A compound essential oil, an anti-corrosion system containing the same, and uses thereof

Abstract

The present invention provides a compound essential oil, an anti-corrosion system containing the same, and their uses. The compound essential oil is composed of the following components: wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and lemon-aldehyde-type camphor leaf essential oil. The compound essential oil has the effect of significantly inhibiting the growth of Propionibacterium acnes, Staphylococcus aureus, and Malassezia for a long time. The present invention also provides a compound anti-corrosion system, which contains the above compound essential oil, 1,2-hexanediol, and caprylyl glycol.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and particularly to a compound essential oil with antibacterial effects, a preservative system containing the same, and their uses. Background Art

[0002] Essential oils are extracted from parts of plants such as leaves, flowers, seeds, fruits, roots, barks, resins, and wood cores by methods such as steam distillation, cold pressing, enfleurage, and supercritical carbon dioxide extraction. Since essential oils are of natural origin, they are widely regarded as closer to nature than chemically synthesized products and are a healthier and more sustainable choice. Essential oils are complex mixtures composed of various chemical components, and the volatile oils, phenols, alcohols, esters, etc. contained in their components endow essential oils with various effects such as antibacterial, anti-inflammatory, antioxidant, and analgesic effects. When essential oils are compounded with preservative-free agents, it belongs to a preservative-free system, which can synergistically enhance the antibacterial effect while reducing the dosage of preservative-free agents to reduce irritation to the skin.

[0003] Internationally, three methods are generally used to evaluate the anti-corrosion and antibacterial performance: determination of the minimum inhibitory concentration (MIC), anti-corrosion challenge test, and linear regression method. Among them, the anti-corrosion challenge test is to add a quantitative amount of microorganisms to the preservative sample. At 0 / 7 / 14 / 21 / 28 days thereafter, the anti-corrosion performance is evaluated by measuring the content of microorganisms in the sample, and the experimental period is as long as 28 days. The anti-corrosion challenge test is required by regulations in many industries (such as cosmetics, medicine, and food), conforms to international standards, and is not only scientific and reliable in evaluating the anti-corrosion and antibacterial performance, but also provides important support for product development, regulatory compliance, and market competition. Summary of the Invention

[0004] One object of the present invention is to provide a compound essential oil with antibacterial effects, which comprises the following components: wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and lemon aldehyde-type camphor leaf essential oil.

[0005] Preferably, the wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and lemon aldehyde-type camphor leaf essential oil of the present invention are respectively prepared by the following steps: the wild chrysanthemum essential oil is extracted from the flowers of the Compositae plant Chrysanthemum indicum ( Chrysanthemum indicum L. ) by steam distillation; the jasmine essential oil is extracted from the flowers of Jasminum sambac ( Jasminum officinalis sambac ) by supercritical carbon dioxide extraction; the patchouli essential oil is extracted from the above-ground part of Pogostemon cablin ( Patchouli ) by steam distillation; the lemon aldehyde-type camphor leaf essential oil is extracted from the fresh leaves of Cinnamomum camphora (L.) Presl var. latilimbum ( Cinnamomum camphora ) by water distillation.

[0006] Preferably, the volume ratio of the wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and lemon aldehyde-type camphor leaf essential oil contained in the compound essential oil of the present invention is 4:1:2:3.

[0007] The present invention has studied the antibacterial effect of the compound essential oil of the present invention. The results show that the antibacterial effect of the compound essential oil of the present invention is higher than that of the single essential oil, and a synergistic effect is produced among the single essential oils. In addition, the compound essential oil overcomes the defect of the narrow antibacterial spectrum of the single essential oil. For example, the antibacterial rate of wild chrysanthemum essential oil against Staphylococcus aureus at a concentration of 50 mg / ml is 68%, and the antibacterial rate against Propionibacterium acnes at the same concentration is close to 95%. However, the compound essential oil of the present invention has little difference in antibacterial effect against the three bacteria, Staphylococcus aureus, Propionibacterium acnes, and Malassezia, within a specific concentration range, all higher than 90%, showing a broad-spectrum antibacterial effect.

[0008] On the other hand, the present invention combines the essential oil as a natural preservative from a plant source with polyols to form a compound preservative composition, namely the "preservative system". The present invention provides a preservative system that includes the above compound essential oil.

[0009] Preferably, the preservative system of the present invention is composed of the above compound essential oil, 1,2-hexanediol, and caprylyl glycol, and the volume ratio of the compound essential oil, 1,2-hexanediol, and caprylyl glycol is (1-10):2:1.

[0010] Preferably, in the preservative system of the present invention, the volume ratio of the compound essential oil, 1,2-hexanediol, and caprylyl glycol is 10:2:1.

[0011] The preservative system of the present invention can reduce the colony by nearly 100% (sterile growth) on the 7th day of co-incubation with bacteria, can effectively kill Escherichia coli and Staphylococcus aureus, and no colony formation is observed during the 28-day experiment; on the 7th day of co-incubation with fungi, the colony can be reduced by nearly 90%, can effectively prevent the growth of Candida albicans, and no colony formation is observed during the 28-day experiment. The above results indicate that the preservative system of the present invention has a strong antibacterial effect.

[0012] On the other hand, the present invention provides the use of the above compound essential oil and preservative system in cosmetics.

[0013] Preferably, the content of the preservative system of the present invention in cosmetics is 0.05%-2%.

[0014] More preferably, the content of the preservative system of the present invention in cosmetics is 0.5%.

[0015] Advantages of the anti-corrosion system of the present invention: The anti-corrosion system of the present invention combines a compound essential oil with antibacterial activity derived from plants and a chemical anti-corrosion component, polyol, to obtain an anti-corrosion system with the synergistic effect of plant essential oil anti-corrosion and chemical anti-corrosion. Selecting the compound essential oil and polyol for compounding and use in a certain proportion can reduce the usage amount of polyol, thereby reducing the risk of irritation to the skin caused by a large amount of polyol usage; at the same time, the active ingredients of the plant essential oil also have functions such as whitening, anti-aging, and repair, making the anti-corrosion system multifunctional. In addition, due to the special fragrance of the plant essential oil, it also has the effect of enhancing fragrance, which highlights the advantages of the essential oil in the application of the additive-free anti-corrosion system in cosmetics. Compounding the compound essential oil and polyol, the additive-free anti-corrosion system after compounding can achieve effects such as improving the safety of use, increasing the antibacterial intensity, and broadening the antibacterial spectrum. Description of the Drawings

[0016] Figure 1 : Bacteriostatic rates of various compound essential oils and their single essential oil components at different concentrations against Staphylococcus aureus.

[0017] Figure 2 : Bacteriostatic rates of various compound essential oils and their single essential oil components at different concentrations against Propionibacterium acnes.

[0018] Figure 3 : Bacteriostatic rates of various compound essential oils and their single essential oil components at different concentrations against Malassezia.

[0019] Figure 4 : Bacteriostatic rates of various compound essential oils at different concentrations against Staphylococcus aureus; among them p < 0.05, p < 0.001.

[0020] Figure 5 : Bacteriostatic rates of various compound essential oils at different concentrations against Propionibacterium acnes.

[0021] Figure 6 : Bacteriostatic rates of various compound essential oils at different concentrations against Malassezia.

[0022] Figure 7 : Inhibitory effects of each anti-corrosion system against Staphylococcus aureus and Escherichia coli.

[0023] Figure 8 : Inhibitory effects of each anti-corrosion system against Candida albicans.

[0024] Figure 9 : GC-MS detection results of wild chrysanthemum essential oil.

[0025] Figure 10 : GC-MS detection results of jasmine essential oil.

[0026] Figure 11: GC-MS detection results of Pogostemon cablin essential oil.

[0027] Figure 12 : GC-MS detection results of Litsea cubeba essential oil of citral type. Detailed implementation manners

[0028] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the scope of the present invention.

[0029] The essential oils used in the following Examples 2-4 are all commercially purchased or extracted by the methods of Example 1 or 2.

[0030] Chrysanthemum indicum essential oil: Purchased from Mujiuye (Shanghai) Technology Co., Ltd., batch number: TCM02611012310, enterprise standard: relative density (20 °C): 0.960 - 1.015; refractive index (20 °C): 1.475 - 1.525. The cosmetic raw material safety information submission code of Chrysanthemum indicum essential oil is 007637-02571-1356.

[0031] Jasmine essential oil: Purchased from Mujiuye (Shanghai) Technology Co., Ltd., batch number: BEO08612012310, enterprise standard: relative density (20 °C): 0.925 - 0.998; refractive index (20 °C): 1.481 - 1.525. The cosmetic raw material safety information submission code of Jasminum sambac essential oil is 004723-02571-4822.

[0032] Pogostemon cablin essential oil: Purchased from Mujiuye (Shanghai) Technology Co., Ltd., batch number: BEO02561012310, enterprise standard: relative density (20 °C): 0.949 - 0.983; refractive index (20 °C): 1.505 - 1.512. The cosmetic raw material safety information submission code of Pogostemon cablin essential oil is 002634-02571-5566.

[0033] Litsea cubeba essential oil of citral type: Purchased from Mujiuye (Shanghai) Technology Co., Ltd., batch number: BEO02241012408, enterprise standard: relative density (20 °C): 0.875 - 0.890; refractive index (20 °C): 1.468 - 1.474. The cosmetic raw material safety information submission code of Litsea cubeba essential oil is: 008504-02571-9021.

[0034]

Example 1

[0035] Instruments used in the steam distillation method: water-sealed distillation kettle, and its equipment composition is:

[0036] Distillation kettle: The overall appearance is cylindrical, with a spherical crown bottom and an upper opening as the feeding port;

[0037] Gooseneck tube: A conical lid with the upper opening of the distillation kettle, connecting to the condenser;

[0038] Condenser: An aluminum tube condenser that condenses steam and cools the distillate;

[0039] Oil-water separator: Made of aluminum, it is both a container for receiving the distillate and a separator for essential oil and water.

[0040] Process flow of extracting essential oil by steam distillation method: Feeding, adding water, distillation, cooling, oil-water separation, essential oil, packaging.

[0041] 1.1 Preparation of wild chrysanthemum essential oil

[0042] Wild chrysanthemum essential oil is a volatile oil extracted by steam distillation method using the flowers of the wild chrysanthemum ( Chrysanthemum indicum L. ), a plant of the Compositae family in Henan Province, China, as raw materials.

[0043] During the full bloom period of wild chrysanthemum from October to December every year, from after the morning dew dries until about 10 am, select wild chrysanthemums when the flower buds start to open but have not fully bloomed for picking. After picking, spread out the wild chrysanthemums for drying in the sun or drying at low temperature. Place the processed wild chrysanthemums evenly in the inner tank of the distillation equipment, and control the filling amount at 70%-80% of the tank volume to ensure that the steam can fully penetrate the flower material layer. Add pure water equal to the volume of the wild chrysanthemums in the outer distillation tank to ensure that steam can be continuously generated and cover the raw materials at the bottom of the inner tank. Turn on the heating device, with the extraction temperature at 90~96°C, the temperature of the condenser tube at 15~25°C, and the distillation time at 60~90 min. When the essential oil content in the distilled condensate significantly decreases, end the heating operation. The condensed mixture flows into the oil-water separator through the condenser tube. The mixture needs to be left standing for 24 hours for natural stratification, and use a dropper or separating funnel to separate the upper wild chrysanthemum essential oil from the lower hydrosol. Filter the separated essential oil through filter paper or filter cloth to remove particles or other impurities suspended in the essential oil to obtain wild chrysanthemum essential oil.

[0044] 1.2 Preparation of jasmine essential oil

[0045] Jasmine essential oil is a volatile oil obtained by supercritical carbon dioxide extraction using the flowers of Chinese jasmine ( Jasminum officinalis sambac ) as raw materials.

[0046] Select jasmine flowers with moderate openness at dusk, avoiding overly young or overly mature flowers. After picking, place the flowers in a dry room with good ventilation and no direct sunlight for 1 - 2 days, and then use a dryer to heat and dry at 45°C. After the flowers are dried, the petals will be in a brittle and hard state without a feeling of moisture. Put the dried jasmine flowers into a crusher for processing, crush them into a coarse powder passing through a 10 - 20 mesh sieve, pack the weighed jasmine flower coarse powder in a sand cloth bag, and put it into an extractor, then seal the top cover. Turn on the power of the constant temperature bath. Open the valve of the carbon dioxide cylinder so that the gas passes through the dryer along the pipeline, the cooling condenser (immersed in an ice - water bath), the plunger pump, and enters the pressure - boosting buffer bottle. After the pressure in the pressure - boosting buffer bottle greatly exceeds the experimental pressure, open valve V1 to raise the pressure of the extractor to the experimental pressure, close valve V1, let the jasmine flowers be pre - infiltrated for 10 min, and perform supercritical CO2 fluid extraction. The flow rate of the CO2 fluid is 25 L / H - 40 L / H, the extraction temperature is 40°C, the extraction pressure is 20 MPa - 25 MPa, the separation temperature is 35°C, the separation pressure is 4.5 MPa - 5.5 MPa, and the extraction time is 1.5 h - 2 h to obtain jasmine essential oil.

[0047] 1.3 Preparation of Pogostemon cablin essential oil

[0048] Pogostemon cablin essential oil is a volatile oil extracted from the aerial parts of Pogostemon cablin ( Patchouli ) by steam distillation.

[0049] During the flowering period of Pogostemon cablin from July to September, select and pick fresh, dry, and uncontaminated whole plants of Pogostemon cablin, and wash them. Disassemble the distiller and clean all parts to ensure there is no oil stain and impurity. Add 2 / 3 to 3 / 4 of the distilled water in the distillation pot and place it on the heating plate, heat the heating plate until the water starts to boil. When the water in the distillation pot starts to boil, put the whole plant of Pogostemon cablin into the distillation pot to ensure it is completely immersed in water. After waiting for the water to start boiling, connect the distiller with a cooling pipe and put the other end of the pipe into the collection bottle. The steam will condense through the cooling pipe and flow into the collection bottle. Wait for 24 hours, the essential oil will float on the upper layer of the water, and use a dropper or a separating funnel to separate it from the water. After collecting the essential oil, filter it with filter paper or filter cloth to remove the particles or other impurities suspended in the essential oil to obtain Pogostemon cablin essential oil.

[0050] 1.4 Preparation of citral - type Cinnamomum camphora leaf essential oil

[0051] Citral - type Cinnamomum camphora leaf essential oil is a volatile oil extracted from the fresh leaves of citral - type Cinnamomum camphora ( Cinnamomum camphora ) by water distillation.

[0052] During the period from June to September, fresh camphor tree leaves were picked from camphor trees of the citral type planted in Yongxiu County, Jiujiang City, Jiangxi Province. The fresh camphor tree leaves were washed and placed on the upper sieve plate of the essential oil extractor. An appropriate amount of water was added to the bottom of the distiller and heated to boiling. The generated steam passed through the sieve plate from bottom to top and reached the position of the fresh camphor tree leaves, causing the components in the fresh camphor tree leaves to flow out, forming an oil-water mixed steam. The oil-water mixed steam entered the oil-water separator through the condenser tube. After distillation for 2 hours, the liquid mixture in the oil-water separator was allowed to stand and layer, and the upper oil phase was taken to obtain the essential oil of camphor tree leaves of the citral type.

[0053]

Example 2

[0054] According to the following formula, each single essential oil was mixed evenly in proportion to obtain Compound Essential Oil 1 to Compound Essential Oil 4.

[0055] Compound Essential Oil 1: Wild chrysanthemum essential oil + Jasmine essential oil + Patchouli essential oil + Essential oil of camphor tree leaves of the citral type, volume ratio 1:4:3:2; that is, the content of wild chrysanthemum essential oil in Compound Essential Oil 1 is 10%, the content of jasmine essential oil is 40%, the content of patchouli essential oil is 30%, and the content of essential oil of camphor tree leaves of the citral type is 20%.

[0056] Compound Essential Oil 2: Wild chrysanthemum essential oil + Jasmine essential oil + Patchouli essential oil + Essential oil of camphor tree leaves of the citral type, ratio 2:3:4:1; that is, the content of wild chrysanthemum essential oil in Compound Essential Oil 2 is 20%, the content of jasmine essential oil is 30%, the content of patchouli essential oil is 40%, and the content of essential oil of camphor tree leaves of the citral type is 10%.

[0057] Compound Essential Oil 3: Wild chrysanthemum essential oil + Jasmine essential oil + Patchouli essential oil + Essential oil of camphor tree leaves of the citral type, ratio 3:2:1:4; that is, the content of wild chrysanthemum essential oil in Compound Essential Oil 3 is 30%, the content of jasmine essential oil is 20%, the content of patchouli essential oil is 10%, and the content of essential oil of camphor tree leaves of the citral type is 40%.

[0058] Compound Essential Oil 4: Wild chrysanthemum essential oil + Jasmine essential oil + Patchouli essential oil + Essential oil of camphor tree leaves of the citral type, ratio 4:1:2:3; that is, the content of wild chrysanthemum essential oil in Compound Essential Oil 4 is 40%, the content of jasmine essential oil is 10%, the content of patchouli essential oil is 20%, and the content of essential oil of camphor tree leaves of the citral type is 30%.

[0059]

Example 3

[0060] This example studied the inhibitory effects of Compound Essential Oils 1 to 4 (Example 2) of the present invention on three bacteria, namely Staphylococcus aureus, Propionibacterium acnes, and Malassezia.

[0061] 3.1 Test materials

[0062] 3.1.1 Test strains: Staphylococcus aureus (ATCC6538), Propionibacterium acnes (SC5314), Malassezia (ATCC14521) were purchased from the Cell Bank of the Chinese Academy of Sciences.

[0063] 3.1.2 Test drugs and reagents: Wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, citral-type camphor leaf essential oil, normal saline, PBS, surfactant HS15, BHI medium, tryptone soy agar medium (TSA), MH broth medium.

[0064] 3.1.3 Consumables and instruments: 10 cm petri dishes, sterilized inoculation loops, 96-well plates, multifunctional microplate readers, anaerobic generation bags, shakers, biochemical incubators, laminar flow hoods.

[0065] 3.2 Test methods

[0066] 3.2.1 Preparation of single essential oil mixed solutions and compound essential oil mixed solutions

[0067] Dissolve single essential oils or Compound Essential Oil 1 - Compound Essential Oil 4 separately into mixed solutions with a mass concentration of 800 mg / ml using a solution containing 2% surfactant HS15: Add 800 mg of single essential oil or Compound Essential Oil 1 - Compound Essential Oil 4 to 1 ml of the solution containing 2% surfactant HS15 and mix well to obtain.

[0068] 3.2.2 Preparation of strains

[0069] 3.2.2.1 Cultivation of Staphylococcus aureus: Before cultivation, take out the cryopreserved strain. After resuscitation, use an inoculation loop to inoculate the bacteria onto a TSA agar plate by the streaking method and culture at 37°C for 24 - 48 h until obvious colonies appear, which is used as the working plate. The working plate is replaced every 20 days to ensure the colony activity and stored at 4°C. To obtain the Staphylococcus aureus bacterial solution in the logarithmic growth phase, pick a small round colony on the working plate each time, place it in 1 ml of TSA liquid medium, and shake the bacteria overnight at a rotation speed of 250 rpm and a temperature of 37°C. Harvest the strain the next day for the experiment.

[0070] 3.2.2.2 Culture of Propionibacterium acnes: Propionibacterium acnes is an anaerobic bacterium and needs to be cultured in an anaerobic environment. An open anaerobic generation bag is placed in an anaerobic culture bag to create an anaerobic environment, and an anaerobic indicator is used to observe the oxygen content in the culture bag. Before culturing, the frozen strain is taken out. After resuscitation, the bacterium is inoculated onto a BHI agar plate using a inoculation loop by the streaking method and placed in an anaerobic environment for culturing at 37 °C for 48 - 72 h until obvious colonies appear, which is used as the working plate. The working plate is replaced every 20 days to ensure the colony activity and stored at 4 °C. To obtain the Propionibacterium acnes bacterial solution in the logarithmic growth phase, each time a small round colony on the working plate is picked and placed in 1 ml of BHI liquid medium, and shaken overnight under the conditions of an anaerobic environment, a rotation speed of 250 rpm, and a temperature of 37 °C. The strain is harvested the next day for experiments.

[0071] 3.2.2.3 Culture of Malassezia: Before culturing, the frozen strain is taken out. After resuscitation, the bacterium is inoculated onto an MH agar plate using a inoculation loop by the streaking method and cultured at 30 °C for 24 - 48 h until obvious colonies appear, which is used as the working plate. The working plate is replaced every 20 days to ensure the colony activity and stored at 4 °C. To obtain the Malassezia bacterial solution in the logarithmic growth phase, each time a small round colony on the working plate is picked and placed in 1 ml of MH liquid medium, and shaken overnight at a rotation speed of 250 rpm and a temperature of 30 °C. The strain is harvested the next day for experiments.

[0072] 3.2.3 Determination of the antibacterial rates of each single essential oil and compound essential oil against Staphylococcus aureus, Propionibacterium acnes, and Malassezia

[0073] The micro-dilution method is used to test the antibacterial rate of plant essential oil samples. The specific steps are as follows:

[0074] (1) Add 100 μl of culture medium to each well of a 96-well plate;

[0075] (2) Add 100 μl of the prepared single essential oil mixed solution or compound essential oil mixed solution to the first row, and then perform two-fold dilution of the essential oil. That is, after adding the single essential oil mixed solution or compound essential oil mixed solution to the first well, use a pipette to blow and mix thoroughly, then aspirate 100 μl and add it to the second well and mix thoroughly again. Repeat this until the last well, and aspirate 100 μl from the last well and discard it. Each essential oil concentration is repeated 3 times. At this time, each test well should contain 100 μl of solution;

[0076] (3) Add 100 μl of the diluted Staphylococcus aureus or Propionibacterium acnes or Malassezia (the concentration of the diluted bacterial solution is 2×10 6 cfu / ml) to each well. After adding, the final concentration of the bacterial solution is 1×10 6cfu / ml, the final dilution factor of the single essential oil mixed solution or the compound essential oil mixed solution in the first well is 4 times;

[0077] (4) Set negative controls (including essential oils and culture media with different dilution factors, removing the drug background OD600) and positive controls (100 µl of culture medium + 100 µl of bacterial solution, the final concentration of the bacterial solution is 1×10 6 cfu / ml) on the same plate;

[0078] (5) After incubating at 37 °C for 24 hours under the culture conditions, measure the OD600 of each well with an enzyme-linked immunosorbent assay (ELISA) reader and calculate the bacterial viability.

[0079] Inhibitory rate (%) = 100% - [OD600 (test) - OD600 (negative)] / [OD600 (positive) - OD600 (negative)]

[0080] 3.3 Test results

[0081] 3.3.1 Bacteriostatic effect of compound essential oil on Staphylococcus aureus

[0082] The bacteriostatic effects of compound essential oils at different concentrations and their single components on Staphylococcus aureus are shown in Table 1 and Figure 1 , and the results show that the compound essential oils and their single components have varying degrees of inhibitory effects on Staphylococcus aureus. Compared with the single essential oils, the inhibitory rates of compound essential oil 4 at each concentration are higher. The inhibitory rate of compound essential oil 4 against Staphylococcus aureus can reach 90% in the concentration range of 25 mg / ml and 50 mg / ml; the inhibitory rate of compound essential oil 1 against Staphylococcus aureus is higher than that of the single essential oil in the concentration range of 25 mg / ml - 200 mg / ml; the inhibitory rates of compound essential oils 2 and 3 against Staphylococcus aureus are both lower than 50% in the concentration range of 6.25 mg / ml - 12.5 mg / ml, and although the inhibitory rates against Staphylococcus aureus are lower than 90% in the concentration range of 25 mg / ml - 100 mg / ml, they are slightly higher than those of the single essential oils.

[0083] Table 1 Bacteriostatic rates of compound essential oils at different concentrations and their single components against Staphylococcus aureus (means, unit: %)

[0084]

[0085] 3.3.2 Bacteriostatic effect of compound essential oil on Propionibacterium acnes

[0086] The bacteriostatic effects of compound essential oils at different concentrations and their single components on Propionibacterium acnes are shown in Table 2 and Figure 2, The results showed that the compound essential oils and their single-component components had varying degrees of inhibitory effects on Propionibacterium acnes. The antibacterial rate of compound essential oil 1 against Propionibacterium acnes reached 90% at a concentration range of 25 mg / ml - 100 mg / ml; the antibacterial rates of compound essential oils 2 and 4 against Propionibacterium acnes were both higher than 90% within the concentration range of 50 mg / ml - 200 mg / ml; the antibacterial rates of compound essential oil 3 against Propionibacterium acnes were all higher than 90% within the concentration range of 100 mg / ml - 200 mg / ml.

[0087] Table 2 Antibacterial rates of compound essential oils with different concentrations and their single-component components against Propionibacterium acnes (mean, unit: %)

[0088]

[0089] 3.3.3 Antibacterial effect of compound essential oils against Malassezia

[0090] The antibacterial effects of compound essential oils with different concentrations and their single-component components against Malassezia are shown in Table 3 and Figure 3 , and the results showed that the compound essential oils and their single-component components had varying degrees of inhibitory effects on Malassezia. Compared with the single essential oils, the antibacterial rates of compound essential oil 4 at each concentration were higher. The antibacterial rates of compound essential oils 1 and 2 against Malassezia were both higher than 50% and lower than 90% within the concentration range of 12.5 mg / ml - 200 mg / ml; the antibacterial rates of compound essential oil 3 against Malassezia were all close to 90% within the concentration range of 6.25 mg / ml - 200 mg / ml; the antibacterial rates of compound essential oil 4 against Malassezia were higher than 50% within the concentration range of 6.25 mg / ml - 25 mg / ml and higher than 90% within the concentration range of 50 - 200 mg / ml.

[0091] Table 3 Antibacterial rates of compound essential oils with different concentrations and their single-component components against Malassezia (mean, unit: %)

[0092]

[0093] 3.3.4 Comparison of antibacterial effects of four compound essential oils against Staphylococcus aureus, Propionibacterium acnes and Malassezia

[0094] The results are shown in Figures 4 - 6 , and the comparison of the two groups of data used an independent samples t test ( t- test). The antibacterial effects of each compound essential oil with different concentrations against Staphylococcus aureus, Propionibacterium acnes and Malassezia had varying degrees of differences. Among them, within the concentration range of 6.25 - 50 mg / ml, the antibacterial effect of compound essential oil 4 against Staphylococcus aureus was significantly enhanced compared with other groups ( p <0.05; p<0.001); There was no significant difference between the antibacterial effects of the four compound essential oils of the present invention against Propionibacterium acnes and those against Malassezia.

[0095] 3.4 Experimental conclusions

[0096] After comprehensive comparison, the antibacterial effect of compound essential oil 4 showed obvious advantages compared with other formulations. Therefore, we selected compound essential oil 4 at a concentration of 50 mg / ml for subsequent experiments.

[0097]

Example 4

[0098] 4.1 Test materials

[0099] 4.1.1 Test strains: Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 25922), Candida albicans (ATCC 10231) were purchased from the Cell Bank of the Chinese Academy of Sciences.

[0100] 4.1.2 Test drugs and reagents: Wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, lemon aldehyde-type camphor leaf essential oil, 1,2-hexanediol, octanediol, physiological saline, PBS, surfactant HS15, tryptone soy agar medium (TSA), Sabouraud dextrose agar medium (SDA).

[0101] 4.1.3 Consumables and instruments: Inoculating loop, culture dish, biochemical incubator, inverted microscope, laminar flow hood.

[0102] 4.2 Test methods

[0103] 4.2.1 Preparation of test bacterial solutions

[0104] Preparation of mixed bacterial solutions of Staphylococcus aureus and Escherichia coli: Take an appropriate amount of Staphylococcus aureus and Escherichia coli cells from the preserved test tube slants (or bacterial strain preservation tubes) and inoculate them on tryptone soy agar (TSA) slants, and incubate at 37 °C for 18 h - 24 h. Use an inoculating loop to pick the first-generation culture, streak it on a TSA plate, and incubate at 37 °C for 18 h - 24 h. Pick the typical colonies from the second-generation culture above, streak them on a TSA agar slant, and incubate at 37 °C for 18 h - 24 h to obtain the third-generation culture. Pipette an appropriate amount of sterile physiological saline into the slant test tube, blow and wash repeatedly to wash off the bacterial lawn. Transfer the washing solution to another sterile test tube and vortex for 20 s. Dilute it with sterile physiological saline to a bacterial suspension of about 1.0×10 8 cfu / mL. The prepared bacterial suspension should be used within 2 h, or stored at 2 °C - 8 °C for no more than 24 h.

[0105] Preparation of Candida albicans bacterial solution: Take an appropriate amount of Candida albicans cells from the well-preserved test tube slant (or strain preservation tube) and inoculate them on a Sabouraud dextrose agar (SDA) slant, and incubate at 28 °C for 18 h - 24 h. Use an inoculation loop to pick the first-generation culture and streak it on an SDA plate, and incubate at 28 °C for 18 h - 24 h. Pick the typical colonies from the above second-generation culture and streak them on an SDA agar slant, and incubate at 28 °C for 18 h - 24 h to obtain the third-generation culture. Pipette an appropriate amount of sterile PBS buffer into the slant test tube, blow and wash repeatedly to wash off the bacterial lawn. Transfer the washing solution to another sterile test tube and vortex for 20 s. Dilute it with PBS buffer to a bacterial suspension of about 1.0×10 7 cfu / mL. The prepared bacterial suspension should be used within 2 h or stored at 2 °C - 8 °C for no more than 24 h.

[0106] 4.2.2 Preparation of the mixed solution of compound essential oil 4 included in the compound preservative system

[0107] Use the mixed solution of 50 mg / ml compound essential oil 4 as the mother liquor of the compound essential oil added in the compound preservative system. Preparation of the mixed solution of 50 mg / ml compound essential oil 4: Add 50 mg of compound essential oil 4 to 1 ml of a solution containing 2% surfactant HS15 and mix well to obtain the mixed solution of 50 mg / ml compound essential oil 4.

[0108] 4.2.3 Compound preservative system

[0109] Compound formulation 1:

[0110] The preservative system is the mixed solution of compound essential oil 4 (1%) + 1,2 - hexanediol (0.2%) + caprylyl glycol (0.1%). Specific operation: Take 20 ml of tryptic soy agar medium (for the mixed bacterial solution of Staphylococcus aureus and Escherichia coli) / Sabouraud dextrose agar medium (for Candida albicans), add 0.2 ml of the mixed solution of compound essential oil 4 prepared in 4.2.2, 0.04 ml of 1,2 - hexanediol, and 0.02 ml of caprylyl glycol, and mix well.

[0111] Compound formulation 2:

[0112] The preservative system is the mixed solution of compound essential oil 4 (0.3%) + 1,2 - hexanediol (0.2%) + caprylyl glycol (0.1%). Specific operation: Take 20 ml of tryptic soy agar medium (for the mixed bacterial solution of Staphylococcus aureus and Escherichia coli) / Sabouraud dextrose agar medium (for Candida albicans), add 0.06 ml of the mixed solution of compound essential oil 4 prepared in 4.2.2, 0.04 ml of 1,2 - hexanediol, and 0.02 ml of caprylyl glycol, and mix well.

[0113] Compound formulation 3:

[0114] The anti-corrosion system is compound essential oil 4 (0.1%) + 1,2 - hexanediol (0.2%) + caprylyl glycol (0.1%). Specific operation: Take 20 ml of tryptone soya agar medium (for the mixed bacterial solution of Staphylococcus aureus and Escherichia coli) / Sabouraud dextrose agar medium (for Candida albicans), add 0.02 ml of the compound essential oil 4 mixed solution prepared in 4.2.2, 0.04 ml of 1,2 - hexanediol, and 0.02 ml of caprylyl glycol, and mix well.

[0115] 4.2.4 Detection of the anti - corrosion efficacy of the compound anti - corrosion system - anti - corrosion challenge test

[0116] The anti - corrosion challenge test evaluates the anti - corrosion and antibacterial ability of a product scientifically by adding specific microorganisms to the product to simulate the risk of microbial contamination during actual use. The test provides quantitative data showing the growth or reduction trend of microorganisms in the product, thereby accurately evaluating the anti - corrosion effect.

[0117] Detection of the anti - corrosion efficacy of compound formulation 1: Add 0.1 ml of the mixed bacterial solution of Staphylococcus aureus and Escherichia coli to 10 ml of the anti - corrosion system of compound formulation 1, and the final concentration of the bacterial suspension is 10 6 cfu / mL; add 0.1 ml of the Candida albicans bacterial solution to 10 ml of the anti - corrosion system of compound formulation 1, and the final concentration of the bacterial suspension is 10 5 cfu / mL. Place them in an incubator at 37°C (for Staphylococcus aureus and Escherichia coli) and 28°C (for Candida albicans) respectively. After culturing for a specific time (7, 14, 21, 28 days), take 0.1 ml of the sample and perform plate counting in 2 sterile petri dishes.

[0118] The detection steps for the anti - corrosion efficacy of compound formulation 2 and compound formulation 3 are the same as those of compound formulation 1.

[0119] 4. 3 Test results

[0120] 4.3.1 Antibacterial activity of the compound anti - corrosion system containing compound essential oil and polyols against Staphylococcus aureus and Escherichia coli

[0121] The results are shown in Table 4 and Figure 7 . It can be seen from the results that in compound formulation 1, the colony reduction was close to 100% (no bacterial growth) on the 7th day, which can effectively kill Escherichia coli and Staphylococcus aureus, and no colony formation was observed during the 28 - day experiment, indicating a strong antibacterial and anti - corrosion effect; in compound formulation 2 and 3 during the 28 - day experiment, as the culture time extended, the colonies in the petri dishes gradually increased. The above results suggest that compound formulation 1 has obvious advantages over compound formulation 2 and 3 in inhibiting bacterial activity.

[0122] Table 4 Colony Concentrations of Staphylococcus aureus and Escherichia coli

[0123]

[0124] 4.3.2 Antifungal Effect of the Compound Preservative System Containing Compound Essential Oil and Polyol on Candida albicans

[0125] The results are shown in Figure 8 . It can be seen from Figure 8 that in the compounding schemes 1, 2 and 3, the fungal colonies decreased by nearly 90% on the 7th day, which can effectively prevent the growth of Candida albicans in the culture medium, and no colony formation was observed during the 28-day experiment, indicating a strong inhibitory effect on fungi. There was no obvious difference among the compounding schemes 1, 2 and 3 in terms of inhibiting fungal activity.

[0126] 4.4 Experimental Conclusions

[0127] In summary, in this experiment, it was found that the preservative system of compounding scheme 1 had a strong antibacterial effect on Escherichia coli, Staphylococcus aureus and Candida albicans, and the preservative systems of compounding schemes 2 and 3 had a strong antifungal effect on Candida albicans.

[0128]

Example 5

[0129] In this example, the quality of each single essential oil of the present invention was detected by GC-MS to control the quality of the raw materials of the compound essential oil of the present invention.

[0130] 5.1 Preparation of Test Samples

[0131] Transfer 0.5 mL of the essential oil to be tested into a 1.5 mL centrifuge tube and centrifuge for 1 min; take 50 μL of the supernatant, weigh it, add ethyl acetate to prepare an essential oil solution with a concentration of 100 mg / mL; add anhydrous sodium sulfate powder according to a mass ratio of 1:10, mix well, let stand, and centrifuge; take 10 μL of the supernatant and dilute it 100 times with ethyl acetate to obtain a test solution with a concentration of 1 mg / mL; centrifuge the test solution for 1 min, take the supernatant, and detect it by GC-MS.

[0132] 5.2 Detection Method

[0133] Chromatographic column: Agilent HP-5 MS UI (30 m×0.25 mm×0.25 μm)

[0134] Carrier gas: High-purity helium

[0135] Flow rate: 1.0 mL / min

[0136] Injection port temperature: 250 °C

[0137] Split ratio: 10 : 1

[0138] Injection volume: 1 μL

[0139] Temperature programming: 50 °C - 180 °C (3 °C / min) - 300 °C (20 °C / min, hold for 5 min)

[0140] Ion source: EI+

[0141] Ion source temperature: 230 °C

[0142] Scanning mode: Full ion scan

[0143] Scanning range: m / z 40 - 600

[0144] Solvent delay: 4 min

[0145] 5.3 Detection results

[0146] The detection results are as Figures 9 - 12 shown. The original data of GC-MS detection was analyzed by Agilent MassHunter Qualitative Analysis (V B.07.00) software to obtain the information of each component peak (retention time, integrated peak area, percentage of total peak area, and peak height).

Claims

1. A compound essential oil, the compound essential oil has antibacterial effects, and is characterized in that, The compound essential oil is composed of the following components: wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and citral-type camphor leaf essential oil; the volume ratio of the wild chrysanthemum essential oil, jasmine essential oil, patchouli essential oil, and citral-type camphor leaf essential oil is 4:1:2:

3. The wild chrysanthemum essential oil is a volatile oil extracted from the flowers of wild chrysanthemum by steam distillation; the jasmine essential oil is a volatile oil obtained by supercritical carbon dioxide extraction from Chinese jasmine flowers; the patchouli essential oil is a volatile oil extracted from the above-ground parts of patchouli by steam distillation; the citral-type camphor leaf essential oil is a volatile oil extracted by water distillation from the fresh leaves of citral-type camphor trees.

2. An anti-corrosion system, characterized in that, It contains the compound essential oil as described in Claim 1.

3. The anti-corrosion system according to claim 2, wherein The anti-corrosion system further contains 1,2-hexanediol and caprylyl glycol.

4. The anti-corrosion system according to claim 3, characterized in that, The volume ratio of the compound essential oil, 1,2-hexanediol, and caprylyl glycol is (1-10):2:

1.

5. The anticorrosion system according to claim 4, characterized in that, The volume ratio of the compound essential oil, 1,2-hexanediol, and caprylyl glycol is 10:2:

1.

6. Use of the compound essential oil as described in Claim 1 or the anti-corrosion system as described in any one of Claims 2-5 in cosmetics.

7. The use according to claim 6, characterized in that, The content of the anti-corrosion system in cosmetics is 0.05%-2%.

8. The use according to claim 7, wherein, The content of the anti-corrosion system in cosmetics is 0.5%.

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