Rapid Purification of Huperzine A and B with Two Polystyrene based Resins by Preparative Low‐Pressure Liquid Chromatography.

A rapid, facile, and environmentally friendly process was developed for purification of huperzine A (HupA) and huperzine B (HupB) from the Chinese traditional herb Huperzia serrata . The process consisted of two steps of preparative low-pressure liquid chromatography (LPLC) on two polystyrene based resins, successively. The first step removed a large amount of impurities and captured HupA and HupB using Amberlite XAD-4 from the herbal extraction prepared by 1% aqueous H 2 SO 4 . This step was more efficient than multi-cycle liquid-liquid extraction as an initial separation step. The second step was able to separate HupA and HupB, employing a novel uniform polystyrene based porous microsphere named PST as the packing material. The PST column demonstrated a better separation and shorter run time compared with a commercial C 18 column. The mobile phases used in both LPLC's simply consisted of water and ethanol that are considered to be of low toxicity. Combination of the XAD-4 and PST chromatography and one step of crystallization enabled purification of HupA and HupB from 0.18% and 0.08% to 98.2% and 98.8%, respectively, with total recoveries of 82.8% and 84.3%. [ABSTRACT FROM AUTHOR]