Traditional Chinese medicine fermented composition for relieving asthenopia and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine fermented composition for relieving asthenopia and a preparation method thereof. The preparation method of the traditional Chinese medicine fermented composition for relieving asthenopia comprises the following steps: carrying out enzymolysis on traditional Chinese medicine components by adopting a compound enzyme, and then fermenting the traditional Chinese medicine components subjected to enzymolysis by virtue of mixed bacteria consisting of lactobacillus rhamnosus, saccharomyces cerevisiae and aspergillus niger, so that the prepared traditional Chinese medicine fermented composition is safe, non-irritant and better in medicine effect according to verification; the massage cream prepared from the traditional Chinese medicine fermented composition has a remarkable effect of relieving asthenopia. The preparation method of the traditional Chinese medicine fermented composition is simple, can solve the problems of complexity, long treatment course and slow effect when the existing traditional Chinese medicine is used for treating asthenopia, and has better clinical application value.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine fermentation composition for relieving visual fatigue and a preparation method thereof. Background Art

[0002] Visual fatigue, also known as asthenopia, refers to the eye fatigue caused by overuse of the eyes when engaged in close-range visual work or study. Its main clinical manifestations include blurred vision, double vision, intolerance to prolonged vision, dry and uncomfortable eyes, soreness and tearing, headache and dizziness, etc. It is a syndrome based on the patient's subjective symptoms of the eyes, in which organic factors of the eyes or the whole body and mental factors are intertwined, so it is also called eye fatigue syndrome and belongs to the category of psychosomatic medicine.

[0003] Western medicine mainly treats asthenopia symptomatically and by correction. Generally, sedatives, nerve nourishing agents are used, and β-blockers are also locally instilled. In addition, surgery, prism correction or visual training are also used to treat asthenopia. Although the above methods have relatively certain curative effects, their general side effects are relatively prominent and their application scope is relatively narrow, unable to meet the prevention and treatment needs of asthenopia syndrome. There are various methods for traditional Chinese medicine to treat asthenopia, and methods such as acupuncture, acupoint massage, traditional Chinese medicine application, and traditional Chinese medicine atomization can all achieve good curative effects. At the same time, because it belongs to the category of external treatment methods of traditional Chinese medicine, it is convenient and flexible to use and has prominent curative effects, which is favored by a large number of asthenopia patients, reflecting the advantages of traditional Chinese medicine in diagnosing and treating asthenopia.

[0004] In recent years, with the advent of the "screen-reading era", people's time of using various electronic products such as mobile phones, tablets, and computers has become increasingly prolonged, resulting in high-load work of the eyes and the exacerbation of the condition of overusing the eyes, greatly increasing the probability of occurrence of eye diseases. However, most of the current traditional Chinese medicine methods for treating asthenopia have the disadvantages of being cumbersome and complex, long course of treatment, and slow onset of effect. Therefore, providing a traditional Chinese medicine product that is convenient and simple to use, and has a short course of treatment and quick effect is an urgent problem to be solved at present. Summary of the Invention

[0005] In view of the above defects or improvement requirements of the prior art, the present invention provides a traditional Chinese medicine fermentation composition for relieving visual fatigue, a preparation method thereof, and a massage cream prepared by using the traditional Chinese medicine fermentation composition, so as to solve the problems of being cumbersome and complex, long course of treatment, and slow onset of effect in the treatment of asthenopia with traditional Chinese medicine.

[0006] The purpose of the first aspect of the present invention is to provide a preparation method of a traditional Chinese medicine fermentation composition for relieving visual fatigue.

[0007] The purpose of the present invention can be achieved by the following technical solutions:

[0008] The first aspect of the present invention provides a preparation method of a traditional Chinese medicine fermentation composition for relieving visual fatigue, including:

[0009] S1. Weigh each raw material in the traditional Chinese medicine composition according to the parts by weight, grind them, and sieve them to obtain a mixed powder; add sterile deionized water to the mixed powder and stir evenly to obtain a mixed solution I.

[0010] S2. Add pectinase and cellulase to the mixed solution I and perform enzymatic hydrolysis assisted by ultrasound; after the enzymatic hydrolysis is completed, raise the temperature for enzyme inactivation treatment to obtain a mixed solution II.

[0011] S3. Activate Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with a liquid medium respectively, and inoculate the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into the mixed solution II for mixed bacteria fermentation.

[0012] S4. After the fermentation is completed, filter, and concentrate the filtrate under reduced pressure to obtain a concentrated solution; the concentrated solution is sterilized at high temperature to obtain the said traditional Chinese medicine fermented composition.

[0013] Preferably, the preservation number of Lactobacillus rhamnosus is CGMCC 1.3724; the preservation number of the said Saccharomyces cerevisiae is CGMCC 2.1527; the preservation number of the said Aspergillus niger is CGMCC 3.316.

[0014] Preferably, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 3 - 15 parts of wolfberry fruits, 1 - 10 parts of chrysanthemums, 6 - 20 parts of Cimicifuga dahurica, 1 - 10 parts of Glossy Privet Fruit, 5 - 16 parts of Rhizoma Ligustici, 5 - 16 parts of safflower, 1 - 10 parts of Coptis chinensis, 1 - 10 parts of Equisetum hiemale, 1 - 10 parts of Gardenia jasminoides, 1 - 10 parts of Tribulus terrestris, 5 - 16 parts of Cuscuta chinensis, 6 - 18 parts of Cassia tora, 1 - 10 parts of Buddleja officinalis, 1 - 10 parts of Paeonia lactiflora, 1 - 10 parts of mulberry leaves.

[0015] More preferably, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 10 parts of wolfberry fruits, 4 parts of chrysanthemums, 13 parts of Cimicifuga dahurica, 5 parts of Glossy Privet Fruit, 10 parts of Rhizoma Ligustici, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum hiemale, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja officinalis, 4 parts of Paeonia lactiflora, 4 parts of mulberry leaves.

[0016] Preferably, in the step S1, the mesh number of sieving is 200 - 300 meshes, and the material - liquid ratio of the traditional Chinese medicine composition mixed powder to sterile deionized water is 1:5 - 7.

[0017] Preferably, in the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 3 - 5 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45 - 55 °C, pH = 4.5 - 5.5, the ultrasonic frequency is 25 - 35 kHz, and the time is 40 - 50 min; the temperature of the enzyme inactivation treatment is 90 - 100 °C, and the time of the enzyme inactivation treatment is 20 - 30 min.

[0018] Preferably, in the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1-2:1:1-1.5.

[0019] More preferably, in the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.2.

[0020] Preferably, the total amount of inoculated bacteria is 5-7% of the volume of the mixed solution II.

[0021] Preferably, in the step S3, the fermentation temperature is 30-34 °C, the fermentation pH = 5.0-6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200-250 rpm, and the fermentation time is 48-72 h.

[0022] Preferably, in the step S4, it is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of concentration under reduced pressure is 40-50 °C, the pressure of concentration under reduced pressure is -0.06 to -0.08 MPa; the sterilization temperature is 121 °C, and the sterilization time is 20-30 min.

[0023] The object of the second aspect of the present invention is to provide a traditional Chinese medicine fermentation composition prepared by the preparation method of the first aspect of the present invention.

[0024] The object of the third aspect of the present invention is to provide a massage cream for relieving visual fatigue containing the traditional Chinese medicine fermentation composition described in the second aspect.

[0025] Preferably, the massage cream contains the following raw materials by weight percentage: traditional Chinese medicine fermentation composition 5.0-20.0%, emulsifier 6.5-8.5%, excipient 3.0-8.0%, emollient 8.0-13.0%, conditioner 0.18-0.68%, suspending agent 0.3-2.3%, covering agent 0.1-1.0%, moisturizer 10.5-14.5%, chelating agent 0.05-0.15%.

[0026] The beneficial effects of the present invention:

[0027] The present invention hydrolyzes the Chinese medicine ingredients by a composite enzyme to release the active substances in the Chinese medicine, and then the Chinese medicine ingredients after the mixed fermentation hydrolysis composed of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger, Aspergillus niger can produce cellulose to help Chinese medicine further release active substances; Lactobacillus rhamnosus is a safe probiotic, and the protein or metabolite secreted by it has multiple potential health benefits such as enhancing immune function and inhibiting bacteria; yeast can cooperate with lactobacillus to breed, complement nutrition, and promote the proliferation of lactobacillus. In order to improve the therapeutic effect of a specific Chinese medicine composition, the inventors screened and verified a large number of experiments in the early stage, and finally obtained a composite bacteria composed of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger. The above bacteria have a good synergistic effect during the fermentation process, so that the prepared Chinese medicine fermentation composition is safe and non-irritating and has better efficacy, and the massage cream prepared by the Chinese medicine fermentation composition has a significant effect of relieving visual fatigue. The preparation method of the Chinese medicine fermentation composition described in the present application is simple, can solve the problems of cumbersome and complex, long course of treatment, and slow effect when the existing Chinese medicine is used to treat visual fatigue, and has good clinical application value. DETAILED DESCRIPTION

[0028] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with embodiments.

[0029] The experimental methods in the following examples without specific conditions are usually carried out under conventional conditions or conditions recommended by the manufacturers. The materials and reagents used can be obtained from commercial sources unless otherwise specified.

[0030] Cellulase was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with an enzyme activity of 50u / mg;

[0031] Pectinase was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with an enzyme activity of 50,000 u / g;

[0032] The viable counts of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger involved in the present invention are all 1×10 7 CFU / mL.

[0033] Example 1

[0034] A method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, the preparation method comprising the following steps:

[0035] S1. Weigh each raw material in the Chinese medicine composition according to weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0036] S2. Add pectinase and cellulase to the mixed solution I, and perform enzymatic hydrolysis assisted by ultrasound; after the enzymatic hydrolysis is completed, raise the temperature for enzyme inactivation treatment to obtain the mixed solution II;

[0037] S3. Activate Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger with a liquid medium respectively, and inoculate the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger into the mixed solution II for mixed bacteria fermentation;

[0038] S4. After the fermentation is completed, filter, and concentrate the filtrate under reduced pressure to obtain a concentrated solution; the concentrated solution is sterilized at a high temperature to obtain the described traditional Chinese medicine fermentation composition;

[0039] Among them, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 3 parts of wolfberry fruit, 1 part of chrysanthemum, 6 parts of Cimicifuga dahurica, 1 part of Glossy Privet Fruit, 5 parts of Rhizoma Ligustici, 5 parts of safflower, 1 part of Coptis chinensis, 1 part of Equisetum hiemale, 1 part of Gardenia jasminoides, 1 part of Tribulus terrestris, 5 parts of Cuscuta chinensis, 6 parts of Cassia tora, 1 part of Buddleja officinalis, 1 part of white peony root, 1 part of mulberry leaf.

[0040] In the step S1, the mesh number of sieving is 200 mesh, and the ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:5.

[0041] In the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 3 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45 °C, pH = 4.5, the ultrasonic frequency is 25 kHz, and the time is 40 min; the temperature of the enzyme inactivation treatment is 90 °C, and the time of the enzyme inactivation treatment is 20 min.

[0042] In the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1:1:1.5; the total inoculum amount is 5% of the volume of the mixed solution II.

[0043] The preservation number of the Lactobacillus rhamnosus is CGMCC 1.3724; the preservation number of the Saccharomyces cerevisiae is CGMCC 2.1527; the preservation number of the Aspergillus niger is CGMCC 3.316.

[0044] In the step S3, the fermentation temperature is 30 °C, the fermentation pH = 5.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200 rpm, and the fermentation time is 48 h.

[0045] In the step S4, it is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of the reduced pressure concentration is 40 °C, the pressure of the reduced pressure concentration is -0.06 MPa; the sterilization temperature is 121 °C, and the sterilization time is 20 min.

[0046] Example 2

[0047] A preparation method of a traditional Chinese medicine fermentation composition for relieving visual fatigue, the preparation method includes the following steps:

[0048] S1. Weigh each raw material in the traditional Chinese medicine composition according to the weight parts, grind them, and sieve them to obtain a mixed powder. Add sterile deionized water to the mixed powder and stir evenly to obtain a mixed solution I.

[0049] S2. Add pectinase and cellulase to the mixed solution I and perform ultrasonic-assisted enzymatic hydrolysis. After the enzymatic hydrolysis is completed, raise the temperature for enzyme inactivation treatment to obtain a mixed solution II.

[0050] S3. Activate Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with a liquid medium respectively, and inoculate the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into the mixed solution II for mixed bacteria fermentation.

[0051] S4. After the fermentation is completed, filter, and concentrate the filtrate under reduced pressure to obtain a concentrated solution. The concentrated solution is sterilized at high temperature to obtain the traditional Chinese medicine fermentation composition.

[0052] Among them, the traditional Chinese medicine composition includes the following raw materials in weight parts: 10 parts of wolfberry fruits, 4 parts of chrysanthemums, 13 parts of Cimicifuga dahurica, 5 parts of Glossy Privet Fruit, 10 parts of Rhizoma Ligustici, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum hiemale, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja officinalis, 4 parts of Paeonia lactiflora, and 4 parts of mulberry leaves.

[0053] In the step S1, the mesh number of sieving is 250 meshes, and the material-liquid ratio of the traditional Chinese medicine composition mixed powder to sterile deionized water is 1:6.

[0054] In the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 4 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 50 °C, pH = 5.0, the ultrasonic frequency is 30 kHz, and the time is 45 min; the temperature of enzyme inactivation treatment is 95 °C, and the time of enzyme inactivation treatment is 25 min.

[0055] In the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger is 1.5:1:1.2; the total inoculum amount is 6% of the volume of the mixed solution II.

[0056] The preservation number of the Lactobacillus rhamnosus is CGMCC 1.3724; the preservation number of the Saccharomyces cerevisiae is CGMCC 2.1527; the preservation number of the Aspergillus niger is CGMCC 3.316.

[0057] In the step S3, the fermentation temperature is 32 °C, the fermentation pH = 5.5, the sterile air ventilation volume is 5 L / min, the rotation speed is 220 rpm, and the fermentation time is 60 h.

[0058] In the step S4, it is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of concentration under reduced pressure is 45 °C, the pressure of concentration under reduced pressure is -0.07 MPa; the sterilization temperature is 121 °C, and the sterilization time is 25 min.

[0059] Example 3

[0060] A preparation method of a traditional Chinese medicine fermentation composition for relieving visual fatigue, the preparation method comprising the following steps:

[0061] S1. Weigh each raw material in the traditional Chinese medicine composition according to the weight parts, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder and stir evenly to obtain a mixed liquid I;

[0062] S2. Add pectinase and cellulase to the mixed liquid I and perform ultrasonic-assisted enzymatic hydrolysis; after the enzymatic hydrolysis is completed, raise the temperature for enzyme inactivation treatment to obtain a mixed liquid II;

[0063] S3. Activate Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger with a liquid medium respectively, and inoculate the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger into the mixed liquid II for mixed bacteria fermentation;

[0064] S4. After the fermentation is completed, filter, and concentrate the filtrate under reduced pressure to obtain a concentrated liquid; the concentrated liquid is sterilized at high temperature to obtain the traditional Chinese medicine fermentation composition;

[0065] Wherein, the traditional Chinese medicine composition comprises the following raw materials in weight parts: 15 parts of wolfberry fruit, 10 parts of chrysanthemum, 20 parts of Cimicifuga dahurica, 10 parts of Glossy Privet Fruit, 16 parts of Ligusticum sinense, 16 parts of safflower, 10 parts of Coptis chinensis, 10 parts of Equisetum hiemale, 10 parts of Gardenia jasminoides, 10 parts of Tribulus terrestris, 16 parts of Cuscuta chinensis, 18 parts of Cassia tora, 10 parts of Buddleja officinalis, 10 parts of white peony root, 10 parts of mulberry leaf.

[0066] In the step S1, the mesh number of the sieve is 300 meshes, and the material-liquid ratio of the traditional Chinese medicine composition mixed powder to sterile deionized water is 1:7.

[0067] In the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 5 wt% of the mass of the mixed liquid I, the enzymatic hydrolysis temperature is 55 °C, pH = 5.5, the ultrasonic frequency is 35 kHz, and the time is 50 min; the temperature of the enzyme inactivation treatment is 100 °C, and the time of the enzyme inactivation treatment is 30 min.

[0068] In the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 2:1:1; the total bacteria inoculation amount is 7% of the volume of the mixed liquid II.

[0069] The preservation number of the Lactobacillus rhamnosus is CGMCC 1.3724; the preservation number of the Saccharomyces cerevisiae is CGMCC2.1527; the preservation number of the Aspergillus niger is CGMCC 3.316;

[0070] In step S3, the fermentation temperature is 34°C, the fermentation pH = 6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 250 rpm, and the fermentation time is 72 h.

[0071] In step S4, it is concentrated under reduced pressure to 80% of the mass of the original filtrate. The temperature of the reduced pressure concentration is 50°C, and the pressure of the reduced pressure concentration is -0.08 MPa; the sterilization temperature is 121°C, and the sterilization time is 30 min.

[0072] Comparative Example 1

[0073] Compared with Example 2, the only difference is that Lactobacillus rhamnosus is not added in Comparative Example 1, and the reduced volume is proportionally distributed to the volumes of Saccharomyces cerevisiae and Aspergillus niger. Other conditions are the same as those in Example 2.

[0074] Comparative Example 2

[0075] Compared with Example 2, the only difference is that Saccharomyces cerevisiae is not added in Comparative Example 2, and the reduced volume is proportionally distributed to the volumes of Lactobacillus rhamnosus and Aspergillus niger. Other conditions are the same as those in Example 2.

[0076] Comparative Example 3

[0077] Compared with Example 2, the only difference is that the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 0.5:1:2. Other conditions are the same as those in Example 2.

[0078] Comparative Example 4

[0079] Compared with Example 2, the only difference is that the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 2.5:1:0.5. Other conditions are the same as those in Example 2.

[0080] Comparative Example 5

[0081] Compared with Example 2, the only difference is that Lactobacillus rhamnosus with the preservation number of ATCC 7469 is used to replace Lactobacillus rhamnosus with the preservation number of CGMCC 1.3724. Other conditions are the same as those in Example 2.

[0082] Comparative Example 6

[0083] Based on Example 2, the only difference is that Saccharomyces cerevisiae with the preservation number of CGMCC 2.3854 is used to replace Saccharomyces cerevisiae with the preservation number of CGMCC 2.1527. Other conditions are the same as those in Example 2.

[0084] Comparative Example 7

[0085] On the basis of Example 2, the difference is only that Aspergillus niger with the preservation number of CGMCC 3.4309 is used to replace Aspergillus niger with the preservation number of CGMCC 3.316, and other conditions are the same as those in Example 2.

[0086] Comparative Example 8

[0087] A preparation method of a traditional Chinese medicine composition for relieving visual fatigue, the preparation method comprising the following steps:

[0088] S1. Weigh each raw material in the traditional Chinese medicine composition according to the weight parts, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder and stir evenly to obtain a mixed solution I;

[0089] S2. Add pectinase and cellulase to the mixed solution I and perform ultrasonic-assisted enzymatic hydrolysis; after the enzymatic hydrolysis is completed, raise the temperature for enzyme inactivation treatment to obtain a mixed solution II;

[0090] S3. Filter the mixed solution II, and concentrate the filtrate under reduced pressure to obtain a concentrated solution, thus obtaining the traditional Chinese medicine composition.

[0091] Among them, the traditional Chinese medicine composition comprises the following raw materials in weight parts: 10 parts of wolfberry fruit, 4 parts of chrysanthemum, 13 parts of Cimicifuga dahurica, 5 parts of Glossy Privet Fruit, 10 parts of Ligusticum sinense, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum hiemale, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja officinalis, 4 parts of white peony root, 4 parts of mulberry leaf.

[0092] In the step S1, the mesh number of sieving is 250 meshes, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:6.

[0093] In the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 4 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 50 °C, pH = 5.0, the ultrasonic frequency is 30 kHz, and the time is 45 min; the temperature of the enzyme inactivation treatment is 95 °C, and the time of the enzyme inactivation treatment is 25 min.

[0094] In the step S3, the reduction is concentrated to 80% of the mass of the original filtrate, the temperature of the reduction concentration is 45 °C, and the pressure of the reduction concentration is -0.07 MPa.

[0095] Effect Example 1 Acute eye irritation test

[0096] Referring to the acute eye irritation test in the "Technical Specifications for Cosmetics Safety" and Patent CN116492383B, the irritations of the compositions prepared in Examples 1-3 and Comparative Examples 1-8 were verified.

[0097] Experimental animals: Adult female New Zealand white rabbits with healthy eyes, weighing 2.0 - 2.5 kg, fed with ordinary-grade rabbit feed, and individually caged under the conditions of a constant temperature of 20 - 26°C without special pathogens and a relative environmental humidity of 40% - 70%.

[0098] The specific method is as follows: Healthy adult New Zealand white rabbits are used. The rabbits need to be acclimatized in the experimental animal house environment for 7 days before the experiment. Select 33 rabbits without eye irritation symptoms, corneal defects, and conjunctival injuries, and randomly divide them into 11 groups, with 3 rabbits in each group, which are respectively used to test the compositions prepared in Examples 1 - 3 and Comparative Examples 1 - 8. Gently pull down the lower eyelid of one eye of the rabbit, and drop 0.1 mL of the test substance (corresponding to the compositions prepared in Examples 1 - 3 and Comparative Examples 1 - 8 respectively) into the conjunctival sac, and passively close the upper and lower eyelids for 1 s to prevent the loss of the test substance. The other eye is not treated as a self-control. Do not rinse the eyes within 24 hours after dropping the test substance. If necessary, rinsing can be carried out at 24 hours. If the above test results show that the test substance is irritating, another 3 rabbits need to be selected for the rinsing effect test, that is, 30 s after dropping the test substance into the rabbit's eye, rinse with a sufficient amount of water flowing at a relatively fast speed but not causing eye damage to the animal for at least 30 s. Check the animal's eyes at 1, 24, 48, 72 h, and on the 4th and 7th days after dropping the composition. If no irritation reaction occurs within 72 h, the test can be terminated.

[0099] Evaluation is carried out according to Table 1. Add up the scores of the corneal, iris, conjunctival, and secretory irritation reactions of each rabbit's eye, which is the total score of the eye irritation reaction of the rabbit. Add up the total scores of the eye irritation reactions of each rabbit's eye and then divide by the number of rabbits, which is the final score of the irritation of the composition to the rabbits.

[0100] Conclusion: As shown in Table 2, when checking the eyes of 11 groups of rabbits at 1 h, 24 h, 48 h, and 72 h, no corneal, iris, or bulbar conjunctival lesions or congestion were observed, and there was no secretion. The total eye irritation score was zero, indicating that the compositions prepared in Examples 1 - 3 and Comparative Examples 1 - 8 are all safe and non-irritating.

[0101] Table 1 Scoring criteria for acute eye irritation

[0102]

[0103]

[0104] Table 2 Integral results of acute eye irritation test

[0105]

[0106]

[0107] Effect Example 2 Pharmacodynamic verification

[0108] The pharmacodynamic verification of the compositions prepared in Examples 1-3 and Comparative Examples 1-8 was carried out, and the specific operation steps are as follows:

[0109] Experimental animals: 112 male and healthy New Zealand white rabbits, weighing 2.0-2.5 kg, were fed with ordinary feed under the condition of constant temperature of 20-26 °C without special pathogens, the environmental relative humidity was 40%-70%, and they were raised in single cages.

[0110] Grouping: 112 New Zealand white rabbits were randomly divided into a blank group, a model group, a positive control group, Example 1-3 groups, and Comparative Example 1-8 groups, a total of 14 groups, 8 in each group, a total of 112.

[0111] Modeling of visual fatigue animals: Except for the blank group, the other groups had a day-night light cycle, with 35 lux (illuminance) during the day and 0 lux at night for 6 consecutive days.

[0112] Grouping and administration: After modeling, at 5 mL / rabbit, the rabbits in Example 1-3 and Comparative Example 1-8 groups were orally administered the corresponding compositions prepared in their groups, the rabbits in the positive control group were orally administered Qiju Dihuang Oral Liquid, and the rabbits in the blank group and the model group were both orally administered an equal amount of normal saline, once a day for 4 consecutive weeks.

[0113] Index detection: 24 hours after the last administration, the experimental animals were sacrificed, and bilateral eyeballs were removed. The ciliary bodies were carefully separated under a surgical microscope. Rinsed with normal saline, blotted dry with filter paper, and weighed. Prepared with an internal cutting tissue homogenizer (homogenization time 10 s / time, interval 30 s, 4 consecutive times, carried out in ice water). The prepared 10% homogenate was centrifuged at about 2000 r / min with a low-temperature and low-speed centrifuge for 10-15 min, and the supernatant was taken. ELISA kits were used to measure the cAMP and cGMP contents in the ciliary bodies of experimental rabbits. All the above kits were purchased from Wuhan Jilide Biotechnology Co., Ltd.

[0114] Data processing: The data statistics were processed using SPSS statistical software. All measurement data were expressed as mean ± standard deviation. Statistical analysis was performed using a t-test, and P < 0.05 was considered statistically significant.

[0115] The experimental results are shown in Table 3. Compared with the blank group, the cAMP content in the ciliary body tissue of the model group was significantly decreased, while the cGMP content was significantly increased, indicating that the modeling was successful.

[0116] Compared with the model group, in Example 1-3 and Comparative Example 1-8 groups fed with the corresponding compositions for 4 weeks, the cAMP in the ciliary body tissue was significantly increased, and the cGMP content was significantly decreased, indicating that the compositions prepared in the examples and comparative examples have a regulatory effect on the abnormal levels of cAMP and cGMP in the ciliary body tissue of visual fatigue animal models.

[0117] Among them, the effects of Examples 1-3 and Comparative Examples 1-7 are better than those of the positive control group, while the effect of Comparative Example 8 is worse than that of the positive control group, indicating that the mixed bacteria fermentation step plays an important role in the release of active ingredients of the traditional Chinese medicine composition, making the prepared traditional Chinese medicine fermentation composition have better medicinal effects. In Comparative Examples 1-2, one of Lactobacillus rhamnosus and Saccharomyces cerevisiae was missing. Compared with the traditional Chinese medicine fermentation composition prepared in Examples 1-3, the improvement effect on the visual fatigue animal model of the above-mentioned comparative examples was significantly weakened; it was shown that the composition of different strains in the composite bacteria directly affected the fermentation result, and experiments showed that Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger could promote and influence each other during fermentation and then produce a synergistic effect, which could effectively improve the therapeutic effect of the composition. Further, in Comparative Examples 3-4, the inoculation ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger exceeded a certain range. Compared with the traditional Chinese medicine fermentation composition prepared in Examples 1-3, the improvement effect on the visual fatigue animal model of the above-mentioned comparative examples was also significantly weakened, indicating that different ratios of the same strain would also affect the fermentation result to a certain extent and have a certain impact on the therapeutic effect of the traditional Chinese medicine fermentation composition; and the inoculation ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger of 1.5:1:1.2 had the best fermentation effect. Finally, in Comparative Examples 5-7, different strains of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger were replaced respectively. The comparison of the above-mentioned comparative examples with the experimental results of the examples showed that even for the same strain, the property differences of different strains would affect the effect of the traditional Chinese medicine fermentation composition; further, in Comparative Example 7, different strains of Aspergillus niger were replaced, and its effect was worse than that of Comparative Examples 5-6, indicating that different strains of Aspergillus niger had a greater impact on the fermentation effect than Lactobacillus rhamnosus and Saccharomyces cerevisiae; furthermore, in Comparative Examples 5 and 6, different strains of Lactobacillus rhamnosus and Saccharomyces cerevisiae were replaced respectively. The comparison of their experimental results with the examples verified that the strains of Lactobacillus rhamnosus and Saccharomyces cerevisiae selected in the present invention were obtained by the inventor through a large number of previous experiments and had the best synergistic effect with the strain of Aspergillus niger.

[0118] To sum up, the present invention uses the mixed bacteria fermentation of traditional Chinese medicine components enzymatically hydrolyzed by Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger. The above-mentioned bacteria have good synergistic effects during the fermentation process, making the prepared traditional Chinese medicine fermentation composition safe and non-irritating through experimental verification and having a significant regulatory effect on the abnormal levels of cAMP and cGMP in the ciliary body tissue caused by the visual fatigue animal model, and can solve the problems of long treatment course and slow effect of existing traditional Chinese medicine in treating visual fatigue.

[0119] Table 3 Results of Pharmacodynamic Verification

[0120] Group cAMP (nmol / L) cGMP (nmol / L) Blank control group 140.80±12.12 31.05±4.76 Model group 53.91±5.05* 144.93±15.18* Positive control group <![CDATA[71.89±9.13 ▲ > <![CDATA[48.21±6.46 ▲ > Example 1 <![CDATA[98.96±10.56 ▲# > <![CDATA[37.23±5.39 ▲# > Example 2 <![CDATA[100.57±8.83 ▲# > <![CDATA[35.77±4.25 ▲# > Example 3 <![CDATA[97.78±9.14 ▲# > <![CDATA[38.59±6.57 ▲# > Comparative example 1 <![CDATA[75.03±8.15 ▲ > <![CDATA[50.11±7.53 ▲ > Comparative example 2 <![CDATA[69.97±9.27 ▲ > <![CDATA[55.67±6.47 ▲# > Comparative example 3 <![CDATA[84.12±10.38 ▲# > <![CDATA[46.91±5.61 ▲ > Comparative example 4 <![CDATA[86.87±8.01 ▲# > <![CDATA[45.43±6.84 ▲ > Comparative example 5 <![CDATA[92.18±9.33 ▲# > <![CDATA[40.26±7.18 ▲# > Comparative example 6 <![CDATA[91.79±11.21 ▲# > <![CDATA[41.92±5.39 ▲# > Comparative example 7 <![CDATA[89.10±10.08 ▲# > <![CDATA[43.79±6.88 ▲# > Comparative example 8 <![CDATA[62.38±7.76 ▲# > <![CDATA[60.24±8.97 ▲# >

[0121] Note: Compared with the blank group: P < 0.05 is indicated by "*"; compared with the model group: P < 0.05 is indicated by "▲ ” indicates that compared with the positive control group: P < 0.05 is represented by " # ".

[0122] Application Example 1

[0123] The production method of the traditional Chinese medicine fermented composition in this application example is the same as that of Example 2. The massage cream is prepared from the following raw materials by weight percentage: 14.62% traditional Chinese medicine fermented composition, 1.5% polyoxyethylene (2) stearyl ether, 2.0% polyoxyethylene (21) stearyl ether, 4.0% glyceryl stearate, 5.0% cetearyl alcohol, 5.0% dicaprylyl carbonate, 3.0% isopropyl myristate, 2.0% polydimethylsiloxane, 0.05% bisabolol, 47.1% water, 5.0% 1,3-propanediol, 0.3% xanthan gum, 1.0% hydroxypropyl starch phosphate, 0.1% disodium edetate, 0.5% betaine, 0.1% allantoin, 0.2% panthenol, 0.5% titanium dioxide, 5.0% glycerin, 2.0% butanediol, 0.03% tetrahydropiperine, 0.1% methylisothiazolinone or iodopropynyl butylcarbamate, 0.1% essence, 0.8% phenoxyethanol or ethylhexylglycerin.

[0124] The massage cream prepared in Application Example 1 was tested for visual function, and the test was carried out with reference to "Methods for the Function Test and Evaluation of Health Foods (2023 Edition)".

[0125] Inclusion criteria: Gender is not limited, age is 18 - 65 years old; those with long-term eye use and easy eye fatigue. Exclusion criteria: Those with internal and external eye diseases such as corneal, crystalline, vitreous, and fundus lesions; those with diseases of the cardiovascular, cerebrovascular, liver, kidney, hematopoietic systems, etc.; pregnant or lactating women, patients with allergic constitution; those who have taken items related to the tested function in the short term and affect the judgment of the results; those who can cooperate with the experiment; those whom the researcher deems unfit to participate in this research. Random grouping, with 63 people in each of the experimental group and the control group. The experimental group used the massage cream prepared in Application Example 1, and the control group used the massage cream prepared from the traditional Chinese medicine composition prepared in Comparative Example 8.

[0126] 1) Usage method: After cleaning the skin, first cover both eyes with a layer of gauze with a specification of 5×6 cm, then take an appropriate amount of the cream and apply it to the entire eye area and gently massage. Then cover it with a layer of plastic wrap, and then cover it with two layers of gauze with a specification of 5×6 cm. After 15 minutes, remove the plastic wrap and gauze, and wipe the remaining cream with a wet cotton or wet towel;

[0127] 2) Improvement of discomfort symptoms: Symptom inquiry, including 8 symptoms related to eye distension, eye soreness, photophobia, blurred vision, eye dryness, foreign body sensation, tearing, and general discomfort related to visual fatigue. If 3 of them are improved and other symptoms do not deteriorate, it is determined that the symptoms are improved. Symptom improvement effective rate (%) = number of cases with symptom improvement / total number of cases × 100%;

[0128] 3) Measurement of the duration of clear vision: The percentage of the clear vision time to the fixation time is called the duration of clear vision. Draw a three-dimensional square pattern like the Chinese character "pin" on the checklist, with each side of the square being 1 cm long and the local illumination being 120 LX. The distance between the checklist and the eyes should be kept at an appropriate distance according to the viewing habit of the subject without moving. It is stipulated that when the subject sees the "pin" image, it is considered clear vision, and when it is an inverted "pin", it is unclear vision. The measurement time is 3 minutes, and the average value is taken after two measurements. Duration of clear vision = (Clear vision time / Total fixation time) × 100%;

[0129] 4) Statistical processing of experimental results: The experimental results are shown in Table 4 - 5. The experimental data are statistically processed using Microsoft Excel 2003, and the results are expressed as mean ± standard deviation. The statistical analysis uses the t - test, and P < 0.05 is considered statistically significant;

[0130] 5) Results: During the trial process, none of the patients showed symptoms such as skin itching, stinging, or redness and swelling. The experimental results are shown in Table 4 - 5.

[0131] Table 4 Improvement results of discomfort symptoms

[0132]

[0133] Table 5 Changes in the duration of clear vision before and after the trial

[0134] Group Before the test After the test Application example 1 56.49±8.76 <![CDATA[71.34±9.85* # > Control group 57.01±7.86 60.37±7.62

[0135] Note: Compared with before the trial: P < 0.05 is indicated by "*"; compared with the control example: P < 0.05 is indicated by " # ".

[0136] The improvement results of discomfort symptoms are shown in Table 4. The application example 1 group had obvious improvements in symptoms such as eye swelling, eye soreness, photophobia, blurred vision, dry eyes, foreign body sensation, tearing, and general discomfort related to visual fatigue, and its improvement effect was significantly better than that of the control group, indicating that the massage cream prepared by the present invention has a significant effect in relieving visual fatigue, and has the advantages of a short treatment course and quick effect; the results of the duration of clear vision are shown in Table 5. The binocular duration of clear vision of application example 1 before and after applying the product was significantly improved and the effect was also better than that of the control group, indicating that the fermentation process of this application significantly improved the release rate of active ingredients, thereby improving the therapeutic effect of the massage cream.

[0137] In summary, the massage cream prepared by the present invention has a significant effect in improving and relieving visual fatigue, has the advantages of a short treatment course and quick effect compared with unfermented traditional Chinese medicine, is safe and non - irritating, and has good application prospects.

[0138] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments with equivalent changes within the scope of the technical solution of the present invention. However, as long as it does not depart from the content of the technical solution of the present invention, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A method for preparing a Chinese medicinal fermentation composition for relieving visual fatigue, characterized in that: The following steps are involved: S1. Weigh each raw material in the Chinese medicine composition according to weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I; S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, the temperature is raised to perform enzyme inactivation treatment to obtain a mixed solution II; S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger with liquid culture medium, inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger into mixed solution II, and performing mixed fermentation; S4, filtering after the fermentation is completed, and concentrating the filtrate under reduced pressure to obtain a concentrated solution; the concentrated solution is sterilized at high temperature to obtain the Chinese medicine fermentation composition; Among them, the Lactobacillus rhamnosus is deposited in CGMCC 1.3724; the Saccharomyces cerevisiae is deposited in CGMCC2.1527; the Aspergillus niger is deposited in CGMCC 3.316; The Chinese medicine composition comprises the following raw materials in parts by weight: 3-15 parts of wolfberry, 1-10 parts of chrysanthemum, 6-20 parts of Xing'an cimicifuga, 1-10 parts of Ligustrum lucidum, 5-16 parts of Ligustrum lucidum, 5-16 parts of safflower, 1-10 parts of coptis chinensis, 1-10 parts of equisetum, 1-10 parts of gardenia, 1-10 parts of tribulus terrestris, 5-16 parts of dodder seeds, 6-18 parts of cassia seed, 1-10 parts of buddlejac, 1-10 parts of white peony root, and 1-10 parts of mulberry leaf.

2. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: The Chinese medicine composition comprises the following raw materials in parts by weight: 10 parts of wolfberry, 4 parts of chrysanthemum, 13 parts of Xing'an cimicifuga, 5 parts of Ligustrum lucidum, 10 parts of Ligustrum lucidum, 10 parts of safflower, 4 parts of coptis chinensis, 5 parts of equisetum, 5 parts of gardenia, 5 parts of tribulus terrestris, 11 parts of dodder seed, 12 parts of cassia seed, 4 parts of buddlejac, 4 parts of white peony root, and 4 parts of mulberry leaf.

3. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: In the step S1, the mesh number of the sieve is 200-300 mesh, and the solid-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:5-7.

4. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: In the step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme dosage is 3-5wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45-55°C, the pH is 4.5-5.5, the ultrasonic frequency is 25-35kHz, and the time is 40-50min; the temperature of the enzyme inactivation treatment is 90-100°C, and the time of the enzyme inactivation treatment is 20-30min.

5. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: In the step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1-2:1:1-1.5; and the total amount of bacteria added is 5-7% of the volume of the mixed solution II.

6. The method for preparing the Chinese medicinal fermentation composition according to claim 5, characterized in that: In step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.

2.

7. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: In step S3, the fermentation temperature is 30-34° C., the fermentation pH is 5.0-6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200-250 rpm, and the fermentation time is 48-72 h.

8. The method for preparing the Chinese medicinal fermentation composition according to claim 1, characterized in that: In the step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of the concentrated under reduced pressure is 40-50° C., the pressure of the concentrated under reduced pressure is -0.06 to -0.08 MPa; the sterilization temperature is 121° C., and the sterilization time is 20-30 min.

9. A massage cream for relieving visual fatigue, characterized in that: The massage cream comprises a Chinese medicinal fermentation composition prepared by the preparation method according to any one of claims 1 to 8.

10. The massage cream for relieving visual fatigue according to claim 9, characterized in that: The massage cream comprises the following raw materials in percentage by weight: 5.0-20.0% of Chinese medicinal fermentation composition, 6.5-8.5% of emulsifier, 3.0-8.0% of excipient, 8.0-13.0% of emollient, 0.18-0.68% of conditioner, 0.3-2.3% of suspending agent, 0.1-1.0% of covering agent, 10.5-14.5% of moisturizing agent and 0.05-0.15% of chelating agent.

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