槐花多糖的分离纯化与抗氧化活性研究
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摘要
本文以中药槐花(Sophora japonica.)为实验对象,对其中多糖的分离纯化及理化特性进行了研究,获得如下结论:
1.采用热水浸提法提取槐花多糖,正交试验得出最佳提取条件为:提取温度80℃、料液比1:25、提取时间6h、提取次数4次。在该工艺条件下多糖的提取率达3.40%。
2.采用酶法提取槐花多糖,正交试验得出最佳提取条件为:提取温度45℃,提取时间1h,酶用量2%,提取pH值4.5。在该工艺条件下多糖的提取率为5.94%。
3.槐花多糖脱除蛋白的最佳方法为胰蛋白酶+Sevag法。此法对蛋白的清除能力最强,蛋白清除率可达85.23%,同时多糖损失率也最小。
4.多糖经DE-52离子交换纤维素柱分离出SJPS1、SJPS2和SJPS3三种组分,其纯度鉴定采用高效液相色谱法和Sepharose CL-6B凝胶柱层析法,结果表明三种组分均为均一组分。
5.气相色谱、理化特性分析结果表明,SJPS1主要由鼠李糖、果糖、阿拉伯糖和木糖组成,分子量为10.85kD,比旋光度+41.6°;SJPS2主要由鼠李糖、果糖和阿拉伯糖组成,分子量为7.72kD,比旋光度+128.7°;SJPS3主要由鼠李糖、果糖组成,分子量为2.88kD,比旋光度+80.1°。
6.抗氧化活性测定结果表明,槐花多糖不仅具有很强的还原能力,而且对超氧阴离子自由基、羟基自由基均有较好的清除能力,且在一定范围内对二者的清除作用呈现良好的量效关系。槐花多糖具有的抗氧化活性使其有望作为天然抗氧化剂和食品功能性成分而得到开发应用,具有较高的研究开发价值。
本实验对槐花多糖的分离效率、得率以及纯化过程进行全面研究,得到了一条分离纯化槐花多糖的详细的工艺路线;对得到的槐花多糖纯品进行了系统全面的理化性质研究,为开发利用槐花多糖提供了基础研究资料,将有助于槐花多糖的各方面深入研究。
In this paper, we mainly studied on Separation and Purification of Polysacchariedes from Sophora japonica. and their physicochemical properties. The results showed as follows:
1.The optimum conditions with the hot-water extraction method obtained by the orthogonal experiment design were as follows: the ratio of sample to water was 1:25, the temperature was 80℃, the time was 6 hours, and the extraction times was 4. Under the conditions, the extraction ratio of the polysaccharides were 3.40%.
2.The optimum conditions with enzymatic extraction method obtained by the orthogonal experiment design were as follows: adding 2% of cellulosase, the extraction time for 1h with temperature at 45℃and pH4.5. In this condition the extraction rate of polysaccharides is 5.94%.
3.The enzymatic+Sevag removal of protein was an effective method, the removal of protein reached 85.23 %.
4.Three fractions named respectively SJPS1, SJPS2, and SJPS3 were isolated and purified from the polysaccharides by the DE-52, the purity were identified by HPLC and Sepharose CL-6B gel filtration Chromatogram and the results showed three fractions were all homogeneous fractions.
5.Gas chromatography analysis, physical and chemical characters showed that SJPS1 is composed of rhamnose, fructose, arabmose and xylose, its molecular weight is 10.85kD and the specific optical rotation is +41.6°; SJPS2 is composed of the rhamnose, fructose, arabmose, its molecular weight is 7.72kD and the specific optical rotation is +128.7°; SJPS3 is composed of the ramnose, fuctose, its molecular weight is 2.88kD and the specific optical rotation is +80.1°.
6.Antioxidant activity results showed that the polysaccharides of Sophora japonica, not only a strong reduction capability, but also better ability to remove superoxide radical and hydroxyl radical, and the scavenging ability related with the concentrations. Polysaccharides of Sophora japonica. with the antioxidant activity are expected to as natural antioxidants and functional food ingredients. Polysaccharides of Sophora japonica, have high value and will be developed and applied.
The experiment has comprehensive studied on separation efficiency, yield and purification process of polysaccharides from Sophora japonica. It provided us a detailed process line purified polysaccharide from Sophora japonica. Then we studied on their physicochemical properties. As a theoretcal reference for the development and production of polysaccharides from Sophora japonica., it also helps all aspects of the in-depth study of polysaccharides from Sophora japonica.
1.采用热水浸提法提取槐花多糖,正交试验得出最佳提取条件为:提取温度80℃、料液比1:25、提取时间6h、提取次数4次。在该工艺条件下多糖的提取率达3.40%。
2.采用酶法提取槐花多糖,正交试验得出最佳提取条件为:提取温度45℃,提取时间1h,酶用量2%,提取pH值4.5。在该工艺条件下多糖的提取率为5.94%。
3.槐花多糖脱除蛋白的最佳方法为胰蛋白酶+Sevag法。此法对蛋白的清除能力最强,蛋白清除率可达85.23%,同时多糖损失率也最小。
4.多糖经DE-52离子交换纤维素柱分离出SJPS1、SJPS2和SJPS3三种组分,其纯度鉴定采用高效液相色谱法和Sepharose CL-6B凝胶柱层析法,结果表明三种组分均为均一组分。
5.气相色谱、理化特性分析结果表明,SJPS1主要由鼠李糖、果糖、阿拉伯糖和木糖组成,分子量为10.85kD,比旋光度+41.6°;SJPS2主要由鼠李糖、果糖和阿拉伯糖组成,分子量为7.72kD,比旋光度+128.7°;SJPS3主要由鼠李糖、果糖组成,分子量为2.88kD,比旋光度+80.1°。
6.抗氧化活性测定结果表明,槐花多糖不仅具有很强的还原能力,而且对超氧阴离子自由基、羟基自由基均有较好的清除能力,且在一定范围内对二者的清除作用呈现良好的量效关系。槐花多糖具有的抗氧化活性使其有望作为天然抗氧化剂和食品功能性成分而得到开发应用,具有较高的研究开发价值。
本实验对槐花多糖的分离效率、得率以及纯化过程进行全面研究,得到了一条分离纯化槐花多糖的详细的工艺路线;对得到的槐花多糖纯品进行了系统全面的理化性质研究,为开发利用槐花多糖提供了基础研究资料,将有助于槐花多糖的各方面深入研究。
In this paper, we mainly studied on Separation and Purification of Polysacchariedes from Sophora japonica. and their physicochemical properties. The results showed as follows:
1.The optimum conditions with the hot-water extraction method obtained by the orthogonal experiment design were as follows: the ratio of sample to water was 1:25, the temperature was 80℃, the time was 6 hours, and the extraction times was 4. Under the conditions, the extraction ratio of the polysaccharides were 3.40%.
2.The optimum conditions with enzymatic extraction method obtained by the orthogonal experiment design were as follows: adding 2% of cellulosase, the extraction time for 1h with temperature at 45℃and pH4.5. In this condition the extraction rate of polysaccharides is 5.94%.
3.The enzymatic+Sevag removal of protein was an effective method, the removal of protein reached 85.23 %.
4.Three fractions named respectively SJPS1, SJPS2, and SJPS3 were isolated and purified from the polysaccharides by the DE-52, the purity were identified by HPLC and Sepharose CL-6B gel filtration Chromatogram and the results showed three fractions were all homogeneous fractions.
5.Gas chromatography analysis, physical and chemical characters showed that SJPS1 is composed of rhamnose, fructose, arabmose and xylose, its molecular weight is 10.85kD and the specific optical rotation is +41.6°; SJPS2 is composed of the rhamnose, fructose, arabmose, its molecular weight is 7.72kD and the specific optical rotation is +128.7°; SJPS3 is composed of the ramnose, fuctose, its molecular weight is 2.88kD and the specific optical rotation is +80.1°.
6.Antioxidant activity results showed that the polysaccharides of Sophora japonica, not only a strong reduction capability, but also better ability to remove superoxide radical and hydroxyl radical, and the scavenging ability related with the concentrations. Polysaccharides of Sophora japonica. with the antioxidant activity are expected to as natural antioxidants and functional food ingredients. Polysaccharides of Sophora japonica, have high value and will be developed and applied.
The experiment has comprehensive studied on separation efficiency, yield and purification process of polysaccharides from Sophora japonica. It provided us a detailed process line purified polysaccharide from Sophora japonica. Then we studied on their physicochemical properties. As a theoretcal reference for the development and production of polysaccharides from Sophora japonica., it also helps all aspects of the in-depth study of polysaccharides from Sophora japonica.
引文
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