohamed S, Mohamed Ekram H
Department of Chemistry, Faculty of Women for Arts, Science and Education, Ain Shams University Cairo Egypt.
Chemistry Department, Faculty of Science, Ain Shams University Abbassia Cairo 11566 Egypt
RSC Adv. 2021 Sep 6;11(47):29797-29806. doi: 10.1039/d1ra05492a. eCollection 2021 Sep 1.
A rapid and selective LC-MS/MS method is described for the simultaneous assay of Avanafil and Dapoxetine in human plasma a protein precipitation (PP) sample preparation technique. Tadalafil was chosen as the internal standard reaching good recovery and reproducibility while diminishing the effects of the matrix. An Agilent Zorbax Eclipse XDB C column (4.6 × 50 mm, 1.8 μm) was used for the chromatographic separation and analysis, while 0.1% formic acid : acetonitrile (60 : 40, v/v) was utilized at a flow rate of 0.5 mL min. It was revealed that 6 min stop time accomplished the best separation. The assay was linear over the range of 10-6000 ng mL for both drugs. The established bio-analytical method validation was demonstrated following US-FDA recommendations including sensitivity, selectivity, linearity, accuracy and precision. Furthermore, other validation parameters were assessed such as the dilution integrity, matrix effect, carryover, and analyte stability during both short- and long-term sample processing and storage. The adopted method was efficaciously applied to a clinical study for the concurrent determination of Avanafil and Dapoxetine in human plasma.
本文描述了一种快速、选择性的液相色谱-串联质谱（LC-MS/MS）方法，用于同时测定人血浆中阿伐那非和达泊西汀，采用蛋白质沉淀（PP）样品制备技术。选择他达拉非作为内标，回收率和重现性良好，同时减少了基质效应。采用安捷伦Zorbax Eclipse XDB C柱（4.6×50 mm，1.8μm）进行色谱分离和分析，以0.1%甲酸∶乙腈（60∶40，v/v）为流动相，流速为0.5 mL/min。结果表明，6分钟的停留时间实现了最佳分离。两种药物在10-6000 ng/mL范围内均呈线性。按照美国食品药品监督管理局（US-FDA）的建议，对建立的生物分析方法进行了验证，包括灵敏度、选择性、线性、准确性和精密度。此外，还评估了其他验证参数，如稀释完整性、基质效应、残留以及短期和长期样品处理及储存过程中分析物的稳定性。所采用的方法有效地应用于一项临床研究，用于同时测定人血浆中的阿伐那非和达泊西汀。
