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荔枝果肉酚类物质大孔树脂分离纯化工艺优化
引用本文:苏东晓,张瑞芬,张名位,黄菲,魏振承,张雁,遆慧慧,邓媛元,唐小俊.荔枝果肉酚类物质大孔树脂分离纯化工艺优化[J].中国农业科学,2014,47(14):2897-2906.
作者姓名:苏东晓  张瑞芬  张名位  黄菲  魏振承  张雁  遆慧慧  邓媛元  唐小俊
作者单位:广东省农业科学院蚕业与农产品加工研究所/农业部功能食品重点实验室;华中农业大学食品科技学院;
基金项目:国家自然科学基金-广东联合基金(U1301211);国家国际合作项目(2012DFA31400);广东省国际合作项目(2011B050400002);广州市应用基础研究项目(11C14070783);广东省主体机构创新能力建设专项(2011)
摘    要:【目的】荔枝是热带亚热带地区重要水果,其果肉中含有丰富的酚类物质。其果肉粗提物具有良好的抗氧化、抗辐射和护肝活性。筛选对荔枝果肉酚类物质具有良好吸附、解吸性能的树脂,并优化分离工艺参数,为建立荔枝果肉多酚分离纯化工艺和鉴定荔枝果肉多酚结构提供理论依据。【方法】比较11种不同极性大孔树脂(HPD-200A、HPD-400A、HPD-100、HPD-500、HPD-600、HPD-722、HPD-826、D4020、NKA-II、NKA-9和AB-8)对新鲜荔枝果肉预冷80%丙酮/水酚类提取物中总酚、总黄酮的静态吸附和解吸性能,筛选分离荔枝果肉总酚和总黄酮均最佳的大孔树脂,考察其静态吸附动力学曲线,优化不同样液浓度(3.2、1.6、0.8和0.4 mg•mL-1)和不同上样流速(4.5、3.0和1.5 BV/h)动态吸附以及不同体积分数洗脱液乙醇浓度(95%、80%和60%)解吸工艺参数;通过与14种酚类物质标准品(儿茶素、没食子酸、绿原酸、香草酸、丁香酸、咖啡酸、表儿茶素、阿魏酸、四甲基邻苯二酚、香豆素、芦丁、槲皮素、白藜芦醇和槲皮素-3-O-芸香糖-7-O-鼠李糖)HPLC保留时间相比较的方法对所得组分酚类物质种类及其含量进行分析。【结果】HPD-826大孔树脂静态吸附和解吸荔枝果肉总酚和总黄酮的效果均较其它树脂好,其静态吸附4 h即可达到吸附饱和,吸附和解吸工艺参数为:荔枝果肉酚类提取物上样浓度0.8 mg•mL-1,上样速度3.0 BV/h,95%乙醇溶液作为洗脱剂,洗脱流速3.0 BV/h。经HPLC分析和鉴定,HPD-826分离纯化荔枝果肉酚类不会造成单体酚组成变化和明显损失(单体酚含量下降15%左右);荔枝果肉酚类物质主要由3,4二羟基苯甲酸、儿茶素、香草酸、咖啡酸、丁香酸、表儿茶素、槲皮素-3-O-芸香糖-7-O-鼠李糖苷、阿魏酸和芦丁等9种单体组成,其中含量较高的依次是槲皮素-3-O-芸香糖-7-O-鼠李糖苷、芦丁和表儿茶素,三者合计占总量的94.37%。【结论】HPD-826型大孔树脂对荔枝果肉总酚和总黄酮有较好的分离纯化效果。

关 键 词:荔枝    总酚    总黄酮    大孔树脂    分离纯化
收稿时间:2014-02-28

Separation and Purification of Polyphenol in Litchi Pulp by Macroporous Resin
SU Dong-Xiao-,ZHANG Rui-Fen-,ZHANG Ming-Wei-,HUANG Fei-,WEI Zhen-Cheng-,ZHANG Yan-,TI Hui-Hui-,DENG Yuan-Yuan-,TANG Xiao-Jun-.Separation and Purification of Polyphenol in Litchi Pulp by Macroporous Resin[J].Scientia Agricultura Sinica,2014,47(14):2897-2906.
Authors:SU Dong-Xiao-  ZHANG Rui-Fen-  ZHANG Ming-Wei-  HUANG Fei-  WEI Zhen-Cheng-  ZHANG Yan-  TI Hui-Hui-  DENG Yuan-Yuan-  TANG Xiao-Jun-
Institution:1、Sericultural and Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences/ Key Laboratory of Functional Food, Ministry of Agriculture, Guangzhou 510610; 2、Department of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070
Abstract:【Objective】Litchi (Litchi chinenesis Sonn.) is an important subtropical to tropical fruit which contain a high amount of phenolics. Previous studies found that litchi exhibits excellent antioxidant, radioprotective and hepatoprotective activities. The aim of this study is to screen a resin which has a good adsorption and desorption performance of phenols of litchi, and optimize the separation process parameters, thus providing a theoretical basis for establishing polyphenols purification technology and identification of polyphenols construction of litchi. The separation and purification process of litchi pulp polyphenol was established by macroporous resin.【Method】The static adsorption and desorption performance of eleven different polarities macroporous resins (HPD-200A, HPD-400A, HPD-100, HPD-500, HPD-600, HPD-722, HPD-826, D4020, NKA-II, NKA-9 and AB-8) to total phenolics and total flavonoids of fresh litchi pulp chilled acetone/water extracts were compared to select suitable resin for purification of phenolic compounds. Kinetic curves of adsorption were studied. The dynamic adsorption and desorption process parameters, including sample concentration (3.2, 1.6, 0.8 and 0.4 mg•mL-1), flow rate (4.5, 3.0 and 1.5 BV/h), and eluent of ethanol concentration (95%, 80% and 60%), of macroporous resin of HPD-826 were optimized. The phenolic compound types and contents of litchi pulp were analyzed by HPLC with the retention time of standard including catechin, gallic acid, chlorogenic acid, vanillic acid, clove acid, caffeic acid, epicatechin, ferulic acid, 4-methylcatechol, coumarin, rutin, quercetin, resveratrol and quercetin-3-O- rutinose-7-O-rhamnose.【Result】The results showed that HPD-826 macroporous resin exhibited the best capability of adsorption and desorption of total phenolics and total flavonoids in litchi pulp. And the macroporous resin reached equilibrium within 4 h. The optimal separating process parameters were as follows: the concentration of litchi pulp extract and the sampling rate were 0.8 mg/mL and 3.0 BV/h, respectively, and the elution concentration and flow velocity were 95% ethanol and 3.0B V/h, respectively. The contents of phenolic compounds of litchi pulp were deduced by about 15% compared to before purification, but the phenolic profiles of litchi pulp were not changed after adsorption and desorption by HPD-826 macroporous resin. Nine phenolic compounds, 3,4 dihydroxybenzoic acid, catechin, vanillic acid, caffeic acid, syringic acid, epicatechin, quercetin 3-O-rutinoside-7-O-α-L- rhamnosidase, ferulic acid and rutin, were preliminary identified by HPLC. The major phenolic profiles were quercetin 3-O- rutinoside-7-O-α-L-rhamnosidase, rutin and epicatechin. The percentage contribution of the three compounds to the total phenolic content was 94.37%.【Conclusion】In conclusion, HPD-826 macroporous resin could be applied to purify total phenolics and total flavonoids in litchi pulp.
Keywords:litchi  total phenolics  total flavonoids  macroporous resin  separation and purification
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