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.
Keywords: Bile; Bile acids; Design of experiments; Serum; Short-chain fatty acids; Stable isotope labeling derivatization; Ultrahigh performance liquid chromatography–tandem mass spectrometry.
Published by Elsevier B.V.