Quantitative Profiling of Bile Acids in Feces of Humans and Rodents by Ultra-High-Performance Liquid Chromatography–Quadrupole Time-of-Flight Mass Spectrometry

Abstract

A simple, sensitive, and reliable quantification and identification method was developed and validated for simultaneous analysis of 58 bile acids (BAs) in human and rodent (mouse and rat) fecal samples. The method involves an extraction step with a 5% ammonium–ethanol aqueous solution; the BAs were quantified by high-resolution mass spectrometry (ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry, UPLC–Q-TOF). The recoveries were 80.05–120.83%, with coefficient variations (CVs) of 0.01–9.82% for three biological species. The limits of detection (LODs) were in the range of 0.01–0.24 μg/kg, and the limits of quantification (LOQs) ranged from 0.03 to 0.81 μg/kg. In addition, the analytical method was used to identify and quantify BAs in end-stage renal disease (ESRD) patients, C57BL/6 mice, and Sprague-Dawley (SD) rats. The fecal BA profile and analysis of BA indices in these samples provide valuable information for further BA metabolic disorder research.

Integration of semi-empirical MS/MS library with characteristic features for the annotation of novel amino acid-conjugated bile acids

“…Fecal samples were extracted using a published method with slight modifications. 18 Briefly, 1 mL 5% ammonia in ethanol was added to 100 mg wet feces. The mixture was shaken for 15 min at 4 °C, sonicated for 15 min, and then centrifuged at 14 000 g at 4 °C for 15 min.…”

Advances in Metabolic Profiling of Biological Samples

“…Other articles have focused on the detection of specific metabolite classes [8][9][10][11], such as bile acids, sphingoid bases, fatty acids, and advanced glycation end products (AGEs), which have been implicated in many diseases. Zhang et al [8] developed and validated a high-throughput method for the comprehensive analysis of bile acids in human and rodent fecal samples. Overall, 58 bile acids were quantified by ultra-high-performance liquid chromatography coupled with mass spectrometry (UPLC-MS).…”