Studies On Chemical Constituents And Morphologic Dissection Of Huperzia Crispate

Huperzine A, a member of a new class of alkaloids, was first isolated from Huperzia serrata(Thunb.ex Murray)Trev in 1986.It is proved that Huperzine A can inhibit acetylcholinesterase, improve the memory of elderly people effectively, and has special effect on Aezheimer's disease(AD).With the aging of population and rising incidence of AD, more and more Huperzine A is needed. By far, there isn't any synthetic Huperzine A whose activity is equal to that of the natural Huperzine A. The amount of H. serrata in the whole of nature can hardly meet the demand of the market. So it is worthwhile discovering the new sources of Huperzine A.There is not any report on the isolation of Huperzine A from other kinds of plants of Huperzia genus except H. serrata. The researches on the determination method of main constituents and relationships between constituents and environment and seasons have also not been reported.H. crispate Ching is a unique species of Huperziaceae family in China. It is mainly distributed in Jiangxi, Hubei, Sichuan, Chongqing, Guizhou , Yunnan, Hunan. Its natural distribution is limited to the damp areas under the trees at an altitude of 900 -2600m. There has not been any report on chemical constituents, pharmacological action and other aspects at present.In this paper, the constituents of H. crispate, including alkaloids, flavonoids, and triterpenes, were studied. Huperzine A was first isolated from it. At the same time, a new compoud-Zhoubian A was found. The methods of determination of Huperzine A, Farrerol, and the total content of flavonoids in H. crispate were established. The relationships between the main constituents and the environment and seasons were studied. The morphologic dissection of H. crispate was also studied . The value of this research resides not only in providing scientific basis of exploring and cultivating the H. crispate ,but also in plugging the gaps in the research of Huperzia genus. The results were as follows:1 Studies on chemical constituents:Through the ethanol extract of H. crispate , Huperzine A (I) , Zhoubian A (II) , n-Triacontanol (III) , Serratenediol-3-acetate(IV) , Serratenediol(V) , B -Sitosterol(VI), Apigenin(VII), Farrerol(VIII) and Luteolin(IX) were separated by column chromatography and identified by phusicochemical and spectroscopic analyse.2 Studies on content of main constituents :1) A method of HPLC was set up to assay the content of Hup. A in H. crispate . Chromatographic conditions were: using Lichrospher Cis column( 150mm x 4.6mm), 40% methanol as mobile phase with 1ml min-1 of flow rate, detecting at 315nm . The column temperature was at 25癈 and injection volume was 20ul. The result showed the content of Hup.A in H. crispate was higher than that in H. serrata written in literature. The content of Hup. A varied in different samples .2 ) A method of HPLC was set up to assay the content of Farrerol in H. crispate . Chromatographic conditions were: using Lichrospher C18 column(150mm x 4.6mm), 30% methanol containing 0.4% phosphorus acid as mobile phase with 0.5ml min-1 of flow rate, detecting at 290nm . The column temperature was at 25C and injection volume was 20ul. The content of Farrerol in samples was different according to different seasons.3 ) The total content of flavonoids in H. crispate from different districts in different seasons were analyzed by spectrophotometryic method. The references were Rutine and Farrerol.The total content of flavonoids showed different numerical values in samples from different districts in different seasons. And the structure of flavonoids in H. crispate in different seasons is also different. In spring, the compounds in H. crispate were mainly flavanones , but in autumn, flavones were dominant.3 Relationship between main constituents and growth environment of H. crispate:The content of quick-acting potassium of the soil at the growing site was the positive correlating factor that correlates with the Hup. A in the plant. The annu...