AB-8大孔树脂纯化绿豆皮黄酮工艺优化及纯化前后抗氧化能力比较
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摘要
采用AB-8大孔树脂初步分离纯化绿豆皮黄酮。分别对上样条件和洗脱条件进行优化,考察上样液质量浓度、上样液pH值、上样流速、以及洗脱剂体积分数、洗脱剂用量、洗脱流速对吸附解吸性能的影响,最终确定AB-8大孔树脂的分离纯化绿豆皮黄酮工艺为上样液质量浓度1.5 mg/m L、上样液pH 5.0、上样流速1.0 m L/min;洗脱液乙醇体积分数70%、洗脱剂用量225 m L、洗脱流速2.0 m L/min,在此条件下分离纯化,绿豆皮黄酮纯度由27.95%提高到62.38%。经紫外-可见光谱扫描,出现黄酮类化合物特征峰带,经红外光谱扫描,光谱具备黄酮类物质特征官能团,验证了黄酮类物质的存在;利用扫描电镜对纯化前后黄酮类物质进行微观分析,得出被包裹的片状和粉粒状颗粒大部分被释放出来,这可能是导致纯化后黄酮类化合物纯度增高的原因。纯化后的绿豆皮黄酮与粗提物相比具有较高的抗氧化能力。
In this study, AB-8 macroporous resin was used to separate and purify flavonoids from mung bean coat. To establish optimized sample loading and the elution conditions, the influence of sample concentration, pH and flow rate as well as eluent concentration, volume and flow rate on the adsorption and desorption efficiencies was investigated. The optimal conditions determined were as follows: sample concentration, 1.5 mg/m L; pH, 5.0; flow rate, 1.0 m L/min; and 225 m L of 70% ethanol as eluent at a flow rate of 2.0 m L/min. Under these conditions, the purity of mung bean coat flavonoids increased from 27.95% to 62.38%. The existence of flavonoids were confirmed by their characteristic peaks in UV-visible spectra and characteristic functional groups identified by infrared spectroscopy. Scanning electron microscopy(SEM) showed that most flaky and powdery particles were released, which might contribute to the increased purity of flavonoids after initial purification. Compared with crude extract, the purified flavonoids had higher antioxidant ability.
In this study, AB-8 macroporous resin was used to separate and purify flavonoids from mung bean coat. To establish optimized sample loading and the elution conditions, the influence of sample concentration, pH and flow rate as well as eluent concentration, volume and flow rate on the adsorption and desorption efficiencies was investigated. The optimal conditions determined were as follows: sample concentration, 1.5 mg/m L; pH, 5.0; flow rate, 1.0 m L/min; and 225 m L of 70% ethanol as eluent at a flow rate of 2.0 m L/min. Under these conditions, the purity of mung bean coat flavonoids increased from 27.95% to 62.38%. The existence of flavonoids were confirmed by their characteristic peaks in UV-visible spectra and characteristic functional groups identified by infrared spectroscopy. Scanning electron microscopy(SEM) showed that most flaky and powdery particles were released, which might contribute to the increased purity of flavonoids after initial purification. Compared with crude extract, the purified flavonoids had higher antioxidant ability.
引文
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[24]华耀祖.超滤技术与应用[M].北京:化学工业出版社,2004.
[25]ZHANG Y,LI S F,WU X W,et al.Macroporous resin adsorption for purification of flavonoids in Houttuynia cordata Thunb[J].Chinese Journal of Chemical Engineering,2007,15(6):872-876.DOI:10.1016/S1004-9541(08)60017-8.
[26]韩博,陈文,景文娟,等.AB-8大孔吸附树脂对甘草总黄酮的吸附[J].南方医科大学学报,2007,27(3):265-267.DOI:10.3321/j.issn.1673-4254.2007.03.005.
[27]FU B Q,LIU J,LI H,et al.The application of macroporous resins in the separation of licorice flavonoids and glycyrrhizic acid[J].Journal of Chromatography,2005,1089(1/2):18-24.DOI:10.1016/j.chroma.2005.06.051.
[28]贾海伦.芹菜黄酮提取及分离纯化工艺研究[D].哈尔滨:黑龙江大学,2008.
[29]骆党委,叶静,黄雅燕,等.A B-8大孔吸附树脂精制芦柑皮总黄酮及黄酮类化合物的分离[J].食品科学,2014,35(6):30-35.DOI:10.7506/spkx1002-6630-201406006.
[30]李侠,邹基豪,王大为.响应面试验优化超声波-酶法提取绿豆皮黄酮类化合物工艺[J].食品科学,2017,38(8):206-212.DOI:10.7506/spkx1002-6630-201708032.
[2]刘笑然.东北三省的红小豆、绿豆生产[J].中国粮食经济,2013,26(9):38-41.DOI:10.3969/j.issn.1007-4821.2013.09.011.
[3]王旭.绿豆中清热解毒成分的提取及稳定性的研究[D].哈尔滨:哈尔滨商业大学,2011.
[4]李昀铮.超声波辅助提取绿豆皮黄酮及其应用研究[D].长春:吉林农业大学,2015.
[5]曾志红,王强,林伟静,等.绿豆的品质特性及加工利用研究概况[J].作物杂志,2011(4):16-19.DOI:10.3969/j.issn.1001-7283.2011.04.004.
[6]庄艳,陈剑.绿豆的营养价值及综合利用[J].杂粮作物,2009,29(6):418-419.DOI:10.3969/j.issn.2095-0896.2009.06.027.
[7]PENG X F,ZHENG Z P,CHENG K W,et al.Inhibitory effect of mung bean extract and its constituents vitexin and isovitexin on the formation of advanced glycation endproducts[J].Food Chemistry,2008,106(2):475-481.DOI:10.1016/j.foodchem.2007.06.016.
[8]ZHANG C Y,SHEN Y,XIAO P,et al.Nondestructive prediction of total phenolics,flavonoid contents,and antioxidant capacity of rice grain using near-infrared spectroscopy[J].Journal of Agricultural and Food Chemistry,2008,56(10):8268-8272.
[9]LU H F,ZHENG H,LOU H Q,et al.Using neural networks to estimate the losses of ascorbic acid,total phenols,flavonoid,and antioxidant activity in asparagus during thermal treatments[J].Journal of Agricultural and Food Chemistry,2010,58(10):2995-3001.
[10]王宇翎,张艳,方明,等.白花蛇舌草总黄酮的免疫调节作用[J].中国药理学通报,2005,21(4):444-447.DOI:10.3321/j.issn:1001-1978.2005.04.016.
[11]BREINHOLT V,NOSSAINI A,SVENDSEN G W,et al.Estrogenic activ ity o f f lav o no id s in mice.Th e imp or tan ce o f es tr o gen receptor distribution,metabolism and bioavailability[J].Food and Chemical Toxicology,2000,33(7):555-564.DOI:10.1016/S0278-6915(00)00046-6.
[12]吕萍,刘景圣,蔡丹,等.超声波法提取绿豆中的黄酮的最佳工艺研究[J].粮油加工,2010,12(8):117-119.
[13]LUO J Q,CAI W X,WU T,et al.Phytochemical distribution in hull and cotyledon of adzuki bean(Vigna angularis L.)and mung bean(Vigna radiate L.),and their contribution to antioxidant,antiinflammatory and anti-diabetic activities[J].Food Chemistry,2016,201:350-360.DOI:10.1016/j.foodchem.2016.01.101.
[14]裴凌鹏,惠伯棣,金宗濂,等.黄酮类化合物的生理活性及其制备技术研究进展[J].食品科学,2004,25(2):203-207.DOI:10.3321/j.issn:1002-6630.2004.02.048.
[15]耿秀芳,孙晓丽,王洪刚,等.银杏叶总黄酮降压作用的实验与临床研究[J].中国中药杂志,2002,27(8):606-608.DOI:10.3321/j.issn:1001-5302.2002.08.017.
[16]刘振国,郑建萍.黄酮类化合物的提取-分离-纯化研究进展[J].黑龙江科技信息,2011(25):20.DOI:10.3969/j.issn.1673-1328.2011.25.035.
[17]唐德智.黄酮类化合物的提取、分离、纯化研究进展[J].海峡药学,2009,21(12):101-104.DOI:10.3969/j.issn.1006-3765.2009.12.044.
[18]关海君,田娟娟,白志明.超滤膜技术在大豆异黄酮生产中的应用[J].中国油脂,2006,31(3):35-36.DOI:10.3321/j.issn.1003-7969.2006.03.012.
[19]李凤林,李青旺,高大威,等.天然黄酮类化合物分离纯化研究进展[J].江苏调味副食品,2008,25(5):20-24.DOI:10.3969/j.issn.1006-8481.2008.05.006.
[20]陈峰,陈剑锋.膜分离-溶剂萃取法联用分离纯化栀子蓝色素[J].食品研究与开发,2013,34(21):33-37.DOI:10.3969/j.issn.1005-6521.2013.21.010.
[21]易克传,岳鹏翔.膜分离技术及其在中草药分离纯化中的应用研究[J].包装与食品机械,2008,26(2):4-7.DOI:10.3969/j.issn.1005-1295.2008.02.002.
[22]HERNANDEZ E,COUTURE R,ROUSEFF R,et al.Evaluation of ultrafiltration and adsorption to debitter grapefruit juice and grapefruit pulp wash[J].Journal of Food Science,1992,57(3):664-666.DOI:10.1111/j.1365-2621.1992.tb08066x.
[23]关海君,田娟娟,白志明.超滤膜技术在大豆异黄酮生产中的应用[J].中国油脂,2006,31(3):35-36.DOI:10.3321/j.issn.1003-7969.2006.03.012.
[24]华耀祖.超滤技术与应用[M].北京:化学工业出版社,2004.
[25]ZHANG Y,LI S F,WU X W,et al.Macroporous resin adsorption for purification of flavonoids in Houttuynia cordata Thunb[J].Chinese Journal of Chemical Engineering,2007,15(6):872-876.DOI:10.1016/S1004-9541(08)60017-8.
[26]韩博,陈文,景文娟,等.AB-8大孔吸附树脂对甘草总黄酮的吸附[J].南方医科大学学报,2007,27(3):265-267.DOI:10.3321/j.issn.1673-4254.2007.03.005.
[27]FU B Q,LIU J,LI H,et al.The application of macroporous resins in the separation of licorice flavonoids and glycyrrhizic acid[J].Journal of Chromatography,2005,1089(1/2):18-24.DOI:10.1016/j.chroma.2005.06.051.
[28]贾海伦.芹菜黄酮提取及分离纯化工艺研究[D].哈尔滨:黑龙江大学,2008.
[29]骆党委,叶静,黄雅燕,等.A B-8大孔吸附树脂精制芦柑皮总黄酮及黄酮类化合物的分离[J].食品科学,2014,35(6):30-35.DOI:10.7506/spkx1002-6630-201406006.
[30]李侠,邹基豪,王大为.响应面试验优化超声波-酶法提取绿豆皮黄酮类化合物工艺[J].食品科学,2017,38(8):206-212.DOI:10.7506/spkx1002-6630-201708032.