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Efficient isolation and purification of five products from microbial biotransformation of cinobufagin by high-speed counter-current chromatography

✍ Scribed by Xiao-Chi Ma; Xiulan Xin; Bao-jing Zhang; Sha Deng; Ji-hong Yao; Chang-yuan Wang; Jian Cui; Yan Tian; Ke-xin Liu

Publisher
John Wiley and Sons
Year
2010
Tongue
English
Weight
203 KB
Volume
33
Category
Article
ISSN
1615-9306

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✦ Synopsis

Abstract

An efficient separation method of using high‐speed counter‐current chromatography was successfully established to directly purify cytotoxic transformed products of cinobufagin by Cordyceps militaris. The two‐phase solvent system composed of n‐hexane–ethyl acetate–methanol–water (4:6:3:4, v/v) was used in high‐speed counter‐current chromatography. A total of 9 mg of 4β,12α‐dihydroxyl‐cinobufagin (1), 15 mg of 12β‐hydroxyl‐cinobufagin (2), 8 mg of 5β‐hydroxyl‐cinobufagin (3), 12 mg of deacetylcinobufagin (4) and 6 mg of 3‐keto‐cinobufagin (5) were obtained in a one‐step separation from 400 mg of the crude extract with purity of 98.7, 97.2, 90.6, 99.1 and 99.4%, respectively, as determined by HPLC. Their chemical structures were identified on the basis of ^1^H‐NMR and ^13^C‐NMR technology. All products (1–5) showed the potent activities against human carcinoma cervicis (Hela) and malignant melanoma (A375) cells in vitro.

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