Rapid analysis of bile acids in different biological matrices using LC-ESI-MS/MS for the investigation of bile acid transformation by mammalian gut bacteria
Rapid analysis of bile acids in different biological matrices using LC-ESI-MS/MS for the investigation of bile acid transformation by mammalian gut bacteria
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DOI：
10.1007/s00216-016-0048-1
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发表时间：
2017-02-01
影响因子：
4.3
通讯作者：
Rohn, Sascha
中科院分区：
文献类型：
--
作者：
Wegner, Katrin;Just, Sarah;Rohn, Sascha
胆汁酸是调节胆固醇、葡萄糖和能量稳态的重要信号分子，因此与代谢紊乱的发展有关。它们的生物利用度受到肠道微生物群的强烈调节，这有助于产生复杂的个体特异性胆汁酸谱。因此，重要的是要有准确的方法来精确测量这些重要的代谢物。在这里，一个快速和灵敏的液相色谱-串联质谱法（LC-MS/MS）的方法，同时识别和定量的初级和二级胆汁酸，以及他们的牛磺酸和甘氨酸共轭物的开发和验证。证明了该方法对哺乳动物组织、生物液体和细胞培养基的适用性。该分析方法主要包括一个简单而快速的液-液萃取过程中存在的氘标记的内标物。实现了所有等压胆汁酸物质的基线分离，并观察到在宽浓度范围内的线性相关性。该方法在日内（1.42- 11.07%）和日间（2.11- 12.71%）分析中显示出可接受的准确度和精密度，并且代表性分析物的回收率良好（83.7- 107.1%）。作为概念验证，该分析方法适用于小鼠组织和生物流体，但特别适用于Coriobacteriaceae肠道细菌体外发酵的样品。所开发的方法表明，该物种的迟缓蛋和Collinsella aerofaciens具有胆盐水解酶的活性，并为第一次，该物种肠杆菌mucosicola是能够在体外解偶联和分解初级胆汁酸。
Bile acids are important signaling molecules that regulate cholesterol, glucose, and energy homoeostasis and have thus been implicated in the development of metabolic disorders. Their bioavailability is strongly modulated by the gut microbiota, which contributes to generation of complex individual-specific bile acid profiles. Hence, it is important to have accurate methods at hand for precise measurement of these important metabolites. Here, a rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous identification and quantitation of primary and secondary bile acids as well as their taurine and glycine conjugates was developed and validated. Applicability of the method was demonstrated for mammalian tissues, biofluids, and cell culture media. The analytical approach mainly consists of a simple and rapid liquid-liquid extraction procedure in presence of deuterium-labeled internal standards. Baseline separation of all isobaric bile acid species was achieved and a linear correlation over a broad concentration range was observed. The method showed acceptable accuracy and precision on intra-day (1.42-11.07 %) and inter-day (2.11-12.71 %) analyses and achieved good recovery rates for representative analytes (83.7-107.1 %). As a proof of concept, the analytical method was applied to mouse tissues and biofluids, but especially to samples from in vitro fermentations with gut bacteria of the family Coriobacteriaceae. The developed method revealed that the species Eggerthella lenta and Collinsella aerofaciens possess bile salt hydrolase activity, and for the first time that the species Enterorhabdus mucosicola is able to deconjugate and dehydrogenate primary bile acids in vitro.