Abstract: A new stationary phase for HPLC was prepared by one-pot reaction method,using natural active ingredient dioscin as raw material and 1,6-diisocyanate as spacer arm.The structure of new stationary phase was characterized by solid-state NMR techniques,infrared spectroscopy,elemental analysis,thermogravimetric analysis and scanning electron microscope (SEM).The results confirmed that dioscin had been immobilized onto the surface of silica gel successfully with suitable thermostability and chemical resistance.The resulting stationary phase contained 127.6 μmol/g dioscin ligand.The chromatographic performance of the new stationary phase were evaluated using different kinds of solute probes,including non-polar compounds,alkaline compounds and acid compounds.The results showed that the new stationary phase had a typical reversed phase chromatographic property,meanwhile it can provide various interactions with solutes,such as hydrogen bonding,dipole-dipole,π-π, etc.It was good on the separation of multi-component natural products.The HPLC fingerprint of PNS (Panax notoginseng saponins) API (Active Pharmaceutical Ingredient) on the dioscin-bonded silica gel stationary phase was established.The mobile phase consisted of acetonitrile and water.The results provided technologic support for the quality control of PNS products.It was a new thinking for preparing stationary phase with natural active ingredient and separation complex ingredient in natural products.

Key words: dioscin-bonded silica gel stationary phase, stationary phase characterization, PNS API, HPLC, fingerprint