An oral liquid for improving sleep disorders and a preparation method thereof

Abstract

The application belongs to the technical field of medicine and health care, and particularly relates to an oral liquid for improving sleep disorders and a preparation method thereof, which is prepared from the following components in parts by weight: 100-120 parts of cereal soup, 0.05-0.07 parts of lactobacillus plantarum L9 inoculum, 0.03-0.05 parts of black tea fungus liquid, 0.03-0.05 parts of acetic acid bacteria, 0.02-0.04 parts of angel yeast SY, 0.02-0.04 parts of angel yeast RW and 5-10 parts of white granulated sugar. The raw materials are Shanbei millet, Chinese wingnut and grape seeds, and based on the microbial fermentation technology, the intestinal flora is adjusted, and the neurotransmitters and beneficial products for improving sleep disorders are enriched.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical and health care technology, specifically to an oral liquid for improving sleep disorders and its preparation method. Background Technology

[0002] Sleep disorders refer to abnormal sleep volume and abnormal behaviors during sleep, and are also phenomena of disruption of the normal sleep-wake rhythm. There are many types of sleep disorders. In 2017, the American Academy of Sleep Medicine published the "Medical Classification of Sleep Disorders, Third Edition," which classifies sleep disorders into seven main categories, including insomnia, sleep-related breathing disorders, central narcolepsy, circadian rhythm-wake disorders, sleep paradoxes, sleep-related motor disorders, and other sleep disorders.

[0003] Sleep is affected by a variety of factors. Sleep disorders can coexist with a variety of diseases and are a potential risk factor for inducing diseases. They can cause a variety of adverse consequences, such as anxiety, cardiovascular diseases, migraines, cognitive impairment, etc., which can harm physical health and easily lead to various safety accidents.

[0004] Current clinical interventions for sleep disorders remain quite limited. Drug treatment for chronic sleep disorders has various adverse reactions and is not suitable as a long-term treatment. In recent years, some scholars have proposed psychological and behavioral interventions supplemented with short-term drug treatment; however, this requires long-term patient cooperation and is costly. Furthermore, there is still a lack of sufficient and accurate understanding of sleep disorders, and misconceptions exist about sleep health. Currently, most products for improving sleep disorders are hormonal products such as melatonin tablets and benzodiazepines. Chemical drugs, such as those containing chemicals, can easily lead to adverse reactions such as endocrine disorders and dependence. Therefore, researching a natural and healthy product to improve sleep disorders has great social significance and value. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides an oral liquid for improving sleep disorders and a method for preparing the same.

[0006] An oral liquid for improving sleep disorders is prepared from the following components in parts by weight: 100-120 parts cereal soup, 0.05-0.07 parts Lactobacillus plantarum L9, 0.03-0.05 parts kombucha liquid, 0.03-0.05 parts acetic acid bacteria, 0.02-0.04 parts Angel yeast SY, 0.02-0.04 parts Angel yeast RW, and 5-10 parts white sugar.

[0007] Preferably, the method for preparing the grain soup is as follows: take 4-6 parts of millet material, 2-4 parts of jujube powder, 1-2 parts of grape seed powder, add 100-120 parts of distilled water, simmer over low heat for 10-15 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0008] Preferably, the method for obtaining the millet material is as follows: take millet, germinate it for 65-72 hours, and obtain the millet material.

[0009] Preferably, the method for obtaining Lactobacillus plantarum L9 is as follows: Take 4-6 parts of millet material, 2-4 parts of jujube powder, and 1-2 parts of grape seed powder, add 100-120 parts of distilled water, simmer for 10-15 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate with 5-10 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 7-9 days to obtain Lactobacillus plantarum L9.

[0010] Preferably, the kombucha liquid is prepared from black tea, green tea, white sugar, water, and kombucha stock solution.

[0011] Preferably, the preparation method of the kombucha liquid is as follows: 0.2-0.5 parts of black tea, 0.2-0.5 parts of green tea, 3-6 parts of white sugar and 100-150 parts of water are mixed and boiled, cooled to obtain mixture one, 20-30 parts of kombucha stock solution are added to mixture one, and fermentation is carried out to obtain kombucha liquid.

[0012] Preferably, the simmering time is 10-15 minutes, and the fermentation conditions are 7-15 days at 24-30℃.

[0013] The method for preparing an oral liquid for improving sleep disorders includes the following steps:

[0014] (8.1) Take 4-6 parts of millet material, 2-4 parts of jujube powder, 1-2 parts of grape seed powder, add 100-120 parts of distilled water, simmer over low heat for 10-15 minutes, transfer to a sterile culture bottle, seal and let cool to obtain grain soup.

[0015] Take 4-6 parts of millet material, 2-4 parts of jujube powder, and 1-2 parts of grape seed powder, add 100-120 parts of distilled water, simmer for 10-15 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate with 5-10 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 7-9 days to obtain Lactobacillus plantarum L9;

[0016] Mix 0.2-0.5 parts black tea, 0.2-0.5 parts green tea, 3-6 parts white sugar and 100-150 parts water and boil until boiling. Let it cool to obtain mixture one. Add 20-30 parts kombucha stock solution to mixture one and ferment to obtain kombucha liquid.

[0017] (8.2) Add 0.05-0.07 parts of Lactobacillus plantarum L9, 0.03-0.05 parts of kombucha liquid, 0.03-0.05 parts of acetic acid bacteria, 0.02-0.04 parts of Angel yeast SY and 0.02-0.04 parts of Angel yeast RW to 4-6 parts of cereal soup, mix well, and perform anaerobic fermentation to obtain fermentation liquid;

[0018] (8.3) Add 5-10 parts of white sugar to the fermentation broth, mix well, sterilize, and obtain the oral liquid.

[0019] Preferably, in (8.2), when preparing the fermentation broth, anaerobic fermentation is carried out at 37°C for 7-9 days.

[0020] Preferably, the sterilization temperature in (8.3) is 121°C.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] Northern Shaanxi millet: A specialty organic crop of Northern Shaanxi. Northern Shaanxi has a semi-arid continental monsoon climate with steep terrain, deep valleys, abundant sunshine and heat resources, and large temperature differences between day and night. This environment is very suitable for the growth of millet, which is conducive to the accumulation of nutrients in millet. As a result, millet is rich in protein, vitamins, minerals, and γ-aminobutyric acid, and has functions such as lowering blood pressure, calming the nerves and aiding sleep.

[0023] Yan'an jujube: A wild variety from the Loess Plateau, pollution-free and harmless, it is a specialty of Yan'an City, Shaanxi Province, and a Chinese National Geographical Indication product. Yan'an jujube has the highest concentration of active ingredients among Chinese jujube varieties. The kernel contains activated charcoal, fatty oils, and other medicinal components, with a total saponin content of 1.1353%, ranking among the top in the country. The fruit is rich in vitamin C, as well as abundant fatty oils, proteins, organic acids, saponins, two types of sterols, and various triterpenoids. Moderate consumption can nourish the heart and calm the nerves, aid digestion, and is a natural dried fruit with both medicinal and edible uses.

[0024] Grape seeds: Grape seeds are also rich in linoleic acid, linolenic acid, proanthocyanidins, and vitamins, which have the effects of preventing cardiovascular diseases, anti-oxidation, scavenging free radicals, and enhancing immunity.

[0025] The strains described in this invention include:

[0026] Lactobacillus plantarum L9: It inhibits the growth of putrefactive bacteria and the production of putrefactive products in the intestine, and regulates the quantity of normal gut flora. For example, sleep disorders such as short sleep duration, low sleep efficiency, and fragmented sleep are related to the abundance of Firmicutes and Bacteroides in the intestine. Lactobacillus plantarum is also a safe and effective strain for producing and enriching GABA, and has significant advantages in improving sleep disorders.

[0027] Kombucha is rich in acetic acid bacteria, mainly acetic acid bacteria, glucosamine, lactic acid bacteria and yeast. These microorganisms can help produce a healthy microbiome and restore the dynamic balance in the intestines. In addition, kombucha also contains organic acids, phenolic compounds, trace nutrients, vitamin C and vitamin B complex, theanine and other nutrients, which have antimicrobial, antioxidant, antiproliferative and anticancer biological activities.

[0028] Acetic acid bacteria are Gram-negative bacteria that can oxidize ethanol to produce acetic acid. These bacteria originate from traditional vinegar-making fermentation products. Their metabolic products contain cellulose, which promotes growth when cultured in symbiosis with yeast. The acetic acid they produce can cleanse the intestines and aid digestion.

[0029] Angel Yeast: Rich in B vitamins, it nourishes nerves and improves sleep. It also regulates the body's pH balance to some extent, enhancing immunity and disease resistance.

[0030] The beneficial effects of this invention include: using Shaanxi millet, jujube and grape seeds as raw materials, based on microbial fermentation technology, regulating intestinal flora, and enriching neurotransmitters and beneficial products that improve sleep disorders.

[0031] The main raw material of this product is natural agricultural products from northern Shaanxi. It has the characteristics of low economic cost, high social benefits, no additives, no dependence, and no side effects. It is suitable for long-term use by patients with sleep disorders, promoting physical health and improving sleep disorders in a subtle and gentle way. Attached Figure Description

[0032] Figure 1 The effect of the compound oral solution on sleep behavior in PCPA-induced sleep deprivation mice is shown in Figure 1, where A represents sleep latency and B represents sleep duration.

[0033] Figure 2 The effect of the compound oral liquid on the emotional behavior of PCPA-induced sleep deprivation mice was investigated, where A represents the number of times the mice crossed the central grid; B represents the number of times the mice groomed themselves; and C represents the number of times the mice defecated.

[0034] Significant difference in the normal control group: **P < 0.01; Significant difference in the model group: #P < 0.05, ##P < 0.01;

[0035] Figure 3 The effects of the compound oral solution on serum inflammatory factors in PCPA-induced sleep deprivation mice are shown in Figure A, where changes in IL-6 levels, B, and C represent changes in IL-1β levels, and C represents changes in TNF-α levels.

[0036] Significant difference in the normal control group: *P<0.05; Significant difference in the model group: #P<0.05;

[0037] Figure 4 The effects of the compound oral liquid on hypothalamic neurotransmitters in PCPA-induced sleep deprivation mice are shown in Figure A, where A represents changes in 5-HT content, B represents changes in GABA content, and C represents changes in Glu content.

[0038] Significant difference in the normal control group: **P<0.01; Significant difference in the model group: #P<0.05, ##P<0.01, ###P<0.001. Detailed Implementation

[0039] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods.

[0040] The millet used in this invention is Shaanxi millet. The Lactobacillus plantarum L9 culture medium is preserved at the China Center for Type Culture Collection (accession number cctcc no: m 2022169). The jujube powder was purchased from Shanxi Shude Agricultural Technology Co., Ltd., the grape seed powder was purchased from Danyang Xunweixuan Food Manufacturing Co., Ltd., the black tea is Lapsang Souchong, the green tea is Anji white tea, the kombucha stock solution is from Shandong Zhishi Weibao Biotechnology Co., Ltd., the acetic acid bacteria were purchased from Jining Yuyuan Biotechnology Co., Ltd., the Angel Yeast SY was purchased from Angel Yeast Co., Ltd., the Angel Yeast RW was purchased from Angel Yeast Co., Ltd., and the Bacillus licheniformis live bacteria capsules were purchased from Northeast Pharmaceutical Group Shenyang First Pharmaceutical Co., Ltd.

[0041] Lactobacillus plantarum L9, classified as Lactobacillus plantarum, was deposited at the China Center for Type Culture Collection on February 28, 2022, with accession number CCTCC No. M2022169.

[0042] Example 1

[0043] An oral liquid for improving sleep disorders is prepared from the following components in parts by weight: 100 parts cereal soup, 0.05 parts Lactobacillus plantarum L9 inoculum, 0.03 parts kombucha liquid, 0.03 parts acetic acid bacteria, 0.02 parts Angel yeast SY, 0.02 parts Angel yeast RW, and 5 parts white sugar.

[0044] The method for preparing the grain soup is as follows: Take 4 parts of millet material, 2 parts of jujube powder, 1 part of grape seed powder, add 100 parts of distilled water, simmer over low heat for 10 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0045] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 65 hours at a germination temperature of 27°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0046] The method for obtaining the Lactobacillus plantarum L9 inoculum is as follows: Take 4 parts of millet material, 2 parts of jujube powder, and 1 part of grape seed powder, add 100 parts of distilled water, simmer for 10 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate 5 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 7 days to obtain Lactobacillus plantarum L9 inoculum.

[0047] The method for preparing the kombucha liquid is as follows: Mix 0.2 parts black tea, 0.2 parts green tea, 3 parts white sugar and 100 parts water, boil for 10 minutes until boiling, let cool to obtain mixture one, add 20 parts kombucha stock solution to mixture one, and ferment at 24℃ for 7 days to obtain kombucha liquid.

[0048] A method for preparing an oral liquid for improving sleep disorders includes the following steps:

[0049] (1) Take 100 parts of the above-prepared grain soup, add 0.05 parts of Lactobacillus plantarum L9 inoculant, 0.03 parts of kombucha liquid, 0.03 parts of acetic acid bacteria, 0.02 parts of Angel yeast SY and 0.02 parts of Angel yeast RW, mix well, and ferment under anaerobic conditions at 37℃ for 7 days to obtain fermentation liquid;

[0050] (2) Add 5 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0051] Example 2

[0052] An oral liquid for improving sleep disorders is prepared from the following components in parts by weight: 110 parts cereal soup, 0.06 parts Lactobacillus plantarum L9 inoculum, 0.04 parts kombucha liquid, 0.04 parts acetic acid bacteria, 0.03 parts Angel yeast SY, 0.03 parts Angel yeast RW, and 7 parts white sugar.

[0053] The method for preparing the grain soup is as follows: Take 5 parts of millet material, 3 parts of jujube powder, 2 parts of grape seed powder, add 110 parts of distilled water, simmer over low heat for 13 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0054] The method for obtaining the millet material is as follows: take millet, clean it, spread it evenly in the cultivation grid of the germination machine, germinate for 70 hours at a germination temperature of 25°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0055] The method for obtaining the Lactobacillus plantarum L9 inoculum is as follows: Take 5 parts of millet material, 3 parts of jujube powder, and 1 part of grape seed powder, add 110 parts of distilled water, simmer for 13 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate 7 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 8 days to obtain Lactobacillus plantarum L9 inoculum.

[0056] The method for preparing the kombucha liquid is as follows: Mix 0.3 parts black tea, 0.3 parts green tea, 4 parts white sugar and 120 parts water, boil for 13 minutes until boiling, let cool to obtain mixture one, add 25 parts kombucha stock solution to mixture one, and ferment at 24℃ for 10 days to obtain kombucha liquid.

[0057] A method for preparing an oral liquid for improving sleep disorders includes the following steps:

[0058] (1) Take 110 parts of the above-prepared grain soup, add 0.06 parts of Lactobacillus plantarum L9 inoculum, 0.04 parts of kombucha liquid, 0.04 parts of acetic acid bacteria, 0.03 parts of Angel yeast SY and 0.03 parts of Angel yeast RW, mix well, and ferment under aseptic conditions at 37°C for 8 days to obtain fermentation liquid;

[0059] (2) Add 7 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0060] Example 3

[0061] An oral liquid for improving sleep disorders is prepared from the following components in parts by weight: 120 parts cereal soup, 0.07 parts Lactobacillus plantarum L9 inoculum, 0.05 parts kombucha liquid, 0.05 parts acetic acid bacteria, 0.04 parts Angel yeast SY, 0.04 parts Angel yeast RW, and 10 parts white sugar.

[0062] The method for preparing the grain soup is as follows: Take 6 parts of millet material, 4 parts of jujube powder, 2 parts of grape seed powder, add 120 parts of distilled water, simmer over low heat for 15 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0063] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 72 hours at a germination temperature of 28°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0064] The method for obtaining the Lactobacillus plantarum L9 inoculum is as follows: Take 6 parts of millet material, 4 parts of jujube powder, and 2 parts of grape seed powder, add 120 parts of distilled water, simmer for 15 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate with 10 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 9 days to obtain Lactobacillus plantarum L9 inoculum.

[0065] The method for preparing the kombucha liquid is as follows: Mix 0.5 parts black tea, 0.5 parts green tea, 6 parts white sugar and 150 parts water, boil for 15 minutes until boiling, let cool to obtain mixture one, add 30 parts kombucha stock solution to mixture one, and ferment at 24℃ for 15 days to obtain kombucha liquid.

[0066] A method for preparing an oral liquid for improving sleep disorders includes the following steps:

[0067] (1) Take 120 parts of the above-prepared grain soup, add 0.07 parts of Lactobacillus plantarum L9 inoculant, 0.05 parts of kombucha liquid, 0.05 parts of acetic acid bacteria, 0.04 parts of Angel yeast SY and 0.04 parts of Angel yeast RW, mix well, and ferment under aseptic conditions at 37°C for 9 days to obtain fermentation liquid;

[0068] (2) Add 10 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0069] Example 4

[0070] An oral liquid for improving sleep disorders is prepared from the following components in parts by weight: 100 parts cereal soup, 0.05 parts Lactobacillus plantarum L9 inoculum, 0.03 parts kombucha liquid, 0.03 parts acetic acid bacteria, 0.02 parts Angel yeast SY, 0.02 parts Angel yeast RW, and 5 parts white sugar.

[0071] The method for preparing the grain soup is as follows: Take 4 parts of millet material, 4 parts of jujube powder, 2 parts of grape seed powder, add 120 parts of distilled water, simmer over low heat for 15 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0072] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 68 hours at a germination temperature of 30°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0073] The method for obtaining the Lactobacillus plantarum L9 inoculum is as follows: Take 6 parts of millet material, 2 parts of jujube powder, and 1 part of grape seed powder, add 100 parts of distilled water, simmer for 10 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate with 8 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 7 days to obtain Lactobacillus plantarum L9 inoculum.

[0074] The method for preparing the kombucha liquid is as follows: Mix 0.4 parts black tea, 0.4 parts green tea, 5 parts white sugar and 130 parts water, boil for 14 minutes until boiling, let cool to obtain mixture one, add 23 parts kombucha stock solution to mixture one, and ferment at 24℃ for 12 days to obtain kombucha liquid.

[0075] A method for preparing an oral liquid for improving sleep disorders includes the following steps:

[0076] (1) Take 100 parts of the above-prepared grain soup, add 0.05 parts of Lactobacillus plantarum L9 inoculum, 0.03 parts of kombucha liquid, 0.03 parts of acetic acid bacteria, 0.02 parts of Angel yeast SY and 0.02 parts of Angel yeast RW, mix well, and ferment under aseptic conditions at 37°C for 7 days to obtain fermentation liquid;

[0077] (2) Add 5 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0078] Comparative Example 1

[0079] An oral liquid is prepared from the following components in parts by weight: 100 parts grain soup, 0.05 parts Bacillus licheniformis, 0.03 parts kombucha liquid, 0.03 parts acetic acid bacteria, 0.02 parts Angel yeast SY, 0.02 parts Angel yeast RW, and 5 parts white sugar.

[0080] The method for preparing the grain soup is as follows: Take 4 parts of millet material, 2 parts of jujube powder, 1 part of grape seed powder, add 100 parts of distilled water, simmer over low heat for 10 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0081] The method for obtaining *Bacillus licheniformis* is as follows: Live *Bacillus licheniformis* capsules are streaked onto LB solid medium and incubated overnight at 37°C. Single colonies are picked and inoculated into 100 mL of LB liquid medium, and cultured with shaking at 150 rpm and 37°C for 4-6 hours. 2% of the inoculum is then inoculated into 200 mL of basal cereal medium and incubated at 37°C for 24 hours to obtain a seed culture. This seed culture is stored at 4°C and used for optimizing the bacterial culture formulation.

[0082] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 65 hours at a germination temperature of 27°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0083] The method for preparing the kombucha liquid is as follows: Mix 0.2 parts black tea, 0.2 parts green tea, 3 parts white sugar and 100 parts water, boil for 10 minutes until boiling, let cool to obtain mixture one, add 20 parts kombucha stock solution to mixture one, and ferment at 24℃ for 7 days to obtain kombucha liquid.

[0084] A method for preparing an oral liquid includes the following steps:

[0085] (1) Mix 0.05 parts of Bacillus licheniformis, 0.03 parts of kombucha liquid, 0.03 parts of acetic acid bacteria, 0.02 parts of Angel yeast SY and 0.02 parts of Angel yeast RW, and ferment under aseptic conditions at 37°C for 7 days to obtain fermentation liquid;

[0086] (2) Add 5 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0087] Comparative Example 2

[0088] An oral liquid is prepared from the following components in parts by weight: 4 parts grain soup, 0.05 parts Lactobacillus plantarum L9 inoculum, 0.03 parts acetic acid bacteria, 0.02 parts Angel yeast SY, 0.02 parts Angel yeast RW, and 5 parts white sugar.

[0089] The method for preparing the grain soup is as follows: Take 4 parts of millet material, 2 parts of jujube powder, 1 part of grape seed powder, add 100 parts of distilled water, simmer for 10 minutes, transfer to a sterile culture bottle, seal and cool, and anaerobic culture at 37°C for 7 days to obtain the grain soup.

[0090] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 65 hours at a germination temperature of 27°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0091] The method for obtaining the Lactobacillus plantarum L9 inoculum is as follows: Take 4 parts of millet material, 2 parts of jujube powder, and 1 part of grape seed powder, add 100 parts of distilled water, simmer for 10 minutes, transfer to a sterile culture bottle, seal and cool to obtain Lactobacillus plantarum grain culture medium, inoculate 5 parts of Lactobacillus plantarum L9 preservation solution, and anaerobic culture at 37℃ for 7 days to obtain Lactobacillus plantarum L9 inoculum.

[0092] A method for preparing an oral liquid includes the following steps:

[0093] (1) Take 4 portions of the above-prepared grain soup, add 0.05 portions of Lactobacillus plantarum L9 inoculum, 0.03 portions of acetic acid bacteria, 0.02 portions of Angel yeast SY and 0.02 portions of Angel yeast RW, mix well, and ferment under aseptic conditions at 37°C for 7 days to obtain fermentation liquid.

[0094] (2) Add 5 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0095] Comparative Example 3

[0096] An oral liquid is prepared from the following components in parts by weight: 6 parts grain soup, 0.03 parts kombucha liquid, 0.03 parts acetic acid bacteria, 0.02 parts Angel yeast SY, 0.02 parts Angel yeast RW, and 5 parts white sugar.

[0097] The method for preparing the grain soup is as follows: Take 4 parts of millet material, 2 parts of jujube powder, 1 part of grape seed powder, add 100 parts of distilled water, simmer over low heat for 10 minutes, transfer to a sterile culture bottle, seal and let cool to obtain the grain soup.

[0098] The method for obtaining the millet material is as follows: take millet, wash it clean, spread it evenly in the cultivation grid of the germination machine, germinate for 65 hours at a germination temperature of 27°C, sprinkle water every half hour to keep the millet moist, and change the water once a day during the period to obtain the millet material.

[0099] The method for preparing the kombucha liquid is as follows: Mix 0.2 parts black tea, 0.2 parts green tea, 3 parts white sugar and 100 parts water, boil for 10 minutes until boiling, let cool to obtain mixture one, add 20-30 parts kombucha stock solution to mixture one, and ferment at 24℃ for 7 days to obtain kombucha liquid.

[0100] A method for preparing an oral liquid includes the following steps:

[0101] (1) Take 4 parts of the above-prepared grain soup, 0.03 parts of kombucha liquid, 0.03 parts of acetic acid bacteria, 0.02 parts of Angel yeast SY and 0.02 parts of Angel yeast RW, mix them well, and ferment them under aseptic conditions at 37°C for 7 days to obtain fermentation liquid.

[0102] (2) Add 5 parts of white sugar to the fermentation liquid, mix well, sterilize, and obtain the oral liquid.

[0103] Sensory evaluations of each embodiment and comparative example are shown in Table 1.

[0104] Table 1 Sensory evaluation of each embodiment and comparative example.

[0105]

[0106] If the pH is too high, it indicates that there is contamination by other bacteria, which inhibits the fermentation of normal flora. As shown in Table 1, the acidity of the example is lower than that of the comparative example, which ensures the fermentation of normal flora.

[0107] Effect verification

[0108] 1. Method

[0109] Grouping: Twenty female C57BL / 6 mice were acclimatized to the laboratory environment for 7 days at a constant temperature of 20±2℃ and humidity, maintaining a 12 / 12h circadian rhythm. After 7 days of acclimatization, they were randomly divided into four groups: a control group, a model group, an oral liquid (KFY) group, and a positive control group receiving diazepam (DZP). Except for the control group, the other three groups underwent sleep deprivation modeling, and the success of the model was determined using a pentobarbital sodium sleep synergistic assay.

[0110] The normal control group and the model group were given distilled water for 14 days. The oral liquid group was given the oral liquid of Example 1 in a water bottle for free drinking for 14 days. The diazepam group was given 2 mL and 10 mg diazepam injection diluted with sodium chloride injection to 0.025 mg / mL and given in a water bottle for free drinking for 14 days. During this period, food and water were replenished in a timely manner.

[0111] Establishment of the insomnia model: The normal control group was injected with physiological saline intraperitoneally from 9:00 to 9:30 every day, while the other groups were injected with parachlorophenylalanine (PCPA) (20 mg / mL, 0.2 mL / 10 g) suspension intraperitoneally once a day at the same time for 2 consecutive days to replicate the mouse sleep deprivation model.

[0112] Sleep behavior assessment: After 14 days of drug administration, mice in each group underwent a pentobarbital sodium sleep synergy test to observe their sleep behavior. Sleep was measured by the disappearance of the righting reflex. When a mouse was placed in a supine position, if it could not right itself for more than 30-60 seconds, the righting reflex was considered to have disappeared, and the mouse had entered sleep. The recovery of the righting reflex was considered wakefulness, and the period from the disappearance to the recovery of the righting reflex was considered sleep time. The sleep latency and sleep duration of each group of mice were recorded, and changes in sleep behavior were compared among the groups.

[0113] Emotional and behavioral experiments: After 14 days of drug administration, mice in each group were tested in an open field test chamber. At the beginning, the mice were placed in the central grid position, and their performance in the open field was recorded by video using a mobile phone. The mice were allowed to adapt for 1 minute, and their behavior was recorded for 6 minutes. The video recordings were then reviewed to record relevant parameters, including the number of times the mice crossed the central grid, the number of times they groomed themselves, and the number of times they defecated. The changes in emotional and behavioral characteristics of the mice in each group were compared.

[0114] Serum inflammatory factor levels in mice were detected: After completing the emotional and behavioral experiments, blood was collected from the eyeballs of mice in each group. The blood was left at room temperature for 1 hour to allow it to clot, then centrifuged at 2000-3000 rpm for 15 minutes at 4°C. The supernatant was aliquoted and stored at -80°C to avoid repeated freeze-thaw cycles. The levels of inflammatory factors IL-6, IL-1β, and TNF-α in the serum of mice in each group were detected using an ELISA kit, and the serum inflammatory factor levels of each group were compared.

[0115] Detection of neurotransmitter changes in the hypothalamus of mice: Mice were euthanized by cervical dislocation after enucleation, decapitation, and brain harvesting. The brain was thoroughly homogenized, centrifuged at 2000-3000 rpm for 20 min at 4℃, and the supernatant was aliquoted and stored at -80℃. ELISA kits were used to detect the levels of 5-HT, GABA, and Glu in brain tissue, and changes in neurotransmitters in different groups of mice were compared.

[0116] Changes in fecal short-chain fatty acids were detected: Mouse feces were collected at 8:30-9:00 AM before modeling, 2 days after modeling, and on days 3, 7, 11, and 14 after drug administration. Feces from the same group on the same day were collected into one EP tube, stored at -80℃, transported on dry ice, and analyzed by gas chromatography-mass spectrometry (GC-MS) at Shanghai Zhongke New Life Biotechnology Co., Ltd. The main short-chain fatty acids detected included acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and hexanoic acid.

[0117] 2. Results

[0118] like Figure 1 As shown, the duration of sleep in mice after sleep deprivation was significantly reduced compared to before deprivation. After 14 days of free access to the oral liquid for improving sleep disorders, the sleep latency was shortened and the duration of sleep was prolonged, indicating that the oral liquid improved the sleep disorders in mice.

[0119] like Figure 2 As shown, after 14 days of oral liquid treatment to improve sleep disorders, the results of open field experiments on mice showed that, compared with the model group, the oral liquid group had significantly reduced grooming frequency and defecation frequency, and the anxiety behavior of the mice was improved.

[0120] like Figure 3 As shown, after sleep deprivation, the IL-6 level in the model group was increased compared with the normal group. After drinking the oral liquid for improving sleep disorders for 14 days, the IL-6 level in the oral liquid group was significantly lower than that in the normal group compared with the model group. Compared with the model group, the IL-1β level in the oral liquid group was decreased and the TNF-α level was increased.

[0121] like Figure 4 As shown, after sleep deprivation, compared with the normal group, the model group had decreased 5-HT and GABA levels and increased Glu levels. After drinking the oral liquid for improving sleep disorders for 14 days, compared with the model group, the model group had increased 5-HT and GABA levels and decreased Glu levels.

[0122] The standard curve equations were obtained by using mass spectrometry signals from standards of different concentrations, as shown in Table 2. The contents of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and hexanoic acid were all within the linear range and showed good correlation.

[0123] Table 2 Standard Curve Equations

[0124] Standard product name Linear regression equation Correlation coefficient Linear range (μg / ml) Acetic acid Y = 0.880161 * X + 1.211652 0.998 0.5～5000 propionic acid Y = 0.850287 * X + 0.185956 0.9985 0.01～100 Isobutyric acid Y = 1.392110 * X + 0.019689 0.9995 0.01～100 butyric acid Y = 2.953959 * X + 0.656305 0.9993 0.01～100 Isovalerate Y = 3.873625 * X + 0.063992 0.9993 0.01～100 Valeric acid Y = 3.912305 * X + 0.06672 0.9992 0.01～100 hexanoic acid Y = 4.426819 * X + 0.027657 0.9985 0.01～100

[0125] It should be noted that when numerical ranges are mentioned in the claims of this invention, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. To avoid redundancy, the present invention describes preferred embodiments.

[0126] Although preferred embodiments of the invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of the invention.

[0127] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. An oral liquid for improving sleep disorders, characterized by, Prepared from the following components by weight: 100-120 parts of cereal soup, 0.05-0.07 parts of Lactobacillus plantarum L9 agent, 0.03-0.05 parts of black tea fungus liquid, 0.03-0.05 parts of acetic acid bacteria, 0.02-0.04 parts of Angelus yeast SY, 0.02-0.04 parts of Angelus yeast RW, 5-10 parts of white granulated sugar; The preparation method of the cereal soup is as follows: taking 4-6 parts of millet material, 2-4 parts of jujube powder and 1-2 parts of grape seed powder, adding 100-120 parts of distilled water, simmering for 10-15 minutes on low heat, transferring to a sterile culture bottle, sealing and cooling, and obtaining the cereal soup; The obtaining method of the Lactobacillus plantarum L9 agent is as follows: 4-6 parts of millet material, 2-4 parts of Chinese jujube powder and 1-2 parts of grape seed powder are taken, 100-120 parts of distilled water is added, and the mixture is decocted on a small fire for 10-15 minutes; then the mixture is transferred to a sterile culture bottle, sealed and cooled to obtain a Lactobacillus plantarum grain culture medium; 5-10 parts of a Lactobacillus plantarum L9 preservative solution is inoculated, and the mixture is anaerobically cultured at 37 DEG C for 7-9 days to obtain the Lactobacillus plantarum L9 agent; the Lactobacillus plantarum L9 is preserved in the China Center for Type Culture Collection on February 28, 2022, and the preservation number is CCTCC NO.M 2022169, and the classification name is Lactobacillus plantarum . The millet material is obtained by taking millet, and germinating for 65-72 hours. The black tea fungus liquid is prepared from black tea, green tea, white granulated sugar, water and black tea fungus stock solution.

2. The oral liquid for improving sleep disorders according to claim 1, characterized by, The preparation method of the black tea fungus liquid is as follows: mixing 0.2-0.5 parts of black tea, 0.2-0.5 parts of green tea, 3-6 parts of white granulated sugar and 100-150 parts of water, simmering to boiling, cooling, obtaining mixture one, adding 20-30 parts of black tea fungus stock solution to mixture one, and fermenting to obtain the black tea fungus liquid. 3.The oral liquid for improving sleep disorders according to claim 2, characterized in that, The simmering time is 10-15 minutes, and the fermentation condition is 24-30℃ for 7-15 days.

4. The method for preparing an oral liquid for improving sleep disorders according to claim 1, characterized in that, Comprising the following steps: Taking 4-6 parts of millet material, 2-4 parts of jujube powder and 1-2 parts of grape seed powder, adding 100-120 parts of distilled water, simmering for 10-15 minutes on low heat, transferring to a sterile culture bottle, sealing and cooling, and obtaining the cereal soup; Taking another 4-6 parts of millet material, 2-4 parts of jujube powder and 1-2 parts of grape seed powder, adding 100-120 parts of distilled water, simmering for 10-15 minutes on low heat, transferring to a sterile culture bottle, sealing and cooling to obtain the Lactobacillus plantarum cereal culture medium, inoculating 5-10 parts of Lactobacillus plantarum L9 stock solution, and anaerobic culturing at 37℃ for 7-9 days to obtain the Lactobacillus plantarum L9 agent; Mixing 0.2-0.5 parts of black tea, 0.2-0.5 parts of green tea, 3-6 parts of white granulated sugar and 100-150 parts of water, simmering to boiling, cooling, obtaining mixture one, adding 20-30 parts of black tea fungus stock solution to mixture one, and fermenting to obtain the black tea fungus liquid; Adding 0.05-0.07 parts of Lactobacillus plantarum L9 agent, 0.03-0.05 parts of black tea fungus liquid, 0.03-0.05 parts of acetic acid bacteria, 0.02-0.04 parts of Angelus yeast SY and 0.02-0.04 parts of Angelus yeast RW to 100-120 parts of cereal soup, mixing, anaerobic fermenting, and obtaining the fermentation liquid; Adding 5-10 parts of white granulated sugar to the fermentation liquid, mixing, sterilizing, and obtaining the oral liquid.

5. The method for preparing an oral liquid for improving sleep disorders according to claim 4, characterized in that, When preparing the fermentation liquid, anaerobic fermenting at 37℃ for 7-9 days.

6. The method for preparing an oral liquid for improving sleep disorders according to claim 4, characterized in that, The sterilizing temperature is 121℃.

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