Scientia Agricultura Sinica ›› 2009, Vol. 42 ›› Issue (5): 1728-1736 .doi: 10.3864/j.issn.0578-1752.2009.05.028

• STORAGE·FRESH-KEEPING·PROCESSING • Previous Articles     Next Articles

Isolation and Purification of Anthocyanins from Blood Oranges by Column Chromatography

  

  1. 华中农业大学食品科技学院
  • Received:2008-07-30 Revised:2009-01-15 Online:2009-05-10 Published:2009-05-10
  • Contact: PAN Si-yi

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Abstract:

【Objective】 Isolation and purification of anthocyanins from blood oranges by column chromatography were made, and then the anthocyanins of blood orange were identified. 【Method】 The behavior of static adsorption and desorption, dynamic adsorption and desorption of twelve kinds of resins was compared. Toyopearl TSK HW-40S column was employed to separate and purify the anthocyanin extracts from blood orange. Then, the anthocyanins of blood orange were identified by HPLC-ESI/MS analysis. 【Result】 The results indicated that NKA-9 macroporous resin was the optimum for the anthocyanins of blood orange purification, and the optimum elution reagent was 50% ethanol with citric acid (pH 2.5). The best separation of Toyopearl TSK HW-40S column was obtained using a mobile phase of 35% methanol with 2% formic acid at a flow-rate of 0.6 ml?min-1. Three major anthocyanins of blood orange were isolated. HPLC-ESI/MS analysis results show that the anthocyanins of blood orange are mainly cyanidin-3-glucoside (35.2%) and cyaniding-3-(6″-malonyl) glucoside (42.9%). Furthermore, cyanidin-3-(3″-malonyl) glucoside, cyanidin 3-(6″-dioxalyl) glucoside and cyanidin-3-glucoside adduct: 4- vinylcatechol were identified in blood orange. 【Conclusion】 The combination of NKA-9 macroporous resin and Toyopearl TSK HW-40S column chromatography for separation and purification of blood orange anthocyanins is an effective method, and HPLC-ESI/MS analysis is a convenient, rapid and effective method for identification of anthocyanins from blood orange.

Key words: blood orange, pigment, anthocyanins, purification, identification

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