Study on Enzymatic Extraction Process of Fritillaria Alkaloids

In recent years, the use of biological enzyme assisted extraction and separation of the active ingredients in natural products has been widely used. As a new technology, its main mechanism of action:mild hydrolysis or degradation of plant cell wall components can accelerate the dissolution of the active ingredients to improve the extraction rate of the target product, and reduce costs through enzyme-catalyzed reaction.In order to optimize fritillaria alkaloid extraction process, center combination design method was used in this paper, to study the ethanol concentration, ultrasonic time, the liquid-solid ratio and their interaction influence on total alkaloid extraction yield. SAS software combined the response surface analysis method was used, to simulate prediction model of the regression equation and its credibility, and through the ridge analysis we get the best extraction conditions:ethanol concentration81.4%, ultrasonic extraction time74.9min, the liquid-solid ratio30.9mL/g. Under these condition, the extraction rate is0.158%.At present, there is almost no literatures about enzyme assisted extraction of fritillaria alkaloids at domestic and foreign. We focused on the influences of papain, cellulase, pectinase, xylanase, amylase enzymes on alkaloids extraction ratio. Alkaloid extraction ratio as index, firstly, we investigated5kinds of enzyme-assisted extraction on the total Alkaloids to determine the range of the conditions of temperature (T), pH value, enzyme amount (mg/g), the reaction time (t). Secondly, to determine the optimal conditions for the five kinds of enzyme assisted extraction Fritillaria alkaloids by orthogonal experiment. And then, the data were compared with traditional extraction methods-ethanol extraction method.The experimental results:papain pH4.5, t=120min, T=55℃, and enzyme quantity20mg/g, when the extraction ratio was highest-0.282%; Cellulose enzyme pH4.5, t=105min, T=55℃, and enzyme quantity17.5mg/g, when the extraction ratio was highest-0.27%; pectinase pH5.0, t=180min, T=52.5℃, and enzyme quantity7.5mg/g, when the extraction ratio was highest-0.278%; Xylanase pH5.0, t=195min, T=52.5℃, and enzyme quantity15mg/g, when the extraction ratio was highest-0.277%; Amylase pH5.5, t=165min, T=42.5℃, and enzyme quantity5mg/g, when the extraction ratio was highest-0.318%.Compared with ethanol extraction method, papain assisted extraction rate improved78.48%, cellulose assisted extraction rate improved70.89%, pectinase assisted extraction rate improved75.95%, xylanase assisted extraction rate improved75.32%, amylase enzyme assisted extraction rate improved101.27%, respectively.