Separation of Non-alkaloid Toxin Lignans and New Flavonoid from Himalayan Mayapple (Podophyllum Hexandrum Royle) by High-Speed Counter-Current Chromatography and Their Anti-inflammatory Activity Evaluation

Abstract

Podophyllum hexandrum Royle (Berberidaceae) is reported from the Himalayan region and China. It is also known as the Himalayan Mayapple and is reported for the treatment of constipation, fever, jaundice, liver disorders, etc. Herein, the isolation of chemical constituents using high-speed counter-current chromatography (HSCCC) from the % Ethanol (EtOH) extract of the rhizomes of Himalayan Mayapple is reported. As a result, kaempferol 3-glucoside (1), quercetin-3-O-β-Dglucopyranoside (2), quercetin 3-O-β-D-glucopyranosyl-(1→6)-3-O-ethyl-β-D-glucopyranoside (3), kaempferol 3-O-β-D-glucopyranoside (4), α-peltatin(5), podophyllotoxin (6), 4'-demethylpodophyllotoxin (7), 4',5'-didemethylpodophyllotoxin (8), and kaempferol (9)were separated. Compounds 6-9 were separated by the normal HSCCC while 1-5 were obtained by the offline-recycling HSCCC using n-hexane, Ehyl Acetate, Methanol and Water % (HEMW at 1:9:4:6, v/v) solvent system. The pure components were tested in lipopolysaccharides-induced mice macrophage cells. Compounds 6 and 7 showed significant inhibition. The nitric oxide production was inhibited by compounds 6 and 7, effectively, with IC₅₀ values of 1.328 x 10^-6 and 2.851 x 10^-6 M, respectively. In this assay, kaempferol (9), a positive inhibitor expressively inhibited lipopolysaccharides-induced nitric oxide production