Institute of Mass Spectrometry, School of Medicine, Grove Building Swansea University Singleton Park Swansea SA2 8PP, UK.
J Lipid Res. 2010 Jan;51(1):23-41. doi: 10.1194/jlr.R001941-JLR200.
The formation of bile acids/bile alcohols is of major importance for the maintenance of cholesterol homeostasis. Besides their functions in lipid absorption, bile acids/bile alcohols are regulatory molecules for a number of metabolic processes. Their effects are structure-dependent, and numerous metabolic conversions result in a complex mixture of biologically active and inactive forms. Advanced methods are required to characterize and quantify individual bile acids in these mixtures. A combination of such analyses with analyses of the proteome will be required for a better understanding of mechanisms of action and nature of endogenous ligands. Mass spectrometry is the basic detection technique for effluents from chromatographic columns. Capillary liquid chromatography-mass spectrometry with electrospray ionization provides the highest sensitivity in metabolome analysis. Classical gas chromatography-mass spectrometry is less sensitive but offers extensive structure-dependent fragmentation increasing the specificity in analyses of isobaric isomers of unconjugated bile acids. Depending on the nature of the bile acid/bile alcohol mixture and the range of concentration of individuals, different sample preparation sequences, from simple extractions to group separations and derivatizations, are applicable. We review the methods currently available for the analysis of bile acids in biological fluids and tissues, with emphasis on the combination of liquid and gas phase chromatography with mass spectrometry.
胆汁酸/胆汁醇的形成对于维持胆固醇的体内平衡具有重要意义。除了在脂质吸收中的作用外，胆汁酸/胆汁醇还是许多代谢过程的调节分子。它们的作用具有结构依赖性，许多代谢转化导致具有生物活性和非活性形式的复杂混合物。需要先进的方法来对这些混合物中的各个胆汁酸进行定性和定量分析。结合这些分析和蛋白质组分析，将有助于更好地了解作用机制和内源性配体的性质。质谱是色谱柱流出物的基本检测技术。采用电喷雾电离的毛细管液相色谱-质谱法在代谢组学分析中具有最高的灵敏度。经典的气相色谱-质谱法灵敏度较低，但提供了广泛的结构依赖性碎裂，增加了对非共轭胆汁酸的等质量异构体分析的特异性。根据胆汁酸/胆汁醇混合物的性质和个体浓度范围，可采用不同的样品制备顺序，从简单的提取到分组分离和衍生化。我们综述了目前用于生物体液和组织中胆汁酸分析的方法，重点是液相和气相色谱与质谱的联用。
