Application of Centrifugal Partition Chromatography for Bioactivity-Guided Purification of Antioxidant-Response-Element-Inducing Constituents from Atractylodis Rhizoma Alba

Abstract

Activity-guided separation of antioxidant response element (ARE)-inducing constituents from the rhizomes of Atractylodis Rhizoma Alba was performed by the combination of centrifugal partition chromatography (CPC) and an ARE luciferase reporter assay. From 3 g of the active n-hexane fraction, one polyacetylene, (6E, 12E)-tetradeca-6,12-dien-8,10-diyne-1,3-diyl diacetate (47.3 mg), and two sesquiterpenes, atractylenolide I (40.9 mg), and selina-4(14), 7(11)-dien-8-one (6.0 mg) were successfully isolated by CPC with n-hexane-ethyl acetate-methanol-water (8: 2: 8: 2, v/v). The chemical structures of the isolated compounds were determined by H-1- and C-13-NMR and ESI-MS. Among the isolated compounds, (6E, 12E)-tetradeca-6,12-diene-8,10-diyne-1,3-diol diacetate and selina-4(14), 7(11)-dien-8-one increased ARE activity 32.9-fold and 16.6-fold, respectively, without significant cytotoxicity, when 5 mu M sulforaphane enhanced ARE activity 27.1-fold. However, atractylenolide I did not increase ARE activity at 100 mu M, and showed cytotoxicity at concentrations over 10 mu M.