Study On The Enzymatic Hydrolysis Of Ginsenosides Based On The Hollow Fiber Liquid Phase Microextraction Techniques

Ginsenosides are the active ingredients in American ginseng and Panax notoginseng.Because of its various molecular structures and different physical properties,ginsenosides have multiple effects on multiple metabolism.Therefore,it is of great significance to extract a variety of ginsenosides simultaneously by simple,fast and low-pollution methods.The main contents and results of this paper are as follows:(1)Four kinds of ginsenosides in American ginseng were extracted and analyzed using hollow fiber mixed solvent bar method followed by HPLC-UV.In the process of liquid phase microextraction of hollow fiber bars,methanol was selected as desorbent,and the solvent type and solvent ratio of mixed solvent bars were mainly selected to extract the effects of four kinds of ginseng saponins.On this basis,other factors affecting the extraction of ginsenosides such as the time of liquid phase microextraction,the temperature of extraction,the number of hollow fiber rods,and the pH of the sample solution were investigated.Under optimal conditions,the good linear relationship for ginsenosides Rg1,Re,Rb1,and Rd was 7.5-37.5?g/mL,30-150?g/mL,75-375?g/mL,and 39.15-195.75?g/mL,respectively(R~2>0.9990).The recovery of extraction varied from 93.91-97.83%,and the relative standard deviation(RSD)was less than 3.37%.This method of simultaneously extracting four ginseng saponins of American ginseng has high extraction efficiency.(2)Based on hollow fiber liquid phase microextraction method proposed in this paper,transforming ginsenoside Rb1 of American ginseng to produce ginsenoside Rd using mixed enzyme hydrolysis was studied.Factors affecting enzymatic hydrolysis such as enzymatic hydrolysis pH,enzymatic hydrolysis time,enzymatic hydrolysis temperature and ratio of mixed enzymes were explored.The response surface method was used to optimize the optimal conditions for the formation of ginsenoside Rd.The optimized condition was as foolows,enzymatic hydrolysis time,91.2 min;the enzymatic hydrolysis temperature,52.5°C;the laccase addition amount,0.27 mL;the acid cellulase,0.4 mL;0.12 M hydrochloric acid,20?L.Under the optimized experiment condition,the transformation recovery of ginsenoside Rd fromginsenoside Rb1 was 0.76%.(3)Based on the hollow fiber liquid phase microextraction technique proposed in this paper,converting ginsenoside Rb1 of Panax notoginseng into ginsenoside Rd using mixed enzyme hydrolysis was studied.Mixed enzymes composed of laccase and acid cellulasewere were used in the experiment.Ginsenoside Rb1 was transformed to improve the extraction efficiency and medicinal value.The optimal conditions for the formation of ginsenoside Rd by the mixed surface enzyme hydrolysis of Panax notoginseng saponins Rb1 were as follows:acidic cellulase was 0.2 mL,laccase was 0.284 mL,enzymatic hydrolysis time was 73.90 min,and enzymatic hydrolysis temperature was55.8?.The extraction efficiency of ginsenoside Rd from Panax notoginseng was 1.13%.