Determination of Malonyl-CoA Levels and Distribution in Rat Brain Tissue Using Liquid-Liquid Extraction and LC/MS/MS

摘要

Our objective was to develop and implement a robust and reproducible assay with a sufficient limit of detection for reporting malonyl-CoA levels in the brain with fast data turn-around in support of discovery efforts. LLE dramatically increased the recovery of malonyl-CoA from brain tissue, a vital improvement in sensitivity that was instrumental in allowing accurate detection of very low levels of malonyl-CoA in brain. As a demonstration of the usefulness of the assay for routine PD screening, a highly potent ACC inhibitor was shown to decrease malonyl-CoA levels not only in peripheral tissues (liver and skeletal muscle) but in brain as well. This method provides a unique opportunity for routinely and rapidly monitoring brain malonyl-CoA levels, a parameter that is important for evaluating therapeutic strategies for the treatment of obesity and metabolic syndrome.