The Preparation Of Novel Stationary Phase For Hydrophilic Interaction Chromatography And Its Effect Of Analysis Chitooligosaccharides

In this paper, hydrophilic interaction liquid chromatography(HILIC) technique was applied to the separation of chitooligosaccharides(COS). Firstly, research status in HILIC area have been summarized, then we introduced the analysis, separation and bioactivities of COS.The experiment was consisted of three parts: first, 3-aminobenzeneboronic acid was selected as function monomer by equimolar continuous change method, then the steps and conditions of synthesis were optimized. The prepared Si O2@Cl and phenylboronic acid-silica were respectively characterized by scanning eletronmicroscopy, infrared spectroscopy, thermogravimetric analyzer and characterization of elements analyzer before the filling of the phenylboronic acid-silica column.In the second part, the retentive properties of the prepared column were investigated under HILIC mode in different mobile phase conditions. The retention time of nucleobases and nucleosides decreased along with increasing the water content in the mobile phase. It is a typical characteristic of the HILIC mode. In addition, the effects of salt concentration, p H and column temperature on the retention of nucleosides and nucleobases were respectively investigated. The results indicated the retention time of nucleosides and nucleobases were increased with salt concentration. The retention time of analytes were almost not affected by p H of the buffer. However, for most analytes, the higher column temperature, the shorter retention time was.At last, the proposed phenylboronic acid-silica column was applied to the separation of COS. When the mobile phase is ACN/water(75/25,v/v)-1.8% TEA with p H=7.8 at a flow of 1.0 m L/min, the column temperature is 25℃with refractive index detection,the COS was successfully separated resulting from 2 to 6 polymerization degree, demonstrating good effects of COS separation of the prepared column.