UPLC-MS/MS测定人血浆中长春西汀和阿朴长春胺酸的含量及其应用

Determination of Vinpocetine and Apovincaminic Acid in Human Plasma by UPLC-MS/MS and Its Application

目的建立和验证UPLC-MS/MS测定人血浆中长春西汀和阿朴长春胺酸的方法,并用于临床样品检测。方法以长春西汀-d5、阿朴长春胺酸-d4为内标,人血浆样品经蛋白沉淀法沉淀处理,色谱柱为Waters ACQUITY UPLC?BEH C18(2.1 mm×50 mm,1.7μm),流动相为乙腈-水(含0.05%甲酸),梯度洗脱,流速0.35 mL·min^(-1);电喷雾离子化源正离子监测模式。结果长春西汀和阿朴长春胺酸线性范围分别为0.04~20.0 ng·mL^(-1)(r=0.9997)、0.50~250 ng·mL^(-1)(r=0.9996),检测限分别为0.01,0.10 ng·mL^(-1),待测物与内标提取回收率均为94.81%~105.0%,基质效应为94.51%~105.0%,RSD均<5%;批内、批间精密度RSD均<10%。结论本法特异性强、快速、准确,重复性好,适用于长春西汀临床生物样品检测及生物等效性研究。

OBJECTIVE To establish and validate a UPLC-MS/MS for the determination of vinpocetine and apovincaminic acid in human plasma and apply to the determination of clinical samples.METHODS Vinpocetine-d5 and apovincaminic acid-d4 were used as internal standards.Human plasma samples were prepared with acetonitrile to precipitate to protein.The chromatographic column was Waters ACQUITY UPLC?BEH C18(2.1 mm×50 mm,1.7μm),the mobile phase was acetonitrile-water(containing 0.05%formic acid).Gradient elution with a flow rate of 0.35 mL·min-1.Mass spectrometry conditions:Electrospray ionization source(ESI),positive ion detection mode.RESULTS The linear range were0.04-20.0 ng·mL^(-1)(r=0.9997)for vinpocetine and 0.50-250 ng·mL^(-1)(r=0.9996)for apovincaminic acid,and the limit of quantification were 0.01,0.10 ng·mL^(-1),respectively.The recoveries of test sample and internal standard were in the range of94.81%-105.0%,and the matrix effect were 94.51%-105.0%,RSDs were<5%.The RSDs of intra-and inter-batch precision were all<10%.CONCLUSION The method is specific,rapid,accurate and reproducible.Therefore,it is suitable for the determination of vinpocetine concentration in plasma and can applied to its bioequivalence study.