Alnouti Yazen, Csanaky Iván L, Klaassen Curtis D
Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE 68198, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Oct 1;873(2):209-17. doi: 10.1016/j.jchromb.2008.08.018. Epub 2008 Sep 3.
The differences among individual bile acids (BAs) in eliciting different physiological and pathological responses are largely unknown because of the lack of valid and simple analytical methods for the quantification of individual BAs and their taurine and glycine conjugates. Therefore, a simple and sensitive LC-MS/MS method for the simultaneous quantification of 6 major BAs, their glycine, and taurine conjugates in mouse liver, bile, plasma, and urine was developed and validated. One-step sample preparation using solid-phase extraction (for bile and urine) or protein precipitation (for plasma and liver) was used to extract BAs. This method is valid and sensitive with a limit of quantification ranging from 10 to 40 ng/ml for the various analytes, has a large dynamic range (2500), and a short run time (20 min). Detailed BA profiles were obtained from mouse liver, plasma, bile, and urine using this method. Muricholic acid (MCA) and cholic acid (CA) taurine conjugates constituted more than 90% of BAs in liver and bile. BA concentrations in liver were about 300-fold higher than in plasma, and about 180-fold higher in bile than in liver. In summary, a reliable and simple LC-MS/MS method to quantify major BAs and their metabolites was developed and applied to quantify BAs in mouse tissues and fluids.
由于缺乏有效且简便的分析方法来定量分析单个胆汁酸（BAs）及其与牛磺酸和甘氨酸的共轭物，目前对于不同的BAs引发不同生理和病理反应的差异仍知之甚少。因此，我们开发并验证了一种简单且灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量小鼠肝脏、胆汁、血浆和尿液中的6种主要BAs及其甘氨酸和牛磺酸共轭物。采用一步固相萃取法（用于胆汁和尿液）或蛋白沉淀法（用于血浆和肝脏）来提取BAs。该方法有效且灵敏，各种分析物的定量限为10至40 ng/ml，具有较宽的动态范围（2500）和较短的运行时间（20分钟）。使用该方法从小鼠肝脏、血浆、胆汁和尿液中获得了详细的BA谱。在肝脏和胆汁中，鼠胆酸（MCA）和胆酸（CA）的牛磺酸共轭物占BAs的90%以上。肝脏中的BA浓度比血浆中高约300倍，胆汁中的BA浓度比肝脏中高约180倍。总之，我们开发了一种可靠且简单的LC-MS/MS方法来定量主要BAs及其代谢物，并将其应用于定量小鼠组织和体液中的BAs。
