Preparative separation of triterpene alcohol ferulates from rice bran oil using a high performance counter-current chromatography

A novel method for the separation of two major triterpene alcohol ferulates from rice bran oil (RBO) was developed using a high performance counter-current chromatography (HPCCC). A two-phase solvent system of n-hexane-acetonitrile (1:1, v/v) was applied to purify cycloartenyl ferulate (CAF) and 24-methylene cycloartanyl ferulate (24-mCAF) from RBO. The yields were 20.50 ± 2.60 mg CAF and 12.62 ± 1.15 mg 24-mCAF from 390 mg RBO through a two-step separation procedure. The purities of the two compounds were 97.97 ± 0.90% and 95.50 ± 0.75%, respectively, as determined by high performance liquid chromatography (HPLC). Their chemical structures were confirmed by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and 1H, 13C and 2D nuclear magnetic resonance (NMR). This represents the first report on direct separation of CAF and 24-mCAF from RBO by HPCCC.