Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 1, January 2008, pp. 81-85
Preparative Isolation of Bufalin and Cinobufagin
from Chinese Traditional Medicine ChanSu
Zhen Yang, Huafei Luo, Hao Wang, and Huimin
Hou
National Pharmaceutical Research Center, Shanghai Institute of
Pharmaceutical Industry, Shanghai, P.R. China
In this investigation, a method is developed for
the isolation and purification of bufadienolides (resibufogenin,
cinobufagin, and bufalin) from the Chinese traditional medicine
ChanSu. The crude ChanSu extract is prepared with solvent refluxing
under optimal conditions. A new chromatographic approach for
separating bufadienolides utilizes silica gel column chromatography
with isocratic elution using cyclohexane–acetone (5:1)
as the mobile phase to separate resibufogenin, followed by reversed-phase
C18 preparative HPLC column using isocratic elution of methanol–water
(72:28) to separate bufalin and cinobufagin. Combining these
two methods, bufalin and cinobufagin could be completely separated
in high purity and recovery, and the amounts obtained were 1.9
and 3.1 g, respectively, from 500 g of ChanSu. These two compounds
have been identified by mass spectrometry and 1H NMR, and their
purities were quantitated by HPLC at 99% and 98%, respectively.
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