FOOD SCIENCE ›› 2020, Vol. 41 ›› Issue (15): 96-104.doi: 10.7506/spkx1002-6630-20190715-193

• Basic Research • Previous Articles     Next Articles

Separation and Purification of Anthocyanins from Mulberry Fruit by High-Speed Counter-Current Chromatography and Their Antioxidant Activity

XUE Hongkun, LI Pengcheng, ZHONG Xue, LIU Chenghai, LI Qian   

  1. (1. College of Engineering, Northeast Agricultural University, Harbin 150030, China; 2. Key Laboratory of Particle & Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, Beijing 100084, China; 3. School of Food Science and Engineering, Jilin Agricultural University, Changchun 130112, China; 4. College of Life Science and Technology, Dalian University, Dalian 116622, China)
  • Online:2020-08-15 Published:2020-08-19

Abstract: In this study, anthocyanins from mulberry fruit were separated and purified by sequential macroporous resin adsorption chromatography and high-speed counter-current chromatography (HSCCC), and were identified by UV-Vis spectroscopy, high performance liquid chromatography-mass spectrometry (HPLC-MS) and nuclear magnetic resonance. We further evaluated the inhibitory effects of the crude extract and the purified components on lipid peroxidation and 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) radical. The HSCCC was performed using a two-phase solvent system composed of n-butanol, methyl tert-butyl ether, acetonitrile, water and trifluoroacetic acid (2:2:1:5:0.01, V/V) with the upper phase as the stationary phase and the lower phase as the mobile phase under the conditions of rotation speed of 850 r/min, flow rate of 2 mL/min and detection wavelength of 254 nm. Finally, four peaks (components I, II, III and IV) were displayed on the HSCCC chromatogram. Component IV was identified as non-anthocyanin, while components I, II and III as delphinidin-3-glucoside, cyanidin-3-glucoside and pelargonidin-3-glucoside with purities of and 92.27%, 94.05% and 90.82%, respectively and their contents in the crude extract were 17.4, 33.7, 9.8 mg/100 mg, respectively. The 50% inhibiting concentration of the crude extract, and components I, II and III were (0.77 ± 0.02), (0.34 ± 0.02), (0.55 ± 0.04) and (0.68 ± 0.01) g/L for inhibiting lipid peroxidation and (0.40 ± 0.01), (0.16 ± 0.01), (0.22 ± 0.01) and (0.35 ± 0.03) g/L for scavenging DPPH radical, respectively.

Key words: high-speed counter-current chromatography, mulberry, anthocyanins, structure identification, antioxidant activity

CLC Number: