Theacrine was prepared

isolated and purified from Camellia assamica var. kucha by highspeed countercurrent chromatography. A twophase solvent system composed of hexanedichloromethanemethanolwater (1∶5∶4∶2

v/v/v/v) was selected with comprehensive analysis of the settling time

the ratio of up and down phases volume and the K value. Five mg of theobromine

389 mg of theacrine and 41 mg of caffeine with their purities over 99% were obtained from 222 g of the crude extract of Camellia assamica var. kucha under the condition of a flow rate of 20 mL/min

800 r/min and the detection wavelength of 278 nm. The obtained fractions were analyzed by high perform liquid chromatography

compared with standard substance and identified by MS

1H NMR and 13C NMR methods. Theacrine was the first time isolated from Camellia assamica var. kucha by HSCCC

which is a simple

rapid and effective method.