Hagio Masahito, Matsumoto Megumi, Ishizuka Satoshi
Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan.
Methods Mol Biol. 2011;708:119-29. doi: 10.1007/978-1-61737-985-7_6.
In recent years, bile acids (BAs) have received much attention as signaling molecules as well as biosurfactants for lipophilic nutrients. To understand exact BA behavior, the precise distribution of BAs in vivo must be determined. However, to date, it has been difficult to know the precise roles of BA due to variations in BA molecules including conjugated forms. Thus, we reconsidered BA extraction methodology and introduced an ultra performance liquid chromatography/electrospray ionization-mass spectrometry (UPLC/ESI-MS) technique for BA analysis. Consequently, we established a rapid and reliable method, using UPLC/ESI-MS, for the analysis of BAs in various biological samples including liver, bile, blood, intestinal contents, and feces. This method enables us to determine the BA profile, including conjugation status, in a single 30 min run. This technique will be a useful tool for the investigation of the roles of BA metabolism in physiological and pathological conditions.
近年来，胆汁酸（BAs）作为信号分子以及亲脂性营养物质的生物表面活性剂受到了广泛关注。为了解胆汁酸的确切行为，必须确定其在体内的精确分布。然而，迄今为止，由于胆汁酸分子（包括共轭形式）的变化，很难了解胆汁酸的确切作用。因此，我们重新考虑了胆汁酸提取方法，并引入了超高效液相色谱/电喷雾电离质谱（UPLC/ESI-MS）技术用于胆汁酸分析。结果，我们建立了一种使用UPLC/ESI-MS的快速可靠方法，用于分析包括肝脏、胆汁、血液、肠内容物和粪便在内的各种生物样品中的胆汁酸。该方法使我们能够在单次30分钟的运行中确定胆汁酸谱，包括共轭状态。这项技术将成为研究胆汁酸代谢在生理和病理条件下作用的有用工具。
