高级检索

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

松栎柱锈菌春孢子总黄酮提取及抗氧化活性

瓮岳太 薛煜 徐硕 张国亮 刘悦 邸雪颖

瓮岳太, 薛煜, 徐硕, 张国亮, 刘悦, 邸雪颖. 松栎柱锈菌春孢子总黄酮提取及抗氧化活性[J]. 北京林业大学学报, 2017, 39(3): 38-47. doi: 10.13332/j.1000-1522.20160269
引用本文: 瓮岳太, 薛煜, 徐硕, 张国亮, 刘悦, 邸雪颖. 松栎柱锈菌春孢子总黄酮提取及抗氧化活性[J]. 北京林业大学学报, 2017, 39(3): 38-47. doi: 10.13332/j.1000-1522.20160269
WENG Yue-tai, XUE Yu, XU Shuo, ZHANG Guo-liang, LIU Yue, DI Xue-ying. Extraction of total flavonoids from Cronartium orientale aeciospores by response surface methodology and its antioxidant activity in vitro[J]. Journal of Beijing Forestry University, 2017, 39(3): 38-47. doi: 10.13332/j.1000-1522.20160269
Citation: WENG Yue-tai, XUE Yu, XU Shuo, ZHANG Guo-liang, LIU Yue, DI Xue-ying. Extraction of total flavonoids from Cronartium orientale aeciospores by response surface methodology and its antioxidant activity in vitro[J]. Journal of Beijing Forestry University, 2017, 39(3): 38-47. doi: 10.13332/j.1000-1522.20160269

松栎柱锈菌春孢子总黄酮提取及抗氧化活性

doi: 10.13332/j.1000-1522.20160269
基金项目: 

“948”国家林业局引进项目 2015-4-35

详细信息
    作者简介:

    瓮岳太。主要研究方向:森林保护学。Email:1150599114@qq.com  地址:150040  黑龙江省哈尔滨市香坊区和兴路26号东北林业大学林学院

    责任作者:

    邸雪颖,教授,博士生导师。主要研究方向:森林防火。Email:dixueying@126.com  地址:同上

  • 中图分类号: S763.15

Extraction of total flavonoids from Cronartium orientale aeciospores by response surface methodology and its antioxidant activity in vitro

  • 摘要: 为了确定松栎柱锈菌春孢子总黄酮超声辅助提取的最佳工艺条件,探索并评价松栎柱锈菌春孢子总黄酮的体外抗氧化活性,使用乙醇溶液提取总黄酮,采用亚硝酸钠-硝酸铝法测定该菌春孢子总黄酮芦丁当量。在单因素探索试验的基础上,通过响应面优化法得出最佳提取工艺条件。采用抗坏血酸(Vc)做阳性对照,对所得松栎柱锈菌春孢子总黄酮进行总抗氧化能力、总还原力以及清除二苯代苦味酰基(DPPH·)、清除羟基自由基(·OH)、清除超氧阴离子(O2-·)活性的测定与评价。研究结果表明:1)松栎柱锈菌春孢子总黄酮最优提取工艺条件:提取温度65℃,料液比1:120,提取时间38min,提取液乙醇体积分数60%,在该提取条件下总黄酮得率为1.382%,与预测值1.416%相差2.46%。2)松栎柱锈菌春孢子总黄酮具有较高的抗氧化能力,其总抗氧化能力与Vc相当。3)该菌春孢子总黄酮还原力高于Vc,量效关系比Vc更明显。4)该菌春孢子总黄酮与Vc对DPPH·的半抑制率(IC50)分别为0.0492和0.0335mg/mL,对DPPH·清除能力低于Vc但是量效关系明显。5)该菌春孢子总黄酮对·OH清除作用显著,IC50仅为0.0129mg/mL,质量浓度为0.0235mg/mL时清除率就能达到84.94%,清除能力远强于Vc。6)尚未查清该菌春孢子总黄酮提取液中对黄嘌呤酶氧化法的干扰因素,无法得知对O2-·的清除能力。利用响应面法得到了可靠的提取工艺,通过对比评价试验表明,该菌春孢子总黄酮具有很好的体外抗氧化活性,为锈菌开发利用和转变森林有害生物防治理念提供科学依据。

     

  • 图  1  全波长扫描结果

    Figure  1.  Results of full wavelength scan

    图  2  芦丁标准曲线

    Figure  2.  Standard curve of rutin

    图  3  乙醇体积分数对黄酮类化合物(CQF)得率的影响

    Figure  3.  Effects of ethanol concentration on extraction rate of flavonoids

    图  4  料液比对CQF得率的影响

    Figure  4.  Effects of solid-liquid ratio on extraction rate of flavonoids

    图  5  提取温度对CQF提取率的影响

    Figure  5.  Effects of extraction temperature on extraction rate of flavonoids

    图  6  提取时间对CQF提取率的影响

    Figure  6.  Effects of extraction time on extraction rate of flavonoids

    图  7  各因素交互作用对CQF得率影响的响应面与等高线图

    Figure  7.  Response surface and contour plots for the interaction effects of various factors on extraction rate of CQF

    图  8  CQF与Vc还原力比较

    Figure  8.  Comparison in reducing power of CQF and Vc

    图  9  受试物清除DPPH·能力的比较

    Figure  9.  Comparison of DPPH· radical scavenging capacity of CQF and Vc

    图  10  受试物总抗氧化能力的比较

    Figure  10.  Comparison in total antioxidant capacity of CQF and Vc

    图  11  受试物清除·OH能力的比较

    Figure  11.  Comparison in ·OH scavenging capacity of CQF and Vc

    图  12  受试物清除O2-·能力的比较

    Figure  12.  Comparison in O2-· scavenging capacity of CQF and Vc

    表  1  建模试验因子水平表

    Table  1.   Factors and levels of flavonoid extraction experiment

    水平
    Level
    影响因子Influencing factor
    温度Temperature(A)/℃ 料液比Solid-liquid ratio(B)/(g·mL-1) 时间Time(C)/min 乙醇体积分数
    Ethanol contentration(D)/%
    -2 35 1:100 20 50
    -1 45 1:120 30 60
    0 55 1:140 40 70
    1 65 1:160 50 80
    2 75 1:180 60 90
    下载: 导出CSV

    表  2  中心组合实验设计与结果

    Table  2.   Design and results of central composite design

    试验号
    Run No.
    温度
    Temperature(A)/℃
    料液比
    Solid-liquid ratio(B)/(g·mL-1)
    时间
    Time(C)/min
    乙醇体积分数
    Ethanol concentration(D)/%
    得率
    Yield(T)/%
    1 0 -2 0 0 1.6329
    2 1 -1 -1 1 1.0071
    3 -1 1 1 1 0.9619
    4 1 1 1 -1 1.4116
    5 2 0 0 0 1.2952
    6 0 0 0 0 1.1149
    7 -1 1 1 -1 1.1555
    8 0 0 0 0 1.1586
    9 1 -1 1 1 1.1196
    10 0 0 0 0 1.0930
    11 1 1 -1 -1 1.3928
    12 0 0 0 2 0.7542
    13 0 0 0 0 1.1477
    14 -1 -1 1 1 0.9697
    15 1 -1 -1 -1 1.2507
    16 -1 -1 -1 -1 1.1524
    17 -1 -1 -1 1 0.8713
    18 1 1 1 1 1.1805
    19 -1 1 -1 1 0.9993
    20 0 2 0 0 1.1453
    21 -1 -1 1 -1 1.1055
    22 0 0 -2 0 1.2133
    23 1 -1 1 -1 1.3959
    24 0 0 2 0 0.1969
    25 1 1 -1 1 1.0555
    26 0 0 0 -2 1.1040
    27 0 0 0 0 1.1258
    28 -2 0 0 0 1.3675
    29 0 0 0 0 1.1477
    30 -1 1 -1 -1 1.2304
    下载: 导出CSV

    表  3  数学模型方差分析

    Table  3.   Parameter estimate of regression model and variance analysis

    方差来源
    Variance source
    平方和Sum
    f squares
    自由度
    df
    均方
    Mean square
    F
    F value
    P
    P value
    显著性
    Significance
    模型Model 1.150 14 0.082 7.469 0.043 1 显著Significant
    温度Temperature(A) 0.062 1 0.062 1.520 0.237 1
    料液比Solid-liquid ratio(B) 0.009 1 0.009 0.210 0.650 1
    时间Time(C) 0.120 1 0.120 2.900 0.109 1
    乙醇体积分数Ethanol concentration(D) 0.290 1 0.290 7.010 0.018 3
    AB 0.000 1 0.000 0.001 0.981 9
    AC 0.013 1 0.013 0.320 0.577 2
    AD 0.004 1 0.004 0.093 0.765 2
    BC 0.005 1 0.005 0.120 0.736 6
    BD 0.000 1 0.000 0.005 0.945 7
    CD 0.004 1 0.004 0.100 0.756 6
    A2 0.095 1 0.095 2.300 0.149 8
    B2 0.150 1 0.150 3.580 0.078 1
    C2 0.260 1 0.260 6.380 0.023 3
    D2 0.048 1 0.048 1.160 0.297 5
    残差Residual 0.620 15 0.041
    失拟项Lack of fit 0.610 10 0.061 0.017 0.149 不显著Not significant
    纯误差Pure error 0.003 5 0.001
    总误差Total error 1.770 29
    下载: 导出CSV
  • [1] 薛煜, 邵力平, 马泽芳, 等.兴凯湖松瘤锈病病、健组织中主要有机成分变化及其对结实的影响[J].东北林业大学学报, 1992, 20(2): 30-34. http://www.cnki.com.cn/Article/CJFD1992-DBLY199202004.htm

    XUE Y, SHAO L P, MA Z F, et al. Change in main organic matter contents between infected and healthy tissues and the effect of Xinkai Lake pine rust to fruiting ability[J].Journal of Northeast Forestry University, 1992, 20(2): 30-34. http://www.cnki.com.cn/Article/CJFD1992-DBLY199202004.htm
    [2] KANEKO S. Cronartium orientale, sp. nov., segregation of the pine gall rust in eastern Asia from Cronartium quercuum[J]. Mycoscience, 2000, 41(2): 115-122. doi: 10.1007/BF02464319
    [3] 邵力平, 薛煜, 王振华, 等.兴凯湖松瘤锈病损失量的研究[J].东北林业大学学报, 1989, 17(2): 1-5. http://d.old.wanfangdata.com.cn/Conference/280810

    SHAO L P, XUE Y, WANG Z H, et al. Study on the losses of Xingkai Lake pine gall rust[J].Journal of Northeast Forestry University, 1989, 17(2): 1-5. http://d.old.wanfangdata.com.cn/Conference/280810
    [4] PIETTA P G. Flavonoids as antioxidants[J]. Journal of Natural Products, 2000, 63(7): 1035-1042. doi: 10.1021/np9904509
    [5] RICE-EVANS C. Flavonoids antioxidants[J]. Current Medicinal Chemistry, 2001, 8(7): 797-807. doi: 10.2174/0929867013373011
    [6] BIRT D F, JEFFERY E. Flavonoids[J]. Advances in Nutrition: An International Review Journal, 2013, 4(5): 576-577. doi: 10.3945/an.113.004465
    [7] YAO L H, JIANG Y M, SHI J, et al. Flavonoids in food and their health benefits[J]. Plant Foods for Human Nutrition, 2004, 59(3): 113-122. doi: 10.1007/s11130-004-0049-7
    [8] RAMAN S T, GANESHAN A K, CHEN C, et al. In vitro and in vivo antioxidant activity of flavonoid extracted from mulberry fruit (Morus alba L.)[J]. Pharmacognosy Magazine, 2016, 12(46): 128-133. doi: 10.4103/0973-1296.177910
    [9] NAEEM I, TASKEEN A, MUBEEN H, et al. Characterization of flavonols present in barks and needles of Pinus wallichiana and Pinus roxburghii[J]. Asian Journal of Chemistry, 2010, 22(1): 41-44. https://www.researchgate.net/publication/281535179_Characterization_of_Flavonols_Present_in_Barks_and_Needles_of_Pinus_wallichiana_and_Pinus_roxburghii
    [10] MAIMOONA A, NAEEM I, SADDIQE Z, et al. Analysis of total flavonoids and Phenolics in different fractions of bark and needles of Pinus roxburghii and Pinus wallichiana [J].Journal of Medicinal Plant Research, 2011, 5(21):5216-5220.
    [11] 冯慧英, 樊金拴, 刘滨, 等.巴山冷杉黄酮的提取鉴定及其抗氧化性分析[J].食品工业, 2016, 37(1): 63-67. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=spgy201601016

    FENG H Y, FAN J S, LIU B, et al.Extraction and identification of flavonoids from Abies fargesii and its antioxidant activity[J].The Food Industry, 2016, 37(1): 63-67. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=spgy201601016
    [12] 李石清, 张春椿, 年慧慧, 等.南方红豆杉枝叶中黄酮组分的含量测定及抗氧化活性研究[J].甘肃中医学院学报, 2012, 29(3): 28-31. doi: 10.3969/j.issn.1003-8450.2012.03.010

    LI S Q, ZHANG C C, NIAN H H, et al. Study on antioxidant activity and determination of different flavonoids components in branches and leaves of Taxus mairei (Lemee et Levl.)[J].Journal of Gansu University College of Traditional Chinese Medicine, 2012, 29(3): 28-31. doi: 10.3969/j.issn.1003-8450.2012.03.010
    [13] 王飞, 樊金拴, 冯慧英, 等.青扦针叶总黄酮超声提取及抗氧化活性[J].西北林学院学报, 2016, 31(1): 243-249. doi: 10.3969/j.issn.1001-7461.2016.01.42

    WANG F, FAN J S, FENG H Y, et al. Ultrasound assisted extraction and antioxidant activities of total flavonoids in the needles of Picea wilsonii[J].Journal of Northwest Forestry University, 2016, 31(1): 243-249. doi: 10.3969/j.issn.1001-7461.2016.01.42
    [14] KOZARSKI M S, KLAUS A S, NIKSIC M P, et al. Polysaccharides of higher fungi: biological role, structure and antioxidative activity[J]. Hemijska Industrija, 2014, 68(3): 305-320. doi: 10.2298/HEMIND121114056K
    [15] 张文州, 许嵘.食药用真菌多糖的研究进展[J].食品工业科技, 2014, 35(15): 395-399. http://d.old.wanfangdata.com.cn/Periodical/smdhx200202022

    ZHANG W Z, XU R. Research progress in edible-medicinal fungi polysaccharide[J].Science and Technology of Food Industry, 2014, 35(15): 395-399. http://d.old.wanfangdata.com.cn/Periodical/smdhx200202022
    [16] FU L, XU B, XU X, et al. Antioxidant capacities and total phenolic contents of 56 wild fruits from south China[J]. Molecules, 2010, 15(12): 8602-8617. doi: 10.3390/molecules15128602
    [17] FREINBICHLER W, BIANCHI L, COLIVICCHI M A, et al. The detection of hydroxyl radicals in vivo[J]. Journal of Inorganic Biochemistry, 2008, 102(5-6): 1329-1333. doi: 10.1016/j.jinorgbio.2007.12.017
    [18] 陈士明, 陆亚蒙, 严小敏, 等.超氧阴离子自由基的产生及其与天然药物的作用[J].复旦学报(自然科学版), 1991, 30(1): 31-36. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=HY000001731385

    CHEN S M, LU Y M, YAN X M, et al. Generation of superoxide anion radical and the effect on natural medicines[J].Journal of Fudan University(Natural Science), 1991, 30(1): 31-37. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=HY000001731385
    [19] ROBAK J, GRYGLEWSKI R J. Flavonoids are scavengers of superoxide anions[J]. Biochemical Pharmacology, 1988, 37(5): 837-841. doi: 10.1016/0006-2952(88)90169-4
    [20] PUPPO A, RIGAUD J, JOB D. Role of superoxide anion in leghemoglobin autoxidation[J]. Plant Science Letters, 1981, 22(4): 353-360. doi: 10.1016/0304-4211(81)90081-X
    [21] 闫琳娜.松针落叶总黄酮的提取、纯化及抗氧化性能分析[D].吉林: 吉林大学, 2009.

    YAN L N. Extraction, purification and antioxidant activity analysis of flavonoids from pine needle[D]. Jilin: Jilin University, 2009.
    [22] 孙琼.杏鲍菇黄酮类化合物提纯及抗氧化活性研究[D].临汾: 山西师范大学, 2015.

    SUN Q.Study on extraction and purification process of flavonoids from Pleurotus eryngii and its antioxidant activities[D]. Linfen: Shanxi Normal University, 2015.
    [23] 黄晓冬, 蔡建秀, 许丽璇, 等.桦褐孔菌总黄酮体内外抗氧化作用[J].食品与生物技术学报, 2012, 31(10): 1062-1068. doi: 10.3969/j.issn.1673-1689.2012.10.009

    HUANG X D, CAI J X, XU L X, et al.Study on antioxidation effect in vitro and in vivo of total flavonoids from Inonotus obliquus[J].Journal of Food Science and Biotechnology, 2012, 31(10): 1062-1068. doi: 10.3969/j.issn.1673-1689.2012.10.009
    [24] BOONSONG S, KLAYPRADIT W, WILAIPUN P. Antioxidant activities of extracts from five edible mushrooms using different extractants[J]. Agriculture and Natural Resources, 2016, 50(2): 89-97. doi: 10.1016/j.anres.2015.07.002
    [25] TREML J, ŠMEJKAL K. Flavonoids as potent scavengers of hydroxyl radicals[J]. Comprehensive Reviews in Food Science and Food Safety, 2016, 15(4): 720-738. doi: 10.1111/1541-4337.12204
  • 加载中
图(12) / 表(3)
计量
  • 文章访问数:  1052
  • HTML全文浏览量:  203
  • PDF下载量:  18
  • 被引次数: 0
出版历程
  • 收稿日期:  2016-08-27
  • 修回日期:  2016-11-17
  • 刊出日期:  2017-03-01

目录

    /

    返回文章
    返回