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Rapid Isolation and Purification of Phorbol Esters from Jatropha curcas by High-Speed Countercurrent Chromatography
journal contribution
posted on 2015-03-18, 00:00 authored by Wan Hua, Huiling Hu, Fang Chen, Lin Tang, Tong Peng, Zhanguo WangIn
this work, a high-speed countercurrent chromatography (HSCCC)
method was established for the preparation of phorbol esters (PEs)
from Jatropha curcas. n-Hexane–ethyl acetate–methanol–water (1.5:1.5:1.2:0.5,
v/v) was selected as the optimum two-phase solvent system to separate
and purify jatropha factor C1 (JC1) with a purity of 85.2%,
as determined by HPLC, and to obtain a mixture containing four or
five PEs. Subsequently, continuous semipreparative HPLC was applied
to further purify JC1 (99.8% as determined by HPLC). In addition,
UPLC–PDA and UPLC–MS were established and successfully
used to evaluate the isolated JC1 and PE-rich crude extract. The purity
of JC1 was only 87.8% by UPLC–UV. A peak (a compound highly
similar to JC1) was indentified as the isomer of JC1 by comparing
the characteristic UV absorption and MS spectra. Meanwhile, this strategy
was also applied to analyze the PE-rich crude extract from J. curcas. It is interesting that there may be more
than 15 PEs according to the same quasi-molecular ion peaks, highly
similar sequence-specific fragment ions, and similar UV absorption
spectrum.