Ginseng pu'er tea for alleviating fatigue caused by staying up late, and preparation method and application thereof

Abstract

The present application provides a ginseng Pu'er tea for relieving fatigue caused by staying up late, a preparation method and application thereof, and belongs to the technical field of health-care tea. The preparation method of the ginseng Pu'er tea comprises sub-stage blanching and soaking of Pu'er raw tea and ginseng and three-stage co-fermentation. The present application activates and inactivates endogenous enzymes through sub-stage blanching, promotes microbial conversion in a directed manner through three-stage temperature control, significantly enriches rare saponins such as Rg3, Rh1 and CK, and the prepared ginseng Pu'er tea can significantly relieve fatigue caused by staying up late. Ginseng and Pu'er tea have a synergistic effect, and can be used for preparing tea bags, loose tea, tea bricks, tea cakes, liquid tea drinks and other foods.

Description

Technical Field

[0001] This invention belongs to the field of health tea technology, and in particular relates to a ginseng Pu-erh tea for relieving fatigue from staying up late, its preparation method and application. Background Technology

[0002] Staying up late has become a widespread phenomenon in modern society. Prolonged sleep deprivation or lack of sleep can lead to a range of health problems, including increased fatigue, decreased cognitive function, weakened immunity, and oxidative stress damage. While various functional foods exist on the market to relieve fatigue, most rely on caffeine or vitamins as their main ingredients, resulting in limited mechanisms of action, short-lived effects, or side effects from excessive intake. Therefore, developing a natural, safe, and sustainable product to relieve fatigue from staying up late is of significant practical importance.

[0003] Pu-erh tea is a unique post-fermented tea variety in my country, renowned for its distinctive pile fermentation process. Studies have shown that ripe Pu-erh tea is rich in active ingredients such as tea polyphenols, theabrownins, and gallic acid, possessing various physiological activities including antioxidant activity, regulation of intestinal flora, and improvement of lipid metabolism. In recent years, the anti-fatigue effects of Pu-erh tea have gradually gained attention. Ginseng, as a traditional food and medicine ingredient, has had its main active ingredient, ginsenosides, proven to have significant anti-fatigue, antioxidant, and neuroprotective effects. Ginsenosides Rb1 and Rg1 can alleviate physical and mental fatigue through multiple pathways, including regulating the hypothalamus-pituitary-adrenal axis, enhancing mitochondrial function, and scavenging free radicals. However, ginseng used alone often suffers from slow onset of action, a bitter taste, and the potential for causing internal heat with long-term use, limiting its widespread application in daily beverages.

[0004] The blending of ginseng and tea has become a hot topic in the development of functional tea drinks in recent years. Existing technologies include directly adding ginseng slices or powder to Pu-erh tea to make tea bags or compressed tea cakes. However, these technologies are limited to simple physical mixing and do not involve deep co-fermentation. Therefore, there is an urgent need to develop a health-promoting ginseng Pu-erh tea that can achieve deep co-fermentation of ginseng and Pu-erh tea, selectively enrich rare saponins, and significantly enhance the effect of relieving fatigue from staying up late. Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide a ginseng Pu-erh tea for relieving fatigue from staying up late, its preparation method and application, which enables ginseng and Pu-erh tea to undergo deep co-fermentation, thereby selectively enriching highly active rare saponins and significantly improving the effect of relieving fatigue from staying up late.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late includes blanching and soaking raw Pu-erh tea and ginseng, followed by co-fermentation. The blanching and soaking process is carried out in stages: first, the raw Pu-erh tea and ginseng are blanched at 55-65℃ for 15-25 minutes, controlling the moisture content to 30%-33%; then, they are blanched at 85-95℃ for 3-5 minutes, controlling the moisture content to 34%-38%. The co-fermentation process is carried out in three stages: core temperature 40-45℃, maintained for 10-15 days; core temperature 50-54℃, maintained for 20-30 days; core temperature 45-48℃, maintained for 10-15 days.

[0007] Preferably, the mass percentage of the raw Pu-erh tea and ginseng is: 70%~95% raw Pu-erh tea and 5%~30% ginseng.

[0008] More preferably, the raw Pu-erh tea is Yunnan large-leaf variety raw Pu-erh tea; and the ginseng is a whole ginseng root that is five years old or younger.

[0009] Preferably, the pile is turned over every 5 to 12 days during the co-fermentation process.

[0010] More preferably, during the co-fermentation process, water is replenished with a solution of 0.05% to 0.1% glucose and 0.02% to 0.05% peptone each time the pile is turned.

[0011] Preferably, it also includes post-processing and shaping: after co-fermentation and maturation, the material is opened and cooled, then dried at a gradient low temperature until the moisture content of the material is 8%~11%, screened and sorted, and aged at room temperature in the dark for more than 15 days.

[0012] More preferably, the gradient low-temperature drying includes: a first stage of drying at 40~45℃ for 12~24h until the moisture content of the material drops to 15%~18%; and a second stage of drying at 30~35℃ for 24~48h until the moisture content of the material drops to 8%~11%.

[0013] This invention also provides ginseng Pu-erh tea prepared by the above method and its application in the preparation of food products that relieve fatigue from staying up late.

[0014] Preferably, the food is a tea bag, loose tea, tea brick, tea cake, liquid tea drink, concentrated liquid, or solid beverage.

[0015] Compared with the prior art, the present invention has the following beneficial effects: (1) By slicing and soaking in water in stages, the endogenous β-glucosidase of ginseng is activated at low temperature, and ginsenoside Rb1 is initially hydrolyzed into Rd and Rg3; at high temperature, the enzyme is inactivated and the bacteria are killed, which not only provides substrate for subsequent microbial transformation, but also avoids non-specific degradation.

[0016] (2) Three-stage temperature-controlled co-fermentation (induction period - conversion period - stabilization period) combined with turning and adding carbon and nitrogen sources to promote the growth of β-glucosidase-producing microorganisms and efficiently convert ginsenosides into rare saponins such as Rg3, Rh1, and CK. The proportion of rare saponins can reach more than 25%.

[0017] (3) The ginseng-pu-erh tea combination has a significant synergistic effect in relieving fatigue caused by staying up late, which is superior to the same dose of ginseng or pu-erh tea alone.

[0018] (4) The product has a mellow taste and does not have the bitterness or heatiness of ginseng alone. It is highly safe and is practically non-toxic. Attached Figure Description

[0019] Figure 1 This invention relates to a pre-fermentation pile of blanched and soaked raw Pu-erh tea and whole ginseng.

[0020] Figure 2 The ginseng Pu-erh tea product of this invention. Detailed Implementation

[0021] This invention provides a method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, comprising the following steps: (1) Raw material selection: In this invention, the preferred raw Pu-erh tea is Yunnan large-leaf Pu-erh tea, and more preferably, it is Yunnan large-leaf Pu-erh tea that is free from mold, odor, and of qualified quality.

[0022] In this invention, the preferred ginseng is a whole ginseng root that is five years old or younger, which is washed before use.

[0023] In this invention, the preferred mass percentage of raw Pu-erh tea and ginseng is: 70%~95% raw Pu-erh tea and 5%~30% ginseng; more preferably, 80%~90% raw Pu-erh tea and 10%~20% ginseng; and even more preferably, 85% raw Pu-erh tea and 15% ginseng.

[0024] This ratio uses raw Pu-erh tea as the main fermentation substrate, providing the carbon source and growth factors needed for microbial metabolism such as tea polyphenols and polysaccharides. At the same time, its rich polyphenols are transformed into functional components such as theaflavins and thearubigins during fermentation. Ginseng is the source of active ingredients. If the amount added is too low, the enrichment effect of rare saponins will not be obvious. If it is too high, it will affect the normal progress of pile fermentation (ginsenosides have certain antibacterial activity, and excessive addition will inhibit the growth of fermenting microorganisms). It will also lead to a bitter taste in the product.

[0025] (2) Blanching and soaking: In this invention, the blanching and soaking process is carried out in two stages: The first stage involves blanching the raw Pu-erh tea and ginseng at 55-65℃ for 15-25 minutes, controlling the moisture content to 30%-33%; preferably, blanching at 60℃ for 20 minutes, controlling the moisture content to 32%. The main purpose of this stage is to activate endogenous hydrolytic enzymes such as β-glucosidase in the ginseng raw material. 55-65℃ is the optimal activation temperature window for these enzymes. At this temperature, ginsenoside Rb1 can be initially hydrolyzed into low-glycosylated saponins such as Rd and Rg3, laying the substrate foundation for the subsequent microbial enzymatic transformation during pile fermentation. Simultaneously, this temperature also helps to initially soften the raw Pu-erh tea leaves, preparing them for the subsequent high-temperature blanching.

[0026] The second stage involves blanching at 85-95℃ for 3-5 minutes, controlling the moisture content to 34%-38%; preferably, blanching at 90℃ for 4 minutes, controlling the moisture content to 35%. The main purpose of this stage is to inactivate enzymes and fix the shape. High-temperature treatment deactivates the endogenous enzymes activated in the first stage, preventing excessive enzymatic reactions that could lead to non-specific degradation of saponins. At the same time, it kills miscellaneous bacteria attached to the surface of the raw materials, reducing competition from harmful microorganisms during pile fermentation. In addition, high-temperature blanching further softens the raw Pu-erh tea leaves and destroys the cell wall structure, which is conducive to the invasion of microbial mycelia and the diffusion of extracellular enzymes during subsequent pile fermentation.

[0027] In this invention, the preferred material-to-liquid ratio is 1:2 to 4, and more preferably 1:3.

[0028] (3) Co-fermentation: In this invention, co-fermentation is carried out in three stages: Phase 1 (Induction Period): Core temperature 40-45℃, maintained for 10-15 days; preferred core temperature 42-43℃, maintained for 12-13 days. This phase is the low-temperature induction period, dominated by mesophilic fungi such as Aspergillus niger. These fungi proliferate rapidly under low-temperature conditions, secreting abundant extracellular enzyme systems, including cellulase, pectinase, protease, and β-glucosidase. In this phase: ① fungal extracellular enzymes gradually hydrolyze ginsenoside Rb1 into Rd and a small amount of Rg3; ② macromolecular polysaccharides and proteins in raw Pu-erh tea are degraded into small soluble substances, providing nutrients for bacterial and yeast growth in subsequent stages; ③ a stable microbial community structure is established. The temperature in this phase should not exceed 45℃, otherwise fungal growth will be inhibited and enzyme production efficiency will decrease.

[0029] The second stage (conversion period): the core temperature is 50-54℃, maintained for 20-30 days; the preferred core temperature is 51-52℃, maintained for 24-26 days. This stage is a mesophilic conversion period. As the core temperature rises, fungi gradually decline, and heat-resistant yeasts and Bacillus become the dominant microbial population. These microorganisms have a stronger ability to hydrolyze saponins and glycosyl groups, further converting the Rd and Rg3 produced in the first stage into highly active rare saponins such as Rh1, Rh2, and compound K (CK). Simultaneously, under the action of microbial polyphenol oxidase, catechins are converted into theaflavins and thearubigins, giving Pu-erh tea its unique color and flavor.

[0030] The third stage (stabilization period): The core temperature is 45-48℃, maintained for 10-15 days; the preferred core temperature is 46-47℃, maintained for 12-13 days. This stage is a cooling and stabilization period, in which the microbial activity is slowed down by lowering the core temperature, preventing further excessive metabolism of rare saponins. This stage: ① stabilizes the already generated rare saponins in a mild environment, preventing further degradation by residual enzymes; ② promotes the formation of molecular complexes between ginsenosides and substances such as tea polyphenols and theabrownins, improving the stability and bioavailability of active ingredients; ③ eliminates off-flavor substances produced during fermentation, making the tea soup taste more mellow and harmonious.

[0031] In this invention, it is preferred that the pile be turned every 5-12 days during co-fermentation; more preferably, it is preferred that the pile be turned every 5-7 days during the first stage (induction period), every 5-7 days during the second stage (conversion period), and every 10-12 days during the third stage (stabilization period). Turning the pile during co-fermentation helps to dissipate heat and lower the temperature, preventing excessively high core temperatures that could lead to microbial community imbalance and thermal degradation of active ingredients; it also replenishes oxygen, providing the necessary oxygen for aerobic and facultative anaerobic microorganisms to maintain fermentation activity; it evens out moisture and temperature, avoiding uneven fermentation caused by localized overheating or overhumidification; and it breaks up clumps, increasing the contact area between the material and air.

[0032] In this invention, it is preferable to replenish water with a solution of 0.05%~0.1% (mass percentage) glucose and 0.02%~0.05% (mass percentage) peptone each time the compost is turned; more preferably, a solution of 0.08% glucose and 0.04% peptone is used for water replenishment; even more preferably, no water replenishment is needed when the compost is nearing the end of fermentation (within 2 days). By introducing low concentrations of carbon and nitrogen sources into the water replenishment during composting, the dominant growth of β-glucosidase-producing microbial strains is promoted in a targeted manner, increasing the overall activity of β-glucosidase in the system, thereby accelerating the targeted conversion of ginsenosides to rare saponins; replenishing the fermentation system with nutrients that can be directly utilized by microorganisms, maintaining fermentation activity, avoiding fermentation stagnation due to nutrient depletion, and maintaining a moisture content of 34%~38%.

[0033] (4) Post-processing molding: In this invention, it is preferred that after the co-fermentation matures, the material is opened and cooled, then dried at a gradient low temperature until the moisture content of the material is 8%~11%, screened and sorted, and aged at room temperature in the dark for more than 15 days.

[0034] In this invention, a further preferred method of gradient low-temperature drying includes: a first stage of drying at 40-45°C for 12-24 hours until the moisture content of the material drops to 15%-18%; and a second stage of drying at 30-35°C for 24-48 hours until the moisture content of the material drops to 8%-11%. Gradient low-temperature drying can effectively avoid the damage of heat-sensitive active ingredients such as rare saponins and theaflavins caused by high temperatures.

[0035] In this invention, it is preferable to remove large pieces of ginseng, coarse old stems and clumps during the screening and sorting process, while retaining high-quality Pu-erh tea and some ginseng rootlets.

[0036] In this invention, it is preferred to age at room temperature in the dark for 15 to 90 days, and more preferably for 15 to 60 days.

[0037] This invention also provides ginseng Pu-erh tea prepared by the above method and its application in the preparation of food products that relieve fatigue from staying up late.

[0038] In this invention, the preferred food products are tea bags, loose tea, tea bricks, tea cakes, liquid tea drinks, concentrated liquids, or solid beverages.

[0039] As one possible implementation method, the tea bag is made by packing the ginseng Pu'er tea of ​​the present invention into a filter paper bag using the conventional tea bag brewing process, and then sealing it.

[0040] As one possible implementation method, loose tea refers to directly packaging the above-mentioned ginseng Pu'er tea of ​​the present invention in a fixed quantity (such as 50g, 100g or 250g per bag).

[0041] As one possible implementation method, the tea bricks / cakes are made by steaming and softening the ginseng Pu'er tea described above, pressing it into square or round tea bricks / cakes, drying it, and then packaging it.

[0042] As one possible implementation method, the liquid tea beverage is prepared by steeping the ginseng Pu'er tea mentioned above in boiling water, filtering to extract the juice, adding appropriate auxiliary materials, sterilizing and bottling to produce a ready-to-drink tea beverage.

[0043] As one possible implementation method, the concentrate is made by concentrating the above-mentioned ginseng Pu'er tea liquid beverage and then bottling it.

[0044] As one possible implementation method, the solid beverage is made by spray drying or freeze drying the above-mentioned ginseng Pu'er tea liquid tea into powder, adding an appropriate amount of excipients, mixing and then bottling.

[0045] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0046] Example 1 A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, the steps are as follows: (1) Raw material selection Pu-erh raw tea: Yunnan large-leaf sun-dried green tea, free from mold and off-odors, of qualified quality, quantity 170kg.

[0047] Ginseng: Whole, five-year-old artificially cultivated ginseng, dosage 30kg.

[0048] (2) Blanching and soaking First stage: Put raw Pu'er tea and ginseng into a blanching tank, add drinking water (material-to-liquid ratio 1:3), and blanch at 60℃ for 20 minutes, stirring twice during the process, controlling the moisture content to 32%.

[0049] Second stage: Quickly raise the temperature to 90℃, continue blanching for 4 minutes, remove and drain, and control the moisture content to 35%.

[0050] (3) Co-fermentation Pit the blanched mixture, such as Figure 1 As shown, the stack height is 50cm, the stack width is 80cm, the workshop temperature is 25±2℃, and the relative humidity is 55±5%.

[0051] Induction period (days 1-12): Core temperature is controlled at 42±1℃, and the reactor is turned over every 6 days. On the 6th day, when turning the reactor over, a makeup water solution containing 0.08% glucose and 0.04% peptone is used to maintain a moisture content of 35%~38%.

[0052] Conversion period (days 13-37): Core temperature is controlled at 52±1℃. The reactor is turned over every 6 days, and water is replenished each time the reactor is turned over to maintain a moisture content of 35%~38%.

[0053] Stabilization period (days 38-50): Core temperature is controlled at 46±1℃. The reactor is turned over every 10 days, and water is replenished during the turning process, with the moisture content gradually reduced to 32%~35%.

[0054] (4) Post-processing molding After fermentation for 50 days, the mixture is removed from the pile and allowed to cool to room temperature.

[0055] Gradient low-temperature drying: the first stage is drying at 42℃ for 18 hours to a moisture content of 16%; the second stage is drying at 32℃ for 36 hours to a moisture content of 9%.

[0056] Sieve the ginseng using a double-layer vibrating sieve (6 mesh and 12 mesh) to remove coarse stems and clumps, and pick out large pieces of ginseng.

[0057] Aged at room temperature in the dark for 45 days, the finished product is obtained. Figure 2 As shown.

[0058] Example 2 A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, the steps are as follows: (1) Raw material selection Pu-erh raw tea: Same as in Example 1, dosage 140kg.

[0059] Ginseng: Whole, five-year-old artificially cultivated ginseng, dosage 60kg.

[0060] (2) Blanching and soaking First stage: Blanch at 55℃ for 25 minutes, controlling the moisture content to 30%.

[0061] Second stage: Blanch at 85℃ for 5 minutes, controlling the moisture content to 34%.

[0062] (3) Co-fermentation Induction period: Core temperature 40±1℃, maintained for 15 days, with the reactor core turned over every 7 days. On the 7th day, a makeup solution containing 0.05% glucose and 0.02% peptone is used for turning over.

[0063] Conversion period: Core temperature 50±1℃, maintained for 30 days, with the reactor core turned over every 7 days, and water replenishment maintained each time the core is turned over.

[0064] Stabilization period: Core temperature 45±1℃, maintained for 15 days, with the reactor being turned over every 12 days and water replenishment maintained.

[0065] (4) Post-processing molding After fermentation for 60 days, the mixture is piled up and spread out to cool.

[0066] Gradient low-temperature drying: the first stage is drying at 40℃ for 24 hours until the moisture content is 18%; the second stage is drying at 30℃ for 48 hours until the moisture content is 11%.

[0067] After screening and aging for 30 days, the finished product is obtained.

[0068] Example 3 A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, the steps are as follows: (1) Raw material selection Pu-erh raw tea: Same as in Example 1, dosage 190kg.

[0069] Ginseng: Whole, five-year-old artificially cultivated ginseng, dosage 10kg.

[0070] (2) Blanching and soaking First stage: Blanch at 65℃ for 15 minutes, controlling the moisture content to 33%.

[0071] Second stage: Blanch at 95℃ for 3 minutes, controlling the moisture content to 38%.

[0072] (3) Co-fermentation Induction period: Core temperature 45±1℃, maintained for 10 days, with the reactor core turned over every 5 days. On the 5th day, a makeup solution containing 0.1% glucose and 0.05% peptone is used for turning over.

[0073] Conversion period: Core temperature 54±1℃, maintained for 20 days, with the reactor core turned over every 5 days, and water replenishment maintained during each turn.

[0074] Stabilization period: Core temperature 48±1℃, maintained for 10 days, with the reactor being turned over once every 10 days. Since the reactor is turned over on the last day, no water replenishment is required.

[0075] (4) Post-processing molding After fermentation for 40 days, the mixture is piled up and spread out to cool.

[0076] Gradient low-temperature drying: the first stage is drying at 45℃ for 12 hours until the moisture content is 15%; the second stage is drying at 35℃ for 24 hours until the moisture content is 8%.

[0077] After screening and aging for 15 days, the finished product is obtained.

[0078] Example 4 A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, the steps are as follows: (1) Raw material selection Pu-erh raw tea: Same as in Example 1, dosage 160kg.

[0079] Ginseng: Five-year-old artificially cultivated ginseng, dosage 40kg.

[0080] (2) Blanching and soaking First stage: Blanch at 58℃ for 22 minutes, controlling the moisture content to 31%.

[0081] Second stage: Blanch at 88℃ for 4 minutes, controlling the moisture content to 36%.

[0082] (3) Co-fermentation Induction period: Core temperature 43±1℃, maintained for 13 days, with the reactor core turned over every 6 days. On the 6th day, a makeup solution containing 0.07% glucose and 0.03% peptone is used for turning over.

[0083] Conversion period: Core temperature 51±1℃, maintained for 25 days, with the reactor being turned over every 6 days. Each turn uses a makeup water solution containing 0.07% glucose and 0.03% peptone.

[0084] Stabilization period: Core temperature 47±1℃, maintained for 12 days, with the pile turned over every 11 days. No water replenishment is required as fermentation is nearing completion.

[0085] (4) Post-processing molding After fermentation for 50 days, the mixture is piled up and spread out to cool.

[0086] Gradient low-temperature drying: the first stage is drying at 43℃ for 20 hours until the moisture content is 17%; the second stage is drying at 33℃ for 30 hours until the moisture content is 10%.

[0087] After screening and aging for 40 days, the finished product is obtained.

[0088] Example 5 A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, the steps are as follows: (1) Raw material selection Pu-erh raw tea: Same as in Example 1, dosage 180kg.

[0089] Ginseng: Five-year-old artificially cultivated ginseng, dosage 20kg.

[0090] (2) Blanching and soaking First stage: Blanch at 62℃ for 18 minutes, controlling the moisture content to 32%.

[0091] Second stage: Blanch at 92℃ for 3.5 minutes, controlling the moisture content to 37%.

[0092] (3) Co-fermentation Induction period: Core temperature 44±1℃, maintained for 11 days, with the reactor being turned over every 5 days. On the 5th day, a makeup solution containing 0.09% glucose and 0.045% peptone is used for turning over.

[0093] Conversion period: Core temperature 53±1℃, maintained for 22 days, with the reactor being turned over every 5 days. Each turn uses a makeup water solution containing 0.09% glucose and 0.045% peptone.

[0094] Stabilization period: Core temperature 46±1℃, maintained for 11 days, with the pile turned over every 10 days. As fermentation is nearing its end, no additional water is needed.

[0095] (4) Post-processing molding After fermentation for 44 days, the mixture is piled up and spread out to cool.

[0096] Gradient low-temperature drying: the first stage is drying at 44℃ for 15 hours to a moisture content of 16%; the second stage is drying at 34℃ for 40 hours to a moisture content of 9%.

[0097] After screening and aging for 50 days, the finished product is obtained.

[0098] Comparative Example 1 Ingredients: Pure raw Pu-erh tea, without added ginseng.

[0099] Treatment: The process was carried out according to the blanching and soaking and three-stage temperature-controlled co-fermentation process in Example 1.

[0100] Comparative Example 2 Raw materials: Same as in Example 1.

[0101] Processing: Skip the blanching step and directly pile the dry materials together for three-stage temperature-controlled co-fermentation.

[0102] Comparative Example 3 Raw materials: Same as in Example 1 Processing: Blanch the Pu-erh tea and ginseng at 90℃ for 5 minutes to control the moisture content to 35%.

[0103] Comparative Example 4 Raw materials: Same as in Example 1.

[0104] Treatment: The co-fermentation was changed to natural heating, with the core temperature controlled at 40~58℃ for a period of 50 days.

[0105] Comparative Example 5 Raw materials: Pu-erh raw tea fermented separately; ginseng ground into fine powder ≤0.15mm.

[0106] Processing: The raw Pu-erh tea was blanched in stages and fermented in three stages under controlled temperature according to Example 1; after fermentation, ginseng powder (15%) was mixed in.

[0107] Experimental Example 1 1. Detection of active ingredients (1) Sample pretreatment: Accurately weigh 1.0g of each component tea powder (pass through a 40-mesh sieve), add 50mL of 70% methanol aqueous solution, and extract by ultrasonic extraction (power 250W, frequency 40kHz) for 30min. After cooling, make up the weight loss, filter through a 0.22μm microporous membrane, and take the filtrate for testing.

[0108] (2) Chromatographic conditions: High-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used for determination. Column: C18 column (4.6 mm × 250 mm, 5 μm); Mobile phase: Acetonitrile (A) - water (B), gradient elution: 0~20 min, 20%~30% A; 20~35 min, 30%~45% A; 35~50 min, 45%~70% A; 50~55 min, 70%~20% A; Flow rate: 1.0 mL / min; Column temperature: 30℃; Injection volume: 10 μL. ELSD parameters: Drift tube temperature: 80℃; Carrier gas (nitrogen) flow rate: 2.0 L / min.

[0109] (3) Standards and quantification: Quantification was performed using external standard method with ginsenosides Rb1, Rd, Rg3, Rh1, and CK standards (purity ≥98%). The total saponin content was the sum of the above five monomeric saponins.

[0110] (4) Test results: As shown in Table 1, the proportion of rare saponins in Examples 1-5 (25.9%-31.0%) was much higher than that in Comparative Examples 2-5 (7.8%-15.8%), proving that the synergistic effect of segmented blanching and three-stage temperature control is the key to the efficient conversion of rare saponins.

[0111] Table 1. Results of detection of active ingredients in different ginseng Pu-erh teas

[0112] Note: Comparative Example 1 was made from pure raw Pu-erh tea fermentation without the addition of ginseng, therefore the above-mentioned ginsenosides were not detected; "-" indicates not detected or below the detection limit. Rare saponin percentage = (Rg3 + Rh1 + CK) / total saponins × 100%.

[0113] 2. Polyphenol conversion status The total amount of tea polyphenols in the samples was determined by the ferric tartrate colorimetric method (GB / T 8313-2018); theaflavins, thearubigins, and theabrownins were determined by systematic analysis (GB / T 30483-2013), and the TF / TR value and the TF+TR+TB ratio were calculated.

[0114] Experimental Results: As shown in Table 2, the total tea polyphenol content of Examples 1-5 (13.5%-15.8%) was significantly lower than that of Comparative Example 1 (pure raw Pu-erh tea fermentation, 22.5%), indicating that co-fermentation promoted the oxidative polymerization of tea polyphenols. The theabrownin content of Examples 1-5 (7.95%-8.35%) was moderate, and the TF / TR ratio (0.105-0.112) was within the range of high-quality ripe Pu-erh tea, with a bright red and rich tea soup. Comparative Example 2 had a higher theabrownin content (9.80%) and a lower theaflavin content, indicating that contamination by miscellaneous bacteria led to over-fermentation. Comparative Example 4 (natural heating) had a lower TF / TR, indicating that temperature fluctuations led to uneven fermentation.

[0115] Table 2. Conversion results of polyphenols in different ginseng Pu-erh teas

[0116] Experimental Example 2 Acute oral toxicity test The limit test was conducted according to GB15193.3-2014, "National Food Safety Standard - Acute Oral Toxicity Test".

[0117] 1. Preparation of test samples: Take the ginseng Pu-erh tea prepared in Example 1, add water and decoct twice (material-to-liquid ratio 1:10, 1 hour each time), combine the filtrates, and concentrate to the maximum feasible concentration of 10 g crude drug / mL. Administer doses of 2.5, 5.0, 10.0, and 20.0 g / kg bw at gavage volumes of 0.1, 0.2, 0.4, and 0.8 mL / 10 g·bw, respectively.

[0118] 2. Experimental animals and grouping: Forty SPF-grade ICR mice, half male and half female, weighing 18-22g, were used. They were acclimatized for 3 days in an environment with a temperature of 22±2℃, relative humidity of 50%-60%, and 12h light-dark alternation.

[0119] The animals were randomly divided into four dosage groups and one solvent control group based on their body weight, with eight animals in each group (four males and four females). The solvent control group was given an equal volume of distilled water.

[0120] 3. Experimental Methods: Animals were fasted for 12 hours prior to the experiment, but water was allowed. The test samples were administered via oral gavage in a single dose, with a gavage volume ≤20 mL / kg body weight (bw). Symptoms of poisoning and mortality were observed and recorded after administration for 14 days. Body weight was measured before administration, on day 7, and on day 14 after administration.

[0121] After the observation period, surviving animals were euthanized and grossly dissected to observe for any abnormalities in major organs such as the heart, liver, spleen, lungs, and kidneys. The organs were weighed, and the organ coefficient (organ weight / body weight × 100%) was calculated. Liver and kidney tissues were fixed in 10% neutral formalin, embedded in paraffin, stained with hematoxylin and eosin (HE), and observed under a light microscope for histopathological changes.

[0122] 4. Test Results (1) Symptoms of poisoning and mortality: No obvious symptoms of poisoning were observed in any of the animals after administration of the drug, and no animals died during the 14-day observation period.

[0123] (2) Changes in body weight: The body weight of animals in each group showed a normal growth trend. There were no statistically significant differences between each dose group and the solvent control group (P>0.05).

[0124] (3) Organ coefficient and pathological examination: Gross anatomical examination revealed no obvious abnormalities in the major organs such as heart, liver, spleen, lungs, and kidneys in the animals of each dosage group. Histopathological examination showed that the liver and kidney tissue cells in each group had normal morphology, the liver lobule structure was intact, and no obvious lesions were found in the glomeruli and renal tubules.

[0125] Under the conditions of this experiment, the maximum tolerated dose (MTD) of ginseng Pu-erh tea in mice via oral acute toxicity was greater than 20.0 g / kg·bw, which, according to the acute toxicity dose grading standard, belongs to the practically non-toxic category.

[0126] Experimental Example 3 Animal experiments to improve fatigue caused by staying up late 1. Test materials Animals: SPF-grade ICR male mice, weighing 18-22g. The animals were acclimatized for 3 days in an environment with a temperature of 22±2℃, relative humidity of 50%-60%, and 12h light-dark alternation.

[0127] Test sample: Ginseng Pu-erh tea extract concentrate prepared in Example 1 (prepared according to the process in Example 1, extracted with water, concentrated and freeze-dried to make extract powder, and prepared with distilled water to the required concentration before use).

[0128] Reference standard: Positive control: Ginsenoside Rb1 standard (purity ≥98%), prepared with distilled water to 20 mg / mL, administered by gavage at a dose of 200 mg / kg·bw; Pu-erh tea extract: Take the finished tea of ​​Comparative Example 1 and extract it using the same process. Then, administer the same dose of tea polyphenols as the ginseng Pu-erh tea group by gavage. Ginseng single-ingredient extract: Five-year-old ginseng was extracted using the same process, and the same dose of ginsenosides as that in the ginseng Pu-erh tea group was administered by gavage.

[0129] 2. Experimental Design (Completely Randomized Grouping) As shown in Table 3.

[0130] Table 3. Animal experimental groupings and treatments for improving fatigue caused by staying up late.

[0131] A total of 8 groups × 12 mice = 96 mice.

[0132] 3. Construction of a model for fatigue caused by staying up late The modified sleep deprivation method (gentle stimulation method) was used: sleep deprivation was carried out for 14 consecutive days from 20:00 to 8:00 the next day.

[0133] Method: The mice were placed in individual cages. Whenever a mouse entered a sleep posture (eyes closed, muscles relaxed), its whiskers / back were gently touched with a soft brush to wake it up. They were allowed to eat and drink freely under normal daylight to simulate a human staying up all night.

[0134] 4. Dosing regimen During sleep deprivation, the medication was administered by gavage once daily at 9:00 AM for 14 consecutive days.

[0135] 5. Testing Indicators (1) Behavioral tests were conducted 1 hour after the last administration. ①Rota-rod experiment The rotator was 3 cm in diameter and rotated at 20 rpm. The time (in seconds) it took for the mouse to fall off the rotator was recorded.

[0136] ② Weighted swimming experiment The water temperature was 25±1℃, the water depth was 30cm, and the mouse's tail base was loaded with lead sheeting equal to 5% of its body weight. The time from entry into the water to exhaustion (sinking underwater for 10 seconds without surfacing) was recorded.

[0137] (2) Biochemical indicators After the final behavioral test, mice were fasted for 12 hours but allowed free access to water. Blood was collected from the retro-orbital venous plexus, and serum was separated. The mice were then euthanized by cervical dislocation, and the liver and gastrocnemius muscle were quickly harvested, rinsed in ice-cold physiological saline, and blotted dry with filter paper. The detection indicators and methods are shown in Table 4.

[0138] Table 4. Detection Indicators and Methods

[0139] 6. Statistical processing Data are expressed as mean ± SD, and statistical analysis was performed using SPSS 26.0 software. One-way ANOVA was used for comparisons among multiple groups. LSD-t tests were used for pairwise comparisons between groups when variances were homogeneous, and Games-Howell tests were used when variances were unequal. A p-value < 0.05 was considered statistically significant.

[0140] 7. Test Results (1) Behavioral test results As shown in Table 5, compared with the normal control group, the time spent on the rotarod and the time to exhaustion during weight-bearing swimming were significantly shorter in the overnight stay model group ( P <0.01 indicates that sleep deprivation successfully established a mouse model of physical fatigue and decreased motor coordination. Compared with the sleep deprivation model group, the positive control group (ginsenoside Rb1) and the single-ingredient groups (Pu-erh tea single-ingredient and ginseng single-ingredient) both showed certain improvement effects. P <0.05%, but the degree of recovery is limited; the low, medium and high dose groups of the composition can significantly prolong the rotator time and swimming exhaustion time ( P <0.01), and the effects of the medium and high dose groups of the composition were superior to those of the equivalent dose groups of Pu'er tea and ginseng alone ( ). P <0.01).

[0141] Table 5. Rotation time and time to exhaustion during weight-bearing swimming in each group of mice.

[0142] Note: Compared with the normal control group, #P<0.05, ##P<0.01; compared with the sleep deprivation model group, P<0.05, P<0.01; compared with the single-ingredient group (Pu-erh tea single-ingredient or ginseng single-ingredient), †P<0.05, ††P<0.01.

[0143] (2) Results of biochemical index testing As shown in Table 6, compared with the normal control group, the serum levels of BLA, BUN, LDH, MDA and cortisol in the mice in the overnight-staying model group were significantly increased. P<0.01%, serum SOD activity, melatonin levels, and liver and muscle glycogen content were all significantly reduced ( P <0.01 indicates that sleep deprivation leads to disordered energy metabolism, oxidative stress damage, and neuroendocrine imbalance in mice.

[0144] Compared with the sleep deprivation model group, the positive control group and the single-drug group showed varying degrees of improvement in the above indicators. P <0.05%, but the improvement effect of each dosage group of the composition was more significant ( P <0.01%. Even at low doses, the composition significantly reduced BLA, BUN, and MDA levels, and increased SOD activity and glycogen reserves. P <0.01); the above indicators were significantly improved in both the medium and high dose groups of the composition ( P <0.01 vs model group), and significantly better than the equivalent dose of Pu'er tea single-ingredient group and ginseng single-ingredient group ( P <0.01).

[0145] Specifically, serum levels of BLA, BUN, MDA, and cortisol were significantly lower in the medium and high dose groups of the composition compared to the single-dose group. P <0.01), SOD activity, liver glycogen, muscle glycogen, and melatonin levels were significantly increased compared to the single-agent group ( P <0.01]. There was no statistically significant difference between the medium and high dose groups of the composition. P The result >0.05 indicates that the medium dose (2.5 g / kg·bw) was close to the maximum effect plateau of the ginseng Pu-erh tea of ​​this invention under this model, and further increasing the dose to 5.0 g / kg·bw did not bring about significant additional improvement in biochemical indicators. The above results suggest that the ginseng Pu-erh tea composition of this invention can improve energy depletion, oxidative stress and HPA axis overactivation caused by staying up late through multi-target synergistic effects.

[0146] Table 6 Serum biochemical indicators of mice in each group

[0147] Note: Compared with the normal control group, #P<0.05, ##P<0.01; compared with the sleep deprivation model group, P<0.05, P<0.01; compared with the single-ingredient group (Pu-erh tea single-ingredient or ginseng single-ingredient), †P<0.05, ††P<0.01.

[0148] Based on the above experimental results, it can be seen that the ginseng Pu-erh tea of ​​the present invention can significantly improve the motor coordination and physical endurance of sleep-deprived mice, reduce serum lactate, urea nitrogen and malondialdehyde levels, and increase superoxide dismutase activity and liver glycogen reserves. Furthermore, the above effects are significantly better in the high-dose group than in the same dose of ginseng or Pu-erh tea alone, indicating that ginseng and Pu-erh tea have a synergistic effect in improving fatigue caused by staying up late.

[0149] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing ginseng Pu-erh tea to relieve fatigue from staying up late, characterized in that, This includes blanching and soaking raw Pu-erh tea and ginseng, followed by co-fermentation. The blanching and soaking process is carried out in stages: first, the raw Pu-erh tea and ginseng are blanched at 55-65℃ for 15-25 minutes, controlling the moisture content to 30%-33%; then, they are blanched at 85-95℃ for 3-5 minutes, controlling the moisture content to 34%-38%. The co-fermentation process is carried out in three stages: core temperature 40-45℃, maintained for 10-15 days; core temperature 50-54℃, maintained for 20-30 days; core temperature 45-48℃, maintained for 10-15 days.

2. The preparation method according to claim 1, characterized in that, The mass percentages of the raw Pu-erh tea and ginseng are: 70%~95% raw Pu-erh tea and 5%~30% ginseng.

3. The preparation method according to claim 1 or 2, characterized in that, The raw Pu'er tea mentioned is Yunnan large-leaf variety raw Pu'er tea; the ginseng mentioned is a whole ginseng root that is five years old or younger.

4. The preparation method according to claim 1, characterized in that, During the co-fermentation process, the pile is turned over every 5 to 12 days.

5. The preparation method according to claim 4, characterized in that, During the co-fermentation process, water is replenished with a solution of 0.05% to 0.1% glucose and 0.02% to 0.05% peptone each time the pile is turned.

6. The preparation method according to claim 1, characterized in that, It also includes post-processing and shaping: after co-fermentation and maturation, the material is opened and cooled, then dried at a gradient low temperature until the moisture content of the material is 8%~11%, screened and sorted, and aged at room temperature in the dark for more than 15 days.

7. The preparation method according to claim 6, characterized in that, The gradient low-temperature drying includes: a first stage of drying at 40-45℃ for 12-24 hours until the material moisture content drops to 15%-18%; and a second stage of drying at 30-35℃ for 24-48 hours until the material moisture content drops to 8%-11%.

8. Ginseng Pu-erh tea prepared by the preparation method according to any one of claims 1 to 7.

9. The application of the ginseng Pu-erh tea according to claim 8 in the preparation of food products that relieve fatigue from staying up late.

10. The application according to claim 9, characterized in that, The food products are tea bags, loose tea, tea bricks, tea cakes, liquid tea drinks, concentrated liquids, or solid beverages.

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