Analysis Of Antibacterial Active Component From Chinese Chives (Allium Tuberosum)

【Objective】In this study, in order to discover the new antibacterials, chinese chive(Allium tuberosum)which to have broad spectrum antibacterial activity were selected. Its active site of antibacterial were sieved by follow-up antibacterial activity, which to obtain the active component, and to analyze. meanwhile, to appraise its stability of active site of antibacterial, It will be to plan to provide on the basis of science which developing new antibacterials and to dig medicine potential of chive.【Method】The different components were extracted from Chinese chive (Allium tuberosum) with water steam distillation and solvent extraction respectively. Antimicrobial activities of different extract were validated by vitro bacteriostasis test, in order to obtain its active site of antibacterial. The minimal inhibitory concentrations (MICs) were determined. At the same time, extract of ethyl acetate of stability were evaluated in accordance with pharmacopoeia of people's republic of china. By silica gel column chromatograph, thin-laver chromatography and Sephadex LH-20 chromatography, antibacterial active components were isolated from the ethyl acetate extract of this plant, and analyzed by GC-MS.【Results】(1)The antimicrobial assay by agar diffusion method showed that extract of ethyl acetate and volatile oil most effectively inhibited the growth of common Pathogenic Bacteria, for E.faecalis ATCC33186, the inhibition zones of the ethyl acetate extract were larger than 1.500 cm, for B.subtilis, P.aeruginosa ATCC27853, S.suis H9801, Shigella dysenteriae, S.aureus ATCC25923, E.coli ATCC25922, S.pullorum, P.vulgaris, B.cereus, S.typhi, E.coli O139, E.coli O2, E.coli O78, E.coli O8, S.choleraesuis, S.gallinarum, Salmonella typhimurium, the inhibition zones were larger than 2.200 cm, Those values for bacterial strains were in the range of 2.5 mg/mL-27 mg/mL depending on extract of ethyl acetate. Depending on volatile oil, MICs were in the range of 2μL/mL-32μL/mL. (2)Drug stability showed that high temperature have an major effect for their bacteriostatic activity, along with the extension handling time, bacteriostatic activity will taper. According a constant temperature test and long-term test, from January to June, bacteriostatic activity of different group have no significant difference(P>0.05), it showed that 4℃±2℃will be used as a best temperature for storage; when it storage in 25℃±2℃and 37℃±2℃, compared to incipient, three months and six months showed significant difference(P<0.05), but not between each other, nine months and twelve months showed significant difference(P<0.01); in simulated gastric fluid and intestinal fluid, compare with undisposed group, their bacteriostatic activity have significant difference(P<0.01)for P.vulgaris, P.aeruginosa ATCC27853, S.pullorum, there isn't significant difference(P>0.05)for others. (3)The analysis of the volatile oils by Gas chromatography mass revealed that the major components and the percentage of volatile oil were 39.308 % diallyl disulphide, 32.763 % disulfide, methyl 2-propenyl, 12.157 % trisulfide, methyl 2-propenyl and 7.330 % trisulfide, di-2-propenyl. Two fraction of distillate were isolated and purified from the ethyl acetate extract, by Gas chromatography mass revealed that the major components were palmitic acid(95.798% and 68.630%).【Conclusion】(1)Extract of ethyl acetate and volatile oil were antibacterial active ingredient in Chinese chives(Allium tuberosum), minimal inhibitory concentration were in the range of 2.5mg/mL-27mg/mL and 2μL/mL-32μL/mL. (2)4℃±2℃was the best storage temperature for extract of ethyl acetate, the stability of antibacterial activity have no significant difference(P>0.05); in simulated gastric fluid and intestinal fluid, compare with undisposed group, their bacteriostatic activity have significant difference(P<0.01) for P.vulgaris, P.aeruginosaATCC27853, S.pullorum, there isn't significant difference(P>0.05) for others. Storage six months in 37℃±2℃, inhibition zone was 2.940±0.032cm, showed the better durative potency. (3)Diallyl disulphide39.308 %, disulfide, methyl 2-propenyl32.763%, trisulfide methyl 2-propenyl 12.157% and palmitinic acid were antibacterial active ingredient in Chinese chives(Allium tuberosum).