基于液质联用技术的中药材多组分在大鼠体内的药代动力学研究

发布时间：2018-06-01 01:43

  本文选题：中药材 + 液质联用　； 参考：《广西大学》2017年硕士论文

【摘要】：中药的疗效是各种活性成分之间相互作用的结果,为阐明其体内作用机制,药代动力学研究是不可或缺的,近些年也越来越受到人们的关注。日趋成熟的液质联用技术基于操作简便、高灵敏度、高效以及特异性强等优势,在中药药代动力学研究中发挥着不可替代的作用。本论文利用UHPLC-MS/MS技术,选取常用中药材白英、灯盏细辛和石见穿为研究对象,分别建立了大鼠血浆中多种活性组分的定性定量分析方法,并探究灌胃给药提取物后大鼠体内的药代动力学特征,为药效机理、新药开发和临床合理用药等相关研究提供科学依据。1.建立了 UHPLC-MS/MS法分析大鼠灌胃给药白英提取物后8种成分(原儿茶酸、绿原酸、咖啡酸、东莨菪内酯、野黄芩苷、芦丁、异绿原酸C和薯蓣皂苷元)的药代动力学。经甲酸酸化后的血浆样品用600 μL的乙酸乙酯-乙腈(3:1,v/v)处理,流动相为含0.1%甲酸的水-甲醇溶液,8种组分和2种内标于15 min内完全分离。日内和日间精密度分别为1.1%～11.4%和1.4%～12.8%,基质效应低,所有组分的提取回收率均大于84.8%。该实验为白英的质量控制、药理学和毒理学研究提供了有效依据。2.建立大鼠血浆中新绿原酸、绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、东莨菪内酯、野黄芩苷、异绿原酸A、灯盏花甲素、异绿原酸C、野黄芩素、木犀草素和芹菜素的UHPLC-MS/MS测定方法,并用于大鼠灌胃灯盏细辛提取物后12种组分在体内的药代动力学研究。用乙酸乙酯和乙腈(95:5,v/v)处理血浆样品,选取0.1%甲酸水和乙腈进行梯度洗脱,流速设定为0.3 mL/min。所有成分在线性范围浓度内线性关系良好,相关系数均大于0.9990。日内和日间准确度分别为-10.43%～9.76%和-10.14%～10.33%,提取回收率在85.0%～111.9%之间,基质效应和稳定性均符合生物样品分析的要求,为灯盏细辛的药代动力学研究和体内作用机理提供参考。3.应用超高效液相色谱-串联质谱联用法,测定大鼠灌胃给予石见穿提取物后丹参素、原儿茶酸、新绿原酸、咖啡酸、芦丁、异槲皮苷、田基黄苷、灯盏花甲素、迷迭香酸和迷迭香酸甲酯的时间-血药浓度。选择0.2%的甲酸水溶液和乙腈以0.3 mL/min流速进行梯度洗脱,采用正负离子同时扫描的多反应离子监测(MRM)对血浆样品进行定量分析,所有成分的相关系数均大于0.9990。10种分析物的定量下限分别为100、3.0、7.0、500、8.0、0.5、0.2、8.0、6.0和1.5ng·mL-1,日内精密度 RSD≤9.6%,日间精密度RSD≤12.1%。该方法简便、灵敏、稳定性好且回收率高,适用于血药浓度监测及药代动力学研究。
[Abstract]:The efficacy of traditional Chinese medicine is the result of interaction between various active components. In order to elucidate the mechanism of its action in vivo, pharmacokinetics research is indispensable, and has been paid more and more attention in recent years. The increasingly mature liquid-mass spectrometry technology, which is based on the advantages of simple operation, high sensitivity, high efficiency and strong specificity, plays an irreplaceable role in the study of pharmacokinetics of traditional Chinese medicine. In this paper, UHPLC-MS/MS technique was used to establish a qualitative and quantitative analysis method of various active components in plasma of rats. The pharmacokinetic characteristics of the rats after intragastric administration of the extract were investigated to provide a scientific basis for the study of pharmacodynamic mechanism, new drug development and rational clinical use. 1. The pharmacokinetics of eight components (protocatechuic acid, Lv Yuan acid, caffeic acid, scopolactone, baicalin, rutin, isoLv Yuan acid C and diosgenin) was determined by UHPLC-MS/MS method. The plasma samples after formic acid acidification were treated with 600 渭 L ethyl acetate and acetonitrile 3: 1 v / v). The mobile phase consisted of 8 components of water-methanol solution containing 0.1% formic acid and two internal standards were completely separated within 15 min. The intra-day and inter-day precision were 1.1% and 1.4%, respectively, and the matrix effect was low. The recovery rate of all components was higher than 84.8%. This experiment provides an effective basis for the quality control, pharmacology and toxicology research. 2. A UHPLC-MS/MS method was established for the determination of new Lv Yuan acid, caffeic acid 1 and 3-dicaffeyl quinine, scopolactone, baicalin, isoLv Yuan acid A, breviscapine, isoLv Yuan acid C, baicalin, luteolin and apigenin in rat plasma. The pharmacokinetics of 12 components of Erigeron breviscapus extract was studied in vivo. The plasma samples were treated with ethyl acetate and acetonitrile 95: 5 v / v), and 0.1% formic acid water and acetonitrile were selected for gradient elution. The flow rate was set at 0.3 mL / min. The linear relationship of all components in the linear range was good, and the correlation coefficient was greater than 0.9990. The intra-day and inter-day accuracy were -10.43% and -10.14%, respectively, and the recovery rate was between 85.0% and 111.9%. The matrix effect and stability met the requirements of biological sample analysis, which provided a reference for pharmacokinetic study and in vivo action mechanism of Erigeron breviscapus. Ultrahigh performance liquid chromatography-tandem mass spectrometry was used to determine Danshensu, protocatechuic acid, new Lv Yuan acid, caffeic acid, rutin, isoquercitrin, field flavin, breviscapine in rats. Time-to-Blood concentration of Rosemary Acid and Methyl Rosemary Acid. 0.2% aqueous solution of formic acid and acetonitrile were selected for gradient elution at a flow rate of 0.3 mL/min. The plasma samples were quantitatively analyzed by simultaneous positive and negative ion scanning multi-reaction ion monitoring (MRM). The correlation coefficients of all components were greater than 0.9990.10. The lower limit of quantification of all kinds of analytes were 100 ~ 3.0 ~ 7.0500 ~ 8.0 ~ (0.2) ~ (0.2) ~ 8.0 ~ (-1) and 1.5ng mL ~ (-1), respectively. The intra-day precision (RSD) 鈮，

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