Purification of Active Component in Wasabi Leafstalk (Wasabia japonica MATSUM.) Extract in Stimulating Bone Calcification in Vitro

Abstract

The active component in wasabi leafstalk (Wasabia japonica MATSUM.), which has a stimulatory effect on bone calcification in mouse calvaria tissue culture in vitro, was purified. Wasabi leafstalk extract was obtained from a homogenate with 20% ethanol. The active component, which was found in ethanol extraction, was purified by gel filtration chromatography with HiLoad 26/80 Superdex 30 pg column and reversed-phase chromatography (RPC) on a resource 15 RPC 3 ml column. The result of ESI mass spectra of the purified active component showed that its material had a molecular weight of 158. The absorption spectra of the material gave the maximum absorption at a wavelength of 221 nm. The material in wasabi leafstalk extract differed from genistein and 17β-estradiol, which can stimulate bone calcification in vitro, and had a comparatively lower molecular weight. This study demonstrates that the material with a low molecular weight of 158 is an active component in wasabi leafstalk which stimulates bone calcification.