A functional beverage for improving sleep and a method of preparing the same

Abstract

The application discloses a functional beverage capable of improving sleep and a preparation method thereof, relates to the technical field of functional beverages, and solves the problem of low extraction rate of active ingredients of existing day lily related beverages. The functional beverage capable of improving sleep comprises a day lily compound enzymolysis concentrated solution and purified water; the day lily compound enzymolysis concentrated solution is prepared from dry day lily through ternary compound enzymolysis, enzyme inactivation centrifugation and reduced-pressure concentration. The application significantly improves the dissolution rate and bioavailability of total flavonoid active ingredients through synergistic enzyme hydrolysis, and does not need to add artificial additives; the product has the effects of soothing emotion, resisting depression and improving sleep, has a refreshing taste and dense bubbles, the process is suitable for large-scale production, and fills the market gap of day lily carbonated functional beverages.

Description

Technical Field

[0001] This invention relates to the field of functional beverage technology, specifically to a functional beverage that helps improve sleep and its preparation method. Background Technology

[0002] With the fast pace of modern life and continuously increasing work and life pressures, anxiety, depression, and other emotional problems, along with accompanying sleep disorders, have become common health concerns. Compared to the side effects and dependence risks of chemically synthesized antidepressants, mood-regulating products derived from natural food ingredients have become a core growth area in the food industry due to their high safety and suitability for long-term daily consumption.

[0003] Daylily is a traditional Chinese food and medicine. Traditional Chinese medicine believes that it has the effects of soothing the liver and relieving depression, calming the mind and promoting sleep. Modern pharmacological research has confirmed that daylily is rich in flavonoids, γ-aminobutyric acid (GABA), plant polysaccharides and other active ingredients. It can play an antidepressant, sleep-improving and mood-soothing role by regulating the level of monoamine neurotransmitters and clearing free radicals in nerve cells. It is a high-quality raw material for developing functional foods for mood regulation.

[0004] Currently, the development of daylily-related beverages still suffers from many technical deficiencies: Firstly, the extraction efficiency of active ingredients is low. Existing processes mostly employ traditional decoction, soaking, or simple enzymatic hydrolysis, which can only break down plant cell walls to release intracellular components. They cannot convert bound flavonoids into free flavonoids with higher bioavailability. The dissolution rate of core active substances such as total flavonoids and GABA is low, resulting in insufficient product efficacy.

[0005] Therefore, developing a functional beverage that can efficiently extract active ingredients and does not rely on artificial additives to help improve sleep is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0006] To address the issues of low extraction rates of active ingredients and limited compatibility of sleep-aiding ingredients in existing daylily-related beverages, this invention proposes a functional beverage that helps improve sleep and its preparation method.

[0007] The specific technical solution of the present invention is as follows: A functional beverage that helps improve sleep, comprising a concentrated extract of daylily compound enzymatic hydrolysate and purified water; The daylily compound enzymatic hydrolysis concentrate is obtained by ternary compound enzymatic hydrolysis of dried daylily, enzyme inactivation centrifugation, and vacuum concentration. The ternary complex enzymatic hydrolysis specifically involves: After crushing dried daylily, add water to prepare a liquid solution, adjust the pH of the liquid solution to 5.0, add cellulase and pectinase, and place it in a 50℃ constant temperature water bath for 1.5 hours for enzymatic hydrolysis. Add β-glucosidase to the solution after the first enzymatic hydrolysis is completed, raise the temperature to 60°C, and continue enzymatic hydrolysis in a constant temperature water bath for 1.5 hours.

[0008] Preferably, based on the total mass of the liquid before enzymatic hydrolysis, the amount of cellulase added is 0.2%, the amount of pectinase added is 0.1%, and the amount of β-glucosidase added is 0.1%.

[0009] Preferably, the ratio of the liquid to the feed solution is 1g:25mL.

[0010] Preferably, the process conditions for enzyme inactivation centrifugation are: incubation at 90°C for 5 min, followed by centrifugation at 4000 rpm for 15 min; The vacuum concentration is carried out at a temperature of 45°C and a vacuum degree of -0.08 to -0.09 MPa.

[0011] Preferably, it also includes γ-aminobutyric acid, jasmine, white sugar, citric acid, taurine, B vitamins, vitamin C, and cold-pressed ginger juice; The ratio of the daylily compound enzymatic hydrolysate concentrate, γ-aminobutyric acid, jasmine, white sugar, citric acid, taurine, B vitamins, vitamin C, and cold-pressed ginger juice is 1 mL: 200 mg: 2 g: 15 g: 0.15 g: 350 mg: 70 mg: 50 mg: 0.05 mL.

[0012] The present invention also provides a method for the biological preparation of the above-mentioned functional beverage that helps improve sleep, comprising the following steps: S1. After removing impurities, drying, and crushing dried daylily buds, add purified water to prepare a liquid and let it stand to swell. S2. After adjusting the pH of the solution, first add cellulase and pectinase for the first stage of isothermal enzymatic hydrolysis; then add β-glucosidase to the solution, raise the temperature, and carry out the second stage of isothermal enzymatic hydrolysis. S3. After enzymatic hydrolysis, the solution is inactivated by enzyme, cooled, centrifuged to remove residue, and the clarified enzymatic hydrolysis supernatant is collected. S4. Concentrate the enzymatic hydrolysis supernatant under reduced pressure to obtain a concentrated enzymatic hydrolysis solution of daylily compound enzyme hydrolysis solution; S5. Mix the concentrate with the remaining components according to the formula ratio, add purified water to make up the volume, and stir until the system is uniform. S6. After filtering the prepared liquid to remove impurities, pasteurize it and then cool it rapidly after sterilization. S7. Food-grade carbon dioxide is introduced into the cooled liquid to complete carbonation. After aseptic filling and sealing, the finished product is obtained.

[0013] Preferably, in step S1, the material is pulverized and then passed through a 60-mesh sieve; the time for static swelling is 30 minutes.

[0014] Preferably, the filtration and impurity removal in step S6 uses a 0.45μm microporous filter membrane, and the pasteurization conditions are 85℃ for 15 minutes.

[0015] This invention addresses the technical shortcomings of existing daylily functional beverages, such as low extraction efficiency of active ingredients, reliance on artificial additives, and limited product forms. Using daylily, a food and medicinal herb, as the raw material, it constructs a synergistic enzymatic hydrolysis system through a ternary compound enzymatic hydrolysis process, coupled with optimized formulation and aseptic filling processes. Compared with existing technologies, the specific beneficial effects of this invention are as follows: This invention employs a two-stage enzymatic hydrolysis strategy involving stepwise temperature control and phased enzyme addition. In the first stage, at 50°C, cellulase and pectinase work synergistically to efficiently degrade the cellulose and pectin components of the daylily cell wall, fully releasing intracellular total flavonoids, plant polysaccharides, and other active substances. In the second stage, after heating to 60°C, β-glucosidase is added to specifically hydrolyze the glycosidic bonds of flavonoids, efficiently converting the bound flavonoids with low bioavailability into free flavonoids that are easily absorbed by the human body, simultaneously achieving high dissolution and high activity of the active ingredients.

[0016] This invention uses daylily, a food and medicine of the same origin, as its core active ingredient. It eliminates the need for artificially synthesized sedative components, regulating the nervous and bodily states through multi-target regulation of natural active substances, thus combining efficacy with food safety. Animal experiments have confirmed that the product has no direct hypnotic effect and does not cause side effects such as drowsiness or fatigue, avoiding disruption to daily activities. Simultaneously, both low and high doses significantly prolong the sleep time induced by sodium pentobarbital in mice and increase the incidence of sleep at subthreshold doses of sodium pentobarbital. It effectively soothes nervous hyperactivity, shortens sleep latency, and improves sleep quality, achieving natural sleep aid through gentle regulation without the risk of drug dependence.

[0017] This invention reduces the grassy and bitter taste of daylily raw materials through a multi-stage enzymatic hydrolysis process. Combined with a small amount of cold-pressed ginger juice and citric acid for flavoring and acidity adjustment, the product presents a natural light golden color, possessing both the herbal fragrance of daylily and the delicate aroma of bellflower, with a refreshing and sweet aftertaste. The addition of γ-aminobutyric acid synergistically enhances the calming and soothing effects; taurine, B vitamins, and vitamin C replenish lost nutrients and relieve fatigue; cold-pressed ginger juice promotes peripheral circulation and soothes muscle tension; and jasmine and sugar-acid components adjust the flavor, effectively masking the grassy and astringent taste of the daylily raw materials. The product has a refreshing and harmonious taste, high drinkability, and achieves a comprehensive conditioning effect across the entire chain of fatigue relief, nerve soothing, and sleep improvement. Combined with a low-temperature segmented carbonation process, the bubbles are dense and long-lasting, with a moderate burnt taste, distinguishing it from the heavy taste of traditional still-type daylily beverages. This invention is the first to combine the functional activity of daylily with the form of carbonated beverage, filling the market gap for ready-to-drink carbonated functional beverages made from daylily. It is suitable for diverse consumption scenarios such as office refreshment, relaxation, and post-meal cleansing, and has a higher acceptance among consumers.

[0018] The functional beverage provided by this invention contains enriched free flavonoids that synergistically target multiple targets with GABA: on the one hand, it can regulate the level of monoamine neurotransmitters, inhibit excessive excitation of the hypothalamus-pituitary-adrenal (HPA) axis, and scavenge oxidative free radicals in nerve cells, achieving a conditioning effect of soothing negative emotions and relieving stress; on the other hand, it can shorten the sleep latency, prolong sleep duration, and improve sleep quality. Animal experiments verifying its sleep-improving properties have shown that this product can significantly prolong the sleep time induced by sodium pentobarbital in mice and increase the incidence of subthreshold dose sleep, without direct hypnotic effects or causing drowsiness. Detailed Implementation

[0019] To make the technical solutions of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. It should be noted that the following embodiments are only used to better understand the technical solutions of the present invention and should not be construed as limiting the present invention.

[0020] Experimental materials: sulfur-free dried daylily (commercially available, moisture content ≤10%); cellulase (enzyme activity ≥10000U / g), pectinase (enzyme activity ≥5000U / g), β-glucosidase (enzyme activity ≥2000U / g).

[0021] Example 1. Take 100g of dried daylily, dry it in an oven at 55℃ for 3 hours, crush it with a grinder and pass it through a 60-mesh sieve. Add purified water at a ratio of 1g:25mL, stir well, seal, and let it stand at low temperature and away from light for 30 minutes to fully swell and obtain the liquid.

[0022] Adjust the pH of the solution to 5.0 with food-grade citric acid. Add 0.2% cellulase and 0.1% pectinase by the total mass of the solution. After stirring evenly, place the solution in a 50°C constant temperature water bath for the first stage of enzymatic hydrolysis, which lasts for 1.5 hours, stirring at low speed every 20 minutes. After the first stage of enzymatic hydrolysis is completed, add 0.1% β-glucosidase to the solution. After stirring evenly, raise the temperature to 60°C and continue the second stage of enzymatic hydrolysis in a constant temperature water bath for 1.5 hours.

[0023] After enzymatic hydrolysis, the solution is rapidly heated to 90°C and kept at that temperature for 5 minutes to inactivate the enzyme. After the solution has cooled to room temperature naturally, it is centrifuged at 4000 rpm for 15 minutes, the bottom residue is discarded, and the clear enzymatic hydrolysis supernatant is collected.

[0024] The enzymatic hydrolysis supernatant was transferred into a rotary evaporator, and the temperature was controlled at 45℃ and the vacuum degree at -0.08~-0.09MPa to obtain a concentrated enzymatic hydrolysis solution of daylily.

[0025] Take 1 mL of the compound enzymatic hydrolysis concentrate, add the basic sugar-acid solution prepared by dissolving 15 g of white sugar and 0.15 g of citric acid in purified water, then add 200 mg of γ-aminobutyric acid, 2 g of jasmine pollen, 350 mg of taurine, 70 mg of B vitamins, 50 mg of vitamin C, and 0.05 mL of cold-pressed ginger juice. Add purified water to a total volume of 260 mL and stir at 200 r / min for 10 min until the system is homogeneous.

[0026] The prepared solution was filtered through a 0.45μm microporous membrane to remove impurities and then pasteurized at 85℃ for 15 minutes. After sterilization, it was rapidly cooled to 0~4℃.

[0027] Food-grade carbon dioxide is introduced into the cooled liquid, and the volume of carbon dioxide is controlled to be 2.5 times the volume of the liquid. The mixture is thoroughly mixed to complete the carbonation. The mixture is then filled and sealed in a Class 100 aseptic environment to obtain the finished compound enzymatic functional beverage.

[0028] Comparative Example 1. Without adding β-glucosidase, single-stage enzymatic hydrolysis was performed using only cellulase and pectinase, with a total hydrolysis time of 3 hours and a temperature of 50°C. The remaining preparation steps were the same as in Example 1.

[0029] Comparative Example 2. Add 0.05% β-glucosidase, and perform single-stage enzymatic hydrolysis using cellulase and pectinase. The total hydrolysis time is 3 hours, and the temperature is 50°C. The remaining preparation steps are the same as in Example 1.

[0030] Example of an effect 1. (1) Test the total flavonoid content in the enzymatic hydrolysis supernatant of each example and comparative example.

[0031] Mix 1 mL of sample extract with 3 mL of distilled water, then add 0.3 mL of 5% [presumably a specific ingredient or solution]. NaNO2. The mixture was allowed to stand at 25°C for 5 min, then 0.3 mL of 10% AlCl3 was added. The reaction mixture was then washed with 2 mL of 1M sodium hydroxide. Distilled water (10 mL) was added to the reaction mixture, and the absorbance was measured at 510 nm. The total flavonoid content was expressed as quercetin equivalents in mg / 100g (dry basis).

[0032] (2) Test the FRAP (Ferric ion reduction antioxidant capacity assay) value in the enzymatic hydrolysate of each embodiment and comparative example.

[0033] Take 150 μL of sample, add 2850 μL of FRAP solution, mix well, and measure the absorbance at a wavelength of 593 nm.

[0034] The reagent preparation method is as follows: 0.28 mol / L acetic acid solution: Take 1.6 mL of glacial acetic acid and add distilled water to bring the volume to 100 mL; Acetic acid buffer solution (pH=3.6): Take 0.16g of sodium acetate, add 100mL of 0.28mol / L acetic acid solution, dissolve and mix well to obtain the solution; 1 mol / L HCl solution: Take 8.33 mL of concentrated hydrochloric acid and add distilled water to make up to 100 mL; 40 mmol / L HCl solution: Take 4 mL of 1 mol / L HCl solution and add distilled water to bring the volume to 100 mL; 10 mmol / L TPTZ solution: Take 0.156 g of TPTZ, add 50 mL of 40 mmol / L HCl solution, dissolve and mix well to obtain the solution; 20 mmol / L FeCl3 solution: Take 0.135 g of FeCl3, add 25 mL of distilled water, dissolve and mix well to obtain the solution; FRAP solution: Mix 25 mL of acetate buffer solution, 2.5 mL of 10 mmol / L TPTZ solution and 2.5 mL of 20 mmol / L FeCl3 solution thoroughly, and heat to 37°C in a water bath before use.

[0035] The absorbance was measured at a wavelength of 593 nm using a spectrophotometer. The results are expressed as micromolar Trolox equivalents (μmol / g, dry weight). The standard curve showed good linearity in the range of 25–800 μmol / g.

[0036] The measured total flavonoid content and FRAP value are shown in Table 1.

[0037] Table 1

[0038] Example of effect 2. Animal experiments verifying the improvement of sleep function: Experimental animals: SPF-grade male ICR mice, weighing 20-22g, were randomly divided into 3 groups after 7 days of acclimatization: blank control group, low-dose group, and high-dose group, with 4-5 mice in each group.

[0039] Test sample: The concentrated enzymatic hydrolysate of daylily-platycodon grandiflorus prepared in Example 1 was administered to the low-dose group at a dose of 0.05 mL / 10 g of body weight and the high-dose group at a dose of 0.2 mL / 10 g of body weight. The blank control group was given an equal volume of purified water and administered orally for 30 consecutive days.

[0040] The following three experiments were conducted: 1. Direct Sleep Experiment: After administering the test substance, observe whether mice fall asleep. Sleep is indicated by the disappearance of the righting reflex. When a mouse is placed in a dorsal position, it should be able to right itself immediately. If it cannot right itself within 30-60 seconds, the righting reflex is considered to have disappeared, and the mouse has entered sleep. The recovery of the righting reflex is considered awakening. The time from the disappearance to the recovery of the righting reflex is considered sleep time. Record the number of animals that fall asleep and the sleep time in the negative control group and the test sample group. The disappearance of the righting reflex (unable to right itself in a dorsal position for 30-60 seconds) is used as the indicator of sleep, and the number of animals that fall asleep and the sleep time are recorded.

[0041] 2. Pentobarbital sodium sleep time prolongation experiment: 10-15 minutes after the last administration, pentobarbital sodium (0.2 mL / 20 g body weight) was injected intraperitoneally, and the time to fall asleep, wake-up time and total sleep time of mice were recorded.

[0042] 3. Subthreshold dose hypnotic experiment of sodium pentobarbital: The maximum subthreshold hypnotic dose was determined in the preliminary experiment. After the last administration, the dose of sodium pentobarbital was injected intraperitoneally, and the number of animals that fell asleep within 30 minutes was recorded and the sleep incidence rate was calculated.

[0043] The experimental results are as follows: 1. In the direct sleep experiment, neither the low-dose nor the high-dose group mice showed direct sleep phenomena, indicating that the test sample had no direct hypnotic effect and would not cause drowsiness as a side effect.

[0044] 2. Effects on sleep duration of threshold dose sodium pentobarbital:

[0045] Compared with the blank control group, both the low- and high-dose groups significantly prolonged the sleep time induced by sodium pentobarbital in mice.

[0046] 3. Effect of subthreshold doses of sodium pentobarbital on the incidence of sleep:

[0047] Compared with the blank control group, both the low-dose and high-dose groups improved the sleep rate of mice within 30 minutes.

[0048] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A functional beverage that helps improve sleep, characterized in that, Includes concentrated enzymatic hydrolysate of daylily and purified water; The daylily compound enzymatic hydrolysis concentrate is obtained by ternary compound enzymatic hydrolysis of dried daylily, enzyme inactivation centrifugation, and vacuum concentration. The ternary complex enzymatic hydrolysis specifically involves: After crushing dried daylily, add water to prepare a liquid solution, adjust the pH of the liquid solution to 5.0, add cellulase and pectinase, and place it in a 50℃ constant temperature water bath for 1.5 hours for enzymatic hydrolysis. Add β-glucosidase to the solution after the first enzymatic hydrolysis is completed, raise the temperature to 60°C, and continue enzymatic hydrolysis in a constant temperature water bath for 1.5 hours.

2. The functional beverage for improving sleep according to claim 1, characterized in that, Based on the total mass of the liquid before enzymatic hydrolysis, the amount of cellulase added is 0.2%, the amount of pectinase added is 0.1%, and the amount of β-glucosidase added is 0.1%.

3. The functional beverage for improving sleep according to claim 1, characterized in that, The ratio of the liquid to the feed solution is 1g:25mL.

4. The functional beverage for improving sleep according to claim 1, characterized in that, The enzyme-inactivating centrifugation process conditions are: incubation at 90℃ for 5 min, followed by centrifugation at 4000 rpm for 15 min; The vacuum concentration is carried out at a temperature of 45°C and a vacuum degree of -0.08 to -0.09 MPa.

5. The functional beverage for improving sleep according to any one of claims 1 to 4, characterized in that, It also includes gamma-aminobutyric acid, jasmine, white sugar, citric acid, taurine, B vitamins, vitamin C, and cold-pressed ginger juice; The ratio of the daylily compound enzymatic hydrolysate concentrate, γ-aminobutyric acid, jasmine, white sugar, citric acid, taurine, B vitamins, vitamin C, and cold-pressed ginger juice is 1 mL: 200 mg: 2 g: 15 g: 0.15 g: 350 mg: 70 mg: 50 mg: 0.05 mL.

6. A method for preparing a functional beverage that helps improve sleep as described in any one of claims 1 to 5, characterized in that, Includes the following steps: S1. After removing impurities, drying, and crushing dried daylily buds, add purified water to prepare a liquid and let it stand to swell. S2. After adjusting the pH of the solution, first add cellulase and pectinase for the first stage of isothermal enzymatic hydrolysis; then add β-glucosidase to the solution, raise the temperature, and carry out the second stage of isothermal enzymatic hydrolysis. S3. After enzymatic hydrolysis, the enzyme in the solution is inactivated, cooled, centrifuged to remove residue, and the clarified enzymatic hydrolysis supernatant is collected. S4. Concentrate the enzymatic hydrolysis supernatant under reduced pressure to obtain a concentrated enzymatic hydrolysis solution of daylily compound enzyme hydrolysis solution; S5. Mix the concentrate with the remaining components according to the formula ratio, add purified water to make up the volume, and stir until the system is uniform. S6. After filtering the prepared liquid to remove impurities, pasteurize it and then cool it rapidly after sterilization. S7. Food-grade carbon dioxide is introduced into the cooled liquid to complete carbonation. After aseptic filling and sealing, the finished product is obtained.

7. The method for preparing the functional beverage that helps improve sleep according to claim 6, characterized in that, The pulverized material in step S1 is passed through a 60-mesh sieve; the static swelling time is 30 minutes.

8. The method for preparing the functional beverage that helps improve sleep according to claim 6, characterized in that, The filtration and impurity removal in step S6 uses a 0.45μm microporous filter membrane, and the pasteurization conditions are 85℃ for 15 minutes.

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