Objective: To study the chemical components of Daphniphyllum calycinum and their anti-cholinesterase activity. Method: The chemical constituents of D.calycinum were isolated and purified by silica gel column chromatography

Sephadex LH-20 column chromatography

MCI column chromatography and preparative HPLC. Their structures were identified according to their physicochemical properties

nuclear magnetic resonance spectroscopy (NMR)

mass-spectrography (MS) and literature analysis. Furthermore

the anti-acetylcholinesterase and anti-butyrylcholinesterase activities of compounds were measured by UPLC-MS/MS method. Result: Eight compounds were isolated from D. calycinum and their structures were identified as 5-oxymaltol (1)

secodaphniphylline (2)

2

6-dimethyl-3-hydroxychromone (3)

deoxycalyciphylline B (4)

calyciphylline A (5)

daphnezzomine M (6)

and deoxyisocalyciphylline B (7). Compound 4 and 7 were showed anti-acetylcholinesterase activity

the IC50 values of compound 4 and 7 were (128.83±21.41) μmol·L-1 and (56.15±11.02) μmol·L-1

respectively. And compound 2

4

5 and 7 were showed anti-butyrylcholinesterase activity

the IC50 values of compound 2

4

5 and 7 were (0.31±0.15) μmol·L-1

(54.53±3.33) μmol·L-1

(811.17±22.49) μmol·L-1 and (8.13±0.78)μmol·L-1

respectively. Conclusion: Compound 1 and 3 were isolated from this plant for the first time. Compound 2 and 7 were showed strong anti-butyrylcholinesterase activity

and were the potential of cholinesterase inhibitors.