Purification of betulinic acid from Eugenia florida (Myrtaceae) by high-speed counter-current chromatography.
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. Purified betulinic acid was characterized by 13C-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.
Frighetto
Nelson
N
Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Universidade Estadual de Campinas, CPQBA-UNICAMP, CP 6171, Brazil. frigheto@cpqba.unicamp.br
Welendorf
Rodolfo Max
RM
Pereira da Silva
Ana Maria
AM
Nakamura
Marcos Jun
MJ
Siani
Antonio Carlos
AC
D003377:Countercurrent Distribution / Q000379:methods*; D053978:Pentacyclic Triterpenes; D018515:Plant Leaves / Q000737:chemistry; D027842:Syzygium / Q000737:chemistry*; D014315:Triterpenes / Q000737:chemistry / Q000302:isolation & purification*; D000094062:Betulinic Acid
D053978:Pentacyclic Triterpenes; D014315:Triterpenes; D000094062:Betulinic Acid