Simultaneously preparative purification of Huperzine A and Huperzine B from Huperzia serrata by macroporous resin and preparative high performance liquid chromatography

Huperzine A (HupA) and Huperzine B (HupB) are natural alkaloids existed in Lycopodium plants. They both have potential clinical application for treating Alzheimer's Disease (AD). For the purpose of better utilizing the limited plant resources, a quick and low cost method to separate and purify HupA and HupB from Huperzia serrata (Thunb. ex Murray) was established in this paper. Low polarity macroporous resin SP850 was selected from eight kinds of resins during initial purification. Trifluoroacetic acid (TFA) was proved to be the best acid modifier reagent among all acids used in our experiment for improving separation. HupA and HupB were baseline separated on a C18 column by preparative high performance liquid chromatography (Preparative HPLC), the optimal gradient mobile phase system contained methanol increasing from 15% (v/v) to 35% (v/v) and 0.1% (v/v) TFA within the water. The purity of HupA and HupB obtained was 99.1% and 98.6%, respectively, and the total recovery for them was 83.0% and 81.8%, respectively.

► We simultaneously purify HupA and HupB by reversed C18 column from Chinese herbs.
► We compare four kinds of acids of their influence to the separation of HupA and HupB.
► Macroporous resins were tested and SP850 was found best for initial purification.
► Trifluoroacetic acid performed best to eliminate the trailing problem.