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Purication of betulinic acid from Eugenia florida (Myrtaceae) by high‐speed counter‐current chromatography
Author(s) -
Frighetto Nelson,
Welendorf Rodolfo Max,
da Silva Ana Maria Pereira,
Nakamura Marcos Jun,
Siani Antonio Carlos
Publication year - 2005
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.860
Subject(s) - chemistry , betulinic acid , countercurrent chromatography , chromatography , ethyl acetate , solvent , aglycone , triterpene , fraction (chemistry) , myrtaceae , glycoside , organic chemistry , high performance liquid chromatography , botany , medicine , genetics , alternative medicine , pathology , biology
A high yield of betulinic acid (up to 17% from the ethanolic extract) was found in the leaves of Eugenia florida collected in south‐eastern Brazil, making this species a potential commercial source of the title compound. Extracts of E. florida were subjected to solvent partition, and rapid high‐speed counter‐current chromatography (HSCCC) was applied to the semi‐crude extracts to afford betulinic acid in high purity. The mobile and stationary phases were derived from the two‐phase solvent system composed of n ‐hexane–ethyl acetate–methanol–water (10:5:2.5:1). The developing solvent system (stationary and mobile phases) for optimum HSCCC separation was chosen by dissolving the fraction to be chromatographed in the proposed solvent mixture and determining the amount of betulinic acid in each phase by densitometric TLC. Puried betulinic acid was characterized by 13 C‐NMR, GC‐MS and co‐injection of its methyl ester with standards in GC‐FID. The HSCCC technique is commonly employed to isolate triterpene glycosides, but is applied in this study to an aglycone. Copyright © 2005 John Wiley & Sons, Ltd.