title: Isolation of high-purity anthocyanin mixtures and monomers from blueberries using combined chromatographic techniques.
abstract: Research on the isolation and preparation of anthocyanins has intensified in recent years because of the requirements of quantitative and bioactive analyses. However, simple and effective methods for the scale purification of pure anthocyanins from natural products are rarely reported. In this study, high-purity anthocyanin mixtures and monomers were successfully isolated from wild blueberries using a combination of column chromatography and semi-preparative HPLC. We established an effective elution system to separate high-purity anthocyanin mixtures with aqueous ethanol containing 0.01% HCl first in an Amberlite XAD-7HP column (ethanol/H2O=35:65) and then in a Sephadex LH-20 column (ethanol/H2O=25:75). Crude anthocyanin extracts were isolated using the Amberlite column, and a purity of 32% was obtained based on UV-vis analysis. Three fractions of anthocyanin mixtures were isolated from the crude extracts using the Sephadex column with purities ranging from 59% to 68%. Three pure monomeric anthocyanins of malvidin-3-O-glucoside, petunidin-3-O-glucoside, and delphinidin-3-O-glucoside were also isolated by semi-preparative HPLC and identified by HPLC-DAD-ESI-MS/MS. The purities of these anthocyanins were determined by analytical HPLC and estimated to be 97.7%, 99.3%, and 95.4%, respectively. The results of this study may help promote the purification of anthocyanins from most blueberry varieties as well as from other plant materials.
authors.affiliation: College of Biological and Agricultural Engineering, Jilin University, Changchun, Jilin, China; Lab of Nutrition and Functional Food, Jilin University, Changchun, Jilin, China.
authors.forename: Erlei
authors.initials: E
authors.lastname: Wang
authors.affiliation: College of Biological and Agricultural Engineering, Jilin University, Changchun, Jilin, China.
authors.forename: Yongguang
authors.initials: Y
authors.lastname: Yin
authors.affiliation: Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, China.
authors.forename: Caina
authors.initials: C
authors.lastname: Xu
authors.affiliation: Lab of Nutrition and Functional Food, Jilin University, Changchun, Jilin, China. Electronic address: ljb168@sohu.com.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Liu
mesh_terms: D000872:Anthocyanins / Q000302:isolation & purification*; D029798:Blueberry Plants / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005638:Fruit / Q000737:chemistry; D005960:Glucosides / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000872:Anthocyanins; D005960:Glucosides; D010936:Plant Extracts; C494120:delphinidin 3-O-glucopyranoside; C455107:petunidin-3-glucoside; C458419:malvidin-3-glucoside
keywords: Anthocyanins; Blueberry; Column chromatography; Semi-preparative HPLC; Separation