Department of Pharmaceutical Analysis, Key Laboratory of Drug Quality control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, PR China.
Department of Pharmaceutical Analysis, Key Laboratory of Drug Quality control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, PR China.
J Pharm Biomed Anal. 2022 Mar 20;211:114596. doi: 10.1016/j.jpba.2022.114596. Epub 2022 Jan 19.
Ultra-high-performance liquid chromatography (UHPLC) hyphenated to tandem mass spectrometric methods using the dimethylpentafluorophenylpropyl stationary phase with polar and nonpolar characteristics of a 2-μm fused-core silica particle packing were evaluated to perform efficient separation for the analysis of donepezil in human plasma. The fused-core silica particle design providing the shorter diffusional mass transfer path for the analytes is less affected in resolving power by increasing mobile-phase velocity for faster chromatographic resolution between the administered compound and multiple interfering peaks from the extracted quality control, calibration standard and study samples following simple protein precipitation extraction procedures. In this work, two UHPLC-MS/MS approaches requiring 1.2 min per sample run time and neither expensive ultra-high-pressure instrumentation nor new laboratory protocols were applied and compared for determination of donepezil in human plasma at sub-nanograms per milliliter region to support a randomized, crossover bioequivalence (BE) study in which healthy volunteers each received a single oral dose of the test and reference formulations of 10 mg donepezil hydrochloride. The column performance in terms of chromatographic separation efficiency, peak asymmetry and resolution and retention time reproducibility was found to be sustainable over a thousand extracted plasma injections. The linear dynamic range was detected over a concentration range of 0.2-50 ng/mL. The intra- and inter-day assay accuracy and precision were within 15% for the analyte in individual biological fluids. Positive correlation coefficients (r) greater than 0.98 and 0.99 for donepezil concentrations in study plasma samplers measured by the proposed UHPLC-MS/MS approaches and another validated HPLC-MS/MS method were observed.
采用具有极性和非极性特性的 2μm 熔融核硅胶颗粒填充的二甲基五氟丙基固定相，将超高效液相色谱 (UHPLC) 与串联质谱联用方法进行了评估，以实现对人血浆中多奈哌齐的有效分离。熔融核硅胶颗粒设计为分析物提供了较短的扩散传质路径，在分辨率方面受流动相速度增加的影响较小，从而可以在给予的化合物与从提取的质控、校准标准和研究样品中获得的多个干扰峰之间实现更快的色谱分辨率，这些样品采用简单的蛋白质沉淀提取程序。在这项工作中，应用了两种 UHPLC-MS/MS 方法，每种方法的样品运行时间为 1.2 分钟，既不需要昂贵的超高压仪器，也不需要新的实验室方案，并对人血浆中纳克级以下的多奈哌齐进行了比较，以支持一项随机、交叉生物等效性 (BE) 研究，其中健康志愿者每人接受 10mg 盐酸多奈哌齐的试验和参比制剂的单次口服剂量。在色谱分离效率、峰不对称性和分辨率以及保留时间重现性方面，该柱性能可持续超过一千次提取的血浆进样。线性动态范围在 0.2-50ng/mL 的浓度范围内检测到。在个体生物液体中，分析物的日内和日间测定准确度和精密度均在 15%以内。通过所提出的 UHPLC-MS/MS 方法和另一种经过验证的 HPLC-MS/MS 方法测量，研究血浆采样器中的多奈哌齐浓度的正相关系数 (r) 大于 0.98 和 0.99。
