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J Sep Sci. 2017 Sep;40(17):3506-3512. doi: 10.1002/jssc.201700471. Epub 2017 Aug 14.

Large-scale isolation of high-purity anthocyanin monomers from mulberry fruits by combined chromatographic techniques.

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School of the Environment and Safety, Jiangsu University, Zhenjiang, China.
School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, China.
School of Food and Biological Engineering, Jiangsu University, Zhenjiang, China.


Anthocyanins have attracted attention over the past several decades because of their beneficial health effects. In this research, a strategy combining column chromatography and high-speed countercurrent chromatography was developed for the separation of high-purity anthocyanin monomers from mulberry fruits. After purification using Amberlite XAD-7HP column with 80% ethanol (0.1% HCl), a fraction of anthocyanins mixtures with a purity of 68.6% was obtained. High-speed countercurrent chromatography with a biphasic solvent system of n-butanol/methyl tert-butyl ether/acetonitrile/water/trifluoroacetic acid (30:10:10:50:0.05, v/v) was used to separate the anthocyanin monomers. Three monomers of delphinidin-3-O-rutinoside, cyanidin-3-O-rutinoside, and cyanidin-3-O-glucoside were obtained, and identified by 1 H and 13 C NMR spectroscopy and high-performance liquid chromatography with electrospray ionization-mass spectrometry. The method developed in this work can be used to conduct large-scale separations of anthocyanin monomers from mulberry fruits and other plants.


anthocyanin monomers; column chromatography; high-speed countercurrent chromatography; large-scale separation; mulberry fruits

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