超声微波协同酶法提取黄精多糖与抗氧化特性分析
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摘要
为探讨超声微波协同酶法提取黄精多糖的工艺条件,采用乙醇分级醇沉制备生黄精和制黄精多糖的不同醇沉组分,研究不同多糖组分的抗氧化特性。试验结果表明,以生黄精为原料时,最佳提取条件为:在超声微波处理下,以料液比1︰23 (g/mL)、纤维素酶添加量0.85%,在温度60℃下酶解20 min,黄精多糖提取率为17.53%。经用乙醇分级沉淀该条件下制备生黄精,获得多糖组分PPs-40, PPs-60和PPs-80,其得率分别为5.61%, 46.11%和48.28%,多糖含量分别为79.66%, 88.92%和99.28%;制备得到的制黄精多糖,获得多糖组分PPPs-40, PPPs-60和PPPs-80,其得率分别为21.08%, 21.08%和57.84%,多糖含量分别为57.94%, 70.18%和75.24%,生黄精和制黄精多糖几乎不含蛋白质和多酚。各多糖组分均具有较好的DPPH·和O_2~-·清除率,其中PPs-40对DPPH·清除能力最强,其IC50值为0.59 mg/m L; PPPs-40对O_2~-·的清除能力最强,其IC50值为0.64 mg/mL。试验结果为分离鉴定具有高抗氧化活性的生黄精和制黄精多糖组分提供科学依据。
The extraction condition of polygonatum polysaccharides(PPs) by ultrasonic-microwave synergistic enzyme was studied, the different alcohol precipitation fractions were prepared by ethanol precipitation, and its antioxidant properties were investigated. The results show that the optimum extraction condition is that the raw material under the ultrasonic microwave treatment, with the ratio of material to water at 1︰23(g/mL), adding 0.85% of cellulase, at the temperature of 60 ℃ for 20 min, the extraction rate of polygonatum polysaccharide reaches 17.53%. The yields of PPs-40, PPs-60, PPs-80 and PPPs-40,PPPs-60, PPPs-80 are 5.61%, 46.11%, 48.28% and 21.08%, 21.08%, 7.84%, respectively, and the contents of polysaccharide are 79.66%, 88.92%, 99.28% and 57.94%, 70.18%, 75.24%, respectively. Almost no protein and polyphenols have been contained. All the polysaccharides fractions have good DPPH· and O2-· clearance, of which PPs-40 is the strongest for DPPH,the IC50 value is 0.59 mg/mL; PPPs-40 has the strongest clearance to O2-·, and its IC50 value is 0.64 mg/mL. The results provide scientific basis for isolation and identification of the polysaccharide fraction with high antioxidant activity from Polygonatum sibiricum.
The extraction condition of polygonatum polysaccharides(PPs) by ultrasonic-microwave synergistic enzyme was studied, the different alcohol precipitation fractions were prepared by ethanol precipitation, and its antioxidant properties were investigated. The results show that the optimum extraction condition is that the raw material under the ultrasonic microwave treatment, with the ratio of material to water at 1︰23(g/mL), adding 0.85% of cellulase, at the temperature of 60 ℃ for 20 min, the extraction rate of polygonatum polysaccharide reaches 17.53%. The yields of PPs-40, PPs-60, PPs-80 and PPPs-40,PPPs-60, PPPs-80 are 5.61%, 46.11%, 48.28% and 21.08%, 21.08%, 7.84%, respectively, and the contents of polysaccharide are 79.66%, 88.92%, 99.28% and 57.94%, 70.18%, 75.24%, respectively. Almost no protein and polyphenols have been contained. All the polysaccharides fractions have good DPPH· and O2-· clearance, of which PPs-40 is the strongest for DPPH,the IC50 value is 0.59 mg/mL; PPPs-40 has the strongest clearance to O2-·, and its IC50 value is 0.64 mg/mL. The results provide scientific basis for isolation and identification of the polysaccharide fraction with high antioxidant activity from Polygonatum sibiricum.
引文
[1]中国药典委员会中华人民共和国药典[M].北京:科学出版社,2000:64-252.
[2]柳威,林懋怡,刘晋杰,等.滇黄精研究进展及黄精研究现状[J].中国实验方剂学杂志,2017(14):226-234.
[3]巫永华,刘恩岐,张建萍,等.黄精多酚的闪式提取及抗氧化活性研究[J].食品科技,2017(8):231-236.
[4]陈克克,强毅.响应面法优化超声波辅助黄精多酚的提取及其抗菌活性[J].陕西师范大学学报(自然科学版),2018(1):91-96.
[5]周宝珍.不同产地黄精薯蓣皂苷元含量的研究[J].陕西农业科学,2017,63(8):42-45.
[6]刘彦东,陈文生,张权,等.不同生长期的黄精中氨基酸含量的测定与分析[J].时珍国医国药,2017(2):444-447.
[7]何杨.黄精多糖的提取、分离纯化及抗病毒活性的研究[D].成都:中国科学院成都生物研究所,1997.
[8]YANG J,WU S,HUANG X,et al.Hypolipidemic activity and antiatherosclerotic effect of polysaccharide of polygonatum sibiricum in rabbit model and related cellular mechanisms[J].Evidence-Based Complementray and Alternative Medicine,2015(3):1-6.
[9]WANG S,WANG B,HUA W,et al.De Novo assembly and analysis of polygonatum sibiricum transcriptome and identification of genes involved in polysaccharide biosynthesis[J]International Journal of Molecular Sciences,2017,18(9):1950.
[10]陈钢,陈红兰,苏伟,等.响应面分析法优化黄精多糖提取工艺参数[J].食品科学,2007(7):198-201.
[11]傅圣斌,钱建鸿,陈乐意,等.黄精多糖的提取及其对小鼠免疫活性的影响[J].中国食品学报,2013,13(1):68-72.
[12]何连军,干雅平,吕伟德,等.高效阴离子交换色谱-脉冲安培检测法测定多花黄精多糖的单糖组成[J].中草药,2017,48(8):1671-1676.
[13]徐世忱,纪耀华.黄精炮制前后粗多糖的提取收率与总糖含量的比较分析[J].中草药,1993(12):629-630.
[14]陈艳,王杰,李慧,等.黄精多糖的闪式提取及对乳酸菌发酵特性的影响[J].食品工业,2017(6):161-166.
[15]马淑凤,陈利梅,徐化能,等.白灵菇多糖的分离纯化及清除自由基研究[J].食品科学,2009,30(19):109-113.
[16]王艾平,周丽明.考马斯亮蓝法测定茶籽多糖中蛋白质含量条件的优化[J].河南农业科学,2014,43(3):152-153.
[17]游见明,曹新志.福林酚法测定茶树中茶多酚的分布水平[J].湖北农业科学,2013,10:2417-2419.
[18]唐仕荣,宋慧,刘全德,等.牛蒡多糖的分离纯化及其抗氧化活性研究[J].徐州工程学院学报(自然科学版)2014(2):30-35.
[19]龚霞,巫永华,刘恩岐,等.山楂叶提取物不同极性组分的体外抗氧化作用[J].食品工业,2016(8):216-219.
[20]曾林燕,宋志前,魏征,等.黄精炮制过程中新产生成分分离及含量变化[J].中草药,2013,44(12):1584-1588.
[2]柳威,林懋怡,刘晋杰,等.滇黄精研究进展及黄精研究现状[J].中国实验方剂学杂志,2017(14):226-234.
[3]巫永华,刘恩岐,张建萍,等.黄精多酚的闪式提取及抗氧化活性研究[J].食品科技,2017(8):231-236.
[4]陈克克,强毅.响应面法优化超声波辅助黄精多酚的提取及其抗菌活性[J].陕西师范大学学报(自然科学版),2018(1):91-96.
[5]周宝珍.不同产地黄精薯蓣皂苷元含量的研究[J].陕西农业科学,2017,63(8):42-45.
[6]刘彦东,陈文生,张权,等.不同生长期的黄精中氨基酸含量的测定与分析[J].时珍国医国药,2017(2):444-447.
[7]何杨.黄精多糖的提取、分离纯化及抗病毒活性的研究[D].成都:中国科学院成都生物研究所,1997.
[8]YANG J,WU S,HUANG X,et al.Hypolipidemic activity and antiatherosclerotic effect of polysaccharide of polygonatum sibiricum in rabbit model and related cellular mechanisms[J].Evidence-Based Complementray and Alternative Medicine,2015(3):1-6.
[9]WANG S,WANG B,HUA W,et al.De Novo assembly and analysis of polygonatum sibiricum transcriptome and identification of genes involved in polysaccharide biosynthesis[J]International Journal of Molecular Sciences,2017,18(9):1950.
[10]陈钢,陈红兰,苏伟,等.响应面分析法优化黄精多糖提取工艺参数[J].食品科学,2007(7):198-201.
[11]傅圣斌,钱建鸿,陈乐意,等.黄精多糖的提取及其对小鼠免疫活性的影响[J].中国食品学报,2013,13(1):68-72.
[12]何连军,干雅平,吕伟德,等.高效阴离子交换色谱-脉冲安培检测法测定多花黄精多糖的单糖组成[J].中草药,2017,48(8):1671-1676.
[13]徐世忱,纪耀华.黄精炮制前后粗多糖的提取收率与总糖含量的比较分析[J].中草药,1993(12):629-630.
[14]陈艳,王杰,李慧,等.黄精多糖的闪式提取及对乳酸菌发酵特性的影响[J].食品工业,2017(6):161-166.
[15]马淑凤,陈利梅,徐化能,等.白灵菇多糖的分离纯化及清除自由基研究[J].食品科学,2009,30(19):109-113.
[16]王艾平,周丽明.考马斯亮蓝法测定茶籽多糖中蛋白质含量条件的优化[J].河南农业科学,2014,43(3):152-153.
[17]游见明,曹新志.福林酚法测定茶树中茶多酚的分布水平[J].湖北农业科学,2013,10:2417-2419.
[18]唐仕荣,宋慧,刘全德,等.牛蒡多糖的分离纯化及其抗氧化活性研究[J].徐州工程学院学报(自然科学版)2014(2):30-35.
[19]龚霞,巫永华,刘恩岐,等.山楂叶提取物不同极性组分的体外抗氧化作用[J].食品工业,2016(8):216-219.
[20]曾林燕,宋志前,魏征,等.黄精炮制过程中新产生成分分离及含量变化[J].中草药,2013,44(12):1584-1588.