Essence oil composition for relieving pressure and helping sleep for ears and application thereof

Abstract

The invention discloses a pressure-reducing sleep-aiding essence oil composition for ears and application thereof, and belongs to the technical field of daily care solutions. The essence oil composition takes rose oil, sandalwood oil, melaleuca alternifolia leaf oil, lavender oil and anthemis nobilis flower oil as core active ingredients, ginger root oil, sweet orange peel oil, an antioxidant and a solvent can also be added, and the melaleuca alternifolia leaf oil is prepared through composite bacillus fermentation and supercritical CO2 extraction. The essence oil composition disclosed by the invention has a remarkable and continuous pressure-reducing and sleep-aiding effect, and the effect is gradually enhanced along with the prolonging of the service time.

Description

Technical Field

[0001] This invention belongs to the technical field of daily care solutions, specifically relating to an essential oil composition for ear use that relieves stress and promotes sleep, and its application. Background Technology

[0002] With the fast pace of modern life and increasing work and life pressures, the incidence of sleep disorders and emotional problems such as insomnia, anxiety, and mental tension is rising year by year. Consumers have an increasingly urgent need for safe and effective stress-relieving and sleep-aiding daily care products. However, existing products have many shortcomings and fail to meet actual usage needs: Limited modes of action and sites of action: Most stress-relieving and sleep-aiding products are concentrated in oral or topical formulations. Oral products need to be absorbed through the gastrointestinal tract, which may increase the metabolic burden on the body and have a slower onset of action. Topical products do not make full use of the physiological characteristics of the ear, such as thin skin, rich capillaries, and easy penetration and absorption of ingredients. Highly effective stress-relieving and sleep-aiding products specifically designed for the ear are relatively scarce, and cannot achieve precise local action to enhance efficacy.

[0003] Poor synergy of ingredients: Existing essential oil-based stress-relieving and sleep-aiding products often use a single essential oil (such as only lavender oil) as the active ingredient, lacking a scientifically designed multi-component combination. This makes it difficult to form a multi-dimensional chain of action, resulting in a single efficacy and an inability to simultaneously address the root causes of stress and sleep quality issues, leading to poor overall results.

[0004] Outdated extraction technology: Traditional essential oil extraction often uses steam distillation, which easily destroys heat-sensitive active ingredients in essential oils (such as citronellol in rose oil and santalol in sandalwood oil), reducing the activity of essential oils; moreover, the extraction efficiency is low and the impurity content is high, which directly affects the stress-relieving and sleep-aiding effects and safety of the product, limiting the product's market competitiveness.

[0005] Therefore, developing a stress-relieving and sleep-aiding essential oil composition that is specifically designed for ear applications, has strong synergistic effects, advanced extraction technology, significant efficacy, and is safe has become an urgent problem to be solved in the current field of daily care products, and has important practical application value and market prospects. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a stress-relieving and sleep-aiding essential oil composition that is specifically designed for ear application, has strong synergistic effects of its ingredients, employs advanced extraction technology, and is highly effective and safe.

[0007] To achieve the above objectives, the present invention discloses the following technical solutions: In a first aspect, the present invention provides an essential oil composition for ear use that reduces stress and promotes sleep, the essential oil composition comprising rose oil, sandalwood oil, Melaleuca alternifolia leaf oil, lavender oil and chamomile flower oil.

[0008] Preferably, the essential oil composition further comprises ginger root oil, sweet orange peel oil, antioxidants, and solvents.

[0009] More preferably, the solvent is at least one of jojoba seed oil, sweet almond oil, and sunflower seed oil.

[0010] Preferably, the essential oil composition contains the following components in parts by weight: 11-15 parts rose oil; 5-10 parts sandalwood oil; 1-2 parts of Melaleuca alternifolia leaf oil; 3-5 parts lavender oil; 0.6-1 part of white chrysanthemum oil.

[0011] More preferably, the method for preparing the Melaleuca alternifolia leaf oil includes the following steps: Step 1.1. Take dried Melaleuca alternifolia leaves, crush and sieve them to obtain Melaleuca alternifolia leaf powder. Then mix the powder with sterile water at a ratio of 1 kg: 1 L to obtain a fermentation substrate, and sterilize it for later use. Step 1.2. Count 10 live bacteria 7-9 Bacillus subtilis culture with CFU / mL and a viable count of 10 7-9 A compound bacterial solution was obtained by mixing CFU / mL of Bacillus licheniformis bacterial suspension; Step 1.3. Ferment and culture the compound bacterial solution with the fermentation substrate for 72-84 hours to obtain the fermented material; Step 1.4. The fermented material is subjected to supercritical CO2 extraction for 3-4 hours to obtain the extract, which is then concentrated under reduced pressure to obtain Melaleuca alternifolia leaf oil.

[0012] More preferably, in step 2, the number of live bacteria is 10. 7-9 Bacillus subtilis culture with CFU / mL and a viable count of 10 7-9 CFU / mL Bacillus licheniformis bacterial suspension was mixed at a volume ratio of (1-4):1.

[0013] More preferably, the fermentation conditions in step 3 are as follows: the amount of compound bacterial solution added is 5-10% of the weight of the fermentation substrate, and the fermentation is carried out at 35-37℃, rotation speed 10-50 r / min, and aeration rate of 0.2-0.3 L per kilogram of fermentation substrate for 72-84 hours.

[0014] More preferably, the parameters for supercritical CO2 extraction in step 4 are: extraction pressure 28-30 MPa, temperature 45-50℃, CO2 flow rate 0.3-0.4 L / min, with anhydrous ethanol as an entrainer at a flow rate of 0.06-0.07 L / min, and extraction time 3-4 h.

[0015] More preferably, the method for preparing the sandalwood oil includes the following steps: Step 2.1. Grind the dried sandalwood and pass it through a 60-mesh sieve to obtain sandalwood powder; Step 2.2. Place sandalwood powder in a supercritical CO2 extraction vessel and adjust the extraction vessel parameters as follows: extraction pressure 23-26 MPa, extraction temperature 50-60℃, CO2 flow rate 0.4-0.5 L / min, and extraction time 3-4 h to obtain sandalwood oil.

[0016] Secondly, the present invention provides the application of the essential oil composition described in the first aspect in the preparation of human care preparations with soothing and stress-relieving effects and sleep-promoting effects.

[0017] In this invention: Rose oil is rich in active ingredients such as citronellol and geraniol, which have both mood-regulating and soothing effects. It can calm anxiety and relieve mental tension, laying a good emotional foundation for falling asleep, and its natural aroma can improve low mood and reduce stress-induced irritability, thus synergistically enhancing the overall stress-reduction effect.

[0018] Sandalwood oil contains components such as α-santalol and β-santalol, which have a prominent sedative effect on the central nervous system. It can slowly release soothing active ingredients, prolong the duration of relaxation, regulate sleep rhythms, and reduce nighttime awakenings. At the same time, its rich aroma can enhance emotional stability and further relieve long-term stress.

[0019] The Melaleuca alternifolia leaf oil prepared by this invention through compound fermentation and supercritical CO2 extraction process fully retains active ingredients such as eucalyptol and α-terpineol. It not only has antibacterial and anti-inflammatory effects, but also improves the skin penetration and absorption efficiency of other essential oils, accelerates the stress relief process, and enhances the overall synergistic effect.

[0020] Lavender oil contains active sleep-aiding components such as linalool and linalyl acetate, which can directly act on the sleep-regulating neural pathways, shorten the sleep latency period, and improve sleep efficiency; its refreshing aroma can also improve the perception of the sleep environment and synergistically enhance the sleep-aiding effect with other essential oils.

[0021] White chrysanthemum flower oil is rich in core active ingredients such as guaiacol lactones, total flavonoids, chlorogenic acid and α-bisabolol, and has excellent anti-inflammatory, repairing, deep calming and gentle soothing effects.

[0022] The beneficial effects of this invention are: 1. The five essential oils provided by this invention—rose oil, sandalwood oil, Melaleuca alternifolia leaf oil, lavender oil, and chamomile flower oil—form a multi-dimensional chain of action: stress reduction, sleep aid, and balance. The absence of any one of these oils leads to a significant decrease in efficacy. Experiments show that composition 4, lacking rose oil, and composition 5, lacking sandalwood oil, resulted in a sleep incidence rate of only 34.29%-38.57% in mice, far lower than the 75.71% of the complete formulations 2 and 3, fully demonstrating the necessity of their synergistic effect.

[0023] 2. The Melaleuca alternifolia leaf oil obtained by the compound fermentation and supercritical CO2 extraction process in this invention reduces raw material impurities and retains heat-sensitive components. Experiments have shown that its activity is superior to direct extraction and enzymatic extraction. The supercritical CO2 extraction of sandalwood oil also improves the yield and purity of santalol compared with the traditional distillation method. Detailed Implementation

[0024] To better illustrate the objectives, technical solutions, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0025] Unless otherwise specified, the test methods used in the examples and comparative examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified; and the percentages mentioned in the examples and comparative examples are mass percentages unless otherwise specified.

[0026] In this invention: Rose oil: Rose (ROSA RUGOSA) flower oil is a vegetable oil obtained by distillation of rose petals and is commercially available. Sandalwood: The heartwood of the sandalwood tree (Santalum album L.) belonging to the genus Santalum of the family Santalaceae; commercially available. Lavender oil: Lavender (LAVANDULA ANGUSTIFOLIA) oil is a plant oil obtained by distillation of lavender and is commercially available. White chamomile flower oil: White chamomile (ANTHEMIS NOBILIS) flower oil is a vegetable oil obtained by distillation of the petals of white chamomile, and is commercially available. Alternate-leaved Melaleuca leaves: Leaves of the plant Melaleuca alternifolia genus in the Myrtaceae family, commercially available; Bacillus subtilis: accession number GDMCC NO.1.21, purchased from Guangdong Provincial Microbial Culture Collection Center; Bacillus licheniformis: accession number GDMCC NO.65046, purchased from Guangdong Provincial Microbial Culture Collection Center.

[0027] Preparation of sandalwood oil Step 1. Place sandalwood in a drying oven and dry it to constant weight. Then, use a high-speed pulverizer to pulverize it and pass it through a 60-mesh sieve to obtain sandalwood powder. Step 2. Place the sandalwood powder in a supercritical CO2 extraction vessel and adjust the extraction vessel parameters as follows: extraction pressure is 24 MPa, extraction temperature is 55℃, CO2 flow rate is 0.45 L / min·kg, and extraction time is 4 h. The CO2 fluid is separated by a separator through a pressure reducing valve to obtain sandalwood oil.

[0028] Preparation of Melaleuca alternifolia leaf oil Alternate leaf Melaleuca alternifolia leaf oil ① Step 1-1. Take the leaves of Melaleuca alternifolia and dry them in a drying oven until constant weight. Then, use a high-speed pulverizer to pulverize them and pass them through a 60-mesh sieve to obtain Melaleuca alternifolia leaf powder. Mix the powder with sterile water at a ratio of 1 kg: 1 L to obtain a fermentation substrate, and sterilize it for later use. Steps 1-2. Activate Bacillus subtilis and Bacillus licheniformis separately to obtain a viable count of 1×10⁻⁶. 8 Bacillus subtilis culture with CFU / mL and a viable count of 1×10⁻⁶ 8 A CFU / mL Bacillus licheniformis bacterial solution was prepared by mixing Bacillus subtilis and Bacillus licheniformis bacterial solutions at a volume ratio of 3:1 to obtain a composite bacterial solution. Steps 1-3. Mix the compound bacterial solution with the fermentation substrate in a solid fermentation tank. The amount of compound bacterial solution added is 7% of the weight of the fermentation substrate. Ferment and culture for 84 hours at 37℃, rotation speed of 15 r / min, and aeration rate of 0.2-0.3 L per kilogram of fermentation substrate to obtain fermented material. Steps 1-4. The fermentation material is loaded into the extraction vessel, and the supercritical CO2 extraction conditions are set: extraction pressure 28 MPa, temperature 45℃, CO2 flow rate 0.34 L / min·kg. Anhydrous ethanol is used as an entrainer with an entrainer flow rate of 0.06 L / min·kg. The extraction system is started for dynamic extraction for 4 hours. The CO2 fluid is separated by a separator to obtain the extract containing the entrainer. The entrainer is removed by vacuum concentration to obtain Melaleuca alternifolia leaf oil ①.

[0029] Alternate leaf Melaleuca alternifolia leaf oil ② Step 2-1. Place the leaves of Melaleuca alternifolia in a drying oven and dry them to constant weight. Then, pulverize them with a high-speed pulverizer and pass them through a 60-mesh sieve to obtain Melaleuca alternifolia leaf powder. Step 2-2. The powdered leaves of Melaleuca alternifolia were loaded into the extraction vessel. The supercritical CO2 extraction conditions were set as follows: extraction pressure 28 MPa, temperature 45℃, CO2 flow rate 0.34 L / min, and anhydrous ethanol was used as an entrainer with an entrainer flow rate of 0.06 L / min. The extraction system was started for dynamic extraction for 4 hours. The CO2 fluid was separated by a separator to obtain the extract containing the entrainer. The entrainer was removed by vacuum concentration to obtain Melaleuca alternifolia leaf oil ②.

[0030] Alternate leaf Melaleuca alternifolia leaf oil ③ Step 3-1. Place the leaves of Melaleuca alternifolia in a drying oven and dry them to constant weight. Then, pulverize them with a high-speed pulverizer and pass them through a 60-mesh sieve to obtain Melaleuca alternifolia leaf powder. Step 3-2. Mix the powdered Melaleuca alternifolia leaves with sterile water at a ratio of 1 kg: 1 L until homogeneous. Then add 10% of the compound enzyme preparation by weight of the total system. The compound enzyme preparation contains protease with an activity of 10%. 4 U / g, cellulase activity 5×10 4 U / g, enzymatic hydrolysis for 12h to obtain enzymatic hydrolysate; Step 3-3. Load the enzymatic hydrolysate into the extraction vessel, set the supercritical CO2 extraction conditions to 28 MPa, 45℃, and 0.34 L / min for CO2 extraction. Simultaneously, use anhydrous ethanol as an entrainer with a flow rate of 0.06 L / min. Start the extraction system and perform dynamic extraction for 4 hours. Separate the CO2 fluid through a separator to obtain the extract containing the entrainer. Remove the entrainer by vacuum concentration to obtain Melaleuca alternifolia leaf oil ③.

[0031] essential oil composition The raw materials were accurately weighed according to the percentage of mass in Table 1, and after compounding and mixing, the essential oil composition was obtained. Compositions 1-3 constitute a complete formulation group; Compositions 4-8 are the default control group: Composition 4 differs from Composition 2 in that it lacks rose oil. The missing amount is made up by the proportions of remaining sandalwood oil, Melaleuca alternifolia leaf①, lavender oil, and white chrysanthemum oil. Composition 5 differs from Composition 2 in that it lacks sandalwood oil. The missing amount is made up by the proportions of remaining rose oil, Melaleuca alternifolia leaf①, lavender oil, and chrysanthemum oil. Composition 6 differs from Composition 2 in that it lacks Melaleuca alternifolia leaves①, and the missing amount is made up by the remaining sandalwood oil, rose oil, lavender oil and white chrysanthemum oil. Composition 7 differs from Composition 2 in that it lacks lavender oil. The missing amount is made up by the proportion of remaining sandalwood oil, Melaleuca alternifolia leaf①, rose oil, and chrysanthemum oil. Composition 8 differs from Composition 2 in that it lacks white chrysanthemum oil. The missing amount is made up by the proportions of remaining sandalwood oil, Melaleuca alternifolia leaf①, lavender oil, and rose oil. Compositions 9-10 serve as a control group for replacing raw materials.

[0032] Table 1. Raw materials for essential oil compositions (unit: %)

[0033] Note: " / " in the table indicates no addition.

[0034] Performance testing The sleep-aiding and stress-relieving effects of the essential oil compositions prepared by compositions 1-10 were tested.

[0035] I. Animal Experiment Data The experimental animals were ICR-grade male BALB / C mice, weighing 20-22g. The experimental environment was 25℃ and 50% humidity. The mice had free access to food and water and were allowed to acclimatize for 5 days before the experiment. The formal experiment began after the mice passed the re-examination.

[0036] 1. Sleep aid effect Referring to the "Implementation Manual of Technical Specifications for Inspection and Evaluation of Health Foods (2003 Edition)," a mouse experiment induced by sodium pentobarbital was conducted to analyze the effect of the essential oil compositions prepared by compositions 1-10 on the incidence of sleep in mice. Sleep was defined as the disappearance of the righting reflex; when mice were placed in a supine position, they could immediately turn over to their right position. If they could not turn over for more than 30-60 seconds, the righting reflex was considered to have disappeared, and the mice had entered sleep.

[0037] Sleep incidence rate = (Number of mice falling asleep / Total number of mice) × 100% Mice were randomly divided into 11 groups of 10 each: a blank group (n=10) and an experimental group (n=100). In the experimental group, 1 mL of the essential oil composition prepared by composition 1-10 was evenly applied to the belly of the corresponding group of mice. In the blank group, 1 mL of a matrix solution composed of 99.8% jojoba seed oil and 0.2% tocopheryl acetate was applied to each mouse. The treatment was carried out once a day for 7 consecutive days. The results are shown in Table 2.

[0038] Table 2

[0039] Results analysis: According to the test results, the number of mice falling asleep within 30 minutes in the blank group remained stable at 1-2 within 7 days, with a sleep incidence rate of only 17.14%. In contrast, the sleep incidence rate of all experimental groups was higher than that of the blank group, reaching as high as 75.71%, proving that the essential oil composition of the present invention has a significant sleep-aiding effect. Combinations 1, 2, and 3, containing the five core essential oils, showed the best sleep-aiding effect. Over seven days, the number of mice falling asleep within 30 minutes remained between 6 and 9, with a sleep incidence rate of 68.57%-75.71%. In contrast, compositions 4-8, lacking any one of the core essential oils, resulted in only 2-5 mice falling asleep within 30 minutes most of the time, with a sleep incidence rate dropping to 27.14%-38.57%. This clearly demonstrates that the synergistic effect of the five core essential oils is crucial for ensuring the sleep-aiding effect, and the absence of any one component leads to a significant reduction in efficacy.

[0040] Composition 2, prepared by combining Bacillus fermentation with supercritical CO2 extraction, showed significantly better sleep-aiding effects than composition 9, which used oil ② obtained by direct supercritical extraction, and composition 10, which used oil ③ obtained by enzymatic hydrolysis extraction. This demonstrates that the fermentation process can effectively enhance the activity of Melaleuca alternifolia leaf oil, thereby strengthening the overall sleep-aiding effect of the essential oil composition.

[0041] 2. Stress Reduction and Tranquility Test 1) Anxiety modeling To establish a chronic stress model, mice were restrained, and several small holes were drilled in a 50mL centrifuge tube. The mice were then inserted into the centrifuge tube, ensuring unobstructed breathing, for 2 hours at a time for 15 days to obtain an anxiety model mouse.

[0042] 2) Experimental Grouping The mice were divided into two groups: a blank control group (n=10) and an experimental group (n=100). The blank control group was coated with 1 mL of a matrix solution composed of 99.8% jojoba seed oil and 0.2% tocopheryl acetate per mouse. The experimental groups 1-10 were coated with 1 mL of the essential oil composition prepared by composition 1-10 per mouse.

[0043] 3) Experimental methods The box with light and shadow (length x width x height: 50cm x 25cm x 30cm) has the same volume as the box with light (each occupying 1 / 2 of the volume).

[0044] The inner wall of the enclosure is white, and a white light source is installed on the top. The illuminance inside the enclosure is adjusted to approximately 200 Lux.

[0045] The inner walls of the darkroom are black, completely blocking out light, with a red bulb at the top serving as a weak light source (for the experimenter to observe).

[0046] The two boxes are separated by a partition with a square passage (6cm x 6cm) at the bottom center of the partition for the mice to move freely. The bottom of the boxes is divided into several squares, and the mice's activities are automatically recorded by video analysis software.

[0047] Mice in each group had the appropriate drug or base solution evenly applied to the shaved area on their abdomen, and were placed back in their cages to rest for 30 minutes. The mice were then gently placed in the center of a brightly lit cage, facing the partition passage. The video recording system was immediately activated to observe and record the time the mice spent in the cage within 5 minutes. This time spent was used as an anxiety indicator; the longer the mice stayed in the cage, the lower their anxiety level.

[0048] After each mouse has been tested, the inside of the test chamber is thoroughly cleaned with 75% alcohol to prevent the odor of the previous mouse from interfering with the next one.

[0049] 4) Statistical analysis: Data are expressed as mean ± standard deviation. A p < 0.05 is considered statistically significant.

[0050] The results are shown in Table 3.

[0051] Table 3 Group Dwell time / s Blank group 48.50±13.32 Composition 1 153.00±15.21 Composition 2 167.90±12.83 Composition 3 173.20±14.44 Composition 4 96.70±17.26*** Composition 5 88.90±21.09*** Composition 6 89.90±14.49*** Composition 7 87.70±15.26*** Composition 8 93.70±17.23*** Composition 9 112.40±14.80*** Composition 10 121.10±18.06*** Note: In the table, "*" indicates a significant difference compared to composition 2, 0.01≤P<0.05; ** indicates 0.001≤P<0.01; *** indicates P<0.001.

[0052] Results analysis: The results showed that the blank group (with the applied matrix solution) of anxiety model mice stayed in the open box for only 48.50 seconds, indicating that the mice were in a state of obvious anxiety and the model was successfully established. In contrast, the experimental group mice stayed in the open box for a significantly longer time, especially the mice in compositions 1-3, which stayed for 153-173 seconds. This proves that the essential oil composition of the present invention can effectively relieve anxiety in mice and has a clear stress-relieving and calming effect.

[0053] II. Subjective Evaluation Test of Human Body To test the stress relief and sleep improvement, 330 individuals experiencing significant stress were randomly divided into 11 groups of 30 each: a matrix group and an experimental group. The matrix group used a matrix solution composed of 99.8% jojoba seed oil and 0.2% tocopheryl acetate, while the experimental groups used essential oil compositions prepared according to compositions 1-10. Before bedtime each day, participants applied 0.5g of the test substance to both auricles and gently massaged until a slight warming sensation was felt. This was continued for 30 days. Follow-up visits were conducted at 15 and 30 days to assess whether stress relief and sleep improvement were observed. The statistical results are shown in Table 4.

[0054] Table 4 Results of stress relief and sleep improvement tests

[0055] Results analysis: The results show that, compared to the matrix group, after using the essential oils of compositions 1-3, 8-10 people experienced stress relief and 12-16 people experienced sleep improvement within 15 days. Within 30 days, the number of people with stress relief increased to 18-24 and the number of people with sleep improvement increased to 19-25. This fully demonstrates that the essential oil compositions of the present invention have significant and continuous stress-relieving and sleep-aiding effects in the human body, and the effects gradually increase with the extension of the usage time.

[0056] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An essential oil composition for ear application to relieve stress and promote sleep, characterized in that, The essential oil composition contains rose oil, sandalwood oil, Melaleuca alternifolia leaf oil, lavender oil, and chamomile flower oil.

2. The essential oil composition according to claim 1, characterized in that, The essential oil composition also includes ginger root oil, sweet orange peel oil, antioxidants, and solvents.

3. The essential oil composition according to claim 2, characterized in that, The solvent is at least one of jojoba seed oil, sweet almond oil, and sunflower seed oil.

4. The essential oil composition according to claim 1, characterized in that, The essential oil composition contains the following components in parts by weight: 11-15 parts rose oil; 5-10 parts sandalwood oil; 1-2 parts of Melaleuca alternifolia leaf oil; 3-5 parts lavender oil; White chrysanthemum oil (0.6-1 part).

5. The essential oil composition according to claim 4, characterized in that, The preparation method of the Melaleuca alternifolia leaf oil includes the following steps: Step 1.

1. Take dried Melaleuca alternifolia leaves, crush and sieve them to obtain Melaleuca alternifolia leaf powder. Then mix the powder with sterile water at a ratio of 1 kg: 1 L to obtain a fermentation substrate, and sterilize it for later use. Step 1.

2. Count 10 live bacteria 7-9 Bacillus subtilis culture with CFU / mL and a viable count of 10 7-9 A compound bacterial solution was obtained by mixing CFU / mL of Bacillus licheniformis bacterial suspension; Step 1.

3. Ferment and culture the compound bacterial solution with the fermentation substrate for 72-84 hours to obtain the fermented material; Step 1.

4. The fermented material is subjected to supercritical CO2 extraction for 3-4 hours to obtain the extract, which is then concentrated under reduced pressure to obtain Melaleuca alternifolia leaf oil.

6. The essential oil composition according to claim 5, characterized in that, In step 2, the viable bacteria count is 10. 7-9 Bacillus subtilis culture with CFU / mL and a viable count of 10 7-9 CFU / mL Bacillus licheniformis bacterial suspension was mixed at a volume ratio of (1-4):

1.

7. The essential oil composition according to claim 5, characterized in that, The fermentation conditions in step 3 are as follows: the amount of compound bacterial solution added is 5-10% of the weight of the fermentation substrate, and the fermentation is carried out at 35-37℃, rotation speed 10-50 r / min, and aeration rate of 0.2-0.3 L per kilogram of fermentation substrate for 72-84 hours.

8. The essential oil composition according to claim 5, characterized in that, In step 4, the parameters for supercritical CO2 extraction are: extraction pressure 28-30 MPa, temperature 45-50℃, CO2 flow rate 0.3-0.4 L / min·kg, with anhydrous ethanol as an entrainer at a flow rate of 0.06-0.07 L / min·kg, and extraction time 3-4 h.

9. The essential oil composition according to claim 4, characterized in that, The method for preparing the sandalwood oil includes the following steps: Step 2.

1. Grind the dried sandalwood and pass it through a 60-mesh sieve to obtain sandalwood powder; Step 2.

2. Place sandalwood powder in a supercritical CO2 extraction vessel and adjust the extraction vessel parameters as follows: extraction pressure 23-26 MPa, extraction temperature 50-60℃, CO2 flow rate 0.4-0.5 L / min·kg, and extraction time 3-4 h to obtain sandalwood oil.

10. The use of the essential oil composition according to any one of claims 1-9 in the preparation of a human care preparation having the effects of soothing stress reduction and promoting sleep.

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