目的 首次建立同时测定藤茶中二氢杨梅素(dihydromyricetin, DMY)和杨梅素(myricetin,MY)的LC-MS/MS,并将其运用到DMY和MY的小鼠体内组织分布研究。方法 ICR小鼠分别灌胃给药144.4 mg·kg-1DMY和MY后处死,采集血、心、肝、脾、肺、肾、脑组织,采用蛋白沉淀法处理样本,Ultimate AQ-C18(3.0 mm×100 mm,3.0 μm)色谱柱进行分离来测定各组织中的药物浓度,并进行数据整理与分析。结果 方法学考察的精密度、准确度、基质效应和萃取回收率等均能符合生物样品分析的要求。组织分布结果表明,DMY和MY体内吸收较快,肝组织中DMY和MY的最大达峰浓度高于其他组织,DMY的血药浓度和组织分布浓度总体上均要高于MY。结论 本实验建立的同时测定各组织中的DMY和MY药物浓度的LC-MS/MS具有快速、稳定、专属性高的特点,其组织分布研究结果为进一步的开发与临床应用DMY和MY提供了参考。
Abstract
OBJECTIVE To establish an LC-MS/MS method for the simultaneous quantitative analysis of dihydromyricetin and myricetin in mice plasma, and for application in tissue distribution study after oral administration dihydromyricetin(DMY) and myricetin(MY). METHODS All groups of mice were sacrificed and the heart, liver, spleen, lung, kidney, brain and plasma were collected after oral administration of DMY and MY at the dose of 144.4 mg·kg-1, respectively. The protein precipitation approach with acetonitrile containing 0.1% formic acid was adopted for the sample preparation. The liquid chromatographic separation process was performed with an Ultimate? AQ-C18(3.0 mm×100 mm,3.0 μm) to determine the concentrations of DMY and MY in all tissue, and then all data were analysed. RESULTS It was proved that the accuracy, precision, matrix effect and extract recovery of this method can meet the requirements of biological sample analysis. The RESULTS indicated that DMY and MY were quickly absorbed in the body. The maximum concentrations of DMY and MY in the liver were higher than those in the other tissues. Moreover,the levels of DMY in plasma and other tissues were higher than MY. CONCLUSION A rapid, reliable, sensitive LC-MS/MS method for the simultaneous quantification of DMY and MY is established in the present research. Meanwhile, the result of the tissue distribution study will provide a reference for the reasonable development and the future clinical applications of DMY and MY.
关键词
液相色谱-二级质谱联用 /
二氢杨梅素 /
杨梅素 /
藤茶 /
组织分布
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Key words
LC-MS/MS /
dihydromyricetin /
myricetin /
Ampelopis grossedentata /
tissue distribution
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中图分类号：
R969.1
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81202985, 81473411,81573686);湖南省教育厅十二五中药学重点学科资助项目
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