Study On The Extraction And Biological Activity Of Active Components From Chinese Chive

In this paper,the above part of the amaranth was used as the research object,andVc was used as the positive control.The Prussian blue method,the phenanthroline-Fe²⁺oxidation method,the pyrogallol autooxidation method,the DPPH method and the ABTS method were used to study the different polarities of the amaranth.The extraction process of volatile oil from amaranth was studied by steam distillation,single factor and orthogonal test.The effect of alliinase on the volatile oil components;the antibacterial effect of some different polar parts and volatile oils on the ground of amaranth was studied by paper method and ultramicro powder agar dilution method.On the basis of the above,the different parts of the aboveground part of amaranth were studied by MTT method.Inhibition of RAW and SiHa cells by sexual sites and volatile oils.The specific results are as follows:（1）Add 75%ethanol solution from the dried powder of the leek to the reflux ratio of1:8（g/mL）and 1:3（g/mL）,and then concentrate the ethanol solution with petroleum ether.Extraction with ethyl acetate and n-butanol gives five different polar sites of ethanol phase,petroleum ether phase,ethyl acetate phase,n-butanol phase and aqueous phase.The five extracts of amaranth have certain in vitro antioxidant capacity.Among them,the order of total reducing capacity is:water phase>ethanol phase≈ethyl acetate phase≈n-butanol phase>petroleum ether phase;·OH scavenging capacity in the order of:ethanol phase>ethyl acetate phase>petroleum ether phase>positive Butanol phase>aqueous phase;O₂^-clearing capacity in the order of:ethanol phase≈petroleum ether phase≈ethyl acetate phase>n-butanol phase>aqueous phase;DPPH·removal capacity sequence:ethanol phase≈n-butanol phase>Ethyl acetate phase>aqueous phase（low concentration）>petroleum ether phase;ABTS⁺·scavenging capacity in the order of:ethyl acetate phase>ethanol phase>n-butanol phase>aqueous phase>petroleum ether phase.The experiment suggested that the ethanol and ethyl acetate phases of amaranth had better scavenging effects,and the antioxidant effects of aqueous phase,petroleum ether phase and n-butanol phase were not stable.（2）The optimum process conditions for extracting volatile oil from fresh amaranth are:material-to-liquid ratio 1:0.5（g/mL）70°C water bath after enzyme digestion,40°C water bath for 30 min,ultrasonic for 40 min,using volatile oil extractor to extract 6 h,the distillate was cooled with ice brine,and then subjected to enzymatic hydrolysis,and the mixture was extracted with dichloromethane,and the reagent was evaporated.Under this condition,the extraction rate of volatile oil from fresh amaranth was 0.0388%.The prepared volatile oil was clear and free of impurities,bright yellow color and strong amaranth smell.The results of GC-MS showed that the sulfur content of the enzymatically treated leek volatile oil（88.18%）was slightly higher than that of the undigested volatile oil（87.52%）,and the main components were dimethyl disulfide and dimethyl trisulfide.（3）Six different polar parts and volatile oils of amaranth are obvious for Acinetobacter baumannii,Streptococcus pneumoniae,Bacillus subtilis,Bacillus thuringiensis,Pseudomonas aeruginosa,Staphylococcus aureus,Escherichia coli and Enterococcus faecalis Inhibition.Ethanol has a good antibacterial effect against Streptococcus pneumoniae,its MIC value is 0.0025g/mL,MBC value is 0.03g/mL;petroleum ether has better antibacterial activity against Streptococcus pneumoniae,Bacillus subtilis and Enterococcus faecalis The effect is that the MIC value can reach0.005g/mL;the petroleum ether phase,the ethyl acetate phase,the n-butanol phase,and the water have the same bactericidal effect against the Enterococcus faecalis,and the MBC value is at least 0.02g/mL;The n-butanol phase and the aqueous phase have good antibacterial effects against Staphylococcus aureus,Streptococcus pneumoniae and Bacillus subtilis,respectively,and their MIC values are both 0.0025 g/mL.The ultramicro powder agar dilution method is simpler and faster than the paper sheet method,and the processing amount is large,which saves the experiment cost.（4）The six extracts of leek have no cytotoxic effect on RAW 264.7 anti-inflammatory cells;they have a certain inhibitory effect on SiHa tumor cells,and show concentration and time dependence.When the 100μg/mL sample solution was applied to SiHa cells for 72 h,the order of inhibition rate was:ethyl acetate phase>ethanol phase>petroleum ether phase>n-butanol phase>aqueous phase>volatile oil,in which ethyl acetate inhibited SiHa cells.The rate reached 29.93%,and the inhibition rate of volatile oil was only 18.57%.In summary,this study compares the antioxidant activities of different polar parts of amaranth,and each phase has certain in vitro antioxidant capacity;optimizes the extraction process of volatile oil from amaranth,and clarifies that the enzymatic hydrolysis of alliinase is beneficial to improve the volatile oil of amaranth.The sulfur content was proved to be antibacterial,and the ethyl acetate phase and n-butanol phase were better.The effects of different extracts of amaranth on SiHa and RAW264.7 cells were investigated,which proved that ethyl acetate had better inhibition than SiHa cells.As a function,each sample had no cytotoxic effect on RAW 264.7cells.