Efficient isolation and purification of five products from microbial biotransformation of cinobufagin by high-speed counter-current chromatography.
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 4beta,12alpha-dihydroxyl-cinobufagin (1), 15 mg of 12beta-hydroxyl-cinobufagin (2), 8 mg of 5beta-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.
Ma
Xiao-Chi
XC
School of Pharmacy, Dalian Medical University, Dalian, P. R. China.
Xin
Xiulan
X
Zhang
Bao-jing
BJ
Deng
Sha
S
Yao
Ji-hong
JH
Wang
Chang-yuan
CY
Cui
Jian
J
Tian
Yan
Y
Liu
Ke-xin
KX
D000970:Antineoplastic Agents / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D001711:Biotransformation; D002018:Bufanolides / Q000737:chemistry / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D032661:Cordyceps / Q000378:metabolism*; D003377:Countercurrent Distribution; D004354:Drug Screening Assays, Antitumor; D006367:HeLa Cells; D006801:Humans; D008968:Molecular Conformation; D010936:Plant Extracts / Q000737:chemistry; D013237:Stereoisomerism
D000970:Antineoplastic Agents; D002018:Bufanolides; D010936:Plant Extracts; C002471:cinobufagin