LC-MS/MS-Based Lipidomics Approach For Type2Diabetes Mellitus

A high-throughput lipidomics analysis method has been implemented by using rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS) with intensive validation of sensitivity, precision and stability. Analysis method based on RRLC-MS/MS has been applied to investigate a large scale of Type2Diabetes Mellitus (T2DM) subjects and many potential lipid biomarkers for diagnosis and treatment have been found.Optimized serum preparation method and RRLC-MS/MS analysis method was applied to investigate T2DM subjects to find biochemically significant lipid biomarkers. After a large scale of serum samples were studied,42potential biomarkers related to diagnosis of T2DM and52potential biomarkers related to the treatment of T2DM were found. Subsequently, high resolution MS and MS/MS analyses, as well as inter-metabolite correlation analysis were performed for the identification of the lipid metabolites of interest. Until now,35and49potential biomarkers have been identified, respectively. The further metabonomic pathway analysis indicated that occurrence and development of T2DM was related to glycerophospholipid, triglyceride, cholesteryl ester and sphingolipid metabolism perturbation.1. Serum preparation and analysis method based on RRLC-MSWe developed and validated a RRLC-MS method with an integrated ionization approach for the determination of endogenous lipid metabolites in human serum, in which the samples were pretreated by lipid extraction with methylene chloride-methanol (2:1, V/V) mixture solvent and introduced to RRLC-MS under intensive optimization of chromatography conditions and mass spectrometry parameters. The method had also been intensively validated for its sentivity, precision and stability, ensuring the method is appropriate for untargeted serum lipidomics study. The lipid extraction solvent was important to lipidomics research. Systematic analysis of the effect of lipid extraction solvent on extraction completion and precision confirmed that methylene chloride-methanol (2:1, V/V) was the optimal lipid extraction solvent. In addition, the methodology of this method was investigated and the results demonstrated that this method had good sensitivity, high precision and high stability, and it could be applied to the lipidomics analysis of serum. Therefore, an accurate untargeted lipidomics based on RRLC-MS was developed to analyze serum samples.2. RRLC-MS/MS-based lipidomics study on T2DMA lipidomics strategy method of untargeted serum metabonomics based on RRLC-MS/MS and multivariate statistics has been implemented to find biologically significant lipid metabolite biomarkers in T2DM. A large number of biological samples including80T2DM patients and28healthy volunteers were analyzed to obtain abundant information and find the potential lipid biomarkers that were closely associated with primary diagnosis and treatment of T2DM. Until now,94potential lipid biomarkers have been found by RRLC-ESIMS/MS in positive and negative ion modes in this lipidomics study, which demonstrating that these abnormal metabolites were related to metabolic disorders in T2DM patients. Up to now,35potential biomarkers related to primary diagnosis of T2DM have been identified by using high resolution MS and MS/MS analyses, as well as fragmentation rules of metabolites, including2fatty acids (FA),6triglycerides (TG),3cholesteryl esters (CE),1sphingomyelin (SM),23glycerophospholipids (GL), et al.49potential biomarkers related to development of T2DM have also been identified, including2fatty acids (FA),1triglyceride (TG),2cholesteryl esters (CE),11sphingomyelins (SM),33glycerophospholipids (GL), et al. The level of GL and TG were significantly increased and the level of CE was apperantly decreased in T2DM serum with the early occurrence of T2DM while the level of SM and GL were dramatically increased in T2DM serum with the development of T2DM. Then the metabolic correlation and pathway analysis have been studied, suggesting occurrence and development of T2DM are closely related with glycerophospholipid, triglyceride, cholesteryl ester and sphingolipid metabolism perturbation.