| Ephedra polysaccharide has significant immune inhibition,and confirmed that it has good therapeutic effect on autoimmune diseases.In view of the important biological activity of Ephedra,the quality evaluation is particularly important.The quality control of polysaccharides was realized by controlling the content of polysaccharide and the distribution of molecular weight at home and abroad.This project take Ephedra polysaccharide as research object,in order to establish an analysis method with UPLC-MS chromatography to achieve the quality control of Ephedra polysaccharide.This paper includes the following main contents:1.Study on direct detection of the non-derivatives Ephedra polysaccharide based on HILIC-LC-MS.To establish a simple,fast,accurate,high sensitivity and good reproducibility method for the determination using the method of hydrophilic interaction liquid chromatography(HILIC)combined with mass spectrometry.This method was proposed to detect the(0,3)-cross ring fragmentation by mass spectrometry,and no pre-column derivatization.This method has the advantages of high specificity,simple,accurate and reliable,and can be used for fast detection of monosaccharides and oligosaccharides content of plant polysaccharide.However,this method has some limitations,when increase the number of target compounds,it can not achieve the purpose of complete separation.So,we try to establish a new UPLC-MS method with PMP derivatization.2.Study on detection of the PMP derivatization Ephedra polysaccharide based on LC-MS.Cortecs C18 is a general reversed-phase high performance liquid chromatography column,which use advance solid core particle and column filling technique,can significantly improve the separation of difficulty carbohydrate compared with HILIC column on the same conditions.Using UPLC-MS method,with 1.6 μm solid particles UPLC-Cortecs column,in the negative ion multiple reaction monitoring(MRM)mode for determination.To establish a UPLC-MS method to simultaneous determination the monosaccharide and oligosaccharide by PMP derivatization.Meanwhile,analysis the composition and content of monosaccharide and oligosaccharide in acid hydrolysis and enzymatic hydrolysis Ephedra samples with this method.This method can be used to predict and control the quality of Ephedra polysaccharide.3.Using multivariate statistical methods such as cluster analysis and principal component analysis,to distinguish and identify four kind of different species and different parts of Ephedra,and the results show that the established method can be divided Ephedra different species and provides a new basis for the quality control of Ephedra.This experiment provide feasible scientific method and the solid theoretical basis for the quality control of Ephedra polysaccharide.At the same time,it also provides reference basis for the quality control and evaluation of traditional Chinese medicine polysaccharides. |
