Department of Neurology, Xuzhou New Health Geriatric Hospital, Xuzhou, Jiangsu Province, China.
Biomed Chromatogr. 2024 Nov;38(11):e6011. doi: 10.1002/bmc.6011. Epub 2024 Sep 19.
Huperzine A is a reversible and selective cholinesterase inhibitor and has been approved for the treatment of Alzheimer's diseases. In this study, we developed a highly sensitive and specific ulta-high-performance liquid chromatography-tandem mass spectrometry method for the determination of Huperzine A in rat plasma. An aliquot of 50 μL of rat plasma sample was pretreated with 200 μL of acetonitrile-methanol (v/v; 1:1) containing 0.2% formic acid followed by solid phase extraction. The resulting sample was separated on a Waters ACQUITY BEH C column using acetonitrile and water containing 0.2% formic acid as mobile phase, at a flow rate of 0.3 mL/min. Multiple-reaction monitoring (MRM) mode was used for quantitative analysis of Huperzine A in positive electrospray ionization. In the concentration range of 0.01-10 ng/mL, Huperzine A showed excellent linearity with correlation coefficient > 0.998. The intra- and inter-day RSD% were less than 9.7%, while the RE% ranged from -6.7% to 10.0%. The mean recovery was >84.5%. The validated method was demonstrated to be selective, sensitive, and reliable, which has been successfully applied to pharmacokinetic study of Huperzine A in rat plasma. Huperzine A displayed a long half-life in rat plasma and high oral bioavailability.
石杉碱甲是一种可逆和选择性的乙酰胆碱酯酶抑制剂，已被批准用于治疗阿尔茨海默病。在这项研究中，我们开发了一种高灵敏度和特异性的超高效液相色谱-串联质谱法，用于测定大鼠血浆中的石杉碱甲。取 50 μL 大鼠血浆样品，用含 0.2%甲酸的乙腈-甲醇（v/v; 1:1）200 μL 预处理，然后进行固相萃取。所得样品在 Waters ACQUITY BEH C 柱上以乙腈和含 0.2%甲酸的水为流动相，以 0.3 mL/min 的流速进行分离。采用正电喷雾电离多反应监测（MRM）模式进行石杉碱甲的定量分析。在 0.01-10 ng/mL 的浓度范围内，石杉碱甲表现出极好的线性关系，相关系数>0.998。日内和日间 RSD%均小于 9.7%，而 RE%的范围为-6.7%至 10.0%。平均回收率>84.5%。验证方法具有选择性、灵敏性和可靠性，已成功应用于大鼠血浆中石杉碱甲的药代动力学研究。石杉碱甲在大鼠血浆中半衰期长，口服生物利用度高。
