PREPARATIVE ISOLATION AND PURIFICATION OF FLAVONE C-GLYCOSIDES FROM THE LEAVES OF FICUS MICROCARPA L.f BY MEDIUM-PRESSURE LIQUID CHROMATOGRAPHY, HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY, AND PREPARATIVE LIQUID CHROMATOGRAPHY

Abstract

Combined with medium pressure liquid chromatography (MPLC) and preparative high performance liquid chromatography (perp-HPLC), high speed countercurrent chromatography (HSCCC) was applied for the separation and purification of flavone C-glycosides from the crude extract of leaves of Ficus microcarpa L.f. HSCCC separation was performed on a two-phase solvent system composed of methyl tert- butyl ether–ethyl acetate –1-butanol–acetonitrile–0.1% aqueous trifluoroacetic acid at a volume ratio of 1:3:1:1:5. Partially resolved peak fractions from HSCCC separation were further purified by preparative HPLC. Four well separated compounds were obtained and their purities were determined by HPLC. The purities of these peaks were 97.28%, 97.20%, 92.23%, and 98.40%. These peaks were characterized by ESI-MSⁿ. According to the reference, they were identified as orientin (peak I), isovitexin-3″-O-glucopyranoside (peak II), isovitexin (peak III), and vitexin (peak IV), and yielded 1.2 mg, 4.5 mg, 3.3 mg, and 1.8 mg, respectively.

Keywords:

• Ficus microcarpa L.f
• flavone C-glycosides
• high speed countercurrent chromatography
• medium pressure liquid chromatography
• preparative high pressure liquid chromatography

Notes

TFA: 0.1% aqueous trifluoroacetic acid, B: 1-butanol, MBE: methyl tert-butyl ether.

ACN: acetonitrile, EA: ethyl acetate.