Simultaneous Isolation And Purification Of Huperzine A And B From Traditional Chinese Herb Huperzia Serrata By Integrated Low-Pressure Liquid Chromatography

The purpose of this study was to develop an efficient low-pressure chromatography process for isolation and purification of natural products from plant, through selection of the chromatographic media for high resolution and low operation pressure, and integration of different chromatographic steps for high purity and recovery of target products. This dissertation described simultaneous isolation and purification of huperzine A and B from traditional Chinese herb Huperzia serrata simply and efficiently by solvent extraction and combination of three steps of column chromatography.This paper started with develepment of a rapid, precise and reproducible routine HPLC assay for keeping track and quantification of the compounds through the separation process, by selecting the Inertsil CN-3 as the HPLC column due to its good selectivity for the, and the solid phase extraction as the sample pretreatment method.A simple and efficient solvent extraction was established. The herb powder of Huperzia serrata was soaked with 1% aqueous H2SO4 twice for 24 h at ambient temperature at the solid-liquid ratio of 1: 10 (w/v) and 1: 5 (w/v), respectively. After extraction, the recoveries of huperzine A and B in the extract solution were 95.2% and 96.8% respectively.Macroporous adsorption resin chromatography was used to capture the huperzines and concentrate the extract solution instead of multi-steps of liquid-liquid extraction. CBJ-23 resin was selected by comparing with other resins for column chromatography. After a step of CBJ-23 adsorption separation, huperzine A and B were purified from 0.188% and 0.079% to 1.72% and 0.734% with the recovery of 92.3% and 93.1%...