水芹中芹菜素的提取纯化与抗氧化活性研究
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摘要
对水芹中芹菜素的提取纯化与抗氧化活性进行研究。首先,以水芹为原料、乙醇为溶剂,黄酮得率为评价指标,通过单因素试验及四因素三水平L_9(3~4)正交试验,确定了粗黄酮的最佳提取条件。同时,通过静态吸附,以树脂对粗黄酮的吸附率和解析率为评价指标,比较了AB-8,X-5及HP-20型这3种大孔吸附树脂对粗黄酮纯化效果,以确定其最佳纯化树脂。采用液相色谱和质谱对纯化后的芹菜素进行分离验证。同时,研究了芹菜素粗提物及单体的清除DPPH自由基、羟基自由基及超氧阴离子自由基的能力。结果表明,粗黄酮的最佳提取条件为料液比1∶35(g∶mL),提取温度80℃,提取时间4 h,乙醇体积分数90%(V/V);AB-8型大孔吸附树脂具有较好的纯化效果。液相色谱结果表明,水芹黄酮粗提物共有4种组分,通过与芹菜素标准品比对,分离出芹菜素单体,并结合质谱图进行鉴定。同时,芹菜素粗提物及单体对DPPH自由基、羟基自由基及超氧阴离子自由基都具有一定的清除能力,其中芹菜素单体的抗氧化性要大于粗提物,且芹菜素单体对超氧阴离子自由基的清除能力较强。
In this paper, the extraction, purification and antioxidant activity of apigenin from water dropwort were studied.Firstly,the yield of flavonoids was evaluated by using water dropwort as raw material and ethanol as solvent. The optimum extraction conditions of crude flavonoids were determined by single factor test and orthogonal test. In the purification stage,three types of macroporous resins including AB-8,X-5 and HP-20 were used for statically adsorbed. Through the calculation of the adsorption rate and desorption rate of crude flavonoids to determine the best purification resin. The purified apigenin was then separated and verified by liquid chromatography and mass spectrometry. At the same time,the ability of crude apigenin extract and monomer to scavenge DPPH·,hydroxyl radicals and superoxide anion radicals was studied. The results showed that the optimal extraction conditions of crude flavonoids were: ratio of material to liquid 1∶35(g∶mL),extraction temperature of 80 ℃,extraction time of 4 h and ethanol concentration of 90%(V/V). At the same time,the results showed that the AB-8 macroporous adsorption resin has a good purification effect. The results of liquid chromatography showed that there were four components in the crude extract of water cress,and the apigenin monomer was isolated by comparison with the apigenin standard and identified by mass spectrometry. At the same time, crude apigenin and monomer had certain scavenging ability to DPPH free radicals, hydroxyl radicals and superoxide anion radicals. The oxidative properties of apigenin monomers were greater than those of crude extracts,and apigenin alone the body had a strong ability to scavenge superoxide anion radicals.
In this paper, the extraction, purification and antioxidant activity of apigenin from water dropwort were studied.Firstly,the yield of flavonoids was evaluated by using water dropwort as raw material and ethanol as solvent. The optimum extraction conditions of crude flavonoids were determined by single factor test and orthogonal test. In the purification stage,three types of macroporous resins including AB-8,X-5 and HP-20 were used for statically adsorbed. Through the calculation of the adsorption rate and desorption rate of crude flavonoids to determine the best purification resin. The purified apigenin was then separated and verified by liquid chromatography and mass spectrometry. At the same time,the ability of crude apigenin extract and monomer to scavenge DPPH·,hydroxyl radicals and superoxide anion radicals was studied. The results showed that the optimal extraction conditions of crude flavonoids were: ratio of material to liquid 1∶35(g∶mL),extraction temperature of 80 ℃,extraction time of 4 h and ethanol concentration of 90%(V/V). At the same time,the results showed that the AB-8 macroporous adsorption resin has a good purification effect. The results of liquid chromatography showed that there were four components in the crude extract of water cress,and the apigenin monomer was isolated by comparison with the apigenin standard and identified by mass spectrometry. At the same time, crude apigenin and monomer had certain scavenging ability to DPPH free radicals, hydroxyl radicals and superoxide anion radicals. The oxidative properties of apigenin monomers were greater than those of crude extracts,and apigenin alone the body had a strong ability to scavenge superoxide anion radicals.
引文
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[20]郭英.HPLC法测定大叶水芹菜中芹菜素含量[J].微量元素与健康研究,2010,27(3):37-38.
[21]王克勤,罗军武,陈静萍,等.高效液相色谱法测定芹菜中芹菜素含量[J].食品与机械,2009, 25(2):74-77.
[22]池玉梅.高效液相-四级杆飞行时间串联质谱分析黄蜀葵花中黄酮醇类化合物[J].分析化学,2009,37(2):227-231.
[23]王健,潘利华.蓝莓花青素的抗氧化活性研究[J].安徽农业科学,2013,41(12):5 487-5 489.
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[25]王二雷.笃斯越桔花青素分离纯化及其抗氧化活性研究[D].长春:吉林大学,2006.
[2]黄正明,杨新波,曹文斌.水芹的本草考证[J].中草药,2001,32(1):59-62.
[3]黄正明,杨新波,曹文斌,等.中药水芹的现代研究与应用[J].解放军药学学报,2001,17(5):266-269.
[4]中华人民共和国卫生部药典委员会.中华人民共和国药典[S].一部.北京:化学工业出版社,2005:附录VA.
[5]董钰明,封士兰,段生玉,等.不同品种芹菜及不同部位中总黄酮含量的测定[J].兰州医学院学报,2002,28(3):32-33.
[6]周毅峰,石开明,艾训儒,等.快速溶剂萃取法提取青蒿素条件研究[J].中药材,2008(2):296-298.
[7]张冬冬,徐迪,洪伟.超声提取柑橘皮中总黄酮的研究[J].农产品加工,2017(17):1-3.
[8]欧阳平,张高勇,康保安.类黄酮提取的基本原理、影响因素和传统方法[J].中国食品添加剂,2003(5):54-57.
[9]毕丽君.水芹中总黄酮类化合物最佳提取工艺的研究[J].食品科学,1999,20(12):35-37.
[10]宫健伟,叶蕾.大孔树脂在中草药及复方提取分离中的应用进展[J].甘肃中医,2007,20(4):55.
[11]Jun L,Lou J,Yi S,et al. A simple method for the simultaneous decoloration and deproteinization of crude levan extract from Paenibacillus polymxa EJS-3 by macroporous resin[J]. Bioresource Technology, 2010(15):6 077-6 083.
[12]王克勤,刘仲华,罗军武,等.XDA-大孔树脂对芹菜黄酮分离纯化的研究[J].食品与机械,2007(3):65-69.
[13]唐巧玉,周毅峰,任甜甜,等.大孔树脂对水芹菜总黄酮的分离纯化研究[J].时珍国医国药,2009,20(10:2 480-2 482.
[14]陈况况,章宏慧,刘东红,等.大孔吸附树脂纯化水芹总黄酮的研究[J].食品工业科技,2013,34(20):246-250.
[15]任荣军.大孔树脂吸附技术在黄酮类成分分离纯化中的应用[J].中国药业,2008,17(16):71-73.
[16]王超,马海乐,王振斌.AB-8大孔树脂分离提纯无花果总黄酮的研究[J].食品研究与开发,2005(4):11-12, 21.
[17]李春美,钟朝辉,窦宏亮,等.大孔树脂分离纯化柚皮黄酮的研究[J].农业工程学报,2006, 22(3):153-157.
[18]周英新.基于黄酮类分子构型的大孔树脂吸附动力学模型研究[D].北京:北京中医药大学,2012.
[19]汪凤山,赵旭伟,方舟,等.高效液相色谱法测定北刘寄奴药材中木犀草素和芹菜素的含量[J].黑龙江医药科学,2015,38(5):70-71.
[20]郭英.HPLC法测定大叶水芹菜中芹菜素含量[J].微量元素与健康研究,2010,27(3):37-38.
[21]王克勤,罗军武,陈静萍,等.高效液相色谱法测定芹菜中芹菜素含量[J].食品与机械,2009, 25(2):74-77.
[22]池玉梅.高效液相-四级杆飞行时间串联质谱分析黄蜀葵花中黄酮醇类化合物[J].分析化学,2009,37(2):227-231.
[23]王健,潘利华.蓝莓花青素的抗氧化活性研究[J].安徽农业科学,2013,41(12):5 487-5 489.
[24]姚钰荣.紫薯花青素的提取纯化、稳定性及抗氧化活性研究[D].保定:河北农业大学,2009.
[25]王二雷.笃斯越桔花青素分离纯化及其抗氧化活性研究[D].长春:吉林大学,2006.