Investigationof Monosaccharides And Disaccharides With 1-Phenyl-3-Methyl-5-Pyrazolone Derivatization Reagent By Mass Spectrometry

Carbohydrate compounds play an important role in the regulation of life activities.With the development of analytical technology,mass spectrometry(MS)has become one of the indispensable tools to characterize the structure of carbohydrates due to its advantages of low sample consumption,high speed,and high sensitivity.However,the disadvantages of low ionization efficiency and incomplete fragment information in mass spectrometry analysis of carbohydrate compounds make its application limited.The method of derivatization can effectively improve the ionization efficiency of mass spectrometry and the accuracy of fine structure analysis.Traditional derivatization methods such as reductive amination require cumbersome desalination steps and are not suitable for the analysis of acid sugars.The PMP(1-phenyl-3-methyl-5-pyrazolone)reagent has the advantages of simple operation,no need for desalting,wide application range and mild reaction conditions for the derivatization of sugar.In this thesis,based on the derivatization of PMP reagents,combined with mass spectrometry,the analysis and application of monosaccharides and disaccharides are systematically studied.1.A reagent pair based on the derivatization reagent PMP and its structural analogue MPMP(1-(4-methylphenyl)-3-methyl-5-pyrazolone)was designed.Combined with the new non-isotopic paired labeling(NIPL)method of liquid chromatography-mass spectrometer,the qualitative and quantitative of 12 kinds of monosaccharides have been realized.This method used PMP-labeled monosaccharides as a one-to-one internal standard,and successfully determined the 12 kinds of monosaccharides in bladder cancer urine samples,and combined with statistical analysis methods,the monosaccharide biomarkers GaINAc and galactose for the diagnosis of bladder cancer were obtained for the first time.2.The multi-stage mass spectrometry results of the PMP derivatization products of 16 disaccharide isomers[M+H]+show that the product ion and fragment intensities will vary with the structure of the disaccharide.This difference combines principal component analysis(PCA)and orthogonal projections to latent structures discrimination analysis(OPLS-DA)in statistical analysis to successfully construct a statistical model.This model can be used to distinguish 16 disaccharide isomers.And this model can be used to distinguish three kinds of rice wine and the relative quantitative analysis of maltose and isomaltose in rice wine.