A simultaneous analysis strategy of glycerophospholipids and lipid mediators based on secondary extraction scheme

Lipid peroxidation plays an important role in physiological and pathophysiological conditions; it occurs mainly in the vicinity of the membrane. Glycerophospholipids are indispensable for normal structure of membranes. Lipid mediators are final products of lipid peroxidation. Glycerophospholipids as a source of lipid mediators are closely related with lipid mediators in lipid peroxidation process. This study developed a strategy for simultaneous analysis of glycerophospholipids and lipid mediators in a sample based on a secondary extraction scheme. Due to differences in physicochemical properties of glycerophospholipids and lipid mediators, we used different solvents and methods to extract glycerophospholipids and lipid mediators based on a sample. In addition, we compared three methods of extracting glycerophospholipids to select the best extraction method. In this study, a UPLC-MRM absolute quantification strategy for three deuterated internal standards of lipid mediator was established. The calibration range of all standards is linear, and the correlation coefficients are greater than 0.999. Quantitative range, precision, and accuracy of all analytes-based analysis method meet requirements of method validation. And the accuracy and precision of this method for glycerophospholipids meet the requirements of method verification. It is satisfactory to apply this method in the analysis of the changes of enzymes in lipid peroxidation during the treatment of asthma with dexamethasone. The quantitative results of phospholipids and lipid mediators in mouse lung tissue were obtained by this method, indicating that dexamethasone may inhibit the activity of phospholipase As enzyme and thus slow down the lipid peroxidation process. This work may be of great significance for exploring the changes of enzymes in lipid peroxidation metabolism.