大麻叶中总黄酮提取工艺的优化及其抗氧化活性测定
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摘要
为优化大麻叶总黄酮提取工艺并评估大麻叶总黄酮抗氧化活性,研究在单因素试验基础上,以提取时间、提取温度、料液比和乙醇体积分数为影响因素,总黄酮浸提量为评价指标,采用响应面试验设计优化提取工艺。采用FRAP总抗氧化能力测定试验、DPPH自由基清除试验以及ABTS自由基清除试验对总黄酮提取物进行抗氧化活性评价。结果表明:提取大麻叶总黄酮的最佳工艺条件为乙醇体积分数50%、料液比1∶20(g/m L)、提取时间2.0 h、提取温度70℃。在该最优提取条件下,大麻叶总黄酮浸提量为14.28 mg/g。抗氧化试验表明,大麻叶总黄酮的总抗氧化能力相当于1.288 mmol/L Fe SO4,对DPPH自由基的清除能力(IC50=3.7μg/m L)低于VC的清除能力(IC50=2.3μg/m L),而对ABTS自由基的清除力相当于0.391 mmol/L的Trolox。该工艺提取大麻叶得到的总黄酮含量较高,且具有一定的抗氧化性,可为未来大麻叶总黄酮的开发提供一定的科学依据。
In order to optimize the extraction process of total flavonoids from hemp( Cannabis sativa L.) and evaluate the antioxidant activity of total flavonoids,the response surface test was used in the research. With the extraction time,extraction temperature,solid-liquid ratio and ethanol volume fraction as the influencing factors,the total flavonoids extraction amount was used as the evaluation index. Meanwhile,the antioxidant activities of total flavonoids were investigated through the measurement of FRAP total antioxidant capacity,scavenging capacities on DPPH radical and ABTS radical. The results showed that the optimal extraction conditions were as follows: extraction time,2. 0 h; ethanol concentration,50%;solid-liquid ratio,1∶ 20( g/m L); temperature,70 °C. Under these conditions,the concentration of total flavonoids could reach 14. 28 mg/g. The total antioxidant capacity was equivalent to 1. 288 mmol/L Fe SO4. The IC50 on scavenging DPPH was 3. 7 μg/m L,which was lower than the scavenging capacities of VC. Meanwhile,the scavenging capacity on ABTS was equivalent to that of 0. 391 mmol/L Trolox. In conclusion,substantial total flavonoids could be obtained under optimal extraction conditions and had a certain antioxidant activity,which would provide reliable scientific instruction on the exploitation of total flavonoids in hemp leaves.
In order to optimize the extraction process of total flavonoids from hemp( Cannabis sativa L.) and evaluate the antioxidant activity of total flavonoids,the response surface test was used in the research. With the extraction time,extraction temperature,solid-liquid ratio and ethanol volume fraction as the influencing factors,the total flavonoids extraction amount was used as the evaluation index. Meanwhile,the antioxidant activities of total flavonoids were investigated through the measurement of FRAP total antioxidant capacity,scavenging capacities on DPPH radical and ABTS radical. The results showed that the optimal extraction conditions were as follows: extraction time,2. 0 h; ethanol concentration,50%;solid-liquid ratio,1∶ 20( g/m L); temperature,70 °C. Under these conditions,the concentration of total flavonoids could reach 14. 28 mg/g. The total antioxidant capacity was equivalent to 1. 288 mmol/L Fe SO4. The IC50 on scavenging DPPH was 3. 7 μg/m L,which was lower than the scavenging capacities of VC. Meanwhile,the scavenging capacity on ABTS was equivalent to that of 0. 391 mmol/L Trolox. In conclusion,substantial total flavonoids could be obtained under optimal extraction conditions and had a certain antioxidant activity,which would provide reliable scientific instruction on the exploitation of total flavonoids in hemp leaves.
引文
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[26]卢赛赛,许凤,王鸿飞,等.杨梅叶中总黄酮提取及其抗氧化能力研究[J].果树学报,2015,32(3):460-468.
[27]刘香萍,王国庆,李国良,等.响应面法优化提取紫花苜蓿叶总黄酮及其抗氧化活性研究[J].中国食品学报,2016,16(4):145-152.
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[2]粟建光,陈基权,谢小美.大麻育种现状与前景[J].中国麻业科学,2006,28(4):212-217.
[3]谭龙涛,喻春明,陈平,等.麻类作物多用途研究现状与发展趋势[J].中国麻业科学,2012:34(2),94-99.
[4]吕咏梅,杨龙,胡万群.大麻产业研究进展---Ⅱ.大麻的综合利用与发展建议[J].中国麻业科学,2012,34(1):38-42.
[5]弓佩含,杨洋,刘玉婷,等.大麻化学成分及药理作用的研究进展[J].中国实验方剂学杂志,2017(13):212-219.
[6]Acker S A B E V,Berg D J V D,Michèl N.J.L.Tromp,et al.Structural aspects of antioxidant activity of flavonoids[J].Free Radical Biology and Medicine,1996,20(3):331-342.
[7]刘莉华,宛晓春,李大祥.黄酮类化合物抗氧化活性构效关系的研究进展[J].安徽农业大学学报,2002,29(3):265-270.
[8]Ren W,Qiao Z,Wang H,et al.Flavonoids:promising anticancer agents[J].Medicinal Research Reviews,2010,23(4):519-534.
[9]黄池宝,罗宗铭,宾丽英.黄酮类化合物抗氧化性与其结构关系的研究[J].广东工业大学学报,2000,17(2):71-75.
[10]刘成伦,唐德容.黄酮类化合物抗氧化性质的研究进展[J].食品研究与开发,2006,27(5):158-160.
[11]姜红波.黄酮类物质的提取和含量测定方法研究进展[J].化工时刊,2010,24(11):45-49.
[12]李加兴,陈选,邓佳琴,等.黄秋葵黄酮的提取工艺和体外抗氧化活性研究[J].食品科学,2014,35(10):121-125.
[13]杨申明,王波,王振吉,等.金雀花总黄酮提取工艺优化及抗氧化性研究[J].浙江农业学报,2015,27(2):278-284.
[14]万忠民,沈玲玲.苦参中黄酮类物质的提取及抗氧化研究[J].食品科学,2009,30(24):137-140.
[15]吕寒,滕杰晖,陈剑,等.枇杷叶总黄酮的纯化工艺及抗氧化活性研究[J].中国现代应用药学,2014(1):40-44.
[16]陈红梅,谢翎.响应面法优化半枝莲黄酮提取工艺及体外抗氧化性分析[J].食品科学,2016,37(2):45-50.
[17]侯学敏,李林霞,张直峰,等.响应面法优化薄荷叶总黄酮提取工艺及抗氧化活性[J].食品科学,2013,34(6):124-128.
[18]千春录,侯顺超,殷健东.响应面试验优化水芹黄酮超声波辅助提取工艺及其抗氧化性[J].食品科学,2016,37(10):76-81.
[19]李婷婷,刘秀芳,蔡光明,等.大孔树脂分离纯化大麻果胶中的总黄酮[J].解放军药学学报,2011,27(3):212-214.
[20]汪秋安,周冰,单杨.天然黄酮类化合物的抗氧化活性和提取技术研究进展[J].化工生产与技术,2004,11(5):29-32.
[21]白生文,汤超,田京,等.沙棘果渣总黄酮提取工艺及抗氧化活性分析[J].食品科学,2015,36(10):59-64.
[22]Hui-Hui L,Hui-Rong C,Lan S,et al.Optimization of Flavonoids Extraction from Anoectochilus Roxburghii by Response Surface Methodology[J].Pharmaceutical Biotechnology,2017,24:321-327.
[23]Ozdemir A,Cho B R.Response surface-based robust parameter design optimization with both qualitative and quantitative variables[J].Engineering Optimization,2017:1-17.
[24]avikin K,ivkovi C'J,Alimpi C'A,et al.Activity guided fractionation of pomegranate extract and its antioxidant,antidiabetic and antineurodegenerative properties[J].Industrial Crops and Products,2018,113:142-149.
[25]Karkouch I,Tabbene O,Gharbi D,et al.Antioxidant,antityrosinase and antibiofilm activities of synthesized peptides derived from Vicia faba protein hydrolysate:A powerful agents in cosmetic application[J].Industrial Crops and Products,2017,109:310-319.
[26]卢赛赛,许凤,王鸿飞,等.杨梅叶中总黄酮提取及其抗氧化能力研究[J].果树学报,2015,32(3):460-468.
[27]刘香萍,王国庆,李国良,等.响应面法优化提取紫花苜蓿叶总黄酮及其抗氧化活性研究[J].中国食品学报,2016,16(4):145-152.
[28]Belhaj D,Frikha D,Athmouni K,et al.Box-Behnken design for extraction optimization of crude polysaccharides from Tunisian,Phormidium versicolor,cyanobacteria(NCC 466):partial characterization,in vitro,antioxidant and antimicrobial activities[J].International Journal of Biological Macromolecules,2017,105(Pt 2):1501-1510.