Design-Expert优化纤维素酶提取一点红黄酮工艺

The Optimization of Cellulase Extraction Process of Flavone from Emilia sonchifolia by Response Surface Methodology

为优化提取一点红黄酮的试验条件,采用响应面优化法考察纤维素酶用量、酶解温度、提取时间和乙醇体积分数对一点红黄酮提取率的影响,通过Design-Expert.V8.0.6.1软件分析,设计四因素三水平响应面试验,建立回归模型,进行方差分析,考察纤维素酶的提取优化结果。一点红黄酮最佳提取条件为纤维素酶酶用量17.02 mg、酶解温度49.7℃、提取时间89.14 min、乙醇体积分数49.21%。试验色谱条件为流动相V_(甲醇)∶V_(0.2%)磷酸=60∶40,总流速0.6 m L/min,检测波长360 nm,进样量10μL,柱温箱最低温度25.0℃、最高温度65.0℃,进样检测分析时间60 min。采用HPLC测一点红黄酮含量,芦丁、槲皮素、山萘酚的含量分别为0.144,0.022和0.146 mg/g。该方法条件温和、操作简便,结果令人满意。

In order to optimize the experimental conditions for extracting flavone from Emilia sonchifolia,the effects of cellulase dosage,enzymatic hydrolysis temperature,extraction time and ethanol concentration on the extraction rate of flavone from Emilia sonchifolia have been investigated by response three-level response surface test,the regression model and analysis variance have been established,and the optimization results of cellulase extraction have been investigated.The optimum extraction condition of flavone from Emilia sonchifolia is as follows:cellulase dosage 17.02 mg,enzymatic hydrolysis temperature 49.7℃,extraction time 89.14 min and ethanol volume fraction 49.21%.The experimental chromatographic condition is as follows:mobile phase of V_(methanol)∶V_(0.2% phosphoric acid)=60∶40,total flow rate 0.6 mL/min,detection wavelength360 nm,injection volume 10μL,minimum temperature of column temperature box 25.0℃,maximum temperature 65.0℃,and sample injection detection and analysis time 60 min.The content of flavone from Emilia sonchifolia is measured by HPLC,and the contents of rutin,quercetin and kaempferol are 0.144,0.022 and 0.146 mg/g,respectively.This method has mild experimental conditions and simple operation.It is a new experimental attempt with satisfactory results.