A saccharomyces boulardii probiotic fermented composition, its preparation method and use

Abstract

The present application relates to a kind of mountain-flower crystal probiotic fermentation composition and its preparation method and application, belong to biological fermentation technical field.The mountain-flower crystal fermentation composition provided by the present application is selected to be fried hawthorn, chrysanthemum, medlar three main raw materials, after probiotic fermentation, can obviously improve the light-aversion, the condition of eye dryness, and relieve visual fatigue, treat dry eye, the effect of improving bright vision duration is significantly enhanced.

Description

Technical Field

[0001] This invention relates to a probiotic fermentation composition of wildflower crystals, its preparation method and application, belonging to the field of bio-fermentation technology. Background Technology

[0002] The eyes are the windows to the soul. In modern society, approximately 80% of the information we receive from the outside world is through our eyes. Therefore, eye health and clear vision are extremely important. With the increasing frequency of use of electronic products such as mobile phones and computers, and the growing severity of environmental pollution, the incidence of eye diseases such as eye strain, dry eye syndrome, and myopia is showing a continuous upward trend.

[0003] Dry eye syndrome, also known as keratoconjunctivitis sicca, is defined as a multifactorial ocular surface disease characterized primarily by tear film homeostasis imbalance, accompanied by ocular discomfort symptoms, according to the second edition of the Dry Eye Consensus series published by the International Tear Film and Ocular Surface Association Working Group. Symptoms of dry eye are diverse, including burning and stinging sensations, soreness, photophobia, transient blurred vision, conjunctival hyperemia, epiphora, and excessive ocular discharge. With the development of technology, people's reliance on video terminals such as televisions, computers, and smartphones is increasing, leading to a rising incidence of dry eye syndrome, which has become a global epidemic. Its prevalence is also increasingly affecting younger people. In my country, the incidence of dry eye syndrome has reached 21%–30%, and the demand for its treatment is increasing year by year.

[0004] Visual fatigue syndrome is a complex syndrome manifested in the eyes by a combination of local, systemic, and psychological factors. It presents with a wide variety of clinical manifestations. Current examination methods can only detect minor lesions or no lesions at all. Although it does not cause blindness, it brings inconvenience and suffering to patients' normal lives.

[0005] As modern medicine deepens its understanding of the eyes, numerous methods for restoring eye health have emerged, with integrated traditional Chinese and Western medicine treatments, as well as drug and non-drug therapies, achieving certain therapeutic effects. However, due to the numerous factors contributing to dry eye syndrome and its complex and variable pathogenesis, further research with greater human and material resources is needed.

[0006] Hawthorn is the dried, ripe fruit of the hawthorn or jujube tree, both belonging to the Rosaceae family. Stir-fried hawthorn is prepared by stir-frying hawthorn berries (without adding any auxiliary ingredients) until they darken in color after impurities and pits have been removed. It has a sour and sweet taste and is slightly warm in nature. The main chemical components of hawthorn include flavonoids, organic acids, triterpenoids, and proanthocyanidins, which have free radical scavenging, antioxidant, anti-cancer, and cancer-preventive effects. Modern research also indicates that it has a certain protective effect on the retina. Hawthorn is also rich in various vitamins and trace elements, including B vitamins, vitamin C, calcium, iron, and zinc; zinc, in particular, can promote the absorption of vitamin A, thus protecting the eyes.

[0007] Chrysanthemum, the dried flower head of the chrysanthemum plant (Chrysanthemum morifolium), belongs to the Asteraceae family. It has a bitter and sweet taste and is slightly cold in nature. It is rich in lutein, flavonoids, triterpenoids, and volatile oils, among other active ingredients. According to traditional Chinese medicine texts, it can be used to treat liver and kidney deficiency, malnourishment of the eyes, blurred vision, and difficulty seeing clearly. Modern medical science has proven that it has anti-diarrheal, anti-inflammatory, vision-improving, blood pressure-lowering, lipid-lowering, and body-strengthening effects.

[0008] Goji berries are the mature fruit of the Lycium barbarum or Lycium chinense plant, belonging to the Solanaceae family. They are sweet in taste and neutral in nature. Rich in polysaccharides, proteins, trace elements, vitamins, betaine, minerals, flavonoids, and other active ingredients, goji berries have a long history of use in traditional Chinese medicine. Traditional Chinese medicine believes they nourish the liver and kidneys, benefit essence and improve eyesight, and are used to treat weakness and deficiency of essence, lower back and knee pain, dizziness and tinnitus, internal heat and thirst, blood deficiency and chlorosis, and blurred vision. Modern medicine has proven that goji berries possess various biological activities, including anti-tumor, anti-aging, anti-fatigue, lipid-lowering, and immune-regulating effects.

[0009] Chinese patent CN103610832A discloses a hawthorn extract oral preparation and its preparation method, the main raw materials of which include 100-200 parts of roasted hawthorn, 50-150 parts of chrysanthemum, and 50-150 parts of wolfberry. The hawthorn extract oral preparation described in this patent undergoes an alcohol precipitation process during extraction, which leads to the loss of a considerable portion of its effective components and nutrients.

[0010] Chinese patent CN109832613A discloses a fermented chrysanthemum product and its preparation method, which is made by directly pulping chrysanthemum as a single medicinal material and then fermenting it. The fermented chrysanthemum product and its preparation method described in this patent focus on the retention of the effective components in chrysanthemum and the antioxidant and free radical scavenging capabilities of the fermented chrysanthemum product.

[0011] Currently, there are no reports on the preparation of fermented Shanhuajing using probiotics. Summary of the Invention

[0012] This invention provides a fermented composition of wildflower crystals, its preparation method, and its application. The fermented composition of wildflower crystals of this invention utilizes probiotic fermentation, altering the active substance composition of traditional wildflower crystal pharmaceutical compositions. Clinical trials have demonstrated significantly enhanced effects in nourishing the liver and kidneys, clearing the liver and improving eyesight, alleviating eye fatigue, treating dry eye syndrome, and improving visual acuity.

[0013] To achieve the above objectives, the present invention employs the following technical solution:

[0014] This invention provides a wildflower crystal fermentation composition, comprising a wildflower crystal composition and probiotics.

[0015] The mountain flower crystal composition, by weight, consists of 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry.

[0016] The mountain flower crystal fermentation composition is prepared by decocting and filtering stir-fried hawthorn, chrysanthemum, and wolfberry, and then adding probiotics for fermentation.

[0017] The probiotics include one or more of the following: Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus salivarius, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus helveticus, Streptococcus acidophilus, or Lactococcus.

[0018] Preferably, the probiotics are Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei.

[0019] The mass ratio of *Lactobacillus plantarum*, *Streptococcus acidophilus*, *Lactobacillus acidophilus*, *Lactococcus lactis*, and *Lactobacillus paracasei* is 1–3:1–3:1–3.

[0020] Preferably, the mass ratio of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei is 1:1:1:1:1.

[0021] This invention also provides a method for preparing a wildflower crystal fermentation composition, comprising the following steps:

[0022] (1) Preparation of Mountain Flower Crystal Solution

[0023] By weight, take 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry, add water and decoct, filter, pasteurize, and cool to 36-40℃ to obtain the original extract of hawthorn flower crystal.

[0024] (2) Fermentation

[0025] Add probiotics and sucrose to the mountain flower crystal stock solution described in step 1), stir well, and then ferment to obtain the mountain flower crystal fermentation composition.

[0026] Preferably, in step (1), 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry are decocted with water 2-3 times, each time for 1-5 hours, filtered, and the filtrates are combined. The mass-volume ratio of the hawthorn crystal composition to the hawthorn crystal stock solution obtained in step (1) is 1:8-30 g / mL. By weight, 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry are decocted with water, filtered, concentrated, cooled, ethanol is added, stirred evenly, precipitated with alcohol, the supernatant is taken, pasteurized, and cooled to 36-40℃ to obtain the hawthorn crystal ethanol precipitation stock solution.

[0027] Preferably, the total number of probiotic colonies added in step (2) is 1×10⁻⁶. 6 ~1×10 8 CFU / g; the amount of sucrose added is 5-20% of the weight of the original extract of the mountain flower crystal, the fermentation temperature is 35-45℃, and the fermentation cycle is 3-15 days.

[0028] The fermented composition of the wildflower crystal of the present invention can be made into oral dosage forms, including granules, tablets, soft capsules, pills, oral liquids, suspensions, emulsions, etc., by adding conventional excipients and using conventional pharmaceutical processes.

[0029] The fermentation composition of the wildflower crystal of the present invention can be made into solid or liquid preparations.

[0030] The conventional excipients include: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, and matrix. Fillers include: starch, pregelatinized starch, lactose, mannitol, chitosan, microcrystalline cellulose, sucrose, etc.; disintegrants include: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, croscarmellose, low-substituted hydroxypropyl cellulose, croscarmellose sodium, etc.; lubricants include: magnesium stearate, sodium lauryl sulfate, talc, silica, etc.; suspending agents include: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, etc.; binders include: starch paste, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; sweeteners include: sodium saccharin, aspartame, sucrose, cyclamate, glycyrrhetinic acid, etc.; flavoring agents include: sweeteners and various flavorings; preservatives include: parabens, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chlorethidium acetate, eucalyptus oil, etc.; matrix includes: PEG6000, PEG4000, insect wax, etc.

[0031] The present invention also provides the application of the above-mentioned mountain flower crystal fermentation composition in the preparation of pharmaceuticals, health foods or ordinary foods that relieve or improve eye fatigue.

[0032] This invention provides the application of the aforementioned mountain flower crystal fermentation composition in the preparation of pharmaceuticals, health foods, or ordinary foods that improve or treat dry eye syndrome.

[0033] This invention provides the application of the aforementioned mountain flower crystal fermentation composition in the preparation of pharmaceuticals, health foods, or ordinary foods that improve the persistence of vision.

[0034] The beneficial effects of this invention are as follows:

[0035] The fermented hawthorn, chrysanthemum and wolfberry provided by this invention are selected as the three main raw materials. After fermentation by probiotics, the composition of active substances in hawthorn is changed, which can significantly improve photophobia and dry eyes, and significantly enhance the effects of relieving eye fatigue, treating dry eye syndrome and improving the duration of clear vision.

[0036] All three main ingredients are food and medicine homologous, with high safety, and can be taken long-term as medicine, health products or food. Detailed Implementation

[0037] The following are specific embodiments of the present invention, which further describe the technical solution of the present invention. However, the scope of protection of the present invention is not limited to these embodiments. Any changes or equivalent substitutes that do not depart from the concept of the present invention are included within the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, and the raw materials and reagents used are commercially available from conventional markets. For example, the *Lactobacillus plantarum*, *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, *Lactobacillus casei*, *Lactobacillus salivarius*, *Bifidobacterium lactis*, *Bifidobacterium longum*, *Streptococcus acidophilus*, *Bifidobacterium infantis*, and *Bifidobacterium breve* used in the embodiments were purchased from Runying Biotechnology (Shanghai) Co., Ltd.; *Bifidobacterium bifidum*, *Lactobacillus paracasei*, *Lactobacillus reuteri*, *Lactobacillus helveticus*, *Lactococcus lactis*, *Lactobacillus gasseri*, and *Streptococcus thermophilus* were purchased from Hebei Yiran Biotechnology Co., Ltd.; and *Bifidobacterium adolescentis* was purchased from Zhenjiang Tianyi Biotechnology Co., Ltd.

[0038] Example 1: Mountain Flower Crystal Probiotic Fermentation Composition

[0039] (1) Preparation of Mountain Flower Crystal Solution

[0040] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0041] (2) Fermentation

[0042] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0043] (3) Filling

[0044] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0045] Example 2: Mountain Flower Crystal Probiotic Fermentation Composition

[0046] (1) Preparation of Mountain Flower Crystal Solution

[0047] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct 3 times, 1 hour each time, filter, combine the filtrates, adjust the final volume to 9000 mL, pasteurize, and cool to 38°C to obtain hawthorn crystal stock solution.

[0048] (2) Fermentation

[0049] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 2:3:1:3:3. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add 20% sucrose (by weight of the original Shanhuajing liquid); stir well, and let it ferment at 35℃ for 15 days to obtain Shanhuajing fermentation liquid.

[0050] (3) Filling

[0051] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0052] Example 3: Mountain Flower Crystal Probiotic Fermentation Composition

[0053] (1) Preparation of Mountain Flower Crystal Solution

[0054] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 5 hours each time, filter, combine the filtrates, adjust the final volume to 2400 mL, pasteurize, and cool to 38°C to obtain hawthorn crystal stock solution.

[0055] (2) Fermentation

[0056] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 3:2:2:1:2. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add 5% sucrose (by weight of the original Shanhuajing liquid); stir well, and let it ferment at 45℃ for 3 days to obtain Shanhuajing fermentation liquid.

[0057] (3) Filling

[0058] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0059] Example 4: Mountain Flower Crystal Probiotic Fermentation Composition

[0060] (1) Preparation of Mountain Flower Crystal Solution

[0061] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0062] (2) Fermentation

[0063] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, Lactobacillus paracasei, Lactobacillus rhamnosus, and Bifidobacterium lactis, with a mass ratio of 1:1:1:1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0064] (3) Filling

[0065] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0066] Example 5: Mountain Flower Crystal Probiotic Fermentation Composition

[0067] (1) Preparation of Mountain Flower Crystal Solution

[0068] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0069] (2) Fermentation

[0070] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Bifidobacterium lactis, with a mass ratio of 1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0071] (3) Filling

[0072] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0073] Example 6: Mountain Flower Crystal Probiotic Fermentation Composition

[0074] (1) Preparation of Mountain Flower Crystal Solution

[0075] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0076] (2) Fermentation

[0077] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, and Lactobacillus acidophilus, added in a mass ratio of 1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0078] (3) Filling

[0079] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0080] Example 7: Mountain Flower Crystal Probiotic Fermentation Composition

[0081] (1) Preparation of Mountain Flower Crystal Solution

[0082] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0083] (2) Fermentation

[0084] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus casei, Lactobacillus salivarius, Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus reuteri, and Lactobacillus helveticus, with a mass ratio of 1:1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0085] (3) Filling

[0086] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0087] Example 8: Mountain Flower Crystal Probiotic Fermentation Composition

[0088] (1) Preparation of Mountain Flower Crystal Solution

[0089] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, concentrate to a certain volume, cool, add 3 times the amount of ethanol, stir well, precipitate with alcohol, take the supernatant, recover the ethanol at 65℃, adjust the final volume to 5000mL, pasteurize, cool to 38℃, and obtain the original ethanol-precipitated hawthorn crystal solution.

[0090] (2) Fermentation

[0091] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a ratio of 5-20% of the original Shanhuajing liquid; stir well, and let it ferment at 35-45℃ for 3-15 days to obtain Shanhuajing fermentation liquid.

[0092] (3) Filling

[0093] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0094] Comparative Example 1: Mountain Flower Crystal Composition

[0095] (1) Preparation of Mountain Flower Crystal Solution

[0096] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0097] (2) Mixing

[0098] Take the mountain flower crystal stock solution obtained in step (1), add sucrose at a weight ratio of 12% of the mountain flower crystal stock solution, stir evenly, and obtain the mountain flower crystal composition product.

[0099] 3) Filling

[0100] The mountain flower crystal composition obtained in step (2) is bottled and filled to obtain oral liquid.

[0101] Comparative Example 2: Mountain Flower Crystal Probiotic Fermentation Composition

[0102] (1) Preparation of Mountain Flower Crystal Solution

[0103] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0104] (2) Fermentation

[0105] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 30℃ for 19 days to obtain Shanhuajing fermentation liquid.

[0106] (3) Filling

[0107] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0108] Comparative Example 3: Mountain Flower Crystal Probiotic Fermentation Composition

[0109] (1) Preparation of Mountain Flower Crystal Solution

[0110] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0111] (2) Fermentation

[0112] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus paracasei, with a mass ratio of 1:5:4:3:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0113] (3) Filling

[0114] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0115] Comparative Example 4: Mountain Flower Crystal Probiotic Fermentation Composition

[0116] (1) Preparation of Mountain Flower Crystal Solution

[0117] By weight, take 150 parts of roasted hawthorn, 75 parts of chrysanthemum, and 75 parts of wolfberry, add water and decoct twice, 3 hours each time, filter, combine the filtrates, adjust the final volume to 5000mL, pasteurize, and cool to 38℃ to obtain hawthorn crystal stock solution.

[0118] (2) Fermentation

[0119] Add a compound bacterial strain to the mountain flower crystal stock solution described in step (1). The compound bacterial strain consists of Bifidobacterium adolescentis, Bifidobacterium infantis, Bifidobacterium breve, Streptococcus thermophilus, and Lactobacillus gasseri, with a mass ratio of 1:1:1:1:1. The total number of compound bacterial colonies is less than 10. 6 ~10 8 Between CFU / g; add sucrose at a weight ratio of 12% of the original Shanhuajing liquid; stir well, and let it ferment at 40℃ for 9 days to obtain Shanhuajing fermentation liquid.

[0120] (3) Filling

[0121] The fermented composition of wildflower crystals obtained in step (2) was bottled and bottled to obtain oral liquid.

[0122] Experimental Example 1: Effect of the fermented composition of the present invention on a dry eye model

[0123] 1. Model preparation and grouping

[0124] Eighty-six SPF-grade male BALB / c mice, weighing between 18 and 20 g, were selected and acclimatized for one week. Six mice were randomly selected as the normal control group, while the other 80 mice were given 5 μL of 0.2% benzalkonium chloride solution in both eyes to establish a mouse model of dry eye. The solution was administered three times daily for seven consecutive days. The success of the model was observed and evaluated using fluorescein staining.

[0125] Except for the normal group (physiological saline), 66 mice that successfully developed the model were randomly selected and divided into 11 groups. The grouping and drug dosage are as follows:

[0126] Normal group (physiological saline), model control group (physiological saline), groups 1-6 (corresponding to the fermented compositions of *Gynostemma pentaphyllum* in Examples 1, 2, 4, 5, 6, and 7 above, respectively), and groups 7-10 (corresponding to comparative examples 1-4 above, respectively).

[0127] The dosage of the drug in groups 1-10 was 24g / kg. The normal group and the model control group were given the same volume of physiological saline. The mice in each group were continuously administered the drug by gavage for 28 days.

[0128] 2. Indicator Measurement

[0129] 2.1 Measurement of tear secretion

[0130] According to the reference method (Wu SQ, Dai Q, Xu JL, et al. Combined effect of traditional Chinese and Western medicine on inflammatory factors in patients with diabetes-induced xerophthalmia[J]. Genetics and Molecular Research, 2016, 15(4): 15-17.), the phenol red cotton thread soaking method was used to detect the amount of tear secretion in mice before and after drug administration, and the length of the cotton thread that turned red was measured and statistical analysis was performed.

[0131] 2.2 Determination of tear film breakup time

[0132] The tear film breakup time of mice in each group before and after drug administration was detected by fluorescein staining method (KIM J,KIM JY,SEO KY, et al. Location and pattern of non-invasive keratographic tear film breakup according to dry eye disease subtypes[J].Acta Ophthalmologica,2019,97(10):324-331.), and the data were recorded and statistically analyzed.

[0133] 3. Test Results

[0134] 3.1 The effects of each group on tear secretion are shown in Table 1.

[0135] Compared with the normal group, the model group showed a highly significant difference in the length of the phenol red cotton thread that turned red, indicating that the model was successfully established.

[0136] Compared with the model control group, the groups in the examples and the comparative examples showed significant or highly significant differences in the length of time the phenol red cotton thread turned red, indicating that all groups could increase tear secretion. However, groups 1-6, especially group 1, showed better results.

[0137] Compared with Group 7 (Comparative Example 1), there were statistically significant differences in the length of the phenol red cotton thread that turned red in each of the groups 1-6 and 8-9 after fermentation of the Shanhuajing composition. This indicates that Shanhuajing kombucha fermentation has a better effect on increasing tear secretion. However, fermentation alone cannot increase tear secretion. Targeted selection of the strains is still needed to achieve the desired effect of increasing tear secretion.

[0138] Table 1. Effects of each group on tear secretion.

[0139]

[0140]

[0141] # Compared with the normal control group, # P<0.05, ## P<0.01, ## P<0.001;

[0142] * Compared with the model control group, * P<0.05, ** P<0.01, *** P<0.001;

[0143] a Compared with group 7, a P<0.05, aa P<0.01, aaa P<0.001;

[0144] 3.2 The effects of each group on tear film breakup time are shown in Table 2.

[0145] Compared with the normal group, the tear film breakup time in the model group showed a highly significant difference, indicating that the model was successfully established.

[0146] Compared with the model control group, the tear film breakup time was significantly or extremely significantly prolonged in the example groups and the comparative groups, indicating that all groups could prolong the tear film breakup time. However, groups 1-6, especially groups 1-3, showed better results.

[0147] Compared with Group 7 (Comparative Example 1), the prolongation of tear film breakup time in Groups 1-6 and 8 after fermentation of the Shanhuajing composition showed significant or highly significant differences, indicating that the probiotic fermentation of Shanhuajing had a better effect on prolonging tear film breakup time. However, probiotic fermentation alone cannot prolong tear film breakup time. It is also necessary to make targeted selections of bacterial strains and their proportions to achieve the effect of increasing tear secretion.

[0148] Table 2. Effects of each group on tear film breakup time

[0149]

[0150] # Compared with the normal control group, # P<0.05, ## P<0.01, ## P<0.001;

[0151] *Compared with the model control group, * P < 0.05, ** P < 0.01, *** P < 0.001;

[0152] a Compared with Group 7, a P < 0.05, aa P < 0.01, aaa P < 0.001;

[0153] The experimental results show that the fermented composition of Shanhuajing of the present invention can significantly increase the tear secretion volume and prolong the tear film break-up time, and has a significant therapeutic effect on dry eye disease.

[0154] Experimental Example 2 Visual Persistence Experiment of the Fermented Composition of Shanhuajing of the Present Invention

[0155] 1. Steps of the Trial Administration Experiment

[0156] (1) Conduct relevant questionnaires for eligible volunteers for preliminary screening.

[0157] (2) Volunteers need to undergo a visual persistence test. Volunteers with a visual persistence ≤ 90% can participate in the trial administration experiment of the fermented composition of Shanhuajing and are determined as subjects.

[0158] (3) The subjects start taking the fermented composition of Shanhuajing prepared in Example 1, three times a day, one bottle each time (150 mL / day), and one treatment course is 10 days.

[0159] (4) After the subjects take three treatment courses, conduct a visual persistence test.

[0160] 2. Steps of the Visual Persistence Test

[0161] Draw a three-dimensional square pattern in the shape of a "pin" on the checklist. Each side of the square is 1 cm long, and the local illumination is 100 LX - 150 LX (a specially made light box can be used). During the measurement, the distance between the checklist and the eyes should be kept at an appropriate distance according to the subject's viewing habit 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 considered unclear vision. The measurement time is 3 minutes.

[0162] During the inspection, let the subject hold a stopwatch that can time intermittently. After the examiner gives the start command, the subject immediately focuses on the pattern in the square (or turns on the light box switch), and at the same time starts the stopwatch in hand to time. When seeing an inverted "pin" during the focusing process, immediately press the pause switch of the stopwatch; when seeing the "pin" image again, start the stopwatch again, and repeat this process. When the subject hears the examiner's command at the end of the specified time of 3 minutes, immediately stop the stopwatch. The reading passed by the stopwatch during this period is the total time for the subject to see the "pin" image, that is, the clear vision time.

[0163] Visual persistence = (visual time / total fixation time) × 100%.

[0164] During the measurement, attention should be paid to the venue and lighting, as well as the subject's level of eye use before the test. These factors should be considered before the experiment.

[0165] 3. The experimental results are shown in Table 3.

[0166] Table 3. Results of the visual persistence test of the mountain flower crystal fermentation composition of the present invention.

[0167]

[0168]

[0169] The experimental results show that among 20 participants, 16 experienced varying degrees of improvement in visual acuity (improvement percentage greater than 10%), resulting in an effectiveness rate of 80.0%. The experimental results indicate that the fermented composition of *Gynostemma pentaphyllum* of this invention has a certain effect on improving visual acuity in individuals with visual fatigue.

[0170] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A fermentation composition for wildflower crystals, characterized in that, The fermented hawthorn syrup composition is made from hawthorn syrup composition and probiotics. The hawthorn syrup composition, by weight, consists of 130-170 parts roasted hawthorn, 50-100 parts chrysanthemum, and 50-100 parts wolfberry. The probiotics include Lactobacillus plantarum, Streptococcus acidophilus, and Lactobacillus acidophilus, with a mass ratio of 1-3:1-3:1-3. The fermented hawthorn syrup composition is prepared by decocting and filtering roasted hawthorn, chrysanthemum, and wolfberry, and then adding probiotics for fermentation.

2. The fermented composition of wildflower crystals as described in claim 1, characterized in that, The probiotics are Lactobacillus plantarum, Streptococcus acidophilus, Lactobacillus acidophilus, Lactococcus paracasei, and Lactobacillus paracasei.

3. The fermented composition of wildflower crystals as described in claim 2, characterized in that, The mass ratio of *Lactobacillus plantarum*, *Streptococcus acidophilus*, *Lactobacillus acidophilus*, *Lactococcus*, and *Lactobacillus paracasei* added is 1:1:1:1:

1.

4. The method for preparing the wildflower crystal fermentation composition as described in claim 1, comprising the following steps: (1) Preparation of Mountain Flower Crystal Original Liquid: By weight, take 130-170 parts of stir-fried hawthorn, 50-100 parts of chrysanthemum and 50-100 parts of wolfberry, add water and decoct, filter, pasteurize and cool to 36-40℃ to obtain Mountain Flower Crystal Original Liquid. (2) Fermentation: Add probiotics and sucrose to the mountain flower crystal stock solution described in step (1), stir evenly, and then ferment to obtain the mountain flower crystal fermentation composition.

5. The method for preparing the wildflower crystal fermentation composition as described in claim 4, characterized in that, Step (1) Take 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry, add water and decoct 2-3 times, 1-5 hours each time, filter, combine the filtrates, the mass-volume ratio of the hawthorn crystal composition to the hawthorn crystal stock solution obtained in step (1) is 1:8-30 g / mL; by weight, take 130-170 parts of roasted hawthorn, 50-100 parts of chrysanthemum, and 50-100 parts of wolfberry, add water and decoct, filter and concentrate, cool, add ethanol, stir evenly, precipitate with alcohol, take the supernatant, pasteurize, cool to 36-40℃, and obtain the hawthorn crystal alcohol precipitation stock solution.

6. The method for preparing the wildflower crystal fermentation composition as described in claim 4, characterized in that, The total number of probiotic colonies added in step (2) is 1×10⁻⁶. 6 ~1×10 8 CFU / g; the amount of sucrose added is 5-20% of the weight of the original extract of the mountain flower crystal, the fermentation temperature is 35-45℃, and the fermentation cycle is 3-15 days.

7. The use of the wildflower crystal fermentation composition according to any one of claims 1-3 in the preparation of health foods or medicines that relieve or improve visual fatigue.

8. The use of the wildflower crystal fermentation composition according to any one of claims 1-3 in the preparation of a medicament for treating dry eye or improving visual persistence.

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