Study On The Composition And Content Of Monosaccharides In Plant Polysaccharides Based On High Performance Anion Exchange Chromatography-High Resolution Mass Spectrometry (HPAEC-HRMS) Technique

Objective:To establish a method for the determination of plant monosaccharide composition and content in polysaccharides by high performance anion-exchange chromatography coupled with quadrupole/electrostatic field orbitrap high resolution mass spectrometry.1.To screen suitable on-line sensitizing reagents to enhance the response value and sensitivity of sugars,in view of the deficiency of low sensitivity of sugars in quadrupole-electrostatic field/orbit trap high resolution mass spectrometry analysis;2.To establish an analytical method for the simultaneous determination of nine monosaccharides by high performance anion-exchange chromatography-quadrupole/electrostatic field orbit trap mass spectrometry and apply it to the determination of monosaccharide composition of polysaccharides in Lonicera japonica flos and lonicerae flos.Methods:1.To compare the sensitization effect of ammonium formate,sodium acetate solution and lithium chloride solution as online sensitization reagents,and to optimize the concentration and flow rate of the sensitization reagents,to investigate the sensitivity changes of nine monosaccharides in quadrupole-electrostatic field/orbitrap high resolution mass spectrometry under different post-column addition solution conditions,and to screen the most suitable sensitization reagents;2.To establish a method for the determination of monosaccharide composition in Lonicera japonica flos and lonicerae flos polysaccharides by high performance anion-exchange chromatography-quadrupole/electrostatic field orbit trap high resolution mass spectrometry(HPAEC-HRMS),the extraction conditions and hydrolysis conditions of Lonicera japonica flos and lonicerae flos polysaccharides were optimized by single-factor experiments,and the monosaccharide composition was analyzed by HPAEC-HRMS using the optimized sensitization conditions in the first part.Results:1.By comparing different post-column addition solutions,a post-column solution condition of 0.2 m M sodium acetate in aqueous methanol(methanol:water=9:1)at a flow rate of 0.04 m L/min was finally selected.2.The optimized post-column addition solution conditions in Part I were used to establish a method for the determination of the monosaccharide composition in the polysaccharides of Lonicera japonica flos and lonicerae flos by high performance anion-exchange chromatography with quadrupole/electrostatic field orbitrap high resolution mass spectrometry.The method showed good linearity(R~2>0.99)in a range of concentrations,with the average recoveries of 91.9-117.4%and intra-day and inter-day precision not exceeding 9.00%;the limits of detection(LODs)were 0.04-2 ng/m L and the limits of quantification(LOQs)were 0.1-5 ng/m L.Conclusion:In this paper,the sensitivity of monosaccharides in high resolution mass spectrometry was greatly improved by adding metal salt solution after the column,which was used to establish a high performance anion-exchange chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry method for the determination of monosaccharide composition of plant polysaccharides in Lonicera japonica flos and lonicerae flos,which is easy to operate,rapid and sensitive.It can not only provide a reference for the structural study of polysaccharides in Lonicera japonica flos and lonicerae flos,but also provide a theoretical basis for the quality control of other products containing polysaccharides.