紫苏叶总三萜超声提取工艺优化及对10种常见致病菌的抑菌作用研究
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摘要
目的:优化紫苏叶总三萜的超声提取工艺,并考察其对10种常见致病菌的抑菌作用。方法:以紫苏叶总三萜提取量为响应值,以料液比、乙醇体积分数、超声时间、超声温度为响应因子,在单因素试验的基础上,采用响应面法优化紫苏叶总三萜的超声提取工艺;采用微量二倍稀释法测定紫苏叶总三萜对10种常见致病菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC),以考察其抑菌作用。结果:优化的紫苏叶总三萜超声提取工艺为料液比1∶30[m(g)/V(mL)],乙醇体积分数85%,超声时间65 min,超声温度75℃。在此条件下的紫苏叶总三萜提取量为37.90 mg/g,与预测值(38.40 mg/g)的相对误差为1.30%,实测值与预测值一致性良好。抑菌作用考察结果表明,紫苏叶总三萜对10种常见致病菌的MIC范围为0.48~15.50 mg/mL,MBC≥0.97 mg/mL,整体抑菌作用较强,尤其对大肠杆菌、金黄色葡萄球菌和铜绿假单胞菌的抑菌作用最强。结论:优化所得紫苏叶总三萜的超声提取工艺稳定、可行;紫苏叶总三萜具有较强的广谱抗菌作用,且对大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌的抗菌作用相当。
OBJECTIVE:To optimize the ultrasonic extraction technology of total triterpenoids from the leaves of Perilla frutescens(TPL) and investigate its antibacterial activities against 10 kinds of common pathogens. METHODS: Using the extraction amount of triterpenoids as response value,the ultrasonic extraction technology of TPL was optimized by response surface methodology on the basis of single factor test with solid-to-liquid ratio,ethanol volume fraction,ultrasonic time and temperature as factors. The MIC and MBC of TPL on 10 kinds of common pathogens were detected by double broth dilution method,so as to investigate antibacterial activities. RESULTS:The optimal ultrasonic extraction technology of TPL included solid-to-liquid ratio of 1 ∶ 30 [m(g)/V(mL)],ethanol volume fraction of 85 %,ultrasonic time of 65 min and ultrasonic temperature of 75 ℃. Under these conditions,the amount of triterpenoids was 37.90 mg/g,relative error of which to predicted value(38.40 mg/g) was 1.30%.Measured value agreed well with predicted value. Results of antibacterial activities study showed that MIC were in the range of 0.48-15.50 mg/mL,and MBC≥0.97 mg/mL. TPL showed significant antibacterial activities, especially to Escherichia coli,Staphylococcus aureus and Pseudomonas aeruginosa. CONCLUSIONS:Optimized ultrasonic extraction technology of TPL is stable and feasible. TPL extracted has strong broad-spectrum antibacterial effect and shows similar antibacterial effecton E.coli,S. anreus and P. aeruginosa.
OBJECTIVE:To optimize the ultrasonic extraction technology of total triterpenoids from the leaves of Perilla frutescens(TPL) and investigate its antibacterial activities against 10 kinds of common pathogens. METHODS: Using the extraction amount of triterpenoids as response value,the ultrasonic extraction technology of TPL was optimized by response surface methodology on the basis of single factor test with solid-to-liquid ratio,ethanol volume fraction,ultrasonic time and temperature as factors. The MIC and MBC of TPL on 10 kinds of common pathogens were detected by double broth dilution method,so as to investigate antibacterial activities. RESULTS:The optimal ultrasonic extraction technology of TPL included solid-to-liquid ratio of 1 ∶ 30 [m(g)/V(mL)],ethanol volume fraction of 85 %,ultrasonic time of 65 min and ultrasonic temperature of 75 ℃. Under these conditions,the amount of triterpenoids was 37.90 mg/g,relative error of which to predicted value(38.40 mg/g) was 1.30%.Measured value agreed well with predicted value. Results of antibacterial activities study showed that MIC were in the range of 0.48-15.50 mg/mL,and MBC≥0.97 mg/mL. TPL showed significant antibacterial activities, especially to Escherichia coli,Staphylococcus aureus and Pseudomonas aeruginosa. CONCLUSIONS:Optimized ultrasonic extraction technology of TPL is stable and feasible. TPL extracted has strong broad-spectrum antibacterial effect and shows similar antibacterial effecton E.coli,S. anreus and P. aeruginosa.
引文
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[4]谢健航,林嘉,雷林生,等.茯苓总三萜抑制小鼠免疫反应及治疗大鼠佐剂性关节炎的实验研究[J].中药药理与临床,2016,32(6):89-92.
[5]王艺玮,徐盛涛,徐进宜.五环三萜羽扇豆烷型天然产物抗肿瘤活性研究进展[J].药学与临床研究,2017,25(4):336-342.
[6]邓鸣,张炜宁.异型南五味子总三萜对类风湿关节炎小鼠抗炎镇痛药效学研究[J].安徽医药,2016,20(8):1469-1472.
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[13]贾瑞博,赵慧,刘凯丽,等.树舌灵芝总三萜提取工艺优化及其抗氧化作用研究[J].应用化工,2016,45(6):1030-1035.
[14]谢勇,张榕,张江湖,等.Box-Behnken试验设计优化刺梨果汁饮料工艺[J].食品研究与开发,2018,39(2):119-126.
[15]曲吴杨,朱文学,刘琛,等.苹果渣果胶提取工艺优化及碱法降酯效果评价[J].食品科学,2014,35(14):87-92.
[16]WEI L,ZHANG W,YIN L,et al.Extraction optimization of total triterpenoids from Jatropha curcas,leaves using response surface methodology and evaluations of their antimicrobial and antioxidant capacities[J].Electron J Biotech,2015,18(2):88-95.
[17]HERRERA RM,PéREZ M,MARTíN-HERRERA DA,et al.Antimicrobial activity of extracts from plants endemic to the Canary Islands[J].Phytother Res,2015,10(4):364-366.
[18]NOSTRO A,GERMANóMP,D’ANGELO V,et al.Extraction methods and bioautography for evaluation of medicinal plant antimicrobial activity[J].Lett Appl Microbiol,2000,30(5):379-384.
[2]冯林慧,李迎秋.紫苏的活性成分及其应用研究[J].中国调味品,2017,42(9):177-180.
[3]BISHAYEE A,AHMED S,BRANKOV N,et al.Triterpenoids as potential agents for the chemoprevention and therapy of breast cancer[J].Front Biosci,2011,16:980-996.
[4]谢健航,林嘉,雷林生,等.茯苓总三萜抑制小鼠免疫反应及治疗大鼠佐剂性关节炎的实验研究[J].中药药理与临床,2016,32(6):89-92.
[5]王艺玮,徐盛涛,徐进宜.五环三萜羽扇豆烷型天然产物抗肿瘤活性研究进展[J].药学与临床研究,2017,25(4):336-342.
[6]邓鸣,张炜宁.异型南五味子总三萜对类风湿关节炎小鼠抗炎镇痛药效学研究[J].安徽医药,2016,20(8):1469-1472.
[7]诸夔妞,吴正凤,蒋翠花,等.三萜类化合物降血糖活性及其作用机制研究进展[J].中国药科大学学报,2015,46(6):764-770.
[8]CHE SULAIMAN IS,BASRI M,FARD MASOUMI HR,et al.Effects of temperature,time,and solvent ratio on the extraction of phenolic compounds and the anti-radical activity of Clinacanthus nutans Lindau leaves by response surface methodology[J].Chem Cent J,2017.DOI:10.1186/s13065-017-0285-1.
[9]贾韶千,吴彩娥,李艳霞.枸杞中黄酮类化合物的超声波强化提取[J].农业机械学报,2009,40(5):130-134.
[10]BAI X,YUE T,YUAN Y,et a1.Optimization of microwave-assisted extraction of polypheno Is from apple pomace using response surface methodology and HPLC analysis[J].J Sep Sci,2010,33(23/24):3751-3758.
[11]李师,雷艳萍,石晓峰,等.雪松松针总皂苷的超声提取工艺优选及体外抗肿瘤活性研究[J].中国药房,2016,27(31):4406-4410.
[12]李峰,王延伟,俞力超,等.四角菱茎多酚的超声提取工艺优化及体外抗肿瘤活性研究[J].中国药房,2016,27(31):4414-4417.
[13]贾瑞博,赵慧,刘凯丽,等.树舌灵芝总三萜提取工艺优化及其抗氧化作用研究[J].应用化工,2016,45(6):1030-1035.
[14]谢勇,张榕,张江湖,等.Box-Behnken试验设计优化刺梨果汁饮料工艺[J].食品研究与开发,2018,39(2):119-126.
[15]曲吴杨,朱文学,刘琛,等.苹果渣果胶提取工艺优化及碱法降酯效果评价[J].食品科学,2014,35(14):87-92.
[16]WEI L,ZHANG W,YIN L,et al.Extraction optimization of total triterpenoids from Jatropha curcas,leaves using response surface methodology and evaluations of their antimicrobial and antioxidant capacities[J].Electron J Biotech,2015,18(2):88-95.
[17]HERRERA RM,PéREZ M,MARTíN-HERRERA DA,et al.Antimicrobial activity of extracts from plants endemic to the Canary Islands[J].Phytother Res,2015,10(4):364-366.
[18]NOSTRO A,GERMANóMP,D’ANGELO V,et al.Extraction methods and bioautography for evaluation of medicinal plant antimicrobial activity[J].Lett Appl Microbiol,2000,30(5):379-384.