目的 建立超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定血浆中长春西汀(VP)及其代谢物阿朴长春胺酸(AVA)的浓度。方法 血浆样品经HCl酸化后用乙酸乙酯液-液萃取,以非那西汀为内标, 采用ACQUITY UPLC BEH C18柱(2.1 mm×100 mm,1.7 μm)分离,以甲醇-水溶液为流动相进行梯度洗脱,流速为0.2 mL·min-1, 通过电喷雾离子源(ESI),采用多反应监测(MRM)正离子方式检测。结果 长春西汀在0.40~149 ng·mL-1(r=0.994 4),阿朴长春胺酸(AVA)在0.80~298 ng·mL-1(r=0.994 9)内线性良好。准确度(RE%)长春西汀为-5.9%~7.3%、阿朴长春胺酸为-7.9%~ 5.6%,长春西汀的日内和日间精密度(RSD)均小于12.9 %,阿朴长春胺酸的日内和日间精密度(RSD)均小于11.1%。结论 所建立的UPLC-MS/MS专属性好,灵敏度高,操作简便,已成功用于长春西汀片剂在大鼠体内的药物动力学研究。
Abstract
To develop a specific,rapid, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of vinpocetine (VP) and its metabolite, apovincaminic acid (AVA) , in rat plasma. METHODS The plasma samples were acidified and extracted with ethyl acetate. Phenacetin was used as internal standard (I.S). The separation was performed on a Waters ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 μm). The mobile phase was composed of methanol-water with gradient elution at the flow rate of 0.20 mL·min-1. The detection was performed on a triple quadrupole tandem mass spectrometer in multiple reaction monitoring mode via electrospray ionization source(ESI)with positive mode. RESULTS The calibration curve was linear in concentration range from 0.40 to 149 ng·mL-1 for vinpocetine and 0.80 to 298 ng·mL-1 for apovincaminic acid. The intra-day and inter-day precision (relative standard deviation) values were within 12.9%,the accuracy (relative error) was from -5.9% to 7.3% for vinpocetine and-7.9% to 5.6% for apovincaminic acid at all quality control levels. CONCLUSION The UPLC-MS/MS method is specific,rapid, sensitive, and is successfully applied to the pharmacokinetic study of VP tables in rat plasma after intragastric administration.
关键词
长春西汀 /
阿朴长春胺酸 /
超高效液相色谱-串联质谱 /
药动学
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Key words
vinpocetine(VP) /
apovincaminic acid(AVA) /
UPLCMS/MS /
pharmacokinetics
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中图分类号:
R917
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参考文献
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脚注
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基金
基金项目:国家自然科学基金资助项目(30873367)
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