A traditional Chinese medicine aromatherapy for relieving anxiety and a preparation method thereof

Abstract

The application belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine aromatherapy for relieving anxiety and a preparation method thereof. The traditional Chinese medicine aromatherapy is prepared from Bupleurum chinense, Radix Paeoniae Alba, Fructus Gardeniae, Cortex Moutan, Albizia julibrissin, Fructus Lycii, Semen Ziziphi Spinosae, Poria cocos, Radix Glycyrrhizae and Fructus Jujubae as raw materials, inoculating them with Lactococcus lactis, Lactobacillus gasseri and Lactobacillus pentosus after sterilization with deionized water, and then performing anaerobic fermentation. The traditional Chinese medicine fermentation composition prepared has excellent effect of relieving anxiety. The traditional Chinese medicine fermentation composition can be applied to the preparation of the traditional Chinese medicine aromatherapy with the effect of relieving anxiety, and can solve the problem of poor effect of relieving anxiety of the existing products. The preparation method of the traditional Chinese medicine aromatherapy is simple in operation, suitable for actual industrial production, and has good clinical application value.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine aromatherapy for relieving anxiety and its preparation method. Background Technology

[0002] Anxiety disorder is a mental illness characterized by anxiety as its core symptom, belonging to the category of psychotic disorders. Its main features include persistent and widespread anxiety or recurrent episodes of panic, often accompanied by a range of autonomic nervous system symptoms and motor tension, such as dizziness, chest tightness, palpitations, shortness of breath, dry mouth, frequent urination, urinary urgency, sweating, and tremors. Currently, anxiety disorders are mainly classified into six types: generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder, obsessive-compulsive disorder, and specific phobias. With rapid societal development and increasing work and study pressures, the incidence of anxiety disorders continues to rise, becoming a hot research area in neuroscience.

[0003] Currently, methods for alleviating daily anxiety mainly include psychological interventions (such as meditation and mindfulness training), physical therapies (such as aromatherapy and music therapy), and some plant-based products (such as herbal teas and essential oils). Among these, aromatherapy has gradually become a popular mood regulation method due to its ease of use, high acceptance, and non-invasiveness. However, most commercially available aromatherapy products are single plant essential oils or simple blends, which have problems such as unclear active ingredients, unclear mechanisms of action, and unstable anxiety-relieving effects. In addition, some products add synthetic fragrances or preservatives to pursue long-lasting aroma, which may cause allergies or respiratory irritation, making it difficult to balance safety and effectiveness.

[0004] Furthermore, although traditional Chinese medicine (TCM) has a long history of application in mood regulation, existing TCM compositions are mostly used for oral treatment of anxiety disorders. There has been no systematic research on applying TCM compound formulas to aromatherapy to achieve mood relief through non-inhalation methods. Therefore, developing a TCM compound formula with a clear effect on relieving anxiety, high safety, and suitability for daily aromatherapy use has become an urgent technical problem to be solved in this field. Summary of the Invention

[0005] In view of the above-mentioned defects or improvement needs of the existing technology, the present invention provides a traditional Chinese medicine aromatherapy for relieving anxiety and its preparation method, which solves the problems of poor efficacy and side effects of existing products.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] In a first aspect, the present invention provides a method for preparing a traditional Chinese medicine fermentation composition for relieving anxiety, comprising the following steps:

[0008] A1. Weigh out each Chinese herbal fermentation raw material according to the weight parts, mix them together, add deionized water, mix evenly, sterilize, and obtain a mixture;

[0009] A2. Inoculate the mixture with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus for anaerobic fermentation to obtain the fermentation product;

[0010] A3. Sterilize and filter the fermentation product, and concentrate the filtrate under reduced pressure to obtain the traditional Chinese medicine fermentation composition.

[0011] The fermented raw materials of traditional Chinese medicine contain the following components by weight: Bupleurum chinense 9-12 parts, Paeonia lactiflora 8-10 parts, Gardenia jasminoides 8-10 parts, Paeonia suffruticosa 7-9 parts, Albizia julibrissin 6-8 parts, Lycium barbarum 7-9 parts, Ziziphus jujuba var. spinosa 10-15 parts, Poria cocos 10-15 parts, Nardostachys jatamansi 6-8 parts, Glycyrrhiza uralensis 4-6 parts, and Ziziphus jujuba 4-6 parts.

[0012] In this invention, Bupleurum has a pungent and bitter taste and enters the liver, gallbladder and lung meridians. It has the effects of harmonizing and reducing fever, soothing the liver and relieving depression, and raising yang qi.

[0013] White peony root is bitter and sour in taste, and enters the liver and spleen meridians. It has the effects of nourishing blood and regulating menstruation, soothing the liver and relieving pain.

[0014] Gardenia has a bitter taste and enters the heart, lung, and triple burner meridians. It has the effects of purging fire and relieving irritability, clearing heat and promoting diuresis, cooling blood and detoxifying.

[0015] Peony bark is bitter and pungent in taste, and enters the heart, liver, and kidney meridians. It has the effects of clearing heat and cooling blood, and promoting blood circulation and removing blood stasis.

[0016] The flowers of the mimosa tree are sweet in taste and enter the heart and liver meridians. They have the effects of relieving depression, calming the mind, promoting blood circulation, and reducing swelling.

[0017] Goji berries are sweet in taste and enter the liver and kidney meridians. They have the effects of nourishing the liver and kidneys, and benefiting the essence and improving eyesight.

[0018] Sour jujube seed is sweet and sour in taste, and enters the heart, liver, and gallbladder meridians. It has the effects of nourishing the heart and calming the mind, astringing sweat and promoting the production of body fluids.

[0019] Poria cocos is sweet and bland in taste, and enters the heart, spleen, and kidney meridians. It has the effects of promoting diuresis and eliminating dampness, strengthening the spleen and calming the mind.

[0020] Nardostachys jatamansi is pungent and sweet in nature, and enters the spleen and stomach meridians. It has the effects of promoting qi circulation, relieving pain, and invigorating the spleen.

[0021] Licorice has a sweet taste and enters the heart, lung, spleen, and stomach meridians. It has the effects of tonifying the spleen and replenishing qi, clearing heat and detoxifying.

[0022] Jujubes are sweet in taste and enter the spleen and stomach meridians. They have the effects of tonifying the middle energizer, replenishing qi, nourishing blood, and calming the mind.

[0023] Preferably, the Lactococcus gasseri has the accession number GDMCC NO.1.965, the Lactobacillus gasseri has the accession number CGMCC NO.1.3396, and the Lactobacillus pentosus has the accession number GDMCC NO.1.524.

[0024] Preferably, the fermented raw materials of traditional Chinese medicine contain the following components by weight: 11 parts Bupleurum chinense, 9 parts Paeonia lactiflora, 9 parts Gardenia jasminoides, 8 parts Paeonia suffruticosa, 7 parts Albizia julibrissin, 8 parts Lycium barbarum, 13 parts Ziziphus jujuba var. spinosa, 13 parts Poria cocos, 7 parts Nardostachys jatamansi, 5 parts Glycyrrhiza uralensis, and 5 parts Ziziphus jujuba.

[0025] Preferably, in step A1, the ratio of the fermentation raw material of traditional Chinese medicine to deionized water is 1g:8-12mL.

[0026] Preferably, in step A2, the total inoculum amount of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 3-6% v / v, and the total viable count of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.0 × 10⁻⁶. 9 -5.0×10 9 CFU / mL.

[0027] Preferably, in step A2, the ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.5-2:1:1.

[0028] More preferably, in step A2, the ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.7:1:1.

[0029] Preferably, the fermentation temperature in step A2 is 32-37℃ and the fermentation time is 36-48h.

[0030] Preferably, the sterilization temperature in step A3 is 115-130℃ and the sterilization time is 15-25 min.

[0031] Preferably, the filtration in step A3 is performed using a diatomaceous earth filter.

[0032] Preferably, the vacuum concentration in step A3 is performed using a rotary evaporator to concentrate to 30-40% of the original volume, and the vacuum concentration temperature is 55-65℃.

[0033] Secondly, the present invention provides the application of the traditional Chinese medicine fermentation composition prepared by the preparation method described in the first aspect in the preparation of traditional Chinese medicine aromatherapy products with anxiety-relieving effects.

[0034] Preferably, the dosage form of the traditional Chinese medicine aromatherapy product is liquid, gel, emulsion or solid.

[0035] Thirdly, the present invention provides an aromatherapy liquid comprising the following raw materials in parts by weight: 9-13 parts of the fermented Chinese medicine composition prepared by the preparation method described in the first aspect, 5-8 parts of plant extract, 25-35 parts of 3-methoxy-3-methylbutanol, 2-4 parts of polyacrylamide, 4-9 parts of glycerol, 4-7 parts of silicone oil, and 0.3-0.7 parts of phenoxyethanol.

[0036] The plant extract is selected from at least one of bergamot extract, ylang-ylang extract, jasmine extract, orange blossom extract, or patchouli extract.

[0037] Fourthly, the present invention provides a method for preparing the aromatherapy liquid described in the third aspect, comprising the following steps:

[0038] S1. Stir 3-methoxy-3-methylbutanol, polyacrylamide, glycerol, and silicone oil at 200-300 rpm for 12-17 min to obtain mixture I;

[0039] S2. Add the traditional Chinese medicine fermentation composition, plant extract and phenoxyethanol to mixture I, and stir at 200-300 rpm for 10-15 min to obtain the aromatherapy liquid.

[0040] The beneficial effects of this invention are:

[0041] The traditional Chinese medicine fermentation composition for relieving anxiety in this invention uses Bupleurum chinense, Paeonia lactiflora, Gardenia jasminoides, Paeonia suffruticosa, Albizia julibrissin, Lycium barbarum, Ziziphus jujuba var. spinosa, Poria cocos, Nardostachys jatamansi, Glycyrrhiza uralensis, and Ziziphus jujuba as raw materials. These materials are prepared by anaerobic fermentation after being mixed with deionized water, sterilized, and inoculated with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus. During fermentation, Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus promote each other, helping to release the active ingredients in the traditional Chinese medicine. This results in a traditional Chinese medicine fermentation composition with excellent anxiety-relieving effects. Applying this traditional Chinese medicine fermentation composition to the preparation of traditional Chinese medicine aromatherapy with anxiety-relieving effects can solve the problem of poor efficacy of existing products. Furthermore, the preparation method of this traditional Chinese medicine aromatherapy is simple to operate, suitable for actual industrial production, and has good clinical application value. Detailed Implementation

[0042] To further illustrate the technical means and effects of the present invention in achieving the intended purpose, the following detailed description of the specific embodiments, structure, features and effects of the present invention, in conjunction with the composition, is provided below.

[0043] For the following compositions, unless otherwise specified, experimental methods are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise specified, all materials and reagents used are commercially available.

[0044] Some of the raw materials and their sources are as follows:

[0045] Bupleurum was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.;

[0046] White peony root was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.;

[0047] Gardenia was purchased from Bozhou Huajunzi Biotechnology Co., Ltd.;

[0048] The peony bark was purchased from Bozhou Huajunzi Biotechnology Co., Ltd.

[0049] The mimosa flowers were purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.

[0050] The goji berries were purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.

[0051] The jujube seed was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.

[0052] Poria cocos was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.

[0053] Gan Song was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.;

[0054] The licorice was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd.

[0055] The jujubes were purchased from Bozhou Huajunzi Biotechnology Co., Ltd.

[0056] Example 1

[0057] A method for preparing a traditional Chinese medicine fermentation composition for relieving anxiety includes the following steps:

[0058] A1. Weigh out each Chinese herbal fermentation raw material according to the weight parts, mix them together, add deionized water, mix evenly, sterilize, and obtain a mixture;

[0059] A2. Inoculate the mixture with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus for anaerobic fermentation to obtain the fermentation product;

[0060] A3. Sterilize and filter the fermentation product, and concentrate the filtrate under reduced pressure to obtain the traditional Chinese medicine fermentation composition.

[0061] The fermented raw materials of traditional Chinese medicine contain the following components by weight: Bupleurum chinense 11 parts, Paeonia lactiflora 9 parts, Gardenia jasminoides 9 parts, Paeonia suffruticosa 8 parts, Albizia julibrissin 7 parts, Lycium barbarum 8 parts, Ziziphus jujuba var. spinosa 13 parts, Poria cocos 13 parts, Nardostachys jatamansi 7 parts, Glycyrrhiza uralensis 5 parts, and Ziziphus jujuba 5 parts.

[0062] The preservation number of Lactococcus gasseri is GDMCC NO.1.965, the preservation number of Lactobacillus gasseri is CGMCC NO.1.3396, and the preservation number of Lactobacillus pentosus is GDMCC NO.1.524.

[0063] In step A1, the ratio of the fermentation raw material of traditional Chinese medicine to deionized water is 1g:10mL.

[0064] In step A2, the total inoculation amount of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 5% v / v, and the total viable count of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 3.0 × 10⁻⁶. 9 CFU / mL, wherein the live count ratio of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.7:1:1;

[0065] The fermentation temperature in step A2 is 35℃, and the fermentation time is 42h.

[0066] The sterilization temperature in step A3 is 121°C, and the sterilization time is 20 minutes.

[0067] The filtration in step A3 uses a diatomaceous earth filter.

[0068] In step A3, the vacuum concentration is performed using a rotary evaporator to concentrate the volume to 35% of its original volume, and the vacuum concentration temperature is 60°C.

[0069] Example 2

[0070] A method for preparing a traditional Chinese medicine fermentation composition for relieving anxiety includes the following steps:

[0071] A1. Weigh out each Chinese herbal fermentation raw material according to the weight parts, mix them together, add deionized water, mix evenly, sterilize, and obtain a mixture;

[0072] A2. Inoculate the mixture with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus for anaerobic fermentation to obtain the fermentation product;

[0073] A3. Sterilize and filter the fermentation product, and concentrate the filtrate under reduced pressure to obtain the traditional Chinese medicine fermentation composition.

[0074] The fermented raw materials of traditional Chinese medicine contain the following components by weight: Bupleurum chinense 9 parts, Paeonia lactiflora 8 parts, Gardenia jasminoides 8 parts, Paeonia suffruticosa 7 parts, Albizia julibrissin 6 parts, Lycium barbarum 7 parts, Ziziphus jujuba var. spinosa 10 parts, Poria cocos 10 parts, Nardostachys jatamansi 6 parts, Glycyrrhiza uralensis 4 parts, and Ziziphus jujuba 4 parts.

[0075] The preservation number of Lactococcus gasseri is GDMCC NO.1.965, the preservation number of Lactobacillus gasseri is CGMCC NO.1.3396, and the preservation number of Lactobacillus pentosus is GDMCC NO.1.524.

[0076] In step A1, the ratio of the fermentation raw material of traditional Chinese medicine to deionized water is 1g:12mL.

[0077] In step A2, the total inoculation amount of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 3% v / v, and the total viable count of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 5.0 × 10⁻⁶. 9 CFU / mL, wherein the live count ratio of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.5:1:1;

[0078] The fermentation temperature in step A2 is 37°C, and the fermentation time is 36 hours.

[0079] The sterilization temperature in step A3 is 130℃, and the sterilization time is 15 minutes.

[0080] The filtration in step A3 uses a diatomaceous earth filter.

[0081] In step A3, the vacuum concentration is performed using a rotary evaporator to concentrate the volume to 30% of its original volume, and the vacuum concentration temperature is 65°C.

[0082] Example 3

[0083] A method for preparing a traditional Chinese medicine fermentation composition for relieving anxiety includes the following steps:

[0084] A1. Weigh out each Chinese herbal fermentation raw material according to the weight parts, mix them together, add deionized water, mix evenly, sterilize, and obtain a mixture;

[0085] A2. Inoculate the mixture with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus for anaerobic fermentation to obtain the fermentation product;

[0086] A3. Sterilize and filter the fermentation product, and concentrate the filtrate under reduced pressure to obtain the traditional Chinese medicine fermentation composition.

[0087] The fermented raw materials of traditional Chinese medicine contain the following components by weight: 12 parts Bupleurum, 10 parts White Peony Root, 10 parts Gardenia, 9 parts Moutan Bark, 8 parts Albizia Flower, 9 parts Lycium Fruit, 15 parts Ziziphus Seed, 15 parts Poria, 8 parts Nardostachys, 6 parts Licorice Root, and 6 parts Jujube.

[0088] The preservation number of Lactococcus gasseri is GDMCC NO.1.965, the preservation number of Lactobacillus gasseri is CGMCC NO.1.3396, and the preservation number of Lactobacillus pentosus is GDMCC NO.1.524.

[0089] In step A1, the ratio of the fermentation raw material of traditional Chinese medicine to deionized water is 1g:8mL.

[0090] In step A2, the total inoculation amount of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 6% v / v, and the total viable count of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.0 × 10⁻⁶. 9 CFU / mL, wherein the live count ratio of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 2:1:1;

[0091] The fermentation temperature in step A2 is 32℃, and the fermentation time is 48h.

[0092] The sterilization temperature in step A3 is 115℃, and the sterilization time is 25 minutes.

[0093] The filtration in step A3 uses a diatomaceous earth filter.

[0094] In step A3, the vacuum concentration is performed using a rotary evaporator to concentrate the volume to 40% of its original volume, and the vacuum concentration temperature is 55°C.

[0095] Comparative Example 1

[0096] The preparation method differs from Example 1 in that Lactococcus gasseri is not inoculated in Comparative Example 1, and the total inoculation amount of Lactobacillus gasseri and Lactobacillus pentosus in step A2 is 5% v / v, with a total viable count of 3.0 × 10⁻⁶ cells. 9 The CFU / mL ratio of viable Lactobacillus gasseri to Lactobacillus pentosus was 1:1, and all other steps and parameters were consistent with those in Example 1.

[0097] Comparative Example 2

[0098] The preparation method differs from Example 1 in that Comparative Example 2 does not inoculate with *Lactobacillus gasseri*, and the total inoculation amount of *Lactococcus gasseri* and *Lactobacillus pentosus* in step A2 is 5% v / v, with a total viable count of 3.0 × 10⁻⁶. 9 The CFU / mL ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus was 1.7:1, and all other steps and parameters were consistent with those in Example 1.

[0099] Comparative Example 3

[0100] The preparation method differs from Example 1 in that Comparative Example 3 does not inoculate with *Lactobacillus pentosus*, and the total inoculation amount of *Lactococcus gasseri* and *Lactobacillus gasseri* in step A2 is 5% v / v, with a total viable count of 3.0 × 10⁻⁶ cells. 9 The CFU / mL ratio of viable Lactococcus gasseri to Lactobacillus gasseri was 1.7:1, and all other steps and parameters were consistent with those in Example 1.

[0101] Comparative Example 4

[0102] The preparation method differs from that of Example 1 in that the ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus in Comparative Example 4 is 1:1:1, while the other steps and parameters are the same as those in Example 1.

[0103] Comparative Example 5

[0104] The preparation method differs from that of Example 1 in that the ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus in Comparative Example 5 is 2.5:1:1, while the other steps and parameters are the same as those in Example 1.

[0105] Comparative Example 6

[0106] The preparation method is the same as that in Example 1, except that the Lactococcus gasseri with accession number GDMCC NO.1.965 in Comparative Example 6 is replaced with Lactococcus gasseri with accession number CGMCC NO.9485. All other steps and parameters are the same as in Example 1.

[0107] Example 1: Behavioral Experiment

[0108] The Chronic restraint stress (CRS) model has been proven to be an effective model of anxiety stress; the open field test for exploratory behavior is used to assess general motor and anxiety-like behaviors; the elevated cross maze test can be used to evaluate animal anxiety behavior, based on the contradictory state of animals wanting to explore new environments but fearing open arms, and the shorter the time mice spend in open arms, the more anxious they are.

[0109] This embodiment uses a restraint stress model and evaluates the anxiety-relieving effects of the compositions prepared in Examples 1-3 and Comparative Examples 1-6 of this invention through open field tests and elevated cross maze tests.

[0110] Experimental animals: 96 male C57BL / 6 mice, weighing 21-25g, were kept under the following conditions: temperature 25±1℃ and relative humidity 50±5%. They were acclimatized for one week before the experiment.

[0111] Grouping: The experimental animals were randomly divided into blank control group, model group, positive control group, and experimental group, for a total of 12 groups, with 8 mice in each group;

[0112] Establishment of the CRS model: Mice in the model group, positive control group, and experimental group were placed in well-ventilated restraint devices for 3 hours a day for 10 consecutive days to induce acute restraint stress, while mice in the blank control group were placed in freely moving home cages during the same period.

[0113] Administration: 30 minutes after the end of restraint stress, the positive control group was given diazepam tablets. The tablets were weighed at a dose of 1 mg / kg, crushed, and dissolved in physiological saline at a gavage volume of 10 mL / kg to prepare a suspension before gavage. The experimental groups were given the compositions prepared in Examples 1-3 and Comparative Examples 1-6, respectively. Each composition was weighed at a dose of 5 mg / kg, dissolved in physiological saline at a gavage volume of 10 mL / kg to prepare a suspension before gavage. The blank control group and the model group were administered an equal volume of physiological saline via gavage. Administration continued for 14 days. Behavioral tests were conducted 30 minutes after the last administration, with one test performed each day.

[0114] Open field test: The experiment was conducted in an open field box measuring 50cm long, 50cm wide, and 50cm deep. The bottom of the open field box was divided into 9 equal squares, with the central area being the central zone and the remaining areas being the peripheral zone. The bottom and sides of the open field box were entirely white, and the camera equipment was placed in the center of this area. Mice were placed in the laboratory 60 minutes before the experiment to allow for acclimatization. The experiment was conducted in a quiet, low-red light environment, ensuring uniform lighting and a quiet environment throughout the experiment. Mice were gently placed in the center of the open field box and allowed to move freely for 10 minutes. The distance traveled was analyzed using Ethovision software (Noldus Information Technology, Sacramento, CA), and the total distance traveled, the proportion of distance traveled in the central zone, and the time spent in the central zone were recorded. One mouse was tested at a time, and after each test, the area was cleaned with 75% alcohol. The next mouse was tested only after the alcohol odor had dissipated and there were no obvious signs of dampness on the bottom of the box. The total distance traveled, the proportion of distance traveled in the central zone, and the proportion of time spent in the central zone were calculated. The specific results are shown in Table 2.

[0115] Elevated cross maze test: The test system consisted of two 35cm×6cm open arms and two 35cm×6cm closed arms. The closed arms were open at the top, and the open arms were open on all four sides. There was a 5cm×5cm relatively open area in the center between the open and closed arms. The maze was 50cm above the ground. Before the test, the test mice were placed in an open area for 5 minutes to acclimatize. Then, the mice were placed in the middle of the cross maze with their heads facing the open arms to start the experiment. The mice were allowed to move freely for 5 minutes. The time it took for the mice to enter the open arms within 5 minutes was recorded using Supermaze software (based on the standard that all four limbs were either inside or outside the arms). One mouse was tested at a time. After the test, the maze was wiped with 75% alcohol. After the alcohol odor dissipated and there were no obvious damp marks on the bottom of the box, the next mouse was tested. The percentage of time the mice spent in the open arms was calculated. The specific results are shown in Table 3.

[0116] Data processing: SPSS statistical software was used for data processing. All measurement data are expressed as mean ± standard deviation. t-test was used for statistical analysis, and P < 0.05 was considered statistically significant.

[0117] Compared with Example 1, Comparative Examples 1-3 were fermented without the addition of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus, respectively; Comparative Examples 4-5 had inoculation ratios of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus that exceeded the scope of this application; and Comparative Example 6 involved replacing Lactococcus gasseri with different preservation numbers.

[0118] As shown in Tables 1-2, the model group and the blank control group showed that, compared with the blank control group, the model group had significantly lower total distance traveled, central region distance traveled, central region time traveled, and time spent in open arms, indicating that CRS induced anxiety-like behavior in mice successfully.

[0119] The data in Tables 1-2, including the total distance traveled, the proportion of distance traveled in the central region, the proportion of time spent in the central region, and the proportion of time spent in the open arm position, show that the effects of Examples 1-3 and Comparative Examples 1-6 are better than those of the model group. This indicates that the traditional Chinese medicine composition prepared from Bupleurum chinense, Paeonia lactiflora, Gardenia jasminoides, Paeonia suffruticosa, Albizia julibrissin, Lycium barbarum, Ziziphus jujuba var. spinosa, Poria cocos, Nardostachys japonica, Glycyrrhiza uralensis, and Ziziphus jujuba var. spinosa, has an anxiety-relieving effect.

[0120] As shown in Tables 1-2, the data from Example 1 and Comparative Examples 1-3 indicate that Example 1 is more effective than Comparative Examples 1-3. This suggests that the composition of the fermentation strains affects the anxiety-relieving effect of the composition. The composition exhibits better anxiety-relieving efficacy only when the fermentation strains simultaneously contain *Lactococcus gasseri*, *Lactobacillus gasseri*, and *Lactobacillus pentosacchari*. This may be because the three fermentation strains mutually promote each other during fermentation, helping to release the active ingredients in the traditional Chinese medicine, thus resulting in the composition's excellent anxiety-relieving effect. As shown in Tables 1-2, the data from Example 1 and Comparative Examples 4-5 indicate that *Lactococcus gasseri*, *Lactobacillus gasseri*, and *Lactobacillus pentosacchari*... The ratio of viable bacteria among the three lactobacilli also affects the anxiety-relieving effect of the composition. When the ratio of viable bacteria of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.5-2:1:1, the composition has the best anxiety-relieving effect. According to the data of Example 1 and Comparative Example 6 in Tables 1-2, different Lactococcus gasseri strains have a certain impact on the fermentation effect, indicating that even for the same species, the differences in the properties of different strains will affect the fermentation effect. The Lactococcus gasseri strain selected in this invention is the best strain that can have a synergistic effect with Lactobacillus gasseri and Lactobacillus pentosus, which was obtained by the inventors through a large number of experiments in the early stage.

[0121] In summary, the traditional Chinese medicine fermentation composition prepared by mixed fermentation of Bupleurum chinense, Paeonia lactiflora, Gardenia jasminoides, Paeonia suffruticosa, Albizia julibrissin, Lycium barbarum, Ziziphus jujuba var. spinosa, Poria cocos, Nardostachys japonica, Glycyrrhiza uralensis, and Ziziphus jujuba var. spinosa can effectively relieve anxiety and solve the problem of poor efficacy of existing products.

[0122] Table 1. Open Field Test Data

[0123] Group Total distance traveled (m) Percentage of distance traveled in the central region (%) Central region time percentage (%) Blank control group <![CDATA[32.48±1.13 ▲ ]]> <![CDATA[14.30±0.86 ▲ ]]> <![CDATA[7.20±0.42 ▲ ]]> Model group <![CDATA[23.96±0.99 * ]]> <![CDATA[7.82±0.34 * ]]> <![CDATA[3.54±0.27 * ]]> Positive control group <![CDATA[30.58±1.46 ▲ ]]> <![CDATA[13.44±0.72 ▲ ]]> <![CDATA[6.12±0.24 ▲ ]]> Example 1 <![CDATA[31.43±1.30 ▲ ]]> <![CDATA[13.39±0.78 ▲ ]]> <![CDATA[6.41±0.39 ▲ ]]> Example 2 <![CDATA[30.29±1.34 ▲ ]]> <![CDATA[12.28±0.69 ▲ ]]> <![CDATA[5.92±0.28 ▲ ]]> Example 3 <![CDATA[30.73±1.09 ▲ ]]> <![CDATA[12.84±0.75 ▲ ]]> <![CDATA[6.08±0.40 ▲ ]]> Comparative Example 1 <![CDATA[25.60±1.56 # ]]> <![CDATA[8.91±0.27 ▲# ]]> <![CDATA[4.32±0.23 ▲# ]]> Comparative Example 2 <![CDATA[26.91±1.24 ▲# ]]> <![CDATA[9.67±0.59 ▲# ]]> <![CDATA[4.62±0.35 ▲# ]]> Comparative Example 3 <![CDATA[26.75±0.92 ▲# ]]> <![CDATA[9.38±0.30 ▲# ]]> <![CDATA[4.57±0.29 ▲# ]]> Comparative Example 4 <![CDATA[27.52±1.50 ▲# ]]> <![CDATA[10.86±0.54 ▲# ]]> <![CDATA[5.11±0.26 ▲# ]]> Comparative Example 5 <![CDATA[27.16±1.37 ▲# ]]> <![CDATA[10.26±0.43 ▲# ]]> <![CDATA[4.95±0.21 ▲# ]]> Comparative Example 6 <![CDATA[28.78±0.64 ▲# ]]> <![CDATA[11.03±0.51 ▲# ]]> <![CDATA[5.23±0.22 ▲# ]]>

[0124] Note:" * "This indicates that compared with the blank control group, P < 0.05;" ▲ "This indicates that compared with the model group, P < 0.05;" # "" indicates that compared with Example 1, Comparative Examples 1-6 have P < 0.05.

[0125] Table 2. Test data of the elevated cross maze

[0126] Group Percentage of time spent in open-arm position (%) Blank control group <![CDATA[25.74±1.04 ▲ ]]> Model group <![CDATA[14.34±0.57 * ]]> Positive control group <![CDATA[24.42±1.53 ▲ ]]> Example 1 <![CDATA[25.11±1.02 ▲ ]]> Example 2 <![CDATA[24.36±1.81 ▲ <!-- 7 -->]]> Example 3 <![CDATA[24.78±1.16 ▲ ]]> Comparative Example 1 <![CDATA[17.73±0.70 ▲# ]]> Comparative Example 2 <![CDATA[19.95±0.87 ▲# ]]> Comparative Example 3 <![CDATA[19.70±0.72 ▲# ]]> Comparative Example 4 <![CDATA[21.53±1.41 ▲# ]]> Comparative Example 5 <![CDATA[21.34±1.36 ▲# ]]> Comparative Example 6 <![CDATA[22.89±1.31 ▲# ]]>

[0127] Note:" * "This indicates that compared with the blank control group, P < 0.05;" ▲ "This indicates that compared with the model group, P < 0.05;" # "" indicates that compared with Example 1, Comparative Examples 1-6 have P < 0.05.

[0128] Application example: An aromatherapy liquid with anxiety-relieving effects.

[0129] The traditional Chinese medicine fermentation compositions of Examples 1-3 were used as raw materials for the aromatherapy liquids with anxiety-relieving effects in Examples 1-3.

[0130] The aromatherapy liquid with anxiety-relieving effects comprises the following raw materials in parts by weight: 11 parts of the traditional Chinese medicine fermentation composition, 1 part of bergamot extract, 1 part of ylang-ylang extract, 2 parts of jasmine extract, 1 part of orange blossom extract, 1 part of patchouli extract, 27 parts of 3-methoxy-3-methylbutanol, 3 parts of polyacrylamide, 7 parts of glycerol, 5 parts of silicone oil, and 0.5 parts of phenoxyethanol.

[0131] The preparation method of the aromatherapy liquid with anxiety-relieving effect specifically includes the following steps:

[0132] S1. Mixture I is obtained by stirring 3-methoxy-3-methylbutanol, polyacrylamide, glycerol and silicone oil at 300 rpm for 15 min.

[0133] S2. Add the traditional Chinese medicine fermentation composition, bergamot extract, ylang-ylang extract, jasmine extract, orange blossom extract, patchouli extract and phenoxyethanol to mixture I, and stir at 300 rpm for 15 min to obtain the aromatherapy liquid.

[0134] Test Example 2: Fragrance Diffusion Ability Test

[0135] This test case refers to the method in CN117752723B to test the diffusion effect and the degree of anxiety relief and relaxation of the aromatherapy liquids prepared in Examples 1-3;

[0136] The specific testing method is as follows:

[0137] Test sample: Aromatherapy liquid prepared using Examples 1-3;

[0138] Test participants: 30 volunteers with scent evaluation abilities and anxiety were divided into 3 groups of 10 people each;

[0139] Diffusion effect test: at 25℃ and 10m respectively 2 In a closed space, four drops of aromatherapy liquid were placed on a diffuser stone. After inhaling the sample for 15 minutes, the testers evaluated the aroma concentration in the room and scored it. The data are presented as average values. The score range is 0-40 points. 39.1-40 points represent good aroma diffusion, 36-39 points represent good aroma diffusion, 32-35.9 points represent acceptable aroma diffusion, 28-31.9 points represent fair aroma diffusion, 24-27.9 points represent passing aroma diffusion, and below 24 points represent failing aroma diffusion.

[0140] Test on the degree of anxiety relief and relaxation: After inhaling the test sample for 30 minutes, the testers rated the degree of anxiety relief and relaxation. The data is presented as an average value. The score range is 0-40 points, with 31-40 points representing obvious effectiveness, 21-30 points representing effectiveness, 11-20 points representing ineffectiveness, and 0-10 points representing increased tension and excitement.

[0141] The specific statistical results are shown in Table 3. The aromatherapy liquids prepared using Examples 1-3 all have excellent diffusion and anxiety-relieving effects.

[0142] Table 3 Experimental Data

[0143] Group Fragrance diffusion effect Relieves anxiety and relaxes the mind and body Application Example 1 40.0 38.7 Application Example 2 38.4 36.2 Application Example 3 39.1 37.5

[0144] The above description is merely a preferred composition of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with preferred compositions, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent compositions without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above compositions based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A method for preparing a traditional Chinese medicine fermentation composition for relieving anxiety, characterized in that, Includes the following steps: A1. Weigh out each Chinese herbal fermentation raw material according to the weight parts, mix them together, add deionized water, mix evenly, sterilize, and obtain a mixture; A2. Inoculate the mixture with Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus for anaerobic fermentation to obtain the fermentation product; A3. Sterilize and filter the fermentation product, and concentrate the filtrate under reduced pressure to obtain the traditional Chinese medicine fermentation composition. The fermented raw materials of the traditional Chinese medicine are composed of the following components by weight: Bupleurum chinense 9-12 parts, Paeonia lactiflora 8-10 parts, Gardenia jasminoides 8-10 parts, Paeonia suffruticosa 7-9 parts, Albizia julibrissin 6-8 parts, Lycium barbarum 7-9 parts, Ziziphus jujuba var. spinosa 10-15 parts, Poria cocos 10-15 parts, Nardostachys jatamansi 6-8 parts, Glycyrrhiza uralensis 4-6 parts, and Ziziphus jujuba 4-6 parts. The preservation number of *Lactococcus gasseri* is GDMCC NO.1.965, the preservation number of *Lactobacillus gasseri* is CGMCC NO.1.3396, and the preservation number of *Lactobacillus pentosus* is GDMCC NO.1.524; the live count ratio of *Lactococcus gasseri*, *Lactobacillus gasseri*, and *Lactobacillus pentosus* is 1.5-2:1:

1.

2. The preparation method according to claim 1, characterized in that, The fermented raw materials of the traditional Chinese medicine are composed of the following components by weight: Bupleurum chinense 11 parts, Paeonia lactiflora 9 parts, Gardenia jasminoides 9 parts, Paeonia suffruticosa 8 parts, Albizia julibrissin 7 parts, Lycium barbarum 8 parts, Ziziphus jujuba var. spinosa 13 parts, Poria cocos 13 parts, Nardostachys jatamansi 7 parts, Glycyrrhiza uralensis 5 parts, and Ziziphus jujuba 5 parts.

3. The preparation method according to claim 1, characterized in that, In step A1, the ratio of the fermentation raw material of traditional Chinese medicine to deionized water is 1g:8-12mL.

4. The preparation method according to claim 1, characterized in that, In step A2, the total inoculation amount of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 3-6% v / v, and the total viable count of Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.0 × 10⁻⁶. 9 -5.0×10 9 CFU / mL.

5. The preparation method according to claim 1, characterized in that, In step A2, the ratio of viable Lactococcus gasseri, Lactobacillus gasseri, and Lactobacillus pentosus is 1.7:1:

1.

6. The preparation method according to claim 1, characterized in that, The fermentation temperature in step A2 is 32-37℃, and the fermentation time is 36-48h.

7. The application of the fermented Chinese medicine composition prepared by any one of claims 1-6 in the preparation of a Chinese medicine aromatherapy product with anxiety-relieving effects, characterized in that, The dosage form of the herbal aromatherapy products is liquid, gel, or solid.

8. An aromatherapy liquid, characterized in that, The raw materials include the following parts by weight: 9-13 parts of the traditional Chinese medicine fermentation composition prepared by the preparation method according to any one of claims 1-6, 5-8 parts of plant extract, 25-35 parts of 3-methoxy-3-methylbutanol, 2-4 parts of polyacrylamide, 4-9 parts of glycerol, 4-7 parts of silicone oil, and 0.3-0.7 parts of phenoxyethanol. The plant extract is selected from at least one of bergamot extract, ylang-ylang extract, jasmine extract, orange blossom extract, or patchouli extract.

9. A method for preparing the aromatherapy liquid as described in claim 8, characterized in that, Includes the following steps: S1. Stir 3-methoxy-3-methylbutanol, polyacrylamide, glycerol, and silicone oil at 200-300 rpm for 12-17 min to obtain mixture I; S2. Add the traditional Chinese medicine fermentation composition, plant extract and phenoxyethanol to mixture I, and stir at 200-300 rpm for 10-15 min to obtain the aromatherapy liquid.