PREPARATIVE SEPARATION AND PURIFICATION OF ALKYLAMIDES FROM ZANTHOXYLUM BUNGEANUM MAXIM BY HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

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Abstract
High-speed counter-current chromatography (HSCCC) was successfully applied to preparative isolation and purification of alkylamides, the tingling-taste components, from pericarp extract of Zanthoxylum bungeanum Maxim (Dahongpao). With the lower phase of n-hexane-ethyl acetate-methanol-water (7:3:5:5, v/v) eluted at a flow rate of 1.5 mL · min−1, and the revolution speed at 850 rpm, 0.19 g of alkylamides in purity of 98.0% by the relative peak area of HPLC was obtained from 1.0 g of extract in one run within 2 hr. On the basis of MS, IR, and 1H NMR analysis, the product of alkylamides was confirmed to be composed of hydroxy-α -sanshool, hydroxy-β-sanshool, and hydroxy-ϵ-sanshool. Furthermore, using the HSCCC product as external standard, alkylamide in the extract was determined to be in total of 21.41%, among which hydroxy-α-sanshool, hydroxy-β-sanshool, and hydroxy-ϵ-sanshool were in 15.69%, 4.76%, and 0.96%, respectively.

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