稳定同位素标记衍生 UHPLC-MS/MS 法检测/定量生物体液中初级/次级胆汁酸的实验设计。
Experimental design of a stable isotope labeling derivatized UHPLC-MS/MS method for the detection/quantification of primary/secondary bile acids in biofluids.
机构信息
College of Pharmaceutical Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan.
Department of Gastroenterology, Japanese Red Cross Nagoya Daini Hospital, 2-9 Myoken-Cho, Showa-Ku, Nagoya 466-8650, Japan.
出版信息
J Pharm Biomed Anal. 2022 Feb 5;209:114485. doi: 10.1016/j.jpba.2021.114485. Epub 2021 Nov 25.
An efficient analytical platform is required to characterize the human metabolome in pathology. For this purpose, ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) combined with chemical derivatization stands out as one of the most powerful techniques. A targeted metabolomics platform for 11 bile acids (BAs) profiling in human serum and bile samples using a stable isotope labeling derivatization (SILD) was applied. For SILD, the design of experiments (DoE) was employed to optimize the reaction conditions such five factors in three levels. The sample preparation built upon a liquid-liquid extraction requiring small volumes (20 μL). In application, the relation between the BA and short-chain fatty acid levels in human serum and bile samples from patients with bile duct diseases were investigated. The proposed method offers significant utility in the large-scale biological analyses of hepato-biliary-pancreatic-related diseases.
需要一个高效的分析平台来对病理学中的人类代谢组进行特征分析。为此，超高效液相色谱与串联质谱（UHPLC-MS/MS）结合化学衍生化成为最强大的技术之一。应用了一种基于稳定同位素标记衍生化（SILD）的针对人血清和胆汁样本中 11 种胆汁酸（BA）的靶向代谢组学平台进行分析。对于 SILD，采用实验设计（DoE）来优化反应条件，例如五个因素三个水平。样品制备基于需要小体积（20 μL）的液-液萃取。在应用中，研究了患有胆管疾病的患者的血清和胆汁样本中 BA 和短链脂肪酸水平之间的关系。该方法在肝胰胆相关疾病的大规模生物分析中具有重要的应用价值。
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