發(fā)布時間：2018-06-01 01:43

  本文選題：中藥材 + 液質(zhì)聯(lián)用��； 參考：《廣西大學(xué)》2017年碩士論文

【摘要】：中藥的療效是各種活性成分之間相互作用的結(jié)果,為闡明其體內(nèi)作用機(jī)制,藥代動力學(xué)研究是不可或缺的,近些年也越來越受到人們的關(guān)注。日趨成熟的液質(zhì)聯(lián)用技術(shù)基于操作簡便、高靈敏度、高效以及特異性強(qiáng)等優(yōu)勢,在中藥藥代動力學(xué)研究中發(fā)揮著不可替代的作用。本論文利用UHPLC-MS/MS技術(shù),選取常用中藥材白英、燈盞細(xì)辛和石見穿為研究對象,分別建立了大鼠血漿中多種活性組分的定性定量分析方法,并探究灌胃給藥提取物后大鼠體內(nèi)的藥代動力學(xué)特征,為藥效機(jī)理、新藥開發(fā)和臨床合理用藥等相關(guān)研究提供科學(xué)依據(jù)。1.建立了 UHPLC-MS/MS法分析大鼠灌胃給藥白英提取物后8種成分(原兒茶酸、綠原酸、咖啡酸、東莨菪內(nèi)酯、野黃芩苷、蘆丁、異綠原酸C和薯蕷皂苷元)的藥代動力學(xué)。經(jīng)甲酸酸化后的血漿樣品用600 μL的乙酸乙酯-乙腈(3:1,v/v)處理,流動相為含0.1%甲酸的水-甲醇溶液,8種組分和2種內(nèi)標(biāo)于15 min內(nèi)完全分離。日內(nèi)和日間精密度分別為1.1%～11.4%和1.4%～12.8%,基質(zhì)效應(yīng)低,所有組分的提取回收率均大于84.8%。該實(shí)驗(yàn)為白英的質(zhì)量控制、藥理學(xué)和毒理學(xué)研究提供了有效依據(jù)。2.建立大鼠血漿中新綠原酸、綠原酸、咖啡酸、1,3-二咖啡�？鼘幩�、東莨菪內(nèi)酯、野黃芩苷、異綠原酸A、燈盞花甲素、異綠原酸C、野黃芩素、木犀草素和芹菜素的UHPLC-MS/MS測定方法,并用于大鼠灌胃燈盞細(xì)辛提取物后12種組分在體內(nèi)的藥代動力學(xué)研究。用乙酸乙酯和乙腈(95:5,v/v)處理血漿樣品,選取0.1%甲酸水和乙腈進(jìn)行梯度洗脫,流速設(shè)定為0.3 mL/min。所有成分在線性范圍濃度內(nèi)線性關(guān)系良好,相關(guān)系數(shù)均大于0.9990。日內(nèi)和日間準(zhǔn)確度分別為-10.43%～9.76%和-10.14%～10.33%,提取回收率在85.0%～111.9%之間,基質(zhì)效應(yīng)和穩(wěn)定性均符合生物樣品分析的要求,為燈盞細(xì)辛的藥代動力學(xué)研究和體內(nèi)作用機(jī)理提供參考。3.應(yīng)用超高效液相色譜-串聯(lián)質(zhì)譜聯(lián)用法,測定大鼠灌胃給予石見穿提取物后丹參素、原兒茶酸、新綠原酸、咖啡酸、蘆丁、異槲皮苷、田基黃苷、燈盞花甲素、迷迭香酸和迷迭香酸甲酯的時間-血藥濃度。選擇0.2%的甲酸水溶液和乙腈以0.3 mL/min流速進(jìn)行梯度洗脫,采用正負(fù)離子同時掃描的多反應(yīng)離子監(jiān)測(MRM)對血漿樣品進(jìn)行定量分析,所有成分的相關(guān)系數(shù)均大于0.9990。10種分析物的定量下限分別為100、3.0、7.0、500、8.0、0.5、0.2、8.0、6.0和1.5ng·mL-1,日內(nèi)精密度 RSD≤9.6%,日間精密度RSD≤12.1%。該方法簡便、靈敏、穩(wěn)定性好且回收率高,適用于血藥濃度監(jiān)測及藥代動力學(xué)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，

本文編號：1962381