An LC-MS/MS method development and validation for quantification of metabolites in sulfur metabolism

Sulfur metabolites play critical roles in cellular function. Changes in the levels and metabolism of sulfur compounds, were associated with cancer, which have led to the need to make precise, accurate and fast determination of the amount of these compounds in the clinic. The concentrations of specific metabolites should be known accurately and precisely, especially when searching for answers to biological questions. Such analyses can be obtained through targeted metabolomic studies. Because of their superior selectivity and sensitivity, LC-MS/MS approaches are widely employed in these studies. A new LC-MS/MS method for the simultaneous analysis of metabolites with a short analysis time (12 min) and metabolites involved in sulfur metabolism without the need for a derivatization step is presented in the thesis study. The method was found linear for metabolites ranging from 0,05 to 10 µg/mL. The developed method was found to be selective, precise, accurate, sensitive, linear, and robust with validation studies. The amounts of sulfur metabolites were determined by applying the developed method to cell lines of human hepatocellular carcinoma (HepG2), human lung adenocarcinoma (A549), human healthy lung epithelium (BEAS-2B), human colorectal adenocarcinoma (Caco-2), and human healthy colon epithelial (FHC).