Phospholipid Metabonomic Research Related With Diabetes Nephropathy

In the dissertation, based on the LC/MS analysis, specific analytical methods for simultaneous determination and quantification of seven major phospholipids classes in human blood ware developed by normal phase high performance liquid chromatography tandem mass spectrometry.Based on the results of determination and quantification comprehensive researches about the relationship of diabetic nephropathy (DN) and phospholipids metabolism disorder have been done. From two different level mogensen phase and TCM diagnosis the relationship of DN and phospholipids metabolism was explained .The research established a new way for mechanism research, diagnosis and treatment of deseases.First of all,a rapid and exact analytical method for simultaneous determination of compound with similar structure was developed by HPLC-ion trap MS. The optimal separation was achieved Isocratic elution method was used for better repeatability and no balance time. The seven major phospholipids classes in human blood that were detected including phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylglycerol (PG) , phosphatidylinositol (PI) phosphatidylcholine (PC), lysophosphatidylcholine (lysoPC) and sphingomyelin (SM).Through researching the fragments of the external standards the phospholipid in blood samples can be determined.Every phospholipids class contains many molecular species which have similar structure. The structure of phospholipids molecular species was identified by ion-trap MSn which produced ion fragments. With this method, hundreds of phospholipids have been identified.The qualification was completed by TOF-MS which shows good accuracy. Through the accurate quantification of one representative phospholipids molecule in each class, a method for simultaneous quantifying hundreds of molecular species in seven major classes was established. The intra-day and inter-day precision and recovery had been investigated in detail. The method satisfied the qualification requirement.Based on the result of qualification,from two different level mogensen phase and TCM diagnosis the Clinical samples have been researched.Through analyzing the differences for different phases ,the variation of concentration of different phospholipids species and total phospholipids have been seen to find out the biological mechanism.The research mode of metabonomics based on PCA was also used in the phospholipid analysis to find out the relationship of DN and phospholipid metabolism.The dissertation provided new ways for mechanism research of of DN and clinical diagnosis.