Nerve-soothing and sleep-aiding dark tea traditional Chinese medicine compound composition and preparation method thereof

Abstract

The invention provides a nerve soothing and sleep aiding dark tea traditional Chinese medicine composite composition and a preparation method thereof, and belongs to the technical field of medicinal tea. The preparation method comprises the following steps: fermenting dark green tea, adding degradation microspheres into fermentation liquor, stirring and degrading to obtain dark green tea ferment without alkaloid, uniformly mixing the dark green tea ferment with a traditional Chinese medicine composition extracted from spina date seed, poria cocos, polygala tenuifolia, liquorice, ginseng and prepared rehmannia root, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxy propyl cellulose and sodium alginate, granulating, and tabletting to obtain the dark green tea tablet. The nerve-soothing and sleep-aiding dark tea traditional Chinese medicine compound composition is prepared. The prepared dark green tea traditional Chinese medicine composite composition for soothing nerves and helping sleep is a product combining low caffeine and high aroma and the slow-release tablets for soothing nerves and helping sleep, the utilization rate of dark green tea resources is increased, good effects of soothing nerves and helping sleep are achieved, and economic benefits are remarkable.

Description

Technical Field

[0001] This invention relates to the field of medicinal tea technology, specifically to a calming and sleep-aiding black tea herbal compound composition and its preparation method. Background Technology

[0002] Insomnia is one of the most common sleep disorders. Indicators of sleep continuity include sleep latency, number of awakenings, number of awakenings after falling asleep, total sleep time, and sleep efficiency. If one or more of these indicators are problematic, it can be called a sleep continuity disorder. With the fast pace of life and various mental stresses, most people are in a sub-healthy state, and the insomnia rate among adults is showing an increasing trend year by year. Insomnia can lead to fatigue, lack of energy, and difficulty concentrating, and can easily trigger diseases such as anxiety, depression, and hypertension. Therefore, improving sleep disorders and enhancing sleep quality is an urgent problem to be solved. At present, clinical treatment of sleep disorders mainly uses sedative-hypnotic drugs such as benzodiazepines and non-benzodiazepines. These chemical drugs are prone to tolerance, dependence, and adverse reactions. Compared with chemical drugs, traditional Chinese medicine treatment for insomnia has fewer toxic side effects, stable efficacy, and does not lead to drug dependence, making it more acceptable to a wide range of patients. However, its use is limited by factors such as long treatment cycles and slow onset of action.

[0003] The existing technology currently suffers from the following main problems:

[0004] 1. Sedative-hypnotic drugs are prone to causing tolerance, dependence, and adverse reactions;

[0005] 2. Traditional Chinese medicine treatments are slow to take effect and have a long treatment cycle;

[0006] 3. Tea products contain stimulating components such as caffeine, which can worsen insomnia. Summary of the Invention

[0007] The purpose of this invention is to propose a sedative and sleep-aiding black tea herbal compound composition and its preparation method. It is a product that combines low-caffeine, high-aroma black tea with a sedative and sleep-aiding sustained-release tablet, thereby improving the utilization rate of black tea resources and having good sedative and sleep-aiding effects, resulting in significant economic benefits.

[0008] The technical solution of this invention is implemented as follows:

[0009] This invention provides a method for preparing a calming and sleep-aiding black tea herbal compound. After fermentation, degradable microspheres are added to the fermentation liquid and stirred to degrade the microspheres, resulting in a black tea fermentation product free of alkaloids. This product is then mixed evenly with a herbal composition extracted from jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate. The mixture is granulated, compressed into tablets, and the calming and sleep-aiding black tea herbal compound is obtained.

[0010] As a further improvement to the present invention, the following steps are included:

[0011] S1. Fermentation of dark tea: Pulverize dark tea leaves, add water, heat and reflux to extract, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid, ferment and culture, filter to obtain dark tea fermentation liquid;

[0012] S2. Degradation of alkaloids: Add degradation microspheres to the fermented black tea liquid, stir to degrade, separate the microspheres with a magnet, freeze dry, and obtain fermented black tea product with alkaloids removed;

[0013] S3. Extraction of the Chinese herbal composition: The jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa were washed, dried, pulverized, mixed, added to an ethanol aqueous solution, heated under reflux for extraction, filtered, and the extraction was repeated. The filtrates were combined, concentrated, and dried to obtain the Chinese herbal composition.

[0014] S4. Preparation of sustained-release tablets: The alkaloid-free fermented black tea, the traditional Chinese medicine composition, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate are mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0015] As a further improvement of the present invention, the bacterial count of the *Aurogonium cristatum* seed solution in step S1 is 10. 8 -10 9 The concentration of cfu / mL and the inoculum amount are 2-4 v / v%. The heating and reflux extraction time is 3-5 h. The solid-liquid ratio of black tea leaves to water is 1:10-20 g / mL. The fermentation culture conditions are pH 5-6, temperature 28-32℃, rotation speed 100-150 r / min, and time 24-36 h.

[0016] As a further improvement of the present invention, the mass ratio of the black tea fermentation liquid to the degradation microspheres in step S2 is 100:1-2, and the preparation method of the degradation microspheres is as follows:

[0017] T1. Preparation of mesoporous TiO2 nanospheres: Tetrabutyl titanate and a pore-forming agent were dissolved in an organic solvent to obtain an oil phase; an emulsifier was added to water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified, the pH of the solution was adjusted, centrifuged, washed, dried, and calcined to obtain mesoporous TiO2 nanospheres;

[0018] T2. Preparation of MoS2 quantum dots / mesoporous TiO2 nanospheres: Mesoporous TiO2 nanospheres were added to water, sodium molybdate was added, the pH of the solution was adjusted, glutathione was added, hydrothermal reaction was carried out, centrifugation was performed, washing was performed, and drying was carried out to obtain MoS2 quantum dots / mesoporous TiO2 nanospheres.

[0019] T3. Preparation of zero-valent iron sulfide: Zero-valent iron powder is evenly spread on the upper stainless steel sieve, sulfur powder is placed in the lower crucible, and under the protection of inert gas, it is heated and calcined, and then cooled to room temperature to obtain zero-valent iron sulfide.

[0020] T4. Polydopamine modification: MoS2 quantum dots / mesoporous TiO2 nanospheres and zero-valent iron sulfide were added to Tris-HCl solution, dopamine hydrochloride was added, the mixture was heated and stirred to react, centrifuged, washed, dried, and ball-milled to obtain the modified composite.

[0021] T5. The seed culture of Pseudomonas stearotherm, obtained after activation, is fermented, lysozyme is added for enzymatic hydrolysis, filtered, and the filtrate is dialyzed to obtain a cell wall broken bacterial culture.

[0022] T6. Preparation of degradable microspheres: N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide were added to the cell wall-broken bacterial solution, stirred and activated, the modified complex was added, the reaction was stirred, separated by magnet, washed, and freeze-dried to obtain degradable microspheres.

[0023] As a further improvement of the present invention, in step T1, the mass ratio of tetrabutyl titanate, pore-forming agent, and emulsifier is 10-15:1-2:2-3, the pore-forming agent is hexadecyltrimethylammonium bromide or hexadecyltrimethylammonium chloride, and the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60, Tween-80, and Tween-85. The calcination temperature is 400-500℃ and the time is 2-4h. In step T2, the mass ratio of mesoporous TiO2 nanospheres, sodium molybdate, and glutathione is 10-15:1-3:1.5-2.5, the pH value of the solution is adjusted to 6.2-6.7, and the hydrothermal reaction temperature is 180-220℃ and the time is 10-15h.

[0024] As a further improvement of the present invention, in step T3, the mass ratio of zero-valent iron powder to sulfur powder is 1-2:0.1-0.3, the heating and calcination temperature is 400-500℃, and the time is 1-2h; in step T4, the mass ratio of MoS2 quantum dots / mesoporous TiO2 nanospheres, zero-valent iron sulfide, and dopamine hydrochloride is 10-14:3-5:4-6, the pH value of the Tris-HCl solution is 8.5-9.5, the heating and stirring reaction temperature is 45-55℃, and the time is 3-5h.

[0025] As a further improvement of the present invention, the bacterial count of the spore liquid in step T5 is 10. 8 -10 9The fermentation conditions are 36-37℃, 200-250 r / min, and 60-72 h. The amount of lysozyme added is 3-5 wt% of the total mass of the system. The dialysis bag used for dialysis has a pore size of 2k-3kDa. In step T6, the mass ratio of the cell wall-breaking bacterial solution, N-hydroxysuccinimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and the modified complex is 100-150:3-5:4-7:15-20.

[0026] As a further improvement of the present invention, the mass ratio of jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa in step S3 is 10-20:5-15:5-15:3-5:2-4:3-5, the concentration of the ethanol aqueous solution is 50-60 wt%, and the heating and reflux extraction time is 3-5 h.

[0027] As a further improvement of the present invention, the mass ratio of the black tea fermentation product with alkaloids removed, the traditional Chinese medicine composition, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate in step S4 is 10-15:8-10:20-30:5-10:4-8:10-20:15-20:5-10.

[0028] This invention further protects a calming and sleep-aiding black tea herbal compound composition prepared by the above-mentioned preparation method.

[0029] The present invention has the following beneficial effects:

[0030] This invention employs a two-step process combining directional fermentation of *Aspergillus cristatus* with magnetic alkaloid degradation microspheres. This process removes stimulating alkaloids such as caffeine, theobromine, and theophylline from fermented black tea products while retaining sleep-promoting active ingredients such as theanine, theabrownin, and γ-aminobutyric acid (GABA), fundamentally eliminating the side effect of tea-induced insomnia. Furthermore, *Aspergillus cristatus* fermentation produces unique "golden flower" metabolites, including polyketides, naphthoquinones, and active oligosaccharides. These metabolites synergistically interact with extracts of traditional Chinese medicine such as jujube seed saponins, polygalactosidone, glycyrrhizic acid, and rare ginsenosides, increasing the expression levels of serotonin, GABA, and brain-derived neurotrophic factor (BDNF), resulting in significant sedative, anti-anxiety, and sleep-improving effects.

[0031] The medicinal components of this invention include Ziziphus jujuba seed, Poria cocos, Polygala tenuifolia, Glycyrrhiza uralensis, Panax ginseng, and Rehmannia glutinosa, wherein:

[0032] The principal herbs are: Ziziphus jujuba seed and ginseng. Ziziphus jujuba seed is neutral in nature, sweet and sour in taste, and enters the liver, gallbladder, and heart meridians. It is a key herb for nourishing the heart and calming the mind. Its astringent properties can astringe yin fluids and nourish heart blood, primarily treating insomnia due to deficiency and restlessness, palpitations, and excessive dreaming; it is the core herb for treating insomnia. Ginseng greatly replenishes vital energy and benefits the qi of the heart and spleen. When qi is abundant, it can generate and circulate blood, forming the basis for replenishing qi to generate blood and nourishing the heart and calming the mind. It is especially suitable for insomnia caused by prolonged illness, overwork, or excessive thinking, resulting in deficiency of both qi and blood.

[0033] Assistant herbs: Rehmannia glutinosa (processed) and Polygala tenuifolia (processed). Rehmannia glutinosa: nourishes Yin and blood, replenishes essence and marrow, and enters the heart, liver, and kidney meridians. Combined with ginseng, it forms a classic "Qi-tonifying and blood-nourishing" combination (such as Ginseng Nourishing Decoction), addressing the root cause of "blood deficiency failing to nourish the heart." Polygala tenuifolia: pungent and bitter, warm in nature, enters the heart and kidney meridians, functions to harmonize the heart and kidneys, resolve phlegm and open the orifices, calm the mind and stabilize the will. Its "ascending to the heart and descending to the kidneys" principle can guide kidney water upwards to nourish heart fire, harmonize Yin and Yang, and improve insomnia caused by heart-kidney disharmony.

[0034] Assistant herb: Poria cocos; Poria cocos: sweet, bland, and neutral in nature, enters the heart, spleen, and kidney meridians, and functions to strengthen the spleen, promote diuresis, calm the mind, and soothe the nerves. On the one hand, it assists ginseng in strengthening the spleen to provide the source of qi and blood production; on the other hand, it assists jujube seed and polygala tenuifolia in calming the mind, enhancing the overall calming effect of the formula, making it an excellent "assistant herb".

[0035] Guiding herb: Licorice; Licorice: harmonizes the effects of other herbs, invigorates qi and harmonizes the middle jiao, relieves spasms and pain. Its sweet and mild nature can moderate the adverse effects of other herbs, making the overall medicinal effect of the formula mild and lasting, thus serving as a "harmonizing guide".

[0036] This combination not only conforms to the traditional Chinese medicine principle of "harmonizing the heart and spleen" in treating insomnia, but is also verified by modern medical theory to have a higher activation rate of γ-aminobutyric acid receptors and a reduced central excitatory effect of caffeine after combination.

[0037] This invention prepares a sustained-release tablet with a hydroxypropyl cellulose K15M-sodium alginate-sodium carboxymethyl cellulose ternary hydrophilic gel framework to achieve a slow release that is "fast at the beginning and stable at the end," avoiding fluctuations in blood drug concentration peaks and troughs, thereby achieving the effects of calming the nerves, promoting sleep for a long time, improving sleep quality, and reducing the number of nighttime awakenings.

[0038] The alkaloid-degrading microspheres prepared in this invention are prepared by the following mechanism:

[0039] When tetrabutyl titanate (oil phase) is added to the aqueous phase, it forms O / W type emulsion droplets under the action of an emulsifier. The emulsifier molecules are adsorbed at the oil-water interface, inhibiting droplet aggregation. Adjusting the pH can regulate the hydrolysis rate of tetrabutyl titanate, avoiding amorphous precipitation caused by excessively rapid hydrolysis and promoting the formation of TiO2. 4+Ti(OH)4 sol is gradually hydrolyzed to form Ti(OH)4 sol. Under the steric hindrance of the porogen, the sol particles self-assemble around the porogen to form an ordered mesoporous structure. After calcination, Ti(OH)4 dehydrates and crystallizes to form mesoporous TiO2 nanospheres with photocatalytic activity, while providing a spatial carrier for subsequent loading of active components.

[0040] Subsequently, when the mesoporous TiO2 nanospheres are dispersed in water, their surface hydroxyl groups (-OH) can adsorb MoO4 from sodium molybdate via hydrogen bonds. 2- Adjust the pH value to avoid MoO4 2- Hydrolysis, glutathione acts as a reducing agent and ligand, its sulfhydryl group reacts with MoO4. 2- A redox reaction occurs, turning Mo 6+ Restored to Mo 4+ Meanwhile, the amino group of glutathione and Mo 4+ Mo-S bonds are formed through coordination, and the MoS2 quantum dots are gradually grown under hydrothermal conditions. The pore structure and surface hydroxyl groups of mesoporous TiO2 can fix the MoS2 quantum dots in situ within the pores and on the surface of the spheres through physical adsorption and coordination, forming a composite heterojunction structure. The semiconductor properties of MoS2 quantum dots and the heterojunction structure formed with TiO2 provide electron transfer channels for subsequent catalytic degradation, enabling them to utilize the visible light region, thereby greatly improving the photocatalytic efficiency of MoS2.

[0041] Under inert gas protection, sulfur powder in the lower crucible is heated and sublimated into sulfur vapor, which diffuses upward to the upper stainless steel screen. Zero-valent iron powder is in a dispersed state and comes into full contact with sulfur vapor, undergoing a gas-solid reaction. The generated FeS adheres to the surface of the zero-valent iron powder, forming a core-shell structure of sulfide zero-valent iron. This structure retains the magnetism and reactivity of zero-valent iron, while the FeS shell inhibits the excessive oxidation of zero-valent iron, which can assist electron transfer and enhance catalytic activity. In the presence of oxygen in water, it can effectively degrade alkaloids, such as caffeine, thereby improving the removal efficiency of alkaloids.

[0042] Dopamine hydrochloride undergoes a self-polymerization reaction under catalytic oxidation, forming a polymer rich in amino and hydroxyl groups. This polymer exhibits strong adhesion and can be coated onto the surface of MoS2 quantum dots / mesoporous TiO2 nanospheres and sulfide zero-valent iron through hydrogen bonding and electrostatic interactions, forming a uniform modified layer. On the one hand, this provides active sites for subsequent coupling of biological enzymes; on the other hand, it can bind the inorganic components of both together to form a complex under ball milling, avoiding functional failure caused by nanoparticle aggregation.

[0043] After activation and fermentation, *Pseudomonas schrenckii* contains a specific enzyme system for degrading tea alkaloids (such as caffeine and theophylline). Treatment with lysozyme hydrolyzes the peptidoglycan structure of the bacterial cell wall, causing cell wall rupture and releasing the intracellular enzyme system. Filtration removes cell wall fragments, and dialysis removes small molecule impurities, ultimately yielding a highly active, cell-wall-broken bacterial solution. This solution provides a bio-enzyme catalytic core for alkaloid degradation. Under the action of N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, the enzyme protein is coupled to the surface of the modified complex, achieving covalent coupling between the enzyme protein and the inorganic carrier. Utilizing the magnetism of zero-valent iron sulfide, the coupling products are rapidly collected via magnetic separation, ultimately forming integrated degradation microspheres. These biodegradable microspheres can achieve visible light catalysis and synergistic catalysis of alkaloid degradation by sulfidated zero-valent iron and compound bacterial enzymes, avoiding problems such as tea polyphenol oxidation and theanine decomposition caused by chemical degradation (such as acid-base hydrolysis and high-temperature oxidation). This enables rapid degradation and rapid recovery under an external magnetic field, allowing for multiple recycling without organic solvent residue, making the process green and environmentally friendly.

[0044] This invention uses a lactose-microcrystalline cellulose system, which has better compressibility, disintegration properties and taste than traditional starch dextrin tablets; and it can mask the bitter taste of traditional Chinese medicine without coating, resulting in high patient compliance.

[0045] The calming and sleep-aiding black tea herbal compound composition prepared by this invention is a product combining low-caffeine, high-aroma black tea with a calming and sleep-aiding sustained-release tablet. It improves the utilization rate of black tea resources, has good calming and sleep-aiding effects, and has significant economic benefits. Attached Figure Description

[0046] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0047] Figure 1 SEM image of the degradable microspheres prepared in Example 1. Detailed Implementation

[0048] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0049] *Pseudomonas schlegelii*, ATCC17588, 100 CFU / g. Activation method: Inoculate the strain into nutrient broth medium and incubate at 37°C and 150 rpm for 18-24 hours to obtain a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution.

[0050] *Eurotium cristatum*, ATCC16468, 100 CFU / g. Activation method: Inoculate the strain into Gao's medium and incubate at 30°C and 100 rpm for 18-24 hours to obtain a bacterial count of 10... 8 -10 9 CFU / mL bacterial seed solution.

[0051] Lysozyme, enzyme activity >20,000 U / mg.

[0052] Preparation Example 1: Degradable Microspheres

[0053] The preparation method is as follows:

[0054] T1. Preparation of mesoporous TiO2 nanospheres: 10g tetrabutyl titanate and 1g hexadecyltrimethylammonium bromide were dissolved in 100mL ethyl acetate to obtain an oil phase; 2g Tween-20 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 500℃ for 2h to obtain mesoporous TiO2 nanospheres;

[0055] T2. Preparation of MoS2 quantum dot / mesoporous TiO2 nanospheres: 10g of mesoporous TiO2 nanospheres were added to 150mL of water, 1g of sodium molybdate was added, the pH of the solution was adjusted to 6.2, 1.5g of glutathione was added, and the mixture was hydrothermally reacted at 180℃ for 10h. After centrifugation, washing, and drying, MoS2 quantum dot / mesoporous TiO2 nanospheres were obtained.

[0056] T3. Preparation of zero-valent iron sulfide: 1g of zero-valent iron powder is evenly spread on the upper stainless steel sieve, and 0.1g of sulfur powder is placed in the lower crucible. Under nitrogen protection, the mixture is heated to 500℃ and calcined for 1 hour. After cooling to room temperature, zero-valent iron sulfide is obtained.

[0057] T4. Polydopamine modification: 10g of MoS2 quantum dot / mesoporous TiO2 nanospheres and 3g of zero-valent iron sulfide were added to 300mL of Tris-HCl solution with a pH of 8.5, and 4g of dopamine hydrochloride was added. The mixture was heated to 45℃, stirred for 3h, centrifuged, washed, dried, and ball-milled to obtain the modified composite.

[0058] T5. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas schrei after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 200 r / min for 60 h, then lysozyme was added at an amount of 3 wt% of the total mass of the system, and enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 2 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0059] T6. Preparation of degradable microspheres: Add 3g of N-hydroxysuccinimide and 4g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 100g of cell wall-broken bacterial solution, stir and activate for 30min, add 15g of modified complex, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres. Figure 1 The image shows a SEM image of the prepared degradable microspheres. As can be seen from the image, the particle size of the microspheres is between 2 and 5 μm.

[0060] Preparation Example 2: Degradable Microspheres

[0061] The preparation method is as follows:

[0062] T1. Preparation of mesoporous TiO2 nanospheres: 15g tetrabutyl titanate and 2g hexadecyltrimethylammonium chloride were dissolved in 100mL ethyl acetate to obtain an oil phase; 3g Tween-40 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 400℃ for 4h to obtain mesoporous TiO2 nanospheres;

[0063] T2. Preparation of MoS2 quantum dots / mesoporous TiO2 nanospheres: 15g of mesoporous TiO2 nanospheres were added to 150mL of water, 3g of sodium molybdate was added, the pH of the solution was adjusted to 6.7, 2.5g of glutathione was added, and the mixture was hydrothermally reacted at 220℃ for 15h. After centrifugation, washing, and drying, MoS2 quantum dots / mesoporous TiO2 nanospheres were obtained.

[0064] T3. Preparation of zero-valent iron sulfide: Spread 2g of zero-valent iron powder evenly on the upper stainless steel sieve, place 0.3g of sulfur powder in the lower crucible, heat to 500℃ under nitrogen protection, calcine for 2h, and cool to room temperature to obtain zero-valent iron sulfide.

[0065] T4. Polydopamine modification: 14g of MoS2 quantum dot / mesoporous TiO2 nanospheres and 5g of zero-valent iron sulfide were added to 300mL of Tris-HCl solution with a pH of 9.5, 6g of dopamine hydrochloride was added, the mixture was heated to 55℃, stirred for 5h, centrifuged, washed, dried, and ball-milled to obtain the modified composite.

[0066] T5. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas stearothermiae obtained after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 250 r / min for 72 h, lysozyme was added at an amount of 5 wt% of the total mass of the system, enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 3 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0067] T6. Preparation of degradable microspheres: Add 5g of N-hydroxysuccinimide and 7g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 150g of cell wall-broken bacterial solution, stir and activate for 30min, add 20g of modified complex, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres.

[0068] Preparation Example 3: Degradable Microspheres

[0069] The preparation method is as follows:

[0070] T1. Preparation of mesoporous TiO2 nanospheres: 12g tetrabutyl titanate and 1.5g hexadecyltrimethylammonium chloride were dissolved in 100mL ethyl acetate to obtain an oil phase; 2.5g Tween-80 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 450℃ for 3h to obtain mesoporous TiO2 nanospheres;

[0071] T2. Preparation of MoS2 quantum dot / mesoporous TiO2 nanospheres: 12g of mesoporous TiO2 nanospheres were added to 150mL of water, 2g of sodium molybdate was added, the pH of the solution was adjusted to 6.5, 2g of glutathione was added, and the mixture was hydrothermally reacted at 200℃ for 12h. After centrifugation, washing, and drying, MoS2 quantum dot / mesoporous TiO2 nanospheres were obtained.

[0072] T3. Preparation of zero-valent iron sulfide: 1.5g of zero-valent iron powder is evenly spread on the upper stainless steel sieve, and 0.2g of sulfur powder is placed in the lower crucible. Under nitrogen protection, the mixture is heated to 450℃ and calcined for 1.5h. After cooling to room temperature, zero-valent iron sulfide is obtained.

[0073] T4. Polydopamine modification: 12g MoS2 quantum dot / mesoporous TiO2 nanospheres and 4g zero-valent iron sulfide were added to 300mL Tris-HCl solution with pH 9, 5g dopamine hydrochloride was added, the mixture was heated to 50℃, stirred for 4h, centrifuged, washed, dried, and ball-milled to obtain the modified composite.

[0074] T5. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas stearothermiae obtained after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 220 r / min for 66 h, lysozyme was added at an amount of 4 wt% of the total mass of the system, enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 2.5 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0075] T6. Preparation of degradable microspheres: Add 4g of N-hydroxysuccinimide and 5.5g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 125g of cell wall-broken bacterial solution, stir and activate for 30min, add 17g of modified complex, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres.

[0076] Comparative Preparation Example 1

[0077] The difference compared to preparation example 3 is that step T2 was not performed.

[0078] The preparation method is as follows:

[0079] T1. Preparation of mesoporous TiO2 nanospheres: 12g tetrabutyl titanate and 1.5g hexadecyltrimethylammonium chloride were dissolved in 100mL ethyl acetate to obtain an oil phase; 2.5g Tween-80 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 450℃ for 3h to obtain mesoporous TiO2 nanospheres;

[0080] T2. Preparation of zero-valent iron sulfide: 1.5g of zero-valent iron powder is evenly spread on the upper stainless steel sieve, and 0.2g of sulfur powder is placed in the lower crucible. Under nitrogen protection, the mixture is heated to 450℃ and calcined for 1.5h. After cooling to room temperature, zero-valent iron sulfide is obtained.

[0081] T3. Polydopamine modification: 12g of mesoporous TiO2 nanospheres and 4g of zero-valent iron sulfide were added to 300mL of Tris-HCl solution with a pH of 9, and 5g of dopamine hydrochloride was added. The mixture was heated to 50℃, stirred for 4h, centrifuged, washed, dried, and ball-milled to obtain the modified composite.

[0082] T4. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas stearothermiae obtained after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 220 r / min for 66 h, lysozyme was added at an amount of 4 wt% of the total mass of the system, enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 2.5 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0083] T5. Preparation of degradable microspheres: Add 4g of N-hydroxysuccinimide and 5.5g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 125g of cell wall-broken bacterial solution, stir and activate for 30min, add 17g of modified complex, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres.

[0084] Comparative Preparation Example 2

[0085] The difference from Preparation Example 3 is that MoS2 quantum dots / mesoporous TiO2 nanospheres were not added in step T4.

[0086] The preparation method is as follows:

[0087] T1. Preparation of zero-valent iron sulfide: 1.5g of zero-valent iron powder is evenly spread on the upper stainless steel sieve, and 0.2g of sulfur powder is placed in the lower crucible. Under nitrogen protection, the mixture is heated to 450℃ and calcined for 1.5h. After cooling to room temperature, zero-valent iron sulfide is obtained.

[0088] T2. Polydopamine modification: 16g of zero-valent iron sulfide was added to 300mL of Tris-HCl solution with pH 9, 5g of dopamine hydrochloride was added, the mixture was heated to 50℃, stirred for 4h, centrifuged, washed, dried, and ball-milled to obtain the modified product.

[0089] T3. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas stearothermiae obtained after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 220 r / min for 66 h, lysozyme was added at an amount of 4 wt% of the total mass of the system, enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 2.5 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0090] T4. Preparation of degradable microspheres: Add 4g of N-hydroxysuccinimide and 5.5g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 125g of cell wall-broken bacterial solution, stir and activate for 30min, add 17g of modifier, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres.

[0091] Comparative preparation example 3

[0092] The difference from Preparation Example 3 is that zero-valent iron sulfide was not added in step T4.

[0093] The preparation method is as follows:

[0094] T1. Preparation of mesoporous TiO2 nanospheres: 12g tetrabutyl titanate and 1.5g hexadecyltrimethylammonium chloride were dissolved in 100mL ethyl acetate to obtain an oil phase; 2.5g Tween-80 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 450℃ for 3h to obtain mesoporous TiO2 nanospheres;

[0095] T2. Preparation of MoS2 quantum dot / mesoporous TiO2 nanospheres: 12g of mesoporous TiO2 nanospheres were added to 150mL of water, 2g of sodium molybdate was added, the pH of the solution was adjusted to 6.5, 2g of glutathione was added, and the mixture was hydrothermally reacted at 200℃ for 12h. After centrifugation, washing, and drying, MoS2 quantum dot / mesoporous TiO2 nanospheres were obtained.

[0096] T3. Polydopamine modification: 16g of MoS2 quantum dot / mesoporous TiO2 nanospheres were added to 300mL of Tris-HCl solution with pH 9, 5g of dopamine hydrochloride was added, the mixture was heated to 50℃, stirred for 4h, centrifuged, washed, dried, and ball-milled to obtain the modified product.

[0097] T4. Preparation of cell wall-broth disrupted bacterial culture: The seed culture of Pseudomonas stearothermiae obtained after activation was inoculated into nutrient broth medium at an inoculation amount of 5 v / v%, fermented at 37℃ and a rotation speed of 220 r / min for 66 h, lysozyme was added at an amount of 4 wt% of the total mass of the system, enzymatic hydrolysis was carried out at 37℃ for 4 h, filtered, and the filtrate was dialyzed through a dialysis bag with a pore size of 2.5 kDa for 12 h to obtain cell wall-broth disrupted bacterial culture;

[0098] T5. Preparation of degradable microspheres: Add 4g of N-hydroxysuccinimide and 5.5g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to 125g of cell wall-broken bacterial solution, stir and activate for 30min, add 17g of modifier, stir and react for 15h, separate by magnet, wash, freeze dry to obtain degradable microspheres.

[0099] Comparative preparation example 4

[0100] The difference compared to preparation example 3 is that steps T5 and T6 were not performed.

[0101] The preparation method is as follows:

[0102] T1. Preparation of mesoporous TiO2 nanospheres: 12g tetrabutyl titanate and 1.5g hexadecyltrimethylammonium chloride were dissolved in 100mL ethyl acetate to obtain an oil phase; 2.5g Tween-80 was added to 200mL water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified at 8000r / min for 15min, the pH of the solution was adjusted to 9.5, centrifuged, washed, dried, and calcined at 450℃ for 3h to obtain mesoporous TiO2 nanospheres;

[0103] T2. Preparation of MoS2 quantum dot / mesoporous TiO2 nanospheres: 12g of mesoporous TiO2 nanospheres were added to 150mL of water, 2g of sodium molybdate was added, the pH of the solution was adjusted to 6.5, 2g of glutathione was added, and the mixture was hydrothermally reacted at 200℃ for 12h. After centrifugation, washing, and drying, MoS2 quantum dot / mesoporous TiO2 nanospheres were obtained.

[0104] T3. Preparation of zero-valent iron sulfide: 1.5g of zero-valent iron powder is evenly spread on the upper stainless steel sieve, and 0.2g of sulfur powder is placed in the lower crucible. Under nitrogen protection, the mixture is heated to 450℃ and calcined for 1.5h. After cooling to room temperature, zero-valent iron sulfide is obtained.

[0105] T4. Polydopamine modification: 12g of MoS2 quantum dot / mesoporous TiO2 nanospheres and 4g of zero-valent iron sulfide were added to 300mL of Tris-HCl solution with a pH of 9, and 5g of dopamine hydrochloride was added. The mixture was heated to 50℃, stirred for 4h, centrifuged, washed, dried, and ball-milled to obtain degraded microspheres.

[0106] Example 1

[0107] This embodiment provides a method for preparing a calming and sleep-aiding black tea herbal compound composition, including the following steps:

[0108] S1. Fermentation of dark tea: Crush 10g of dark tea leaves, add 100mL of water, heat and reflux for 3h, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid, inoculation amount is 2v / v%, pH value is 5, temperature is 28℃, rotation speed is 100r / min, fermentation culture for 24h, filter, and obtain dark tea fermentation liquid.

[0109] S2. Degradation of alkaloids: 1g of the degradation microspheres prepared in Example 1 were added to 100g of black tea fermentation liquid, stirred and degraded for 30min, the microspheres were separated by magnet, and freeze-dried to obtain black tea fermentation product with alkaloids removed.

[0110] S3. Extraction of the traditional Chinese medicine composition: 10g of Ziziphus jujuba seed, 5g of Poria cocos, 5g of Polygala tenuifolia, 3g of Glycyrrhiza uralensis, 2g of Panax ginseng, and 3g of Rehmannia glutinosa were washed, dried, pulverized, mixed, and added to 500mL of 50wt% ethanol aqueous solution. The mixture was heated and refluxed for 3 hours, filtered, and the extraction was repeated 3 times. The filtrates were combined, concentrated, and dried to obtain the traditional Chinese medicine composition.

[0111] S4. Preparation of sustained-release tablets: 10g of alkaloid-free fermented black tea, 8g of traditional Chinese medicine composition, 20g of lactose, 5g of microcrystalline cellulose, 4g of magnesium stearate, 10g of sodium carboxymethyl cellulose, 15g of hydroxypropyl cellulose K15M, and 5g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0112] Example 2

[0113] This embodiment provides a method for preparing a calming and sleep-aiding black tea herbal compound composition, including the following steps:

[0114] S1. Fermentation of dark tea: 10g of dark tea leaves are crushed, added to 200mL of water, heated and refluxed for 5h, cooled to room temperature, sterilized, and inoculated with Aspergillus cristatus seed liquid at an inoculation amount of 4v / v%, pH value of 6, temperature of 32℃, rotation speed of 150r / min, fermented for 36h, filtered, and dark tea fermentation liquid is obtained.

[0115] S2. Degradation of alkaloids: 2g of the degradation microspheres prepared in Example 2 were added to 100g of black tea fermentation liquid, stirred and degraded for 30min, the microspheres were separated by magnet, and freeze-dried to obtain black tea fermentation product with alkaloids removed.

[0116] S3. Extraction of the traditional Chinese medicine composition: 20g of Ziziphus jujuba seed, 15g of Poria cocos, 15g of Polygala tenuifolia, 5g of Glycyrrhiza uralensis, 4g of Panax ginseng, and 5g of Rehmannia glutinosa were washed, dried, pulverized, mixed, and added to 500mL of 60wt% ethanol aqueous solution. The mixture was heated and refluxed for 5h, filtered, and the extraction was repeated 3 times. The filtrates were combined, concentrated, and dried to obtain the traditional Chinese medicine composition.

[0117] S4. Preparation of sustained-release tablets: 15g of alkaloid-free fermented black tea, 10g of traditional Chinese medicine composition, 30g of lactose, 10g of microcrystalline cellulose, 8g of magnesium stearate, 20g of sodium carboxymethyl cellulose, 20g of hydroxypropyl cellulose K15M, and 10g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0118] Example 3

[0119] This embodiment provides a method for preparing a calming and sleep-aiding black tea herbal compound composition, including the following steps:

[0120] S1. Fermentation of dark tea: Crush 10g of dark tea leaves, add 150mL of water, heat and reflux for 4h, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid at an inoculation amount of 3v / v%, pH value of 5.5, temperature of 30℃, rotation speed of 125r / min, ferment for 30h, filter to obtain dark tea fermentation liquid.

[0121] S2. Degradation of alkaloids: 1.5g of the degradation microspheres prepared in Example 3 were added to 100g of black tea fermentation liquid, stirred and degraded for 30min, the microspheres were separated by magnet, and freeze-dried to obtain black tea fermentation product with alkaloids removed;

[0122] S3. Extraction of the traditional Chinese medicine composition: 15g of Ziziphus jujuba seed, 10g of Poria cocos, 10g of Polygala tenuifolia, 4g of Glycyrrhiza uralensis, 2g of Panax ginseng, and 4g of Rehmannia glutinosa were washed, dried, pulverized, mixed, and added to 500mL of 55wt% ethanol aqueous solution. The mixture was heated and refluxed for 4h, filtered, and the extraction was repeated 3 times. The filtrates were combined, concentrated, and dried to obtain the traditional Chinese medicine composition.

[0123] S4. Preparation of sustained-release tablets: 12g of alkaloid-free fermented black tea, 9g of traditional Chinese medicine composition, 25g of lactose, 7g of microcrystalline cellulose, 6g of magnesium stearate, 15g of sodium carboxymethyl cellulose, 17g of hydroxypropyl cellulose K15M, and 7g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0124] Comparative Example 1

[0125] The difference from Example 3 is that the degradable microspheres were prepared by Comparative Preparation Example 1.

[0126] Comparative Example 2

[0127] The difference from Example 3 is that the degradable microspheres were prepared by Comparative Preparation Example 2.

[0128] Comparative Example 3

[0129] The difference between Example 3 and Example 4 is that the degradable microspheres were prepared using Comparative Preparation Example 3. The degradable microspheres were separated by centrifugation.

[0130] Comparative Example 4

[0131] The difference from Example 3 is that the degradable microspheres were prepared by Comparative Preparation Example 4.

[0132] Comparative Example 5

[0133] The difference from Example 3 is that no *Aspergillus cristatus* seed culture was inoculated for fermentation in step S1.

[0134] The preparation method is as follows:

[0135] S1. Extraction of dark tea: Crush 10g of dark tea leaves, add 150mL of water, heat under reflux for 4 hours, cool to room temperature, filter, and obtain dark tea extract;

[0136] S2. Degradation of alkaloids: 1.5g of the degradation microspheres prepared in Preparation Example 3 were added to 100g of black tea extract, stirred and degraded for 30min, the microspheres were separated by magnet, and freeze-dried to obtain black tea extract with alkaloids removed.

[0137] S3. Extraction of the traditional Chinese medicine composition: 15g of Ziziphus jujuba seed, 10g of Poria cocos, 10g of Polygala tenuifolia, 4g of Glycyrrhiza uralensis, 2g of Panax ginseng, and 4g of Rehmannia glutinosa were washed, dried, pulverized, mixed, and added to 500mL of 55wt% ethanol aqueous solution. The mixture was heated and refluxed for 4h, filtered, and the extraction was repeated 3 times. The filtrates were combined, concentrated, and dried to obtain the traditional Chinese medicine composition.

[0138] S4. Preparation of sustained-release tablets: 12g of black tea extract with alkaloids removed, 9g of traditional Chinese medicine composition, 25g of lactose, 7g of microcrystalline cellulose, 6g of magnesium stearate, 15g of sodium carboxymethyl cellulose, 17g of hydroxypropyl cellulose K15M, and 7g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0139] Comparative Example 6

[0140] The difference from Example 3 is that step S2 was not performed.

[0141] The preparation method is as follows:

[0142] S1. Fermentation of dark tea: Crush 10g of dark tea leaves, add 150mL of water, heat and reflux for 4h, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid at an inoculation amount of 3v / v%, pH value of 5.5, temperature of 30℃, rotation speed of 125r / min, ferment for 30h, filter to obtain dark tea fermentation liquid.

[0143] S2. Extraction of the traditional Chinese medicine composition: 15g of Ziziphus jujuba seed, 10g of Poria cocos, 10g of Polygala tenuifolia, 4g of Glycyrrhiza uralensis, 2g of Panax ginseng, and 4g of Rehmannia glutinosa were washed, dried, pulverized, mixed, and added to 500mL of 55wt% ethanol aqueous solution. The mixture was heated and refluxed for 4h, filtered, and the extraction was repeated 3 times. The filtrates were combined, concentrated, and dried to obtain the traditional Chinese medicine composition.

[0144] S3. Preparation of sustained-release tablets: 12g of fermented black tea, 9g of traditional Chinese medicine composition, 25g of lactose, 7g of microcrystalline cellulose, 6g of magnesium stearate, 15g of sodium carboxymethyl cellulose, 17g of hydroxypropyl cellulose K15M, and 7g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

[0145] Comparative Example 7

[0146] The difference from Example 3 is that no traditional Chinese medicine composition was added in step S4.

[0147] The preparation method is as follows:

[0148] S1. Fermentation of dark tea: Crush 10g of dark tea leaves, add 150mL of water, heat and reflux for 4h, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid at an inoculation amount of 3v / v%, pH value of 5.5, temperature of 30℃, rotation speed of 125r / min, ferment for 30h, filter to obtain dark tea fermentation liquid.

[0149] S2. Degradation of alkaloids: 1.5g of the degradation microspheres prepared in Example 3 were added to 100g of black tea fermentation liquid, stirred and degraded for 30min, the microspheres were separated by magnet, and freeze-dried to obtain black tea fermentation product with alkaloids removed;

[0150] S3. Preparation of sustained-release tablets: 21g of alkaloid-free fermented black tea, 25g of lactose, 7g of microcrystalline cellulose, 6g of magnesium stearate, 15g of sodium carboxymethyl cellulose, 17g of hydroxypropyl cellulose K15M, and 7g of sodium alginate were mixed evenly, granulated, and compressed into tablets to obtain a soothing and sleep-aiding black tea composition.

[0151] Test Example 1

[0152] Healthy Wistar rats, weighing 180-220g, of any sex, were randomly divided into a normal group, a model group, a positive control group, Example 1-3 groups, and Comparative Examples 1-7 groups, with 10 rats in each group. Except for the normal group, the other groups received an intraperitoneal injection of 300mg / kg of p-chlorophenylalanine. After 28-30 hours, their diurnal rhythm disappeared, and they remained active during the day, with significantly reduced total sleep, almost all reaching complete insomnia. Example 1-3 and Comparative Examples 1-7 groups received 5g / kg of the corresponding product via gavage daily. The normal and model groups received an equal amount of water. The positive control group received 1mg / kg of diazepam injection via intraperitoneal injection. This treatment continued for 7 days. The rats were euthanized by decapitation, the skull was removed, and the brain tissue was fully exposed. The medulla oblongata was quickly severed at the foramen magnum, and the brain tissue was extracted to detect the levels of serotonin, 5-hydroxyindoleacetic acid, norepinephrine, and dopamine.

[0153] The results are shown in Table 1.

[0154] Table 1

[0155]

[0156] Note: * indicates P < 0.05 compared to the normal group; # indicates P < 0.05 compared to the model group.

[0157] As shown in the table above, the calming and sleep-aiding black tea herbal compound composition prepared in Examples 1-3 of this invention can significantly increase the content of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid, and decrease the content of norepinephrine and dopamine.

[0158] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a dark tea-based herbal compound composition for calming the nerves and aiding sleep, characterized in that, After fermentation, degradable microspheres are added to the fermentation liquid and stirred to degrade the alkaloids, resulting in fermented black tea product with alkaloids removed. This product is then mixed evenly with a traditional Chinese medicine composition extracted from jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate. The mixture is then granulated, compressed into tablets, and a calming and sleep-aiding black tea traditional Chinese medicine compound composition is obtained.

2. The preparation method according to claim 1, characterized in that, Includes the following steps: S1. Fermentation of dark tea: Pulverize dark tea leaves, add water, heat and reflux to extract, cool to room temperature, sterilize, inoculate with Aspergillus cristatus seed liquid, ferment and culture, filter to obtain dark tea fermentation liquid; S2. Degradation of alkaloids: Add degradation microspheres to the fermented black tea liquid, stir to degrade, separate the microspheres with a magnet, freeze dry, and obtain fermented black tea product with alkaloids removed; S3. Extraction of the Chinese herbal composition: The jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa were washed, dried, pulverized, mixed, added to an ethanol aqueous solution, heated under reflux for extraction, filtered, and the extraction was repeated. The filtrates were combined, concentrated, and dried to obtain the Chinese herbal composition. S4. Preparation of sustained-release tablets: The alkaloid-free fermented black tea, the traditional Chinese medicine composition, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate are mixed evenly, granulated, and compressed into tablets to obtain a calming and sleep-aiding black tea traditional Chinese medicine compound composition.

3. The preparation method according to claim 2, characterized in that, The bacterial count of the *Eurotium cristatum* seed solution in step S1 is 10. 8 -10 9 The concentration of cfu / mL and the inoculum amount are 2-4 v / v%. The heating and reflux extraction time is 3-5 h. The solid-liquid ratio of black tea leaves to water is 1:10-20 g / mL. The fermentation culture conditions are pH 5-6, temperature 28-32℃, rotation speed 100-150 r / min, and time 24-36 h.

4. The preparation method according to claim 2, characterized in that, In step S2, the mass ratio of the black tea fermentation liquid to the degradation microspheres is 100:1-2. The preparation method of the degradation microspheres is as follows: T1. Preparation of mesoporous TiO2 nanospheres: Tetrabutyl titanate and a pore-forming agent were dissolved in an organic solvent to obtain an oil phase; an emulsifier was added to water to obtain an aqueous phase; the oil phase was added dropwise to the aqueous phase, emulsified, the pH of the solution was adjusted, centrifuged, washed, dried, and calcined to obtain mesoporous TiO2 nanospheres; T2. Preparation of MoS2 quantum dots / mesoporous TiO2 nanospheres: Mesoporous TiO2 nanospheres were added to water, sodium molybdate was added, the pH of the solution was adjusted, glutathione was added, hydrothermal reaction was carried out, centrifugation was performed, washing was performed, and drying was carried out to obtain MoS2 quantum dots / mesoporous TiO2 nanospheres. T3. Preparation of zero-valent iron sulfide: Zero-valent iron powder is evenly spread on the upper stainless steel sieve, sulfur powder is placed in the lower crucible, and under the protection of inert gas, it is heated and calcined, and then cooled to room temperature to obtain zero-valent iron sulfide. T4. Polydopamine modification: MoS2 quantum dots / mesoporous TiO2 nanospheres and zero-valent iron sulfide were added to Tris-HCl solution, dopamine hydrochloride was added, the mixture was heated and stirred to react, centrifuged, washed, dried, and ball-milled to obtain the modified composite. T5. The seed culture of Pseudomonas stearotherm, obtained after activation, is fermented, lysozyme is added for enzymatic hydrolysis, filtered, and the filtrate is dialyzed to obtain a cell wall broken bacterial culture. T6. Preparation of degradable microspheres: N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide were added to the cell wall-broken bacterial solution, stirred and activated, the modified complex was added, the reaction was stirred, separated by magnet, washed, and freeze-dried to obtain degradable microspheres.

5. The preparation method according to claim 4, characterized in that, In step T1, the mass ratio of tetrabutyl titanate, pore-forming agent, and emulsifier is 10-15:1-2:2-3. The pore-forming agent is hexadecyltrimethylammonium bromide or hexadecyltrimethylammonium chloride, and the emulsifier is selected from at least one of Tween-20, Tween-40, Tween-60, Tween-80, and Tween-85. The calcination temperature is 400-500℃, and the time is 2-4 hours. In step T2, the mass ratio of mesoporous TiO2 nanospheres, sodium molybdate, and glutathione is 10-15:1-3:1.5-2.

5. The pH of the solution is adjusted to 6.2-6.7, and the hydrothermal reaction temperature is 180-220℃, and the time is 10-15 hours.

6. The preparation method according to claim 4, characterized in that, In step T3, the mass ratio of zero-valent iron powder to sulfur powder is 1-2:0.1-0.3, and the calcination temperature is 400-500℃ for 1-2 hours. In step T4, the mass ratio of MoS2 quantum dots / mesoporous TiO2 nanospheres, zero-valent iron sulfide, and dopamine hydrochloride is 10-14:3-5:4-6, the pH of the Tris-HCl solution is 8.5-9.5, and the heating and stirring reaction temperature is 45-55℃ for 3-5 hours.

7. The preparation method according to claim 4, characterized in that, The bacterial count of the inoculum solution mentioned in step T5 is 10. 8 -10 9 The fermentation conditions are 36-37℃, 200-250 r / min, and 60-72 h. The amount of lysozyme added is 3-5 wt% of the total mass of the system. The dialysis bag used for dialysis has a pore size of 2k-3kDa. In step T6, the mass ratio of the cell wall-breaking bacterial solution, N-hydroxysuccinimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and the modified complex is 100-150:3-5:4-7:15-20.

8. The preparation method according to claim 4, characterized in that, In step S3, the mass ratio of jujube seed, poria cocos, polygala tenuifolia, licorice, ginseng, and rehmannia glutinosa is 10-20:5-15:5-15:3-5:2-4:3-5, the concentration of the ethanol aqueous solution is 50-60 wt%, and the heating and reflux extraction time is 3-5 h.

9. The preparation method according to claim 4, characterized in that, The mass ratio of the black tea fermentation product with alkaloids removed, the traditional Chinese medicine composition, lactose, microcrystalline cellulose, magnesium stearate, sodium carboxymethyl cellulose, hydroxypropyl cellulose K15M, and sodium alginate in step S4 is 10-15:8-10:20-30:5-10:4-8:10-20:15-20:5-10.

10. A sleep-aiding dark tea herbal compound composition prepared by the preparation method according to any one of claims 1-9.

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