High-speed countercurrent chromatography (HSCCC) has been successfully used for the preparative isolation of flavonoids from the ethyl acetate extracts of black currant leaves. The HSCCC separation was performed with a two-phase solvent system composed of n-hexane/EtOAc/MeOH/H2O (1:10:1:10, v/v) at a flow rate of 1.5mL/min. When the flow rate was increased from 1.0 to 3.0mL/min, the retention of stationary phase decreased from 60.3% to 39.7% resulting in a loss of peak resolution, while the stationary phase retention is stable with an increase in sample size from 25 to 200mg. From 100mg of the crude sample, HSCCC separation yielded 4.0mg of kaempferol 3-O-galactoside, 6.0mg of kaempferol 3-O-glucoside, and 9.0mg of fraction I containing a mixture of hyperoside and isoquercitrin. Then, from 18mg of fraction I, 3.0mg of hyperoside and 11.0mg isoquercitrin were separated by preparative HPLC by successive sample injection at every 100min interval. Chemical structures of all these compounds were confirmed by MS and NMR.
Chinese Acad Sci, Xinjiang Tech Inst Phys & Chem, Xinjiang Key Lab Plant Resources & Nat Prod Chem, Urumqi 830011, Peoples R China;Shihezi Univ, Sch Life Sci, Shihezi, Peoples R China;Chinese Acad Sci, Grad Univ, Beijing, Peoples R China;NHLBI, Bioseparat Technol Lab, Biochem & Biophys Ctr, NIH, Bethesda, MD 20892 USA
He Dajun,Huang Yun,Ayupbek Amatjan,et al. SEPARATION AND PURIFICATION OF FLAVONOIDS FROM BLACK CURRANT LEAVES BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY AND PREPARATIVE HPLC[J]. JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES,2010,33(5):615-628.