CN102618386A - Method for preparing chenopodium vulvaria volatile oil and application of chenopodium vulvaria volatile oil - Google Patents
Method for preparing chenopodium vulvaria volatile oil and application of chenopodium vulvaria volatile oil Download PDFInfo
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Abstract
The invention relates to application of chenopodium vulvaria volatile oil to bacteriostatic/bactericidal and antioxidative products. The chenopodium vulvaria volatile oil can be obtained by a water steam distillation or CO2 supercritical extraction method, and main ingredients of the chenopodium vulvaria volatile oil are identified by gas chromatograph/mass spectrum. The chenopodium vulvaria volatile oil has an obvious effect of killing or inhibiting bacteria such as staphylococcus, streptococcus, enterobacter, salmonella, shigella, pseudomonas, bacillus, escherichia coli and the like and fungi such as candida, cryptococcus, penicillin, aspergillus, mucor, microsporon, trichophyta, epidermophyton and the like. A diphenyl picryl phenylhydrazine (DPPH) method proves that the chenopodium vulvaria volatile oil has the high antioxidant activity, can be widely applied to industry of food, medicines, cosmetics, health-care products and the like, and is used for preparing the bacteriostatic/bactericidal and antioxidative products.
Description
Technical field
Antibacterial, the sterilization and the oxidation-resistance effect of preparation and the application of this volatile oil, especially this volatile oil of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf have been the present invention relates to.
Background technology
Vegetables oil has its unique composition and biological activity, all has been widely used at aspects such as spices, medicine, antibiotic, makeup, additive, Ecotourisms.Plants essential oil is the intravital secondary metabolites of plant, is made up of the simple compounds that molecular weight is less relatively, has certain aromatic odour, at normal temperatures can evaporable oily liquids material.The contained chemical ingredients more complicated of plants essential oil can be divided into aliphatics, aromatic series and terpene 3 big compounds and their containing oxygen derivative like alcohol, aldehyde, ketone, acid, ether, ester, lactone etc. by chemical structure, also has nitrogenous and compound sulfur-bearing in addition.
Vast territory and abundant resources in China, and floristics is various, and aromatic oil plant distributes extensively, the plants essential oil aboundresources.The research of spice berry derived essential oil; Generally used gas chromatograph-mass spectrometer (GC-MS) fast and accurately; Multiple in addition chromatographic technique and wave spectrometer use, and make the research level of China's spice berry derived essential oil obtain improving very soon, increasing plants essential oil by people cognition.
The plants essential oil resource is Biological resources that are rich in potentiality, and it can be used as important source material, in aspect widespread uses such as essence, spices, medicine, healthcare products, pest control, organic synthesis, feed and food.Although to plants essential oil itself and use, exploitation does a lot of work, relevant work is done still inadequately, still there is space widely the deep understanding and the exploitation side of plants essential oil.
The fragrant lamb's-quarters of chrysanthemum leaf, another name chrysanthemum leaf thorn lamb's-quarters, Nu Mari, anaesthetic name Wu Nurite-Nuo Yile, formal name used at school:
Chenopodium foetidumSchrad. the morphological specificity of plant is an annual herb, and is high 20～60 centimetres.Complete stool tool body of gland and band joint are dredged and are given birth to glandular hairs, and overpowering odor is arranged.Stem is upright, and the green vitta of tool has branch.Single leaf alternate, tool handle, long 0.5～l centimetre; The blade square is circular, and is long 2～6 centimetres, wide 1.5～3.5 centimetres; Edge pinniform is shallow to be split to the pinniform drastic crack, and tip is blunt or sharp, has mucro sometimes; The base portion wedge shape, (top no hair or young hairiness slightly when tender have the pubescence of tool joint below and have the particulate state body of gland of yellow stockless concurrently by pubescence and pale brown look gland point on the two sides; Seldom be bordering on and do not have hair), the long 2-10 millimeter of petiole.It is interior or terminal that Hua Dan is born in the sprig armpit, forms dichasium, the big panicle of integrated turriform again; The flower both sexes, perianth diameter 1-1.5 millimeter, 5 drastic cracks, tepal 5, ovum shape lanceolar has narrow membranous edge, and by yellow gland point and thorn-like projection, the edge is membranous, carries out during fruit; Stamen 5, filigree is flat, and flower pesticide is subsphaeroidal, does not expose; The utricle spheroid, pericarp is membranous; Seed grows wild, spheroid, and 0.5～0.8 millimeter of diameter, glossy, black or sorrel, tool fine-structure mesh texture; The embryo semisphere is around endosperm.7～September of florescence, really 9～October of phase.
The fragrant lamb's-quarters of chrysanthemum leaf is born in border meadow, ditch bank, river bank, near other, farmland, canal limit and roadside.Be distributed in China Liaoning, the Inner Mongol, Shanxi, Shaanxi, Gansu, Qinghai, Sichuan, Yunnan, Tibet.All herbal medicine,
Cure mainly:, headache, measles without adequate eruption, gold is hindered, skin pruritus etc.Make a general survey of Chinese and foreign documents, the research of the fragrant lamb's-quarters volatile oil component of chrysanthemum leaf does not appear in the newspapers.Antibacterial, sterilization and anti-oxidant activity to this volatile oil do not appear in the newspapers yet.In view of the good pharmacologically active of this medicine, the present invention has carried out detailed introduction to its composition and in the activity aspect antibacterial, sterilization and the oxidation-resistance.
Summary of the invention
Technical problem:The purpose of this invention is to provide the preparation and the application thereof of the fragrant lamb's-quarters volatile oil of a kind of chrysanthemum leaf, analyze the composition of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf, activity introduction and the application of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf aspect antibacterial, sterilization and oxidation-resistance is provided.
Technical scheme:The preparation method of the fragrant lamb's-quarters volatile oil of a kind of chrysanthemum leaf of the present invention adopts the steam distillation preparation, and the fragrant lamb's-quarters of the fresh chrysanthemum leaf of gathering is added in the extractor; Add water up to soaking sample fully, open the oil bath heating, oil bath temperature is transferred to 100～250 degrees centigrade; The reaction kettle top adds a cover condensing works, collects refrigerative steam with Erlenmeyer flask, then the steam condensate of collecting is put into to leave standstill in the separating funnel and treats layering; Wait the oil reservoir that is divided no longer increase promptly no longer fuel-displaced till, oil reservoir take out is collected, use siccative dry (like anhydrous calciumsulphate) then; Obtain weak yellow liquid, i.e. the fragrant lamb's-quarters volatile oil of chrysanthemum leaf.
Adopt CO
2Supercritical extraction obtains, with the fragrant lamb's-quarters of the fresh chrysanthemum leaf of gathering, and flushing, chopping, in the extractor of packing into, control CO
2Flow, pressure 11～40MPa, temperature 308～330K, time 1～4h collects volatile oil.
The application of the fragrant lamb's-quarters volatile oil of prepared chrysanthemum leaf: this volatile oil is applied to food, medicine as natural antibacterial, sterilization, inhibitor.Described this volatile oil is antibacterial, sterilization is a bacterium, or fungi.
Described bacterium is Staphylococcus, streptococcus, enterobacter, Salmonella genus, Shigella, Rhodopseudomonas, bacillus, escherichia coli genus, especially subtilis, bacillus cereus, enterococcus faecalis, Salmonellas, intestinal bacteria, Pseudomonas aeruginosa, pneumobacillus, dysentery bacterium, pseudomonas aeruginosa or streptococcus acidi lactici.
Described fungi is Candida, genera cryptococcus, penicillium mould, Aspergillus, Mucor, microsporum, trichophyton, Epidermophyton, especially Fusarium oxysporum, black mold, fusarium culmorum, saccharomyces cerevisiae, tobacco brown spot pathogen, Candida albicans, Microsporum canis, trichophyton or withered mould.
Described this volatile oil is used for industries such as food, medicine, makeup, healthcare products as natural inhibitor, replace the synthetized oxidation preventive agent of toxic side effect.
Beneficial effect:Antibacterial, the fungicidal activity of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf
Through evidence, the fragrant lamb's-quarters volatile oil of chrysanthemum leaf has broad-spectrum sterilization and bacteriostatic action, and the fragrant lamb's-quarters volatile oil of chrysanthemum leaf belongs to Staphylococcus, streptococcus, enterobacter, Salmonella genus, Shigella, Rhodopseudomonas, bacillus, escherichia coli; Fungies such as Candida, genera cryptococcus, penicillium mould, Aspergillus, Mucor, microsporum, trichophyton, Epidermophyton have certain effect of killing, suppressing to grow.Especially common bacteria subtilis, bacillus cereus, enterococcus faecalis, Salmonellas, intestinal bacteria, Pseudomonas aeruginosa, pneumobacillus, dysentery bacterium, pseudomonas aeruginosa, streptococcus acidi lactici, Fusarium oxysporum, black mold, fusarium culmorum, saccharomyces cerevisiae, tobacco brown spot pathogen, cryptococcus neoformans, Candida albicans, acrothesium floccosum, Microsporum canis, trichophyton, withered mould had the very strong effect of killing and suppressing to grow.Therefore, the fragrant lamb's-quarters volatile oil of chrysanthemum leaf can be applicable to industries such as food, medicine, healthcare products, makeup, replaces the synthetic antibacterial agents of toxic side effect.
Embodiment
1. the preparation method of the volatile oil of the fragrant lamb's-quarters of chrysanthemum leaf of the present invention is following:
Fragrant lamb's-quarters 5kg is added in the reaction kettle with the fresh chrysanthemum leaf of gathering, and adds water up to soaking sample fully, opens the oil bath heating, and oil bath temperature is transferred to about 130 degree; Reaction kettle top adds a cover condensing works, collects refrigerative steam with Erlenmeyer flask, and the steam that will collect then is put into to leave standstill in the separating funnel treats layering, treat no longer fuel-displaced till; Afterwards oil reservoir is taken out and collect, use anhydrous sodium sulfate drying then, obtain weak yellow liquid; Flavor is fragrant, has fresh and sweet and is being with the fragrance of a flower, and fragrance is strong and lasting.This technology is to be example with the wet distillation, can also be CO
2Supercritical extraction obtains: adopt CO
2Supercritical extraction obtains, with the fragrant lamb's-quarters of the fresh chrysanthemum leaf of gathering, and flushing, chopping, in the extractor of packing into, control CO
2Flow, pressure 11～40MPa, temperature 308～330K, time 1～4h collects volatile oil.
2. the volatile oil component analysis of the fragrant lamb's-quarters of chrysanthemum leaf of the present invention.
Gas chromatography and mass spectromentry coupling technique gained mass spectrum has been confirmed the chemical composition of part in the fragrant lamb's-quarters volatile oil of chrysanthemum leaf through computingmachine mass-spectrometric data library searching.Gas phase phase chromatographic condition:
(1) GC conditions
Chromatographic column: CP-8 (30 meters * 0.25 millimeter * 0.25 micron) mass spectrum post, instrument model VARIAN CP-3800,1079 injection ports.Split stream sampling, injector temperature: 250 degree, electronic flow control.Temperature programming: post starting temperature 40 degree, kept 1 minute, with temperature rise rate 3.5 degree/minute, be warmed up to 200 and spend, kept 1 minute.Carrier gas: helium, flow velocity: 1 ml/min.Sample size: 1 μ L.Splitting ratio 1:10.
(2) mass spectrum condition
Mass spectrum model: VARIAN Saturn 2200.Ionization mode: EI.Electron energy is 70eV.Ion source temperature: 200 degree.Interface temperature: 280 degree.Mass scanning scope: 50-550, unit mass resolving power.
Analysis and Identification result shows; The fragrant lamb's-quarters volatile oil of chrysanthemum leaf main chemical compositions is that aliphatics, aromatic series and terpene 3 big compounds and their containing oxygen derivative are like alcohol, aldehyde, ketone, acid, ether, ester, lactone etc.; Identify 64 compositions wherein, accounted for more than 94% of whole essential oils.Wherein main compound is 3-carene (3.6877%), double-heptene [2.2.1]-heptane; 2,2-dimethyl--1-methylene radical (7.0838%), D-PC 560 (7.2210%), double-heptene [2.2.1]-heptane-2 ketone-1.3.3-trimethylammonium (7.3754%), beta-elemene (3.9964%), DL-Camphene-1 (10.5999%), DL-Camphene-2 (5.7631%), ledol (4.1679%), xanthophyll (3.0702%) etc.
The volatile oil component result for retrieval of the fragrant lamb's-quarters of chrysanthemum leaf is seen table 1.1.
The fragrant lamb's-quarters Essential Oil Chemistry of table 1.1 chrysanthemum leaf composition result for retrieval
| Peak number | RT | The compound title | Molecular formula | Relative content |
| 1 | 7.734min | Bicyclo[2.2.1]hept-2-ene,1,7,7-trimethyl- | C 10H 16 | 1.3053% |
| 2 | 8.847min | 3-Carene | C 10H 16 | 3.6877% |
| 3 | 9.115min | 2-Methyl-1-nonene-3-yne | C 10H 16 | 0.0858% |
| 4 | 9.460min | Bicyclo[2.2.1]heptane,2,2-dimethyl-3-methylene- | C 10H 16 | 7.0838% |
| 5 | 10.669min | 1R-.alpha.-Pinene | C 10H 16 | 0.1520% |
| 6 | 12.125min | .alpha.-Phellandrene | C 10H 16 | 0.3102% |
| 7 | 12.590min | .(+)-4-carene | C 10H 16 | 0.9476% |
| 8 | 13.088min | Benzene,1-methyl-3-(1-methylethyl)- | C 10H 14 | 0.6275% |
| 9 | 13.184min | D-limonene | C 10H 16 | 7.2210% |
| 10 | 13.407min | Santolina triene | C 10H 16 | 0.0352% |
| 11 | 14.420min | 1,4-Cyclohexadiene,1-methyl-4-(1-methylethyl)- | C 10H 16 | 1.0258% |
| 12 | 15.158min | 2-Cyclohexen-1-ol,1-methyl-4-(1-methylethyl)-,trans- | C 10H 18O | 0.1464% |
| 13 | 15.504min | Cyclohexene,1-methyl-4-(1-methylethylidene)- | C 10H 16 | 1.3119% |
| 14 | 15.934min | Bicyclo[2.2.1]heptan-2-one,1,3,3-trimethyl- | C 10H 16O | 7.3754% |
| 15 | 17.435min | trans-2-Pinanol | C 10H 18O | 0.8762% |
| 16 | 17.579min | 3-Cyclohexene-1-ol,1-methyl-4-(1-methylethyl)- | C 10H 18O | 2.0411% |
| 17 | 18.053min | 3-Cyclohexene-1-ol,1-methyl-4-(1-methylethyl)- ,cis | C 10H 18O | 0.5041% |
| 18 | 18.521min | 2-Cyclohexen-1-ol,1-methyl-4-(1-methylethyl)-,cis | C 10H 18O | 0.1023% |
| 19 | 20.068min | Terpineol,cis-beta- | C 10H 18O | 0.5099% |
| 20 | 20.956min | Fechyl acetate | C 12H 20O 2 | 2.2641% |
| 21 | 23.735min | Bornyl acetate | C 12H 20O 2 | 0.9010% |
| 22 | 25.324min | .gamma.-Elemene | C 15H 24 | 0.2569% |
| 23 | 25.610min | Cyclohexene, 4-ethenyl-4-methyl-3-(1-mehylethenyl)-1-(1-methylethyl)-,(3R-trans)- | C 15H 24 | 0.1835% |
| 24 | 26.069min | .alpha.-Cubebene | C 15H 24 | 0.4779% |
| 25 | 26.916min | Ylangene | C 15H 24 | 0.0540% |
| 26 | 27.210min | Copaene | C 15H 24 | 0.7657% |
| 27 | 27.482min | Caryophyllene | C 15H 24 | 0.2246% |
| 28 | 27.561min | Bicyclo[4.3.0]nonane,7-methylene-2,4,4-trimethyl-2-vinyl- | C 15H 24 | 0.1528% |
| 29 | 27.734min | Unidentified | C 15H 24 | 0.2767% |
| 30 | 27.912min | .beta.-Elemene | C 15H 24 | 3.9964% |
| 31 | 28.853min | Humulen-(v1) | C 15H 24 | 1.0952% |
| 32 | 29.362min | 1H-Cyclopenta[1,3]cyclopropa[1,2]enzzene,octahydro-7-methyl-3-methylene-4-(1-methylethyl)-,[3aS-(3a.alpha.,4.beta.,7.alpha.,7 aS*)]- | C 15H 24 | 0.3619% |
| 33 | 29.761min | Naphthalene,1,2,3,4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethyl)- | C 15H 24 | 0.0638% |
| 34 | 29.887min | 1H-Cyclolprop[e]azulene,decahydro-1,1,7-trimethyl-4-methylene-, | C 15H 24 | 0.0657% |
| 35 | 30.095min | .alpha.-Cubebene | C 15H 24 | 0.5497% |
| 36 | 30.285min | Azulene,1,2,3,3a,4,5,6,7-octahydro-1,4-dimethyl-7-(1-methenyl)-,[1R-(1.alpha.,3a.beta.,4.alpha.,7.beta.)]- | C 15H 24 | 0.3808% |
| 37 | 30.801min | (+)-Epi-bicyclosesquiphellandrene | C 15H 24 | 0.8548% |
| 38 | 30.913min | Naphthalene,1,2,4a,5,8,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)- | C 15H 24 | 0.5369% |
| 39 | 31.117min | Naphthalene,1,2,3,4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethyl)-,(1.alpha.,4a.alpha.,8a.alpha.)- | C 15H 24 | 1.3282% |
| 40 | 31.388min | 1H-Cyclopenta[1,3]cyclopropa[1,2]benzene,octahydro-7-methyl-3-methylene-4-(1-methylethyl)- | C 15H 24 | 2.8815% |
| 41 | 31.595min | Naphthalene,1,2,3,4,4a,5,6,8a-octahydro-1,6-dimethyl-4-(1-methylethyl)- | C 15H 24 | 0.3241% |
| 42 | 31.706min | Patchoulene | C 15H 24 | 1.2758% |
| 43 | 31.820min | Longifolene-(V4) | C 15H 24 | 0.7088% |
| 44 | 31.869min | Isoledene | C 15H 24 | 1.1489% |
| 45 | 32.011min | .alpha.-Muurolene | C 15H 24 | 2.8987% |
| 46 | 32.203min | .beta.-Guaiene | C 15H 24 | 0.4018% |
| 47 | 32.331min | Unidentified | C 15H 24 | 0.0610% |
| 48 | 32.582min | Naphthalene,1,2,4a,5,8,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-,[1S-(1.alpha.,4a.beta.,8a.alpha.)]- | C 15H 24 | 10.5999% |
| 49 | 32.661min | Naphthalene,2,3,4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethyl)-,(1.alpha.,4a.beta.,8a.alpha.)- | C 15H 24 | 5.7631% |
| 50 | 32.850min | 7-Tetracyclo[6.2.1.0(3.8)0(3.9)]undecanol,4,4,11,11-tetramethyl- | C 15H 24O | 0.0045% |
| 51 | 32.946 | Cyclohexane,1-ethenyl-1-methyl--(1-methylethenyl)-4-(1-methylidene)- | C 15H 26O | 5.1971% |
| 52 | 33.103min | Ledol | C 15H 26O | 4.1679% |
| 53 | 33.392min | Cedrene | C 15H 24 | 0.8920% |
| 54 | 33.518min | .alpha.-Muurolene | C 15H 24 | 1.3484% |
| 55 | 33.807min | Unidentified | ? | 0.1038% |
| 56 | 34.094min | Cyclohexanemethanol,4-ethenyl-,.alpha.,4-trimethyl-3-(1-methylethenyl)- | C 15H 26O | 0.5998% |
| 57 | 35.123min | Cedran-diol,8S,14- | C 15H 26O 2 | 0.0938% |
| 58 | 35.219min | 1R,4S,7S,11R-2,2,4,8-Tetramethyltricyclo[5.3.1.0(4,11)]undec-8-ene | C 15H 24 | 0.3168% |
| 59 | 35.677min | 6,9,12,15-Docosatetraenoic acid, methyl ester | C 23H 38O 2 | 0.0859% |
| 60 | 35.911min | Diethyl Phthalate | C 12H 14O 4 | 0.4912% |
| 61 | 36.274min | Cubenol | C 15H 26O | 0.2219% |
| 62 | 36.663min | Naphthalene,decahydro-4a-methyl-1-methylene-7-(1-methylethylidene)-,(4aR-trans)- | C 15H 24 | 0.0788% |
| 63 | 36.872min | .gamma.-Himachalene | C 15H 26O | 0.3943% |
| 64 | 37.401min | Bicyclo[4.4.0]dec-1-ene,2-isopropyl-5-methyl-9-methylene- | C 15H 26O | 2.7615% |
| 65 | 37.548min | .tau.-Cadinol | C 15H 26O | 1.3078% |
| 66 | 37.882min | .alpha.-Muurolol | C 15H 26O | 2.5556% |
| 67 | 39.171min | Carotol | C 15H 26O | 3.0702% |
| 68 | 44.013min | Unidentified | ? | 0.1056% |
Embodiment one: the volatile oil of the fragrant lamb's-quarters of chrysanthemum leaf suppresses bacteriological action
(1) bacterial classification and substratum
Bacterial classification is provided by Wuxi City, Jiangsu Province Southern Yangtze University Foodstuffs Academy, tabulates as follows:
| Belong to | Plant name |
| Staphylococcus | Streptococcus aureus |
| Streptococcus | Streptococcus pneumoniae, streptococcus acidi lactici |
| Enterobacter | Enteroaerogen |
| Salmonella belongs to | Salmonellas |
| Shigella | Dysentery bacterium |
| Rhodopseudomonas | Pseudomonas aeruginosa |
| Bacillus | Subtilis, bacillus cereus |
| Escherichia coli belongs to | Intestinal bacteria, enterococcus faecalis |
Beef-protein medium: Carnis Bovis seu Bubali cream 5 g, peptone 10 g, sodium-chlor 5 g, water 1000mL, agar 18g, pH7.2～7.4,121 ℃ sterilization 30min.
(2) experimentation
2.1 preparation bacteria suspension
Provoke certain thalline in the test tube that sterilized water is housed with transfering loop, process concentration and be about 10
5～10
6The bacteria suspension of cfu/mL.
2.2 the mensuration of volatile oil bacteriostatic action.
The Oxford agar diffusion method: the various bacteria suspensions that tried are got 1.0mL respectively in sterilized petridish; Pour the about 15mL of substratum that melts about postcooling to 60 ℃ into, shake up, after putting level attitude and solidifying; Place 6 in aseptic Oxford cup in each petridish moderate distance; Use liquid-transfering gun to draw respectively to use concentration that anhydrous methanol is made into material 200uL injection to be detected Oxford cup, make blank, do positive control with penbritin with anhydrous methanol as 0.1ug/uL.In 37 ℃ of cultivation 24h, observe the thalli growth situation, measure antibacterial circle diameter, represent fungistatic effect with the size of antibacterial circle diameter.D representes bacteriostatic activity with antibacterial circle diameter.The fungistatic effect judgement criteria is: D≤8 mm are insensitive, and < D≤13 mm are low responsive to 8 mm, and 13 mm < D≤19mm is a medium sensitivity, D>19mm is extremely sensitive.
2.3 the mensuration of minimal inhibitory concentration MIC and MBC MBC adopts Microdilution plate method
The test bacterial concentration is 10
6Cfu/ml.Good bacterium liquid is added in the 96 porocyte culture plates to draw dilution; Every hole 100uL; Every kind of medicine 100uL; Contrast dilution 10 holes make each hole Chinese traditional medicine concentration be respectively 100 mg/ml, 50 mg/ml, 25mg/ml, 12.5 mg/ml, 6.25 mg/ml, 3.125 mg/ml, 1.56 mg/ml, 0.78 mg/ml, 0.39mg/ml, 0.195 mg/ml successively.Last hole does not add medicine (only adding substratum and bacterium) and adds dilution bacterium liquid 100uL, is the bacterial growth control wells.Stay a row hole not add bacterium (only adding substratum and medicine) and make the medicine control wells, positive control is a penbritin, does doubling dilution by explanation.Place the Imin that vibrates on the vibrator, make in the hole behind the abundant mixing of solution, microwell plate is added a cover and is placed in the square enamel tray that is covered with wet gauze, in 37 ℃ of incubators, hatches 18h, observes the contained lowest drug concentration in no bacterial growth hole and is minimal inhibitory concentration.
The mensuration of MBC: get >=liquid culture (0. Iml) of MIC drug level is coated on the corresponding nutrient agar; Hatched 18 hours in 37 ℃ of incubators; The survive counting of bacterium colony is judged MBC value (colony count is the MBC value less than the pairing lowest drug concentration of 5 culture).
2.4 experimental result
The fragrant lamb's-quarters volatile oil of table 3 chrysanthemum leaf is to the inhibition zone size (mm) of various bacteriological actions
| Bacterium | The fragrant lamb's-quarters volatile oil of chrysanthemum leaf | Blank | Positive control |
| Streptococcus aureus | 19.76 | 8.27 | 28.65 |
| Streptococcus pneumoniae | 18.28 | 5.39 | 25.28 |
| Streptococcus acidi lactici | 16.23 | 5.02 | 24.35 |
| Enteroaerogen | 18.37 | 6.58 | 27.15 |
| Salmonellas | 19.05 | 8.29 | 28.29 |
| Dysentery bacterium | 15.36 | 5.89 | 20.07 |
| Pseudomonas aeruginosa | 14.27 | 6.71 | 21.59 |
| Subtilis | 18.85 | 7.92 | 29.01 |
| Bacillus cereus | 16.29 | 5.38 | 24.68 |
Volatile oil minimal inhibitory concentration (MCI) and MBC (the MBC) (unit: mg/mL) of the fragrant lamb's-quarters of table 4 chrysanthemum leaf
| Bacterium | MCI | MBC |
| Streptococcus aureus | 0.039 | 0.313 |
| Streptococcus pneumoniae | 0.039 | 0.156 |
| Streptococcus acidi lactici | 0.156 | 0.625 |
| Enteroaerogen | 0.313 | 1.25 |
| Salmonellas | 0.078 | 0.313 |
| Dysentery bacterium | 0.625 | 2.50 |
| Pseudomonas aeruginosa | 0.156 | 0.625 |
| Subtilis | 0.039 | 0.313 |
| Bacillus cereus | 0.078 | 0.625 |
Embodiment two: the volatile oil of the fragrant lamb's-quarters of chrysanthemum leaf suppresses the fungi effect
(1) bacterial classification and substratum
Bacterial classification is provided by Wuxi City, Jiangsu Province Southern Yangtze University Foodstuffs Academy, tabulates as follows:
| Belong to | Plant name |
| Candida | Candida albicans |
| Genera cryptococcus | Cryptococcus neoformans |
| Penicillium mould | Withered mould |
| Aspergillus | Black mold |
| Mucor | Mucor racemosus |
| Trichophyton | Trichophyton |
| Microsporum | Microsporum canis |
| Epidermophyton | Acrothesium floccosum |
Substratum adopts perfect medium: yeast extract paste 10g, peptone 10g, glucose 20g, agar 20g, water 1000ml, natural pH.Substratum and Oxford cup, petridish are all sterilized.
(2) experimentation
Bacteria suspension, experimentation such as enforcement process one are said
(3) experimental result
The fragrant lamb's-quarters volatile oil of table 5 chrysanthemum leaf is to the inhibition zone size (mm) of various bacteriological actions
| Fungi | The fragrant lamb's-quarters volatile oil of chrysanthemum leaf | Blank | Positive control |
| Candida albicans | 18.20 | 6.48 | 19.57 |
| Cryptococcus neoformans | 17.36 | 8.59 | 17.66 |
| Withered mould | 16.23 | 5.84 | 24.35 |
| Black mold | 15.47 | 5.59 | 17.05 |
| Mucor racemosus | 14.28 | 6.29 | 18.04 |
| Trichophyton | 18.75 | 6.91 | 19.01 |
| Microsporum canis | 19.05 | 8.62 | 20.29 |
| Acrothesium floccosum | 15.86 | 7.29 | 19.60 |
Volatile oil minimal inhibitory concentration (MCI) and MBC (the MBC) (unit: mg/mL) of the fragrant lamb's-quarters of table 6 chrysanthemum leaf
| Fungi | MCI | MBC |
| Streptococcus aureus | 0.039 | 0.313 |
| Streptococcus pneumoniae | 0.078 | 0. 625 |
| Streptococcus acidi lactici | 0.078 | 0.625 |
| Enteroaerogen | 0.156 | 1.25 |
| Salmonellas | 0.078 | 0.313 |
| Dysentery bacterium | 0.625 | 2.50 |
| Pseudomonas aeruginosa | 0.078 | 0.625 |
| Subtilis | 0156 | 0.313 |
| Bacillus cereus | 0.078 | 0.625 |
We can find through above two concrete experiments: the fragrant lamb's-quarters volatile oil of chrysanthemum leaf has the good restraining effect for fungi and bacterium.Fungies such as bacteriums such as Staphylococcus, streptococcus, enterobacter, Salmonella genus, Shigella, Rhodopseudomonas, bacillus, escherichia coli genus and Candida, genera cryptococcus, penicillium mould, Aspergillus, Mucor, microsporum, trichophyton, Epidermophyton had the obvious suppression function.Its restraining effect is compared with penbritin a little less than most Pseudomonas parts omitted are micro-, but its restraining effect can phase trans-substitution, for example cryptococcus and trichophyton in some fungi.In general, the volatile oil of the fragrant lamb's-quarters of chrysanthemum leaf has important use and is worth, and can be used as natural antiseptic-germicide and be used for industries such as food, medicine, replaces the synthetic antibacterial agents of toxic side effect.
4. the anti-oxidant activity of the volatile oil of the fragrant lamb's-quarters of chrysanthemum leaf
Implement bill DPPH method and measure the oxidation-resistance of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf
4.1.1DPPH principle
The DPPH radical is a kind of highly stable; The radical that can preserve for a long time; Often be used as test oxidation-resistance reagent. when it ran into the material that can discharge proton or is reduced, radical was eliminated, compound solution color generation noticeable change; Solution takes off to faint yellow from purple. and add of the absorbancy variation of institute test sample article front and back through measuring, can try to achieve the clearance rate of sample DPPH at 517 nm.
4.1.2 experimentation
In 10 mL test tubes of the DPPH ethanol solution of the 75umol/L concentration that 2.5 mL are housed; Add quantitative essential oil sample solution; Add a certain amount of absolute ethyl alcohol again, making TV is 3mL, behind room temperature held 30 min of concussion back; Measure the absorbance of 517 nm wavelength, by the computes clearance rate:
Clearance rate=[A
0-(A-A
B)]/A
0* 100%
The blank absorbency value is 0.303 (75 umol/LDPPH solution absorbance 0.590) under the Ao:517 nm.
A:517 nm adds the absorbance A value of the DPPH of sample down;
A
b: the absorbance of sample itself under 517 nm (not adding DPPH)
We add the clearance rate that 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL volatile oil record separately then respectively and see the following form:
| Volatilization oil mass (mL) | Clearance rate (%) |
| 0.2 | 58.9 |
| 0.3 | 89.6 |
| 0.4 | 91.9 |
| 0.5 | 93.0 |
| 0.6 | 93.3 |
Can find out when the volatilization oil mass that adds is less by above data; Clearance rate is directly proportional with the volatilization oil mass of adding; When reaching certain value along with the volatilization oil mass that adds, the variation of clearance rate is no longer linear with the variation of volatilization oil mass, and clearance rate increases slower.Can find that to the clearance rate of DPPH the fragrant lamb's-quarters volatile oil of chrysanthemum leaf has good oxygenizement through measuring the fragrant lamb's-quarters volatile oil of chrysanthemum leaf; Can be used as natural inhibitor and be used for industries such as food, medicine, makeup, healthcare products, replace the synthetized oxidation preventive agent of toxic side effect.For the fragrant lamb's-quarters volatile oil of chrysanthemum leaf Study on oxidation resistance can be to use the DPPH method, also can be method for distinguishing, for example ultra-oxygen anion free radical, hydroxyl radical free radical method etc.
Can find that through above three kinds of instances the fragrant lamb's-quarters volatile oil of chrysanthemum leaf has excellent antibiotic, fungicidal activity, also has good antioxygenation simultaneously.Exploitation for the fragrant lamb's-quarters of chrysanthemum leaf has important practice significance and application prospect.
Claims (7)
1. the preparation method of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf is characterized in that, adopts the steam distillation preparation; The fragrant lamb's-quarters of the fresh chrysanthemum leaf of gathering is added in the extractor, adds water, open the oil bath heating up to soaking sample fully; Oil bath temperature is transferred to 100～250 degrees centigrade, and the reaction kettle top adds a cover condensing works, collects refrigerative steam with Erlenmeyer flask; Then the steam condensate of collecting is put into to leave standstill in the separating funnel and treats layering, wait the oil reservoir that is divided no longer increase promptly no longer fuel-displaced till, oil reservoir take out is collected; Use siccative dry then, obtain weak yellow liquid, be i.e. the fragrant lamb's-quarters volatile oil of chrysanthemum leaf.
2. the preparation method of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf is characterized in that adopting CO
2Supercritical extraction obtains, with the fragrant lamb's-quarters of the fresh chrysanthemum leaf of gathering, and flushing, chopping, in the extractor of packing into, control CO
2Flow, pressure 11～40MPa, temperature 308～330K, time 1～4h collects volatile oil.
3. application like claim 1 or the fragrant lamb's-quarters volatile oil of 2 prepared chrysanthemum leaves is characterized in that this volatile oil is applied to food, medicine as natural antibacterial, sterilization, inhibitor.
4. the application of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf as claimed in claim 3 is characterized in that described this volatile oil is antibacterial, sterilization is a bacterium, or fungi.
5. the application of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf as claimed in claim 4; It is characterized in that described bacterium is Staphylococcus, streptococcus, enterobacter, Salmonella genus, Shigella, Rhodopseudomonas, bacillus, escherichia coli genus, especially subtilis, bacillus cereus, enterococcus faecalis, Salmonellas, intestinal bacteria, Pseudomonas aeruginosa, pneumobacillus, dysentery bacterium, pseudomonas aeruginosa or streptococcus acidi lactici.
6. the application of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf as claimed in claim 4; It is characterized in that described fungi is Candida, genera cryptococcus, penicillium mould, Aspergillus, Mucor, microsporum, trichophyton, Epidermophyton, especially Fusarium oxysporum, black mold, fusarium culmorum, saccharomyces cerevisiae, tobacco brown spot pathogen, Candida albicans, Microsporum canis, trichophyton or withered mould.
7. the application of the fragrant lamb's-quarters volatile oil of chrysanthemum leaf as claimed in claim 3 is characterized in that described this volatile oil is used for industries such as food, medicine, makeup, healthcare products as natural inhibitor, replaces the synthetized oxidation preventive agent of toxic side effect.
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| CN107502450A (en) * | 2017-08-28 | 2017-12-22 | 广西科技大学 | Wooden grape essential oil and its application |
| CN114052046A (en) * | 2021-11-24 | 2022-02-18 | 中国科学院成都生物研究所 | Application of chenopodium quinoa linn in preventing and treating root rot |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782667A (en) * | 2014-01-20 | 2015-07-22 | 中国农业大学 | Use of Chenopodium vulvaria plant volatile oil in prevention and treatment of plant mites |
| CN106538526A (en) * | 2016-09-27 | 2017-03-29 | 中国中医科学院中药研究所 | The application of Fusarium oxysporum antibacterial and its application process and caryophyllene |
| CN107502450A (en) * | 2017-08-28 | 2017-12-22 | 广西科技大学 | Wooden grape essential oil and its application |
| CN114052046A (en) * | 2021-11-24 | 2022-02-18 | 中国科学院成都生物研究所 | Application of chenopodium quinoa linn in preventing and treating root rot |
| CN114052046B (en) * | 2021-11-24 | 2023-01-24 | 中国科学院成都生物研究所 | Application of chenopodium quinoa linn in preventing and treating root rot |
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