| 柱层析法分离纯化血橙花色苷 |
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| 引用本文： | 曹少谦,潘思轶,姚晓琳,傅虹飞.柱层析法分离纯化血橙花色苷[J].中国农业科学,2009,42(5):1728-1736. |
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| 作者姓名： | 曹少谦 潘思轶 姚晓琳 傅虹飞 |
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| 作者单位： | 华中农业大学食品科技学院,武汉,430070 |
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| 基金项目： | 农业部农业结构调整重大技术研究专项项目，湖北省自然科学基金，公益性行业(农业)科研专项基金 |
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| 摘 要： | 【目的】研究适合分离纯化血橙花色苷的柱层析法，并对血橙中的花色苷进行初步鉴定。【方法】分别采用12种不同类型树脂对血橙花色苷静态和动态吸附解吸试验进行比较，优化了大孔树脂提取分离血橙花色苷的工艺，采用Toyopearl TSK HW-40S柱层析对血橙花色苷提取物进一步分离纯化。同时，利用高效液相色谱-电喷雾质谱联用（HPLC-ESI/MS）对血橙花色苷进行定性分析。【结果】大孔树脂NKA-9对血橙花色苷的分离效果最好，最佳工艺条件为：50%乙醇用柠檬酸调pH为2.5时洗脱效果最好;Toyopearl TSK HW-40S柱层析分离纯化条件为：35%甲醇（经2%甲酸酸化）为洗脱液，流速0.6 ml?min-1，可得到3个单一花色苷组分和1个混合花色苷组分。HPLC-ESI/MS分析表明，血橙花色苷主要是为矢车菊素-3（3″-丙二酰）葡萄糖苷和矢车菊素-3（6″-二乙二酰）葡萄糖苷（组分1），矢车菊素-3-葡萄糖苷（组分2），矢车菊素-3（6″-丙二酰）葡萄糖苷（组分3）及矢车菊素-3-葡萄糖苷加成物4-乙烯基儿茶酚（组分4）。【结论】NKA-9大孔树脂与Toyopearl TSK HW-40S凝胶柱层析相结合，可以有效分离纯化血橙中花色苷，HPLC-ESI/MS分析可以方便、快捷、有效地为花色苷的鉴定提供依据。
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| 关 键 词： | 血橙 色素 花色苷 纯化 鉴定 |
| 收稿时间： | 2008-7-30 |
Isolation and Purification of Anthocyanins from Blood Oranges by Column Chromatography |
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| CAO Shao-qian,PAN Si-yi,YAO Xiao-lin,FU Hong-fei.Isolation and Purification of Anthocyanins from Blood Oranges by Column Chromatography[J].Scientia Agricultura Sinica,2009,42(5):1728-1736. |
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| Authors: | CAO Shao-qian PAN Si-yi YAO Xiao-lin FU Hong-fei |
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| Affiliation: | College of Food Science and Technology, Huazhong Agricultural University |
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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. |
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| Keywords: | blood orange pigment anthocyanins purification identification |
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