金缕半枫荷多酚提取及其抗氧化抗菌活性研究
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摘要
为进一步开发和利用金缕半枫荷,该研究以超声时间、液料比、乙醇体积分数和超声温度为考察因素,用金缕半枫荷总多酚提取率作为评价指标,采用单因素和正交试验优化提取工艺确定最佳工艺,并以最佳工艺制备金缕半枫荷多酚;进一步采用DPPH·和羟自由基清除法评价金缕半枫荷多酚的抗氧化能力,以抑菌圈和最小抑菌浓度(MIC)为指标,采用牛津杯和肉汤稀释法考察其抗菌活性。结果表明:最佳提取工艺为乙醇浓度70%,料液比为1∶16,提取时间为60 min,提取温度为60℃。金缕半枫荷多酚清除DPPH·自由基的IC_(50)为5.22μg·mL~(-1),与阳性对照维生素C的IC_(50)值4.31μg·mL~(-1)无显著差异,羟自由基清除率IC_(50)为105μg·mL~(-1),显著低于维生素C的IC_(50)值(180μg·mL~(-1))。对金黄色葡萄球菌和大肠杆菌具有抑制作用,抑菌圈直径分别为(13.7±1.2) mm和(10.0±1.3) mm,MIC分别为0.393和0.785 mg·mL~(-1)。这表明金缕半枫荷多酚具有抗菌和显著抗氧化能力,为开发利用金缕半枫荷资源提供了理论依据。
In order to develop and utilize Semiliquidambar cathayensis,orthogonal design was used to explore the effects of ethanol content,liquid to solid ratio,extraction time and temperature. Moreover,the DPPH· and hydroxyl radical scavenging activities of the polyphenols were also investigated. The inhibition zone and minimal inhibitory concentration( MIC) were used to detect the antibacterial activities,and determined by Oxford Cup method and broth dilution method. The results were as follows: the best extraction condition was ethanol content of 70%,the solid to liquid ratio of 1∶16,the extraction time of 60 min,and the extraction temperature of 60 ℃. In addition,there was no significant difference of the IC_(50) value of DPPH· free radical scavenging between 5.22 μg·m L~(-1) polyphenols and 4.31 μg·m L~(-1) ascorbic acid. And the IC_(50) value 105 μg·m L~(-1) of hydroxyl free radical scavenging of the polyphenols was significantly lower than that of ascorbic acid 180 μg·m L~(-1). The results also showed that the polyphenols had antibacterial activity against Staphyloccocus aureus and Escherichia coli with the antibacterial circle were (13.7 ± 1.2) mm and (10.0 ± 1.3)mm,respectively. And the MIC were 0.393 and 0.785 mg·m L~(-1),respectively. This research provides the theoretical information for further development and utilization of Semiliquidambar cathayensis resources.
In order to develop and utilize Semiliquidambar cathayensis,orthogonal design was used to explore the effects of ethanol content,liquid to solid ratio,extraction time and temperature. Moreover,the DPPH· and hydroxyl radical scavenging activities of the polyphenols were also investigated. The inhibition zone and minimal inhibitory concentration( MIC) were used to detect the antibacterial activities,and determined by Oxford Cup method and broth dilution method. The results were as follows: the best extraction condition was ethanol content of 70%,the solid to liquid ratio of 1∶16,the extraction time of 60 min,and the extraction temperature of 60 ℃. In addition,there was no significant difference of the IC_(50) value of DPPH· free radical scavenging between 5.22 μg·m L~(-1) polyphenols and 4.31 μg·m L~(-1) ascorbic acid. And the IC_(50) value 105 μg·m L~(-1) of hydroxyl free radical scavenging of the polyphenols was significantly lower than that of ascorbic acid 180 μg·m L~(-1). The results also showed that the polyphenols had antibacterial activity against Staphyloccocus aureus and Escherichia coli with the antibacterial circle were (13.7 ± 1.2) mm and (10.0 ± 1.3)mm,respectively. And the MIC were 0.393 and 0.785 mg·m L~(-1),respectively. This research provides the theoretical information for further development and utilization of Semiliquidambar cathayensis resources.
引文
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YANG L,WANG YQ,LIU SZ,et al.,2016.Research progress of chemical constituents and pharmacological activities from three commonly used Ban-feng-he medicinal plants[J].Chin J Exp Trad Med Form,22(22):192-196.[杨丽,王雅琪,刘升长,等,2016.三种常用半枫荷类药用植物的化学成分与生物活性研究概况[J].中国实验方剂学杂志,22(22):191-196.]
YANG WL,YAO ZS,LUO XQ,et al.,1999.Analgesic and anti-inflammatory effects of the ethanol extracts of jinlu ban fenghe(Semiliquidambar cathayensis)root[J].Jiangxi Sci,17(3):176-179.[杨武亮,姚振生,罗小泉,等,1999.金缕半枫荷的镇痛和抗炎作用[J].江西科学,17(3):176-179.]
ZHANG SY,ZHUANG YB,LUO LK,et al.,2017.Antioxidant and inhibitory effect on nitrosation of extracts from Balanophora arlandii[J].Nat Prod Res Dev,29(12):2081-2086.[张声源,庄远杯,罗铝铿,等,2017.红冬蛇菰提取物抗氧化及抑制亚硝化作用研究[J].天然产物研究与开发,29(12):2081-2086.]
ZHANG Y,WU YT,ZHENG W,et al.,2017.The antibacterial activity and antibacterial mechanism of a polysaccharide from Cordyceps cicadae[J].J Funct Foods,38:273-279.
ZHOU GX,YANG YC,SHI JG,et al.,2002.Studies on chemical constituents from Semiliquidambar cathayensis[J].Chin Trad Herbal Drugs,33(7):589-591.[周光雄,杨永春,石建功,等,2002.金缕半枫荷化学成分研究[J].中草药,33(7):589-591.]
ZHU YX,SONG H,ZHANG X,et al.,2017.Recovery of flavonoids from walnuts de-pellicle wastewater with macroporous resins and evaluation of antioxidant activities in vitro[J].J Food Proc Eng,40(1):e12275.
FLOYD RA,TOWNER RA,HE T,et al.,2011.Translational research involving oxidative stress and diseases of aging[J].Free Radical Biol Med,51(5):931-941.
GONZALEZ N,ELISSETCHE J,PEREIRA M,et al.,2017.Extraction of polyphenols from Eucalyptus nitens and Eucalyptus globulus:Experimental kinetics,modeling and evaluation of their antioxidant and antifungical activities[J].Ind Crops Products,109:737-745.
HUANG DT,PANG SH,LIANG QP,et al.,2012.Research status of Yao medicine resources in Guangxi Province[J].JMed Pharm Chin Minorities,3:68-69.[黄东挺,庞声航,梁琼评,等,2012.广西瑶药资源的现状调查[J].中国民族医药杂志,3:68-69.]
KUPFER R,DWYER-NIELD LD,MALKINSON AM,et al.,2002.Lung toxicity and tumor promotion by hydroxylated derivatives of 2,6-di-tert-butyl-4-methylphenol(BHT)and 2-tertbutyl-4-methyl-6-iso-propylphenol:Correlation with quinone methide reactivity[J].Chem Res Toxicol,15(8):1106-1112.
LU HX,WU ZL,LIANG WJ,et al.,2015.Chemical constituents from Semiliquidambar cathayensis roots[J].JChin Med Mat,38(12):2543-2546.[卢海啸,吴卓玲,梁伟江,等,2015.半枫荷根的化学成分研究[J].中药材,38(12):2543-2546.]
MIAO WL,ZHAO SQ,ZHANG JL,et al.,2017.Study on antibacterial activity and toxicity of main related substance in doxycycline hydrochloride[J].Chin J Pharm Anal,37(12):2202-2208.[缪文玲,赵述强,张锦琳,等,2017.盐酸多西环素主要有关物质体外抗菌活性及毒性的研究[J].药物分析杂志,37(12):2202-2208.]
TODARO L,RUSSO D,CETERA P,et al.,2017.Effects of thermo-vacuum treatment on secondary metabolite content and antioxidant activity of poplar(Populus nigra L.)wood extracts[J].Ind Crops Products,109:384-390.
WEI CL,LU HX,SHI HM,et al.,2017.Extraction technology research on orthogonal excogitation to optimize the polyphenol of Semiliquidambar cathayensis[J].J Yulin Norm Univ,38(2):75-78.[韦春玲,卢海啸,施海敏,等,2017.正交设计优选半枫荷多酚提取工艺研究[J].玉林师范学院报,38(2):75-78.]
WEI H,YAN XJ,YANG JW,et al.,2017.Determination of total phenols content and antioxidant capacity of walnut leaf[J/OL].Guihaia,:1-10.http://kns.cnki.net/kcms/detail/45.1134.Q.20170706.1641.002.html.[魏欢,颜小捷,杨建文,等,2017.核桃叶多酚含量的测定和抗氧化能力的研究[J/OL].广西植物,:1-10.http://kns.cnki.net/kcms/detail/45.1134.Q.20170706.1641.002.html.]
XIA YJ,SUN XM,ZHANG J,et al.,2014.Simultaneous determination of five polyphenolic substances in the leaves of olea europaea by high performance liquid chromatography[J].Chin J Anal Lab,33(7):766-770.[夏雅俊,孙小明,张佳,等,2014.HPLC法同时测定油橄榄叶中的5种多酚类化合物含量[J].分析试验室,33(7):766-770.]
XIE Q,WANG W,CHEN QX,2014.Comparative study on three different methods for the determination of total phenolics in Chinese olive[J].Food Sci,35(8):204-207.[谢倩,王威,陈清西,2014.橄榄多酚含量测定方法的比较[J].食品科学,35(8):204-207.]
YANG L,WANG YQ,LIU SZ,et al.,2016.Research progress of chemical constituents and pharmacological activities from three commonly used Ban-feng-he medicinal plants[J].Chin J Exp Trad Med Form,22(22):192-196.[杨丽,王雅琪,刘升长,等,2016.三种常用半枫荷类药用植物的化学成分与生物活性研究概况[J].中国实验方剂学杂志,22(22):191-196.]
YANG WL,YAO ZS,LUO XQ,et al.,1999.Analgesic and anti-inflammatory effects of the ethanol extracts of jinlu ban fenghe(Semiliquidambar cathayensis)root[J].Jiangxi Sci,17(3):176-179.[杨武亮,姚振生,罗小泉,等,1999.金缕半枫荷的镇痛和抗炎作用[J].江西科学,17(3):176-179.]
ZHANG SY,ZHUANG YB,LUO LK,et al.,2017.Antioxidant and inhibitory effect on nitrosation of extracts from Balanophora arlandii[J].Nat Prod Res Dev,29(12):2081-2086.[张声源,庄远杯,罗铝铿,等,2017.红冬蛇菰提取物抗氧化及抑制亚硝化作用研究[J].天然产物研究与开发,29(12):2081-2086.]
ZHANG Y,WU YT,ZHENG W,et al.,2017.The antibacterial activity and antibacterial mechanism of a polysaccharide from Cordyceps cicadae[J].J Funct Foods,38:273-279.
ZHOU GX,YANG YC,SHI JG,et al.,2002.Studies on chemical constituents from Semiliquidambar cathayensis[J].Chin Trad Herbal Drugs,33(7):589-591.[周光雄,杨永春,石建功,等,2002.金缕半枫荷化学成分研究[J].中草药,33(7):589-591.]
ZHU YX,SONG H,ZHANG X,et al.,2017.Recovery of flavonoids from walnuts de-pellicle wastewater with macroporous resins and evaluation of antioxidant activities in vitro[J].J Food Proc Eng,40(1):e12275.