CN115487227B - Antibacterial composition and application thereof - Google Patents
Antibacterial composition and application thereof Download PDFInfo
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- CN115487227B CN115487227B CN202211282590.2A CN202211282590A CN115487227B CN 115487227 B CN115487227 B CN 115487227B CN 202211282590 A CN202211282590 A CN 202211282590A CN 115487227 B CN115487227 B CN 115487227B
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- antibacterial
- essential oil
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- bacteria
- staphylococcus
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- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 82
- 239000000203 mixture Substances 0.000 title claims abstract description 73
- 239000000341 volatile oil Substances 0.000 claims abstract description 81
- 241000894006 Bacteria Species 0.000 claims abstract description 15
- 241000191967 Staphylococcus aureus Species 0.000 claims description 18
- 240000002262 Litsea cubeba Species 0.000 claims description 14
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- 244000223760 Cinnamomum zeylanicum Species 0.000 claims description 12
- 235000016639 Syzygium aromaticum Nutrition 0.000 claims description 12
- 244000223014 Syzygium aromaticum Species 0.000 claims description 12
- 235000007303 Thymus vulgaris Nutrition 0.000 claims description 12
- 235000017803 cinnamon Nutrition 0.000 claims description 12
- 239000001585 thymus vulgaris Substances 0.000 claims description 12
- 230000000845 anti-microbial effect Effects 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 7
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- 230000002335 preservative effect Effects 0.000 claims description 6
- 239000004599 antimicrobial Substances 0.000 claims description 5
- 241000191940 Staphylococcus Species 0.000 claims description 4
- 239000002537 cosmetic Substances 0.000 claims description 4
- 239000006210 lotion Substances 0.000 claims description 4
- 240000007436 Cananga odorata Species 0.000 claims description 3
- 241000588722 Escherichia Species 0.000 claims description 3
- 230000001815 facial effect Effects 0.000 claims description 3
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
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- KJPRLNWUNMBNBZ-QPJJXVBHSA-N (E)-cinnamaldehyde Chemical compound O=C\C=C\C1=CC=CC=C1 KJPRLNWUNMBNBZ-QPJJXVBHSA-N 0.000 description 1
- QMVPMAAFGQKVCJ-SNVBAGLBSA-N (R)-(+)-citronellol Natural products OCC[C@H](C)CCC=C(C)C QMVPMAAFGQKVCJ-SNVBAGLBSA-N 0.000 description 1
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
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- 229920000742 Cotton Polymers 0.000 description 1
- AERBNCYCJBRYDG-UHFFFAOYSA-N D-ribo-phytosphingosine Natural products CCCCCCCCCCCCCCC(O)C(O)C(N)CO AERBNCYCJBRYDG-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
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- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
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- 150000001491 aromatic compounds Chemical class 0.000 description 1
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- 125000003118 aryl group Chemical group 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
- JGQFVRIQXUFPAH-UHFFFAOYSA-N beta-citronellol Natural products OCCC(C)CCCC(C)=C JGQFVRIQXUFPAH-UHFFFAOYSA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
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- 210000005056 cell body Anatomy 0.000 description 1
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- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- KJPRLNWUNMBNBZ-UHFFFAOYSA-N cinnamic aldehyde Natural products O=CC=CC1=CC=CC=C1 KJPRLNWUNMBNBZ-UHFFFAOYSA-N 0.000 description 1
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- 150000002148 esters Chemical class 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
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- AERBNCYCJBRYDG-KSZLIROESA-N phytosphingosine Chemical compound CCCCCCCCCCCCCC[C@@H](O)[C@@H](O)[C@@H](N)CO AERBNCYCJBRYDG-KSZLIROESA-N 0.000 description 1
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- FCBUKWWQSZQDDI-UHFFFAOYSA-N rhamnolipid Chemical compound CCCCCCCC(CC(O)=O)OC(=O)CC(CCCCCCC)OC1OC(C)C(O)C(O)C1OC1C(O)C(O)C(O)C(C)O1 FCBUKWWQSZQDDI-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
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- A—HUMAN NECESSITIES
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- A61K36/13—Coniferophyta (gymnosperms)
- A61K36/15—Pinaceae (Pine family), e.g. pine or cedar
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/54—Lauraceae (Laurel family), e.g. cinnamon or sassafras
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- A61K36/61—Myrtaceae (Myrtle family), e.g. teatree or eucalyptus
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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- Oil, Petroleum & Natural Gas (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of antibacterial compositions, and particularly relates to an antibacterial composition and application thereof. The components in the antibacterial composition provided by the invention are synergistic with each other, and the antibacterial effect is obviously higher than that of a single essential oil. The antibacterial composition can effectively act on cell membranes of bacteria and fungi, change the permeability of the cell membranes and influence the integrity of the cell membranes, and simultaneously, the antibacterial composition can also damage cell walls of the bacteria and the fungi and damage cell structures, thereby effectively inhibiting the growth of the bacteria.
Description
Technical Field
The invention belongs to the technical field of antibacterial compositions. And more particularly to an antibacterial composition and application thereof.
Background
The plant essential oil is also called essential oil, volatile oil or aromatic oil, has the American term of liquid gold, mainly refers to a class of volatile secondary metabolites which exist in aromatic plants and are not mutually miscible with water, and is externally expressed as oily liquid substances with strong fragrance and smell. The plant essential oil is widely distributed in nature and can be derived from branches and leaves, flowers, roots, pericarps and seeds of plants of Zingiberaceae, verbenaceae, umbelliferae, labiatae, rutaceae and Myrtaceae. The chemical components are very complex and generally comprise hundreds of organic compounds, and can be mainly divided into four types according to the chemical components: terpenes (including monoterpene derivatives, sesquiterpene derivatives, diterpene derivatives and oxygenated terpenes derivatives), aliphatic compounds, aromatic compounds (mainly including phenols, aldehydes, ketones, esters and related derivatives), and nitrogen-containing and sulfur-containing compounds.
Plant essential oils are widely used for their ability to effectively inhibit or retard the growth of these microorganisms, bacteria, yeasts and molds. For example, cinnamaldehyde and cinnamon oil have certain antibacterial effect on gram negative bacteria shigella, salmonella and coliform. However, the antibacterial ability of a single essential oil is limited, and the expected value of people cannot be achieved. Researches show that the recombined compound essential oil has synergistic effect among the essential oil components, can widen the antibacterial spectrum of the recombined compound essential oil, and has better antibacterial effect than any single essential oil in the components, so that the recombined essential oil is widely applied to bacteriostasis. As disclosed in chinese patent application, an essential oil composition with antibacterial and disinfectant effects is prepared by compounding various essential oils, and the obtained essential oil composition can effectively kill bacteria or viruses, but the antibacterial effect of the essential oil composition is still to be further improved.
Disclosure of Invention
The invention aims to overcome the defect and the defect that the antibacterial effect of the traditional essential oil composition needs to be further improved, and provides an antibacterial composition with good antibacterial capability.
It is a further object of the present invention to provide an antimicrobial composition for use in the preparation of an antimicrobial product or preservative.
The object of the present invention is to provide a pharmaceutical composition.
It is another object of the present invention to provide a preservative.
It is another object of the present invention to provide a cosmetic composition.
The above object of the present invention is achieved by the following technical solutions:
an antibacterial composition comprises thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, lysimachia christinae hance leaf essential oil and ylang flower essential oil.
The composition creatively adopts thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, lysimachia christinae hance leaf essential oil and ylang flower essential oil to compound, has excellent antibacterial effect, has obviously better efficacy than that of each component which is used independently and lacks one or increases the effect of one of the components, and has obvious synergistic effect.
Preferably, the essential oil in the antimicrobial composition is obtained by water extraction.
More preferably, the water extraction method comprises the steps of:
crushing thyme, clove, cinnamon, litsea cubeba, longhairy antenoron leaf and ylang flower respectively by a medicinal material powdering machine, respectively adding the crushed thyme, the clove, the cinnamon, the litsea cubeba, the longhairy antenoron leaf and ylang flower into an essential oil distillation device together with water (the feed-liquid ratio is 1:6-10), heating to a boiling state, starting to count, adjusting the temperature to keep the liquid medicine in a low boiling state for 2-6 hours, collecting essential oil, drying by anhydrous sodium sulfate, respectively preparing thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, longhairy antenoron leaf essential oil and ylang flower essential oil, and storing at 4 ℃ for standby.
Preferably, the thyme essential oil, the clove essential oil, the cinnamon essential oil, the litsea cubeba essential oil, the lysimachia christinae hance leaf essential oil and the ylang-ylang flower essential oil are in a weight part ratio of 1-3: 1 to 3.5: 1-2: 1 to 3:1 to 3.5:1 to 3.
Preferably, the antibacterial composition has antibacterial activity against bacteria.
Preferably, the bacterium is a staphylococcus bacterium or an escherichia bacterium.
More preferably, the staphylococcus bacterium is staphylococcus aureus or staphylococcus albus.
More preferably, the escherichia bacteria are escherichia coli.
The invention further protects the application of the antibacterial composition in preparing antibacterial products.
Preferably, the antibacterial product is an antibacterial hand sanitizer, an antibacterial wet wipe, an antibacterial sachet, an antibacterial drug, an antibacterial facial cleanser, an antibacterial shampoo, an antibacterial body wash, an antibacterial facial mask, an antibacterial toning lotion or an antibacterial cleansing lotion.
The invention further provides a medicinal composition which comprises the antibacterial composition and medicinal auxiliary materials.
Preferably, the pharmaceutical excipients comprise phytosphingosine, rhamnolipid, plumeria rubra, pomelo peel, citronellol, nerol and water.
The invention further provides a preservative which comprises the antibacterial composition and preservative auxiliary materials.
Preferably, the antiseptic auxiliary materials comprise 1.3 to 5.5 percent of emulsifying thickener, 0.3 to 0.5 percent of chitosan, 1 to 7 percent of sodium alginate, 1 to 1.5 percent of calcium chloride, 1 to 5 percent of carrageenan and 0.3 to 0.5 percent of citric acid.
The present invention further provides a cosmetic composition comprising the above-described antibacterial composition and an antioxidant.
Preferably, the cosmetic composition further comprises a stabilizer, a solubilizer, a vitamin, a pigment or a fragrance.
The invention has the following beneficial effects:
the components in the antibacterial composition provided by the invention are synergistic with each other, and the antibacterial effect is obviously higher than that of a single essential oil. The antibacterial composition can effectively act on cell membranes of bacteria and fungi, change the permeability of the cell membranes and influence the integrity of the cell membranes, and simultaneously, the antibacterial composition can also damage cell walls of the bacteria and the fungi and damage cell structures, thereby effectively inhibiting the growth of the bacteria.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless otherwise indicated, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 1 example of antibacterial composition antibacterial experiments
1. Preparation of essential oils
Crushing thyme, clove, cinnamon, litsea cubeba, longhairy antenoron leaf and ylang flower respectively by a medicinal material powdering machine, respectively adding the crushed thyme, clove, cinnamon, litsea cubeba, longhairy antenoron leaf and ylang flower into high-efficiency essential oil distillation equipment together with water (the feed-liquid ratio is 1:8), heating to boiling, starting timing, adjusting the temperature to keep the liquid medicine in a low-boiling state for 4 hours, collecting essential oil, drying by anhydrous sodium sulfate, respectively preparing thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, longhairy antenoron leaf essential oil and ylang flower essential oil, and preserving at 4 ℃ for standby.
Preparation of LB broth: 5.0g of LB broth culture medium powder (formula: tryptone 10.0g/L, yeast powder 5.0g/L, sodium chloride 10.0 g/L) is taken, deionized water is added to 200mL, stirred and dissolved, and the mixture is placed in an autoclave for sterilization at 121 ℃ for 30min, and then taken out for cooling for standby.
3. Preparation of bacterial suspension: e.coli and staphylococcus aureus which are respectively picked up on a bacterial plate for one night are placed in 5mL of LB liquid culture medium, then the mixture is placed in a constant temperature shaking table for shaking culture at 37 ℃ and 170rpm for overnight, the absorbance (OD) value of the bacterial liquid cultured overnight is measured at 562nm by an enzyme-labeled instrument, and the concentration when the LB liquid culture medium is diluted to the OD value of 0.5 is used as a standard for standby.
4. Minimum inhibitory concentration test
The experimental method comprises the following steps: the single essential oil or the antibacterial composition (the components and the proportion are shown in Table 1) are subjected to multiple dilution on an aseptic 96-well plate by adopting a micro broth dilution method through an LB culture medium, so that the final concentration of each essential oil in the aseptic 96-well plate is 20, 10, 5, 2.5, 1.25, 0.625 and 0.313 mu L/mL respectively; adding 100 mu L of escherichia coli or staphylococcus aureus suspension with an OD value of 0.5 into each hole by using a pipetting gun; the negative control well was the same volume of LB medium without added bacterial suspension, and the positive control well was added with 100. Mu.L of LB medium and 100. Mu.L of bacterial suspension. After the 96-well plate is placed in a constant temperature incubator at 37 ℃ for culturing for 24 hours, the 96-well plate is taken out for observation, the essential oil concentration corresponding to the non-growing holes of the tested bacteria is taken as the lowest inhibitory concentration, three groups of parallel control are arranged for experiment, and the average value is obtained, and the results are shown in Table 2.
TABLE 1 antibacterial composition ingredients and proportions (parts)
Experimental results:
TABLE 2 minimum inhibitory concentration results for Staphylococcus aureus and Escherichia coli
The experimental results are shown in table 2: the bacteriostatic effect of both composition 1 and composition 2 was better than the other groups.
Calculation of FIC index: FIC index = MIC (combination of group a)/MIC (single use of group a) +mic (combination of group B)/MIC (single use of group B), when the FIC index is less than 0.5, the two drugs are synergistic; when the FIC index is 0.5-1, the two medicines are added; when the FIC index is greater than l and less than 2, the two drugs are irrelevant; when the FIC index is greater than 2, the two drugs are antagonism.
The MIC ratio of composition 1 and composition 2, i.e., FIC, was calculated to be between 0 and 0.5, which is a synergistic effect. The FIC value of the rest composition is 0.5-1, and the rest composition has additive effect. The data show that the composition comprising thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, largeflower leaf essential oil and ylang-ylang flower essential oil in specific proportion has good synergistic effect.
5. Antibacterial zone test
The testing method comprises the following steps:
preparation of LB solid medium: weighing 5.0g of LB broth culture medium powder and 3.0g of nutrient agar powder, supplementing deionized water to a total volume of 200mL, heating, stirring, dissolving, placing in an autoclave, sterilizing at 121 ℃ for 30min, and taking out for later use.
Cutting the filter paper into 6mm round pieces by using a puncher, placing the round pieces into a 50mL conical flask, plugging the bottle mouth by cotton, placing the round pieces into an autoclave, sterilizing the round pieces at 121 ℃ for 30min, and taking out the round pieces for later use.
Preparation of agar plates: after the LB solid culture medium is slightly cooled after sterilization, transferring a proper amount of LB solid culture medium into a sterilized culture dish near an alcohol lamp, standing and cooling until the LB solid culture medium is solidified, and covering a dish cover and placing the dish cover in a refrigerator at 4 ℃ for later use.
And (3) sucking 100 mu L of the overnight cultured escherichia coli by a pipette, inoculating the escherichia coli onto an agar plate, uniformly smearing bacterial liquid by holding a coating rod, clamping a sterilized and blow-dried filter paper sheet onto the agar plate inoculated with the escherichia coli by using forceps, fixing the position of the filter paper sheet by using a light filter-press paper sheet, wherein the process needs to ensure that the distance between the centers of the two filter papers exceeds 2cm and the distance between the filter papers is as consistent as possible, respectively dripping 3 mu L of essential oil antibacterial composition into the centers of the filter paper sheets, taking 3 mu L of litsea cubeba essential oil with the strongest antibacterial effect in a lowest antibacterial concentration test, taking 3 mu L of 25mg/mL chloramphenicol as a positive control, and setting three parallel controls for each different treatment. After essential oil and antibiotics are added dropwise, the culture dish is covered tightly, the cover is fixed by a sealing film, all agar plates are inverted and placed in a 37 ℃ incubator for culturing for 24 hours, and the diameter of a bacteriostasis ring is used for representing the bacteriostasis effect of each group. Staphylococcus aureus is coated on an agar plate in the same way, after filter paper sheets are placed in the same way, 3 mu L of essential oil antibacterial composition is respectively dripped in the center of the filter paper sheets, 3 mu L of litsea cubeba essential oil with the strongest antibacterial effect in the lowest antibacterial concentration test is dripped, 3 mu L of 50mg/mL of ampicillin is used as positive control, and three parallel controls are arranged for each different treatment. The antibacterial effect of the compound essential oil on staphylococcus aureus is evaluated according to the diameter of the antibacterial ring, and the result is shown in table 3.
Experimental results
Table 3: results of zone of inhibition test of bacteriostatic compositions
As shown in table 3: the diameter of the inhibition zone of the composition 1 is the largest, and the inhibition capacity is the strongest.
Example 2 example of the Effect of antibacterial compositions on bacterial cell membranes
The cell membrane is used as a natural barrier, can regulate and control the entry and exit of nutrient substances and metabolic products inside and outside the cell, and is a guarantee for maintaining normal life activities of the cell. When the permeability of the cell membrane is changed and the bacterial cell membrane is destroyed, small molecular substances in the bacterial cell escape from the membrane, and the final result is that the conductivity value of the bacterial liquid is increased. The present experiment will evaluate the effect of an antimicrobial composition on the permeability of the cell membrane of staphylococcus aureus by measuring the change in conductivity of the bacterial fluid after addition of the antimicrobial composition.
Adjusting the concentration of the bacterial liquid of staphylococcus aureus to an OD value of 0.5, adding an antibacterial composition, and taking the bacterial liquid which is not subjected to any treatment as a blank control, wherein the final concentration is 1/2 multiplied by the minimum antibacterial concentration; culturing the experimental group and the blank control group together in a constant temperature shaking table at 37 ℃ under 170r/min, sampling 1.5mL after culturing for 4 hours, setting the collected sample bacterial liquid in a refrigerated centrifuge at 4 ℃ under 4000r/min, centrifuging for 10min, taking supernatant, diluting to 50 times, and measuring the conductivity. The results are shown in Table 4.
Table 4: effect of antimicrobial composition on conductivity of Staphylococcus aureus fluid
As shown in table 4: the composition 1 has the highest bacterial liquid conductivity, which means that the intracellular electrolyte is released most and the antibacterial effect is best.
The nucleic acid of staphylococcus aureus has maximum absorption at the wavelength of 260nm, and the OD value of bacterial suspension at 260nm can effectively reflect the nucleic acid leakage condition of staphylococcus aureus. The cell membrane of the thallus can not pass through the macromolecular substance of nucleic acid under normal conditions, so that the leakage of the nucleic acid can effectively reflect the integrity of the cell membrane of staphylococcus aureus in a sample to be tested. The concentration of staphylococcus aureus bacterial liquid is regulated to OD value of 0.5, then the antibacterial composition is added and shaken uniformly, the final concentration is 1X the minimum antibacterial concentration, and bacterial liquid without any treatment is set as a blank control. After all samples were placed in a shaker and cultured at 37℃at 170r/min for 4h, each sample was sampled 1.5mL in a centrifuge tube, the samples were placed in a refrigerated centrifuge and centrifuged at 4℃at 10000r/min for 10min, the supernatant was collected and 200. Mu.L was sampled and placed in a sterile 96-well plate, the absorbance of the supernatant was measured at a wavelength of 260nm using a microplate reader, which was the nucleic acid leakage amount, and the nucleic acid leakage condition of the test bacteria was evaluated, and the results were shown in Table 5.
Table 5: effect of antimicrobial compositions on nucleic acid leakage from Staphylococcus aureus
As shown in table 5: the most severe nucleic acid leakage of composition 1 represents the best bacteriostatic effect of the composition.
Example 3 Effect of antibacterial combinations on bacterial cell walls
Alkaline phosphatase is usually present between the cell membrane and the cell wall of bacteria, and normally does not leak outside the cell body, and the alkaline phosphatase has maximum absorption at a wavelength of 510 nm. Thus, leakage of alkaline phosphatase can be tested to reflect the integrity of the staphylococcus aureus cell wall in the test sample.
After the overnight cultured staphylococcus aureus culture solution is diluted to a certain concentration, 15mL of the culture solution is absorbed into an aseptic centrifuge tube, the antibacterial composition is added and then shaken uniformly, so that the final concentration of the culture solution is one time of the lowest antibacterial concentration of the antibacterial composition, bacterial suspension without the antibacterial composition is used as a blank control, three parallel controls are arranged in two groups, two groups of total 8 bacterial liquid samples are placed in a shaking table together for culturing at 37 ℃ and 170r/min, after 4 hours of culturing, 200 mu L of each sample of each experimental group bacterial liquid and each blank control group bacterial liquid is transferred onto an aseptic 96-hole plate, and an enzyme-labeling instrument is used for measuring absorbance under the condition with the set wavelength of 510nm, and the results are shown in table 6.
Table 6: effect of antimicrobial compositions on Staphylococcus aureus cell walls
As can be seen from table 6: the alkaline phosphatase activity of the composition 1 and the composition 2 is low, the alkaline phosphatase activity of the composition 2 is lowest, the inhibition degree of the composition on the cell wall is highest, and the antibacterial effect is best.
The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (8)
1. An antibacterial composition, which is characterized by comprising thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, largeflower leaf essential oil and ylang-ylang flower essential oil;
wherein, the weight part ratio of thyme essential oil, clove essential oil, cinnamon essential oil, litsea cubeba essential oil, largeflower leaf essential oil and ylang flower essential oil is 1.5:2:1.5:1.5:2:1.5, 1.5:2.5:1.5:2:2:1.5; the antibacterial composition has antibacterial activity against bacteria.
2. The composition of claim 1, wherein the bacteria are staphylococcus or escherichia bacteria.
3. The composition of claim 2, wherein the staphylococcus bacteria are staphylococcus aureus or staphylococcus albus.
4. Use of an antimicrobial composition according to any one of claims 1 to 3 for the preparation of an antimicrobial product.
5. The use according to claim 4, wherein the antibacterial product is an antibacterial hand wash, an antibacterial wet wipe, an antibacterial sachet, an antibacterial agent, an antibacterial facial cleanser, an antibacterial shampoo, an antibacterial body wash, an antibacterial mask, an antibacterial lotion or an antibacterial cleansing lotion.
6. A pharmaceutical composition comprising the antibacterial composition of any one of claims 1 to 3 and a pharmaceutically acceptable adjuvant.
7. A preservative comprising the antimicrobial composition of any one of claims 1 to 3 and a preservative adjuvant.
8. A cosmetic composition comprising the antibacterial composition of any one of claims 1 to 3 and an antioxidant.
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| CN108042614A (en) * | 2017-12-19 | 2018-05-18 | 江西中医药大学 | Volatile clove oil is for sterilization and/or antibacterial, antimicrobial agent application |
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