Preparation and Properties of Hydrogenated Rosin Ester Bonded Silica Gel Column

Hydrogenated rosin glycol acrylate(EGHRA) was bonded to the surface of thioglycolated silica gel(SH@SiO₂) by click chemical reaction to prepare a novel hydrogenated rosin acrylate ethylene glycol ester bonded silica gel(EGHRA@SiO₂) stationary phase. The synthesized stationary phase was characterized by elemental analysis, infrared spectroscopy, thermogravimetric analysis, N₂ adsorption-desorption experiments, and particle size distribution tests. The results showed that the EGHRA@SiO₂ stationary phase had an increased C content of 10.72 percentage points compared to SiO₂, a specific surface area of 270.06 m²/g, an average pore diameter of 6.16 nm, and an average particle size of 10 μm. A 250 mm×30 mm preparation column was filled by homogenization method, and a series of chromatographic evaluations were performed, demonstrating good flowability typical reversed-phase chromatographic characteristic as well as excellent stability and reproducibility of the EGHRA@SiO₂ preparation column. Tanaka tests showed that the spatial selectivity of EGHRA@SiO₂ column was superior to that of C18 column owing to the phenanthrene-like skeleton of the bonding monomer. Under reversed-phase chromatographic mode, EGHRA@SiO₂ preparation column exhibited ideal separation performance for the crude extracts of Taxus bark. Using the screened acetonitrile-water(volume ratio of 47∶53) mobile phase at a flow rate of 15 mL/min, the purity of paclitaxel(PTX), 10-deacetylbaccatin III(10-DAT), 7-xylosyl-10-deacetylbaccatin III(7-xyl-10-DAT) and cephalomannine(CEP) in the crude extract were increased by 91.52, 83.24, 62.99 and 55.85 percentage points after a single purification step of 1.0 mL injection sample, respectively.