白花蛇舌草中黄酮、多酚、多糖提取工艺及抗氧化研究
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摘要
本研究利用水浴-回流法提取,分光光度法分析了云南不同产地白花蛇舌草样品中黄酮、多酚、多糖含量;同时,分别研究了不同乙醇体积分数、液料比、提取温度、时间对样品中这3类物质提取效率的影响;此外,利用清除DPPH·自由基的方法,分析了提取液的抗氧化活性,以期为白花蛇舌草资源的利用提供理论和实践指导。结果表明,白花蛇舌草中这3类物质最佳提取条件为乙醇体积分数60%、液料比1:35、温度80℃、时间2. 5 h;不同产地23个样品提取液抗氧化活性IC_(50)(DPPH半数清除率浓度)在0. 18～0. 58 mg·mL~(-1)之间,平均值为0. 35 mg·mL~(-1);相关性分析表明,这些样品中黄酮、多酚含量和IC_(50)值之间呈极显著负相关,相关系数分别为-0. 852和-0. 886(p<0. 01),多糖含量与IC_(50)无显著相关性。以上结果表明,水浴-回流法可同时提取白花蛇舌草中黄酮、多酚、多糖三类物质;不同产地样品中这3类物质含量存在差异,这可能与生态因素不同有关;白花蛇舌草中最主要的抗氧化活性物质为黄酮和多酚。
In the present study,by using water bath-reflux method and spectrophotometry,we investigated the contents of flavonoids,polyphenols and polysaccharides in the Hedyotis diffusa Willd from different areas of Yunnan province. In addition,we studied the effects of different ethanol volume fraction,liquid ratio,extraction temperature and time on extraction efficiency of these 3 compounds.Furethermore,we evaluated the antioxidant activity of these extracts by determining the radical scavenging activity of DPPH·,aiming at provide theoretical and practical guidance for utilization of Hedyotis diffusa Willd. The results showed that,the optimal extraction conditions of these 3 compounds were ethanol volume fraction: 60%,liquid ratio: 1 ∶ 35,extraction temperature:80 ℃,time 2. 5 h,respectively.Moreover,the antioxidant activity( concentration of half scavening rate of DPPH,IC_(50)) of these extracts of the 23 samples were ranged from 0. 18 ～ 0. 58 mg·mL~(-1)and averaged 0. 35 mg·mL~(-1).Addiyionally,correlation analysis showed that the contents of flavonoids and polyphenols and IC_(50) value had negative correlation,which crrrelation rate were-0. 852and-0. 886( p<0. 01),respectively. However,the content of polysaccharides was not correlated with IC_(50). We concluded that water bath-reflux method can be used to analyze the contents of flavonoids,polyphenols and polysaccharides simultaneously in the Hedyotis diffusa Willd plants.Furthermore,the contents of these 3 compounds in the plants from different areas of Yunnan were varied,which can be ascribed to the different ecological factors in different areas.Additionally,flavonoids and polyphenols were primary anoxidant compounds in Hedyotis diffusa Willd.
In the present study,by using water bath-reflux method and spectrophotometry,we investigated the contents of flavonoids,polyphenols and polysaccharides in the Hedyotis diffusa Willd from different areas of Yunnan province. In addition,we studied the effects of different ethanol volume fraction,liquid ratio,extraction temperature and time on extraction efficiency of these 3 compounds.Furethermore,we evaluated the antioxidant activity of these extracts by determining the radical scavenging activity of DPPH·,aiming at provide theoretical and practical guidance for utilization of Hedyotis diffusa Willd. The results showed that,the optimal extraction conditions of these 3 compounds were ethanol volume fraction: 60%,liquid ratio: 1 ∶ 35,extraction temperature:80 ℃,time 2. 5 h,respectively.Moreover,the antioxidant activity( concentration of half scavening rate of DPPH,IC_(50)) of these extracts of the 23 samples were ranged from 0. 18 ～ 0. 58 mg·mL~(-1)and averaged 0. 35 mg·mL~(-1).Addiyionally,correlation analysis showed that the contents of flavonoids and polyphenols and IC_(50) value had negative correlation,which crrrelation rate were-0. 852and-0. 886( p<0. 01),respectively. However,the content of polysaccharides was not correlated with IC_(50). We concluded that water bath-reflux method can be used to analyze the contents of flavonoids,polyphenols and polysaccharides simultaneously in the Hedyotis diffusa Willd plants.Furthermore,the contents of these 3 compounds in the plants from different areas of Yunnan were varied,which can be ascribed to the different ecological factors in different areas.Additionally,flavonoids and polyphenols were primary anoxidant compounds in Hedyotis diffusa Willd.
引文
[1]姚志红,周先强,温丽荣,等.UPLC指纹图谱结合化学计量学的白花蛇舌草真伪鉴别研究[J].中药材,2016,39(4):737-741.
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[9]蒋剑平,许海顺,卢烨琳,等.响应面分析法优化白花蛇舌草水溶性多糖的提取工艺[J].浙江中医药大学学报,2012,36(2):187-191.
[10]赵楠楠,朱晓冉,李德海,等.红松松壳多酚、黄酮和多糖含量及抗氧化功能相关性研究[J].现代食品科技,2017,33(12):1-5.
[11]Hosu A,Cristea V M,Cimpoiu C.Analysis of total phenolic,flavonoids,anthocyanins and tannins content in Romanianred wines:Prediction of antioxidant activities and classification of wines using artificial neural networks[J].Food Chem,2014,150(4):113
[12]杨家楷,杨新周,李学娇.高速匀浆法提取辣蓼茎中总黄酮[J].云南化工,2014,41(6):23-25.
[13]Aspe E,Fernandez K. The effect of different extraction techniques on extraction yield,total phenolic,and antiradical capacity of extracts from Pinus radiate bark[J].Ind Crop Prod,2011,34(1):838-844
[14]杨新周,郝志云,朱以常.辣蓼不同部位抗氧化活性研究[J].江苏农业科学,2014,42(2):284-285.
[15]穆怀雪,杨新周,李学森,等.不同提取方法对五味子清除DPPH自由基作用的影响研究[J].时珍国医国药,2012,23(6):1454-1455.
[16]杨新周,郝志云,杨子仙.佛手花抗氧化活性研究[J].黑龙江农业科学,2014,2:72-74.
[17]蒲月华,邓旗,何锦锋,等.水解珍珠体外抗氧化活性的研究[J].化学研究与应用,2017,29(4):555-559.
[2]侯山岭.中药白花蛇舌草化学成分及药理活性研究进展[J].中医临床研究,2018,(6):140-141.
[3]杨新周,郝志云,朱以常.不同溶剂和提取方法对白花蛇舌草提取物抗氧化活性的影响[J].贵州农业科学,2014,42(2):43-45.
[4]于亮,王芳,郭琪,等.白花蛇舌草的化学成分及其药理活性研究进展[J].沈阳药科大学学报,2017,34(12):1104-1114.
[5]国家药典委员.中华人民共和国药典[M].第一部.北京:中国医药科技出版社,2015.
[6]张俊霞,王应玲,姜宁,等.白花蛇舌草总黄酮提取工艺及其抗氧化活性研究[J].湖北农业科学,2017,56(18):3523-3527.
[7]李芳,杨培民,曹广尚,等.Box-Behnken设计-效应面法优化白花蛇舌草总黄酮的提取工艺[J].中国实验方剂学杂志,2015,21(8):21-24.
[8]林珊,陈达鑫,蔡巧燕,等.Folin-Ciocaheu比色法测定白花蛇舌草总酚的含量[J].福建中医药,2016,47(5):31-32.
[9]蒋剑平,许海顺,卢烨琳,等.响应面分析法优化白花蛇舌草水溶性多糖的提取工艺[J].浙江中医药大学学报,2012,36(2):187-191.
[10]赵楠楠,朱晓冉,李德海,等.红松松壳多酚、黄酮和多糖含量及抗氧化功能相关性研究[J].现代食品科技,2017,33(12):1-5.
[11]Hosu A,Cristea V M,Cimpoiu C.Analysis of total phenolic,flavonoids,anthocyanins and tannins content in Romanianred wines:Prediction of antioxidant activities and classification of wines using artificial neural networks[J].Food Chem,2014,150(4):113
[12]杨家楷,杨新周,李学娇.高速匀浆法提取辣蓼茎中总黄酮[J].云南化工,2014,41(6):23-25.
[13]Aspe E,Fernandez K. The effect of different extraction techniques on extraction yield,total phenolic,and antiradical capacity of extracts from Pinus radiate bark[J].Ind Crop Prod,2011,34(1):838-844
[14]杨新周,郝志云,朱以常.辣蓼不同部位抗氧化活性研究[J].江苏农业科学,2014,42(2):284-285.
[15]穆怀雪,杨新周,李学森,等.不同提取方法对五味子清除DPPH自由基作用的影响研究[J].时珍国医国药,2012,23(6):1454-1455.
[16]杨新周,郝志云,杨子仙.佛手花抗氧化活性研究[J].黑龙江农业科学,2014,2:72-74.
[17]蒲月华,邓旗,何锦锋,等.水解珍珠体外抗氧化活性的研究[J].化学研究与应用,2017,29(4):555-559.