AbstractOne new fawcettimine‐type alkaloid (1), one new miscellaneous‐type alkaloid (2), four new lycodine‐type alkaloids (3–6), and eight known ones (7–14) were isolated from the whole plants of Huperzia serrata. Their structures and absolute configurations were elucidated based on spectroscopic data, X‐ray diffraction, ECD calculation and Mosher's method. Compound 1 was a rare C18N2‐type Lycopodium alkaloid, possessing serratinine skeleton with an amide side chain in C‐5. The absolute configuration of the 18‐OH of compounds 4–6 were first determined by Mosher's method. Moreover, compounds 1–14 were assayed anti‐acetylcholinesterase effect in vitro, and compound 7 showed significant anti‐acetylcholinesterase activity with an IC50 value of 16.18±1.64 μM.