Ultrasonic Assisted Extraction Process of Flavonoids from Houttuynia cordata Thunb and Its Antioxidant Activity
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摘要: 为优化超声辅助酶法提取鱼腥草根总黄酮的工艺，以黄酮提取量为指标，通过单因素实验筛选出适宜的液料比、粒径、乙醇浓度、酶解时间、加酶量，通过Box-Behnken响应面优化鱼腥草黄酮提取最佳条件，并对黄酮抗氧化活性进行研究。结果表明，最佳提取工艺为液料比28:1 mL/g、粒径<250 μm、乙醇浓度54%、酶解时间22 min，在该条件鱼腥草黄酮提取量为22.197 mg/g，产品干燥后纯度为26.63%。所得鱼腥草黄酮对DPPH·、·OH、ABTS+·有良好的清除能力，EC50值分别为0.097、2.250、0.384 mg/mL。本研究为鱼腥草根总黄酮提取提供了理论依据，为进一步实现鱼腥草资源的高值化利用提供了参考。Abstract: To optimize the process of ultrasound-assisted enzymatic extraction of total flavonoids from the roots of Houttuynia cordata, the appropriate liquid to material ratio, particle size, ethanol concentration, enzymatic digestion time, and enzyme addition was screened out by single-factor experiments, and the optimal conditions for the extraction of the flavonoids from Houttuynia cordata were optimized by Box-Behnken response surface. Meanwhile, the antioxidant activity of flavonoids was investigated. The results showed that the optimum extraction process was liquid to material ratio of 28:1 mL/g, particle size of <250 μm, ethanol concentration of 54%, and enzymatic digestion time of 22 min. The extraction amount of flavonoids was 22.197 mg/g under the optimized conditions, and the purity reached 26.63% by concentration under reduced pressure. The obtained flavonoids showed good scavenging ability for DPPH·, ·OH, and ABTS+· with EC50 values of 0.097, 2.250, and 0.384 mg/mL, respectively. The results of the study would provide a theoretical basis for the extraction of total flavonoids from the roots of Houttuynia cordata and a reference for further realization of the utilization of Houttuynia cordata.
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Key words:
- Houttuynia cordata Thunb /
- flavonoid /
- extraction process /
- antioxidant activity
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图 2 液料比对鱼腥草黄酮提取量的影响
Figure 2. Effect of liquid-material radio on extraction amount of flavonoids from Houttuynia cordata
图 3 乙醇浓度对鱼腥草黄酮提取量的影响
Figure 3. Effect of ethanol concentration on extraction amount of flavonoids from Houttuynia cordata
图 4 酶解时间对鱼腥草黄酮提取量的影响
Figure 4. Effect of enzymolysis time on extraction amount of flavonoids from Houttuynia cordata
图 5 加酶量对鱼腥草黄酮提取率的影响
Figure 5. Effect of enzyme dosage on the yield of the flavonoids from Houttuynia cordata
图 6 各因素交互作用对鱼腥草黄酮提取量的影响
Figure 6. Effect of interaction on extraction amount of flavonoids from Houttuynia cordata
图 8 鱼腥草黄酮对·OH清除率
Figure 8. ·OH scavenging effect of flavonoids from Houttuynia cordata
图 9 鱼腥草黄酮对ABTS+·清除率
Figure 9. ABTS+· scavenging effect of flavonoids from Houttuynia cordata
表 1 响应面试验因素水平表
Table 1. Factors and levels of Box-Behnken test
水平 因素 A液料比（mL/g） B粒径（μm） C乙醇浓度（%） D酶解时间（min） ?1 20:1 <425 50 10 0 30:1 <250 60 20 1 40:1 <180 70 30 
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表 2 响应面试验设计及结果
Table 2. Test design and result of response surface methodology
实验号 A B C D 提取量（mg/g） 1 1 0 ?1 0 16.866 2 1 0 0 ?1 18.534 3 0 0 ?1 1 21.431 4 ?1 0 0 ?1 16.690 5 ?1 0 ?1 0 19.392 6 0 1 0 ?1 16.953 7 ?1 ?1 0 0 17.305 8 0 1 ?1 0 19.236 9 1 0 0 1 16.778 10 ?1 1 0 0 15.022 11 0 1 1 0 14.934 12 0 0 1 1 17.392 13 0 1 0 1 14.671 14 1 ?1 0 0 18.27 15 0 ?1 ?1 0 18.236 16 0 ?1 0 1 18.973 17 0 0 0 0 22.133 18 1 0 1 0 19.675 19 ?1 0 1 0 12.827 20 0 0 1 ?1 19.324 21 0 ?1 0 ?1 18.183 22 0 ?1 1 0 19.324 23 0 0 ?1 ?1 20.904 24 ?1 0 0 1 18.183 25 0 0 0 0 21.708 26 0 0 0 0 22.336 27 1 1 0 0 13.354 
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表 3 响应面二次模型方差分析
Table 3. Analysis of variance of response surface quadratic model
方差来源 平方和 自由度 均方差 F值 P值 显著性 模型 156.31 14 11.16 15.55 < 0.0001 *** A 1.37 1 1.37 1.91 0.1921 B 21.66 1 21.66 30.15 0.0001 *** C 13.21 1 13.21 18.39 0.0011 ** D 0.8321 1 0.8321 1.16 0.3029 AB 1.73 1 1.73 2.41 0.1463 AC 21.97 1 21.97 30.59 0.0001 *** AD 2.64 1 2.64 3.67 0.0794 BC 7.26 1 7.26 10.11 0.0079 ** BD 2.36 1 2.36 3.28 0.095 CD 1.51 1 1.51 2.1 0.1725 A2 57.02 1 57.02 79.4 < 0.0001 *** B2 50.41 1 50.41 70.18 < 0.0001 *** C2 7.54 1 7.54 10.5 0.0071 ** D2 10.15 1 10.15 14.14 0.0027 ** 残差 8.62 12 0.7182 失拟项 8.41 10 0.8413 8.19 0.1136 不显著 纯误差 0.2054 2 0.1027 总误差 164.92 26 R2=0.9477， R2Adj=0.8868 注：“*”表示P<0.05，影响显著；“**”表示P<0.01，影响较显著；“***”表示P<0.001，影响极显著。 
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