响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定

doi:10.7506/spkx1002-6630-201524010

食品科学

响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定

陈建中1,2，葛水莲1，昝立峰1，邢浩春1，付静1，王更先1

1.邯郸学院生命科学与工程学院，河北邯郸 056005；2.冀南太行山区野生资源植物研发中心，河北邯郸 056005

出版日期:2015-12-25 发布日期:2015-12-24
基金资助:
河北省科技计划项目（13222907）；邯郸市科技局项目（1422104057-2）；邯郸学院校级项目（15202）

Optimization of Aqueous Two-Phase Extraction of Total Flavonoids from Farfugium japonicum (L. f.) Kitam by Response Surface Methodology and Antibacterial Activity

CHEN Jianzhong1,2, GE Shuilian1, ZAN Lifeng1, XING Haochun1, FU Jing1, WANG Gengxian1

1. College of Life Science and Engineering, Handan College, Handan 056005, China;
2. Wild Resources Plant Research Center of South Hebei Mt. Taihang, Handan 056005, China

Online:2015-12-25 Published:2015-12-24

摘要/Abstract

摘要：

目的：优化大吴风草总黄酮（total flavonoids of Farfugium，TFF）双水相萃取体系并研究其抑菌活性。方法：超声波辅助C2H5OH-(NH4)2SO4双水相萃取TFF，依据Box-Behnken试验设计原理，采用三因素三水平响应面分析法，以TFF萃取率为响应值进行方差分析，获取多元二次线性回归方程；采用K-B纸片扩散法测定6 种供试菌种的抑菌圈直径，对比最小抑菌浓度（minimum inhibitory concentration，MIC）和最小杀菌浓度（minimumbactericidal concentration，MBC）确定其抑菌活性。结果：24% C2H5OH-18% (NH4)2SO4双水相萃取体系条件下TFF萃取率最高，响应面试验方差分析表明，粗提液质量分数显著影响TFF萃取率（P=0.023 9＜0.05），而pH值和NaCl质量分数对萃取率的影响不显著；最佳双水相萃取条件为粗提液质量分数20%、pH 7.64、NaCl质量分数2.68%，萃取率模型预测最大值为96.366 7%（P=0.994）；TFF对枯草芽孢杆菌抑制作用最强，高剂量作用下抑菌率可达98.67%，MIC为1.56 mg/mL；对沙门氏菌、金黄色葡萄球菌和大肠杆菌抑制作用次之，对青霉和黑曲霉抑制作用最弱。结论：粗提液质量分数对萃取率影响较大，pH值和NaCl质量分数对萃取率的影响较小；TFF对6 种供试菌种均表现出较强的抑菌活性，抑菌率与TFF质量浓度呈正相关，TFF对细菌的抑制效果相对强于真菌。

关键词: 响应面优化, 双水相萃取, 黄酮类物质, 大吴风草, 抑菌活性

Abstract:

Objective: To optimize the aqueous phase extraction system for total flavonoids (TFF) from the aboveground
parts of Farfugium japonicum and to study the antibacterial activity of the extract. Methods: Ultrasonic was used to assist
C2H5OH-(NH4)2SO4 aqueous two-phase extraction of TFF, and the multivariate quadratic regression model using TFF yield
as the response variable was established based on a Box-Behnken experimental design involving three factors at three levels
each and subjected to analysis of variance (ANOVA) and response surface analysis. The Kirby-Bauer disk diffusion method
was used to determine the diameters of inhibition zone of six tested strains and the minimum inhibitory concentration (MIC)
and minimum bactericidal concentration (MBC) values were compared to study their antibacterial activities. Results: The
24% C2H5OH-18% (NH4)2SO4 aqueous two-phase extraction system was found to be optimal to extract TFF. The results
of ANOVA showed that mass fraction of TFF in crude ethanolic extracts significantly affected the extraction rate of TFF
(P = 0.023 9 < 0.05), but the effects of pH and NaCl concentration in the aqueous two-phase system were not significant.
The optimal extraction conditions were determined as follows: crude extract concentration, 20%; pH, 7.64; and NaCl
concentration, 2.68%. Under these conditions, the maximum extraction rate of 96.366 7% (P = 0.994) was obtained. The
inhibitory effect of TFF on Bacillus subtilis was the strongest with a percentage inhibition of 98.67% at high dose and an
MIC of 1.56 mg/mL, followed by Salmonella, Staphylococcus aureus and Escherichia coli, and the inhibitory effects on
Penicillium and Aspergillus niger were the weakest. Conclusions: The flavonoid concentration of crude extracts had greater
influence on the extraction rate of TFF than pH and NaCl. TFF showed obvious antibacterial activity against the six tested
strains. The percentage inhibition was positively correlated to TFF concentration. The antibacterial activity of TFF was
relatively stronger against bacteria than fungi.

Key words: response surface methodology, aqueous two-phase extraction, total flavonoids, F. japonicurn (L. f.) Kitam, antimicrobial activity

中图分类号:

TS202.3

陈建中，葛水莲，昝立峰，邢浩春，付静，王更先. 响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定[J]. 食品科学, doi: 10.7506/spkx1002-6630-201524010.

CHEN Jianzhong1,2, GE Shuilian1, ZAN Lifeng1, XING Haochun1, FU Jing1, WANG Gengxian1. Optimization of Aqueous Two-Phase Extraction of Total Flavonoids from Farfugium japonicum (L. f.) Kitam by Response Surface Methodology and Antibacterial Activity[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201524010.

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响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定

Optimization of Aqueous Two-Phase Extraction of Total Flavonoids from Farfugium japonicum (L. f.) Kitam by Response Surface Methodology and Antibacterial Activity

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