Study On The Extraction Of Total Flavonoids And EUG From Eucommia Ulmoides Leaves By Microwave Ultrasonic Surfactant Assisted Method Combined With Fungus Cascade

Eucommia ulmoides Olive.It belongs to perennial deciduous tree,which is divided into eucommiaceae,eucommiaceae,eucommiaceae,Eucommia ulmoides Olive.Modern scientific research shows that Eucommia ulmoides gum(EUG)contains a lot of flavonoids.The traditional extraction solvent of flavonoids is ethanol or acetone and water,which has the potential risk of environmental pollution.At present,EUG extraction is often hexane or petroleum ether as the main solvent,organic solvent consumption,high cost,environmental pollution,safety hazards and other problems.Moreover,there has been no report on the process of green extraction of flavonoids and EUG at the same time,so it is urgent to upgrade the process.Based on the above reasons,this paper firstly selected the anionic surfactant with better solubility for flavonoids in eucommia ulmoides leaves as the extraction solvent,and used microwave ultrasound to enhance the extraction of flavonoids.At the same time,it showed a strong removal effect on the cuticle layer on the surface of eucommia ulmoides leaves to facilitate the cultivation of microorganisms.On this basis,green trichoderma was used as the strain to ferment and cultivate the residue of eucommia ulmoides leaves after extracting total flavonoids,and the cellulose was metabolized to destroy the structure of eucommia ulmoides leaves,improve the contact area between the eontent of eucommmoides gum and the extraction solvent,so as to improve the extraction efficiency of eucommia ulmoides gum and save the solvent.The main research contents and results are as follows:1.The total flavonoids in the leaves of eucommia ulmoides were extracted with the surfactant sodium dodecyl benzene sulfonate micellar as the extraction medium,and the cuticles on the surface of eucommia ulmoides were removed by microwave ultrasonic assisted extraction.Response surface optimization was used to determine the optimal extraction process of secondary flavonoids:the surfactant sodium dodecyl benzene sulfonate concentration was 1.5%,the extraction time was 30 min,the extraction times were 2 times,the solid-liquid ratio was 1:50 and the microwave power was 700 W.Under these conditions,the flavonoids yield of eucommia ulmoides wasl.45%,and the extraction yield reached 73.97%,which was about 4 times higher than that of pure water(0.36%).At the same time,the extraction of eucommia ulmoides leaf surface before and after corneous layer Sudan Ⅲ staining showed that after extraction of basic remove cuticle,cortex eucommiae leaf surface for subsequent green trichoderma viride fermentation culture.2.With flavonoids extracted and cuticle removed from the surface as raw materials,eucommia ulmoides leaf residue was metabolized by green trichoderma liquid fermentation to improve the content of eucommia ulmoides gum.The optimal fermentation conditions were determined by orthogonal experiment method:the suspension pH of eucommia ulmoides leaves residue was 6,the solid phase content of eucommia ulmoides leaves was 9.9%(solid to liquid ratio 1:10),the inoculation amount of trichoderma gellini was 20%(calculated by suspension volume,the strain concentration was about 5 106 CFU/mL),the fermentation time was 5 d,and the enzyme activity of trichoderma gellaria was 19.88 U/mL.Under these conditions,the weight loss rate of eucommia ulmoides leaf residue was 43.81%,and the content of eucommia ulmoides gum increased from 1.7%to 4.36%.At the same time,the content of cellulose and hemicellulose in the residue of eucommia ulmoides leaves decreased from 25.93%and 19.33%to 14.81%and 8.35%respectively,while the content of lignin was almost unchanged,indicating that the main degradation substrate of green trichoderma was syncellulose.Through visual observation and SEM scanning electron microscopy combined with the changes of cellulose before and after comprehensive analysis,it can be concluded that green trichoderma can effectively damage the leaf structure of eucommia ulmoides.3.Using green trichophyllum fermentation to remove eucommia ulmoides leaves as raw materials and petroleum ether as solvent to extract EUG,compared with the original leaves of eucommia ulmoides,the solvent consumption per unit mass of EUG decreased from 2.74 mL/g to 0.70 mL/g,and the solvent saving rate was 74.5%.Green solvent turpentine for EUG extract,unit quality EUG solvent consumption by.2.91 mL/g decreased to 0.96 mL/g,and the solvent saving rate was 67.01%.On this basis,petroleum ether was used to wash EUG,and the final purity was 96.6%EUG.The results of IR spectrum analysis showed that the structure of EUG extracted from turpentine after green trichoderma fermentation was consistent with the traditional one.