摘要
目的建立HPLC-MS法测定血浆中多种胆汁酸的浓度,比较天然牛黄和安宫牛黄丸中胆汁酸在小鼠体内的药代动力学过程,从药代动力学角度分析其药效差异的原因。方法用微量采血法采集小鼠全血20μL,全血样品乙腈直接沉淀蛋白后用HPLC-MS法检测全血中各种胆汁酸的浓度;组织分布研究取小鼠肺、脑组织,匀浆离心后取上清液直接沉淀蛋白后检测。结果各种胆汁酸的线性范围为4～4000mg/L,最低检测浓度为4mg/L,日间和日内精密度均小于10.0%。灌胃后,安宫牛黄丸中的牛磺胆酸、牛磺鹅去氧胆酸在小鼠体内的曲线下面积(AUC)及其在肺组织中的分布明显高于天然牛黄,胆酸、牛磺胆酸在脑组织中的分布明显高于天然牛黄。结论该法灵敏度高,特异性好,适用于胆汁酸的体内和体外药代动力学研究,安宫牛黄丸中含有的某些非牛黄成分可促进天然牛黄中胆汁酸的吸收与组织分布。
Objective To establish a high performance liquid chromatography-mass spectrometry （HPLC- MS） for determining the plasma concentrations of multiple bile acids, comparing the pharmacokinetic course of bile acid of natural Calculus Bovis or Angongniuhuang Pills in mice, and to analyze the cause of different effects. Methods The capillary blood collection method was used to collect the whole blood samples （20 μL） from mice, and then the protein in the samples were deposited directly with acetonitrile, in which the plasma concentrations of multiple bile acids were detected by HPLC-MS. The tissue of lung and brain were taken, and then homogenized and centrifuged to obtain supematant for detecting tissue distribution after protein precipitation. Results The linear range of multiple bile acids was at 4 - 4 000 mg/L, the lowest detective concentration was 4 mg/L, and inter-day precision and intra-day precision were all lower than 10.0%. After mice were fed, the area under curves （AUC） and distribution in lung tissue of taurocholic acid and taurochenodeoxyeholic acid in Angongniuhuang Pills were higher than those in natural Calculus Bovis in mouse body. The distribution of bile acid and taurocholic acid in brain tissue was higher than that of natural Calculus Bovis. Conclusion The method has a higher sensitivity and specificity, which can be used in the study of pharmacokinetics of bile acids in vitro and in vivo. Some constituents of non- Calculus Bovis in Angongniuhuang Pills can improve the absorption and tissue distribution of bile acid of natural Calculus Bovis.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2009年第5期344-348,共5页
Journal of Beijing University of Traditional Chinese Medicine
