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Antipyretic and Detoxication Components Extracted from Mung Bean and Study on Stability

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Tutor: LiJian
School: Harbin University of Commerce
Course: Of Food Science
Keywords: Mung bean,Antibacteria,Antipyretic and Detoxication,Active ingredient
CLC: TQ461
Type: Master's thesis
Year:  2011
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Mung bean was extracted with ethanol,and crude extraction were extracted successively by different solvents anti-microbial. activity anti-endotoxin Activity antipyretic Activities were studied with different extractions. Different compositions of extractions were isolated and characterized.The preparation of anti-endotoxin from mung bean with ethanol extraction was studied. The extraction was determined endotoxin clearance by the Limulus test and then optimizing the extraction process by the standard of clearance rate.As the same time,the ethanol extraction extracted with petroleum ether, chloroform, ethyl acetate and n-butanol through two-phase, and each extraction was tested by the Limulus test. The results indicated that, mung bean extraction was provided with the effect of anti-endotoxin in vitro, the optimal conditions of extraction of anti-endotoxin from mung bean were:ethanol concentration was 70% temperature 60¡æ, extraction time of 2 hours, liquid ratio was 1:20.With the two-phase extracting, anti-endotoxin from mung bean were concentrated in the n-butanol extraction, Clearance rate 78.90%.Antibacterial experiment. Antibacterial effect was determined by filter paper method. Study on the different total momordicosides, different extractions had defferent antibacterial effect on Bacillus subtilis, Staphylococcus, E. coli, Penicillium. The result indicated that the effective components existed in the extraction of n-butano. The MIC were Bacillus subtilis 0.85mg/ml¡¢Staphylococcus 0.79 mg/ml¡¢E. coli 0.59 mg/ml¡¢Penicillium 35.32mg/ml. Total momordicosides had better antimicrobial stability under 30¡æ¡«70¡æand pH 7.0¡«8.0. It was effected by ultraviolet irradiation time.Study on effect of antipyretic and detoxication. The content of n-butanol in the n-butanol extraction was determined.It was 0.0236%and in line with the standards of Pharmacopoeia. 10mg/ml, 100mg/ml butanol extraction can significantly reduce the mortality rate in mice and prolong survival time.It was significant different (P<0.01) compared with LPS group; 1mg/ml extraction was different with endotoxin group (P<0.05). SOD activity and MDA levels of serum¡¢liver and brain from the survivals were determined.The results showed that the 10mg/ml, 100mg/ml butanol extraction can significantly increased SOD activity in mice and reduce the MDA levels compared with LPS group (P<0.05).1mg/ml butanol extraction can significantly decreased MDA levels (P<0.05); antipyretic experiment were studied on mices leaded to fever by endotoxin. Anorectal temperature were measured when butanol extraction was given in 1 hour.1.0%and 0.6%butanol extraction can lower the temperature.It showed significant differences compared with model group(P<0.05).0.2%n- butanol extraction group was significant with the model group (P<0.05) after 2h.The compositions of butanol extraction were separated and prepared. The butanol extraction was separated into 5 compositions with Ethyl acetate:n-butanol:water:acetic acid= 4:1:1:1. F01 (yield 20.12%) and F02 (yield 15.35%) are respectively eluted with ethyl acetate: butanol=4:1¡¢ethyl acetate acetate:butanol:acetic acid= 20:5:3.Residue was eluted by polyamide column. F03 (yield 12.56%) was eluted by 60%ethanol. Residue were prepared for further through and abtain F05 (yield16.05%)each component was divided into a single component by thin layer chromatography. the melting point:n-butanol 247.3-261.5, F01, 259.8-265.9, F02,281.6-287.5, F03,284.1¡«291.3, F05,196.0¡«201.6. Five components were separated by Thin Layer Chromatography, defining a flavonoids and three saponins.Stability test. Butanol extraction were easily Deliquesce. F01, F02, F04 components were influenced by temperature extremely, when the temperature exceeds 70¡æ, levels drop sharply. And the content remained unchanged only below 50¡æ. F03 was destroyed, when the temperature over 100¡æ.But characteristics did not change. Content of each component has nothing to do with ultraviolet light. And effect ultraviolet radiation from each component in the range of pH values of 5.0-9.0 stable. Activity of F01 is very strong between pH 5.0-9.0.
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