Abstract: Piperine was separated from crude ethanolic pepper extract by high-speed countercurrent chromatography (HSCCC) with a solvent system composed of n-hexane-acetic ether-methanol-water (1:1:1:1, V/V). The method allowed the separation of 1.58 g of individual piperine with a purity of 98.72% from 5 g of crude ethanolic pepper extract. The separated piperine was structurally identified by ESI-MS, 1H NMR and 13C NMR. The method is of high sample loading capacity, high separation efficiency, and great importance for piperine applications in the food and medical fields.

Key words: high-speed countercurrent chromatography (HSCCC), piperine, pepper