A precise and selective reversed phase high-performance liquid chromatographic method was developed for quantifying huperzine A (HupA) in samples of the Huperziaceae in China. This method was used to quantify the levels of HupA in samples of Huperzia serrata collected from a single population at different times of the year, in different organs of the same H. serrata plant, and from different geographical locations of H. serrata plants in China. For different species of Huperziaceae, the highest content of HupA was found in Phlegmariurus carinatus. Members of the genus Phlegmariurus possessed higher levels of HupA than Huperzia species. H. serrata plants growing in humid forests contained significantly more HupA than plants growing in less humid environments. Finally, HupA content varied significantly by season, with the highest levels being found in mid fall and the lowest levels in early spring, suggesting that HupA is turned over in the plant.