Extraction, Separation, Antioxidant Activities And Anti-microbial Activities Of Effective Component Rfom Equisetum Hiemale L.

Equisetum hiemale L. is the dry herba of Equisetaceae plant，it is widely distributedin China. It has obvious effect on decreasing the serum levels of total cholesterol，analgesia，protection of vascular endothelial cells and antitumor. At home and abroad,reports about extraction and purification of its active ingredients are few. In this paper，we use Equisetum hiemale L. as raw material to study the extraction separation andpurification process of Equisetum hiemale L.. Its chemical composition (flavonoids,polysaccharides and alkaloids) are analyzed by the qualitative and quantitative assay.All these results provide a theory basic for the further separation of flavonoids, alkaloidsand polysaccharides from Equisetum hiemale L.. The main contents and results are asfollows:1、The extraction yield of total flavonoids is1.16%under these optimal extractionconditions:45%ethanol aqueous solution as the extraction solvent, material-to-liquidratio1∶30(g/mL), ultrasonic treatment time30min, and extraction temperature60℃.The total flavonoids can scavenge hydroxyl free radicals in a concentration-dependentfashion and has stronger inhibitory effect against lipid peroxidation in tea seed oil thanin lard.2、The application of D-101macroporous resin for the adsorption and desorption oftotal flavonoids is investigated. The adsorption equilibrium and adsorption kinetics arediscussed in detail. The data obtained from kinetic and isotherm experiments can bewell described using pseudo-second-order rate model and Langmuir isotherm equation,respectively. The practicable technical parameters for adsorption/desorption of totalflavonoids by D-101resin are2.0mg/mL flavonoids solution concentration, pH value2.1, amount (D-101resin weight ratio/desorption solvent volume)1∶20, standing time60min and eluting with50mL90%ethanol, under which the desorption rate could reach99.01%. The purity of total flavonoids increased from10.54%to60.01%.3、Polysaccharide is extracted by ultrasonic-assisted method. The influence factorsare investigated based on One-factor-at-a-time and orthogonal array design methods.The optimal technological conditions are twice,1∶25(g/mL) material-to-liquid ratio,60min ultrasonic extracting time and65℃ultrasonic temperature. Under the optimalconditions, yield of Polysaccharide is1.014%. Results showed that the best technicalparameters for adsorption/desorption of polysaccharide by D-101resin are2.0mg/mLpolysaccharide solution concentration, pH value6.0, amount (D-101resin weight ratio/adsorptions solvent volume)1∶10and eluting with50mL90%ethanol, under whichthe adsorption rate can reach99.01%. The adsorption process insolution exhibitesFreundlich behavior. The value of the enthalpy, free energy and entropy indicate thatthe adsorption is an endothermic process. The datas obtained from kinetic experimentscan be well described using pseudo-first-order rate model. 4、Polysaccharide is extracted and purified from Equisetum hiemale L.. Theantioxidant activity of crude and pure polysaccharides is investigated by testingscavenging effects of·OH, O2-·, NO2-and against the peroxidation of oils and fats. Theresults showed that crude and pure polysaccharides of extract exhibites a strong abilityto remove·OH，O2-·，NO2-and good inhibitory effect to peroxidation of oils and fats. Inaddition, the antioxidant activity of crude polysaccharides is stronger than that of purepolysaccharides. The results also suggest that the polysaccharides extract has strongantioxidant ability. Furthermore, the antioxidant activity is increased with concentrationof polysaccharides extract increasing.5、The extraction methods and antibacterial activity of total alkaloids are studied.The single factor experiment and orthogonal test are conducted to find out the optimalextracting conditions and determine the minimum in vitro inhibitory concentration(MIC) and diameter of inhibition zone of total alkaloids extracted against Proteusmirabilis, Escherichia coli, Micrococcus lutes, Aspergillus niger, Bacillus subtilis,Bacillus aerogenes, Penicillium and Staphylococcus aureus. The results showed that9.53%total alkaloids can be obtained when extraction condition are controlled asfollows:1.25%of HCl，1∶14of solid-liquid ratio，90℃of water bath temperature andreaping3times. The total alkaloids have strong antibacterial effect on Proteus mirabilis,Escherichia coli, Micrococcus luteus and Aspergillus niger.