Aim: Objective of this study is to develop a robust multi-matrix LC-MS/MS for the quantitation of endogenous short-chain fatty acids (SCFA) biomarkers in human plasma and urine. Methods: Developed method utilizes stable isotope-labeled internal standards, high-throughput derivatization procedure for sample preparation and LC-MS/MS analysis using multiple reaction monitoring transitions in positive electrospray ionization mode. Results: Surrogate matrix method was used for quantitation. Accuracy, precision, parallelism, curve linearity, derivatization efficiency, stability and recovery were all evaluated, and the results were well within the acceptable criteria. Conclusion: SCFA levels in human plasma and urine of inflammatory bowel disease patients versus non-disease subjects were quantified and compared by LC-MS/MS.
Keywords: 2-methylbutyric acid; 3-methylvaleric acid; biomarker; butyric acid; caproic acid; derivatization; human plasma and urine; isobutyric acid; isocaproic acid; isovaleric acid; liquid–liquid extraction; propionic acid; reversed-phase LC–MS/MS; short-chain fatty acid; valeric acid.