An Analytical Perspective on Liquid Chromatography-Mass Spectrometry Methods for the Analysis of Bile Acids in Human Blood Samples

Bile acids (BA) are traditionally recognized as detergents that facilitate lipid and glucose digestion and homeostasis. More recently, they have emerged as signaling molecules with the ability to influence metabolic processes via the gut-brain axis. BA are implicated in a wide range of diseases and conditions, including gastrointestinal, hepatobiliary, metabolic, cardiovascular, and neurological disorders. Accurate quantification of total bile acids (TBA) and individual BA species is essential for understanding their roles in health and disease. Liquid chromatography coupled with mass spectrometry (LC-MS) offers the sensitivity and selectivity required to detect these metabolites at trace levels in human blood samples. This systematic review critically examines validated LC-MS methodologies for BA analysis in human blood, focusing on studies published between January 2010 and April 2024. It highlights experimental designs, validation criteria, and methodological differences, aiming to inform the development of standardized analytical protocols. By addressing current gaps and emphasizing comprehensive analytical validation, this review seeks to enhance the reliability of BA quantification and support future biomarker discovery and clinical applications.胆汁酸（Bile acids, BA）历来被视作促进脂质与葡萄糖消化、维持体内稳态的去垢剂。近年来，其作为信号分子的功能被重新认知，可通过肠-脑轴（gut-brain axis）调节代谢进程。胆汁酸与多种疾病及病理生理异常密切相关，涵盖胃肠道、肝胆、代谢、心血管及神经系统病症。精准定量总胆汁酸（Total bile acids, TBA）及各单体胆汁酸组分，对于阐明其在健康与疾病中的调控作用至关重要。液相色谱-质谱联用技术（Liquid chromatography coupled with mass spectrometry, LC-MS）具备所需的灵敏度与选择性，可在人体血液样本中实现这类代谢物的痕量检测。本系统综述对2010年1月至2024年4月期间发表的、经验证的人体血液胆汁酸分析液相色谱-质谱联用方法开展批判性评估。综述重点梳理了实验设计方案、验证标准及方法学差异，旨在为标准化分析规程的开发提供参考依据。通过厘清当前研究空白并强调全面的分析验证工作，本综述旨在提升胆汁酸定量结果的可靠性，为后续生物标志物发现及临床应用提供支撑。