响应面法优化苦丁茶熊果酸提取工艺及其抑菌活性研究
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摘要
为优化苦丁茶熊果酸的提取工艺,并探讨其抑菌活性。在单因素试验基础上,通过响应面分析法,研究了液料比、提取温度、乙醇浓度对苦丁茶熊果酸得率的影响。以金黄色葡萄球菌、枯草芽孢杆菌、生孢梭菌、铜绿假单胞菌、大肠杆菌、白色念珠菌、黑曲霉为供试菌,探究了苦丁茶熊果酸的抑菌活性及最小抑菌浓度(Minimum inhibitory concentration,MIC)。结果表明,最优提取工艺为:液料比为17∶1(mL/g)、乙醇浓度为83%、提取温度83℃;抑菌实验表明,苦丁茶熊果酸对7种菌均有一定的抑制效果,对大肠杆菌、铜绿假单胞菌的MIC为6. 25 mg/mL,对生孢梭菌、金黄色葡萄球菌、枯草芽孢杆菌的MIC为12. 5 mg/mL,对白色念珠菌、黑曲霉的MIC为25 mg/mL。
In order to optimize extraction of ursolic acid( UA) from Ilex kudingcha C. J. Tseng and investigate its antibacterial activity. Based on the results of single-factor tests,the ratio of liquid to solid,extraction temperature and ethanol concentration were optimized by response surface analysis. Staphylococcus aureus,Bacillus subtilis,Clostridium sporogenes,Pseudomonas aeruginosa,Escherichia coli,Candida albicans,Aspergillus niger were used as tested bacterial strains in this experiment to explore antibacterial activity of UA from Ilex kudingcha C. J. Tseng and its MIC. The results demonstrated that the process of optimum extraction were the ratio of liquid to solid of 17∶ 1( mL/g),ethanol concentration of 83%,extraction temperature of 83 ℃. The antibacterial experiments showed that UA had a certain inhibition on 7 strains. The MIC was as following:6. 25 mg/mL to Escherichia coli and Pseudomonas aeruginosa,12. 5 mg/mL to Staphylococcus aureus,Bacillus subtilis,Clostridium sporogenes,25 mg/mL to Candida albicans and Aspergillus niger.
In order to optimize extraction of ursolic acid( UA) from Ilex kudingcha C. J. Tseng and investigate its antibacterial activity. Based on the results of single-factor tests,the ratio of liquid to solid,extraction temperature and ethanol concentration were optimized by response surface analysis. Staphylococcus aureus,Bacillus subtilis,Clostridium sporogenes,Pseudomonas aeruginosa,Escherichia coli,Candida albicans,Aspergillus niger were used as tested bacterial strains in this experiment to explore antibacterial activity of UA from Ilex kudingcha C. J. Tseng and its MIC. The results demonstrated that the process of optimum extraction were the ratio of liquid to solid of 17∶ 1( mL/g),ethanol concentration of 83%,extraction temperature of 83 ℃. The antibacterial experiments showed that UA had a certain inhibition on 7 strains. The MIC was as following:6. 25 mg/mL to Escherichia coli and Pseudomonas aeruginosa,12. 5 mg/mL to Staphylococcus aureus,Bacillus subtilis,Clostridium sporogenes,25 mg/mL to Candida albicans and Aspergillus niger.
引文
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11 Li HY(李宏杨),Liu F(刘飞),Zhang FQ(张凤琴),et al.Supercritical fluid CO2extraction and purification of urosolic acid from Ilex kudingcha C.J.Tseng[J].J South Chin Univ Trop Agric(热带生物学报),2012,3:258-260.
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13 Zhang Y(张艳),Li YZ(李永哲).Response surface methodology and its application in pharmacy domain[J].J Jilin Inst Chem Tech(吉林化工学院学报),2012,29(7):21-26.
14 Xing YL(邢雅丽),Bi LW(毕良武),Zhao ZD(赵振东),et al.Optimization of ultrasonic extraction of ursolic acid from Paulownia elongata leaves by response surface methodology[J].Nat Prod Res Dev(天然产物研究与开发),2015,27:301-305.
15 Sun Y,Xu WQ,Zhang WQ,et al.Optimizing the extraction of phenolic antioxidants from kudingcha made from Ilex kudingcha C.J.Tseng by using response surface methodology[J].Sep Purif Technol,2011,78:311-320.
16 Cai J(蔡娟),Huang MT(黄敏桃),Huang YF(黄云峰),et al.Study on antimicrobial activity of different parts of guangxi Kudingcha[J].Chin Tradit Pat Med(中成药),2014,36:198-201.
2 Yu ZL(于志龙),Zhang C(张曾),Zhu RX(祝瑞雪),et al.Studies on the antioxidant activity of Ligustrum robustum(Rxob.)blume extracts[J].J Chin Inst Food Sci Tech(中国食品学报),2017,17:234-240.
3 Zhu KX(朱科学),Zhao SF(赵书凡),Zhu HY(朱红英),et al.A comparative study on the hypolipidemic activities of various solvent extracts derived from Ilex kudingcha C.J.Tseng[J].Sci Technol Food Ind(食品工业科技),2017,38:330-334.
4 Zhao X(赵欣),Yi RK(易若琨),Sun P(孙鹏),et al.Improvement effects of Kuding tea flavonoids extracts on D-galactose induced mice aging[J].Sci Technol Food Ind(食品工业科技),2017,38:303-308.
5 Zhu K,Li GJ,Sun P,et al.In vitro and in vivo anti-cancer activities of Kuding tea(Ilex kudingcha C J Tseng)against oral cancer[J].Exp Ther Med,2014,7:709-715.
6 Wolska KI,Grudniak AM,Fiecek B,et al.Antibacterial activity of oleanolic and ursolic acids and their derivatives[J].Cent Eur J Biol,2010,5:543-553.
7 Vasconcelos MA,Royo VA,Ferreira DS,et al.In vivo analgesic and anti-Inflammatory activities of ursolic acid and oleanoic acid from Miconia albicans(Melastomataceae)[J].ZNaturforsch,C:Biosci,2015,61:477-482.
8 Jian L(简磊),Gao MJ(高明杰),Xia X(夏旋).Ursolic acid attenuate myocardial ischemia reperfusion injury in diabetic rats[J].Nat Prod Res Dev(天然产物研究与开发),2017,29:843-848.
9 Gu XF(顾学芳),Tian S(田澍),Huang D(黄丹).Extraction and determination of ursolic acid and oleanolic acid in Kudingcha[J].Food Res Dev(食品研究与开发),2008,29(5):96-98.
10 Liu HM,Yang CH,Zeng XJ,et al.Solid-phase extraction of ursolic acid from herb using beta-cyclodextrin-based molecularly imprinted microspheres[J].J Sep Sci,2008,31:3573-3580.
11 Li HY(李宏杨),Liu F(刘飞),Zhang FQ(张凤琴),et al.Supercritical fluid CO2extraction and purification of urosolic acid from Ilex kudingcha C.J.Tseng[J].J South Chin Univ Trop Agric(热带生物学报),2012,3:258-260.
12 Zhang M(张明),Liu SX(刘树兴).Studies on enzymatic extraction technology of ursolid acid in Ilex kudingcha C.J.Tseng[J].Food Ind(食品工业),2008,2:3-5.
13 Zhang Y(张艳),Li YZ(李永哲).Response surface methodology and its application in pharmacy domain[J].J Jilin Inst Chem Tech(吉林化工学院学报),2012,29(7):21-26.
14 Xing YL(邢雅丽),Bi LW(毕良武),Zhao ZD(赵振东),et al.Optimization of ultrasonic extraction of ursolic acid from Paulownia elongata leaves by response surface methodology[J].Nat Prod Res Dev(天然产物研究与开发),2015,27:301-305.
15 Sun Y,Xu WQ,Zhang WQ,et al.Optimizing the extraction of phenolic antioxidants from kudingcha made from Ilex kudingcha C.J.Tseng by using response surface methodology[J].Sep Purif Technol,2011,78:311-320.
16 Cai J(蔡娟),Huang MT(黄敏桃),Huang YF(黄云峰),et al.Study on antimicrobial activity of different parts of guangxi Kudingcha[J].Chin Tradit Pat Med(中成药),2014,36:198-201.