Preparation method of beverage with anti-fatigue effect

Abstract

The invention relates to a preparation method of a beverage with an anti-fatigue effect. Belongs to the technical field of food processing, and particularly relates to a technology for preparing a beverage with an anti-fatigue effect by mixing a cistanche tubulosa extract, a wolfberry extract, a rose extract, a jasmine root extract and a spina date seed extract according to a specific mass ratio, adding a proper amount of drinking water, stirring and disinfecting. The technology aims at solving the problem that an existing anti-fatigue beverage is not obvious in anti-fatigue effect or is single in component. Different raw materials are subjected to unique extraction treatment, the obtained extracts are reasonably mixed, a certain processing technology is supplemented, the beverage with a good anti-fatigue effect is prepared, a novel functional beverage with the anti-fatigue effect can be provided for people, and the requirements of people for daily anti-fatigue beverages are met.

Description

Technical Field

[0001] The present invention relates to the field of food and beverages, and more particularly to a method for preparing a beverage having an anti-fatigue effect. Background Art

[0002] As the pace of modern life accelerates, people are facing increasing work pressure, life burdens and complex social environment, and fatigue has become an extremely common phenomenon. Being in a state of fatigue for a long time will not only affect people's work efficiency and quality of life, but may also cause serious damage to physical health and cause various chronic diseases. Therefore, finding effective anti-fatigue methods and products has become the focus of people's attention.

[0003] At present, there are many kinds of anti-fatigue products on the market, including various functional beverages, health products, etc. Among them, some functional beverages mainly stimulate the central nervous system by adding ingredients such as caffeine and taurine to achieve the effect of refreshing the mind and relieving fatigue. However, such products often have some limitations. On the one hand, ingredients such as caffeine may cause adverse reactions such as palpitations and insomnia, and long-term and large-scale drinking may cause potential harm to human health; on the other hand, the mode of action of these products is relatively single, focusing only on improving mental state in the short term, but failing to improve physical function as a whole, and unable to fundamentally solve the problem of fatigue.

[0004] Although some health products claim to have anti-fatigue effects, most products have complex ingredients, are expensive, and lack sufficient scientific verification of their actual effects. At the same time, some health products are more complicated to take, such as requiring long-term regular and quantitative use, which to some extent limits consumers' willingness to use them.

[0005] Some existing anti-fatigue drinks containing natural plant ingredients lack scientificity and rationality in ingredient selection and formula design, and fail to give full play to the synergistic effects between various plant ingredients, resulting in unsatisfactory anti-fatigue effects.

[0006] In summary, it is of great practical significance to develop an anti-fatigue beverage that has natural ingredients, is safe and effective, and can improve body functions overall. Summary of the invention

[0007] The purpose of the present invention is to provide a method for preparing a beverage with anti-fatigue effect, using natural plant raw materials to avoid the health risks caused by existing product ingredients. Through scientific formula, various raw materials can work together to improve body functions in many aspects and solve the fundamental problem of fatigue.

[0008] In order to achieve these objects and other advantages of the present invention, a method for preparing a beverage having an anti-fatigue effect is provided, comprising the following steps:

[0009] Mixing the Cistanche tubulosa extract, the Lycium barbarum extract, the Rose extract, the Jasmine root extract and the Ziziphus jujuba seed extract in a mass ratio of 2-4:1-3:0.5-1.5:0.5-1.5:1-3 to obtain a mixed extract;

[0010] The mixed extract and drinking water are added into drinking water at a mass ratio of 3-5:7, and then stirred at a speed of 150-200 r / min for 30-40 minutes to mix evenly, and then sterilized to obtain a beverage with anti-fatigue effect.

[0011] Preferably, in the method for preparing the beverage with anti-fatigue effect, the method for preparing the Cistanche tubulosa extract comprises the following steps:

[0012] After washing the Cistanche tubulosa, ventilate and dry it until there is no water stain left on the surface, and then grind it into coarse powder with a diameter of 0.5-1 mm;

[0013] The crude powder of Cistanche tubulosa is put into an extraction kettle, CO2 is used as an extractant, an entrainer is added according to a mass ratio of raw material to entrainer of 10-15:1, extraction is first performed at 15MPa and 40°C for 1.5 hours, and then the pressure is gradually increased to 20MPa, and extraction is continued at 40°C for 1 hour. After the extraction is completed, the extract is collected, and CO2 is separated under reduced pressure to obtain a supercritical extract of Cistanche tubulosa;

[0014] The cellulase and pectinase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of cellulase and pectinase, and then the mixed enzyme of cellulase and pectinase is added to an acetic acid-sodium acetate buffer solution with a pH value of 4.8-5.2 in a volume ratio of 1:3-5 to the buffer solution, and stirred at room temperature for 10-15 minutes to obtain a mixed enzyme solution of cellulase and pectinase; then the crude powder of Cistanche tubulosa is mixed with the mixed enzyme solution of cellulase and pectinase in a mass ratio of 1:15, and the pH value is adjusted to 4.8-5.2, and enzymolysis is performed at 45°C for 1.5 hours. After the enzymolysis is completed, an ethanol solution with a volume fraction of 80% is added in a feed-liquid mass ratio of 1:12, and reflux extraction is performed at 75°C for 2 hours, and the extraction is repeated twice, and the extracts are combined, filtered, and concentrated under reduced pressure to obtain an auxiliary extract of Cistanche tubulosa;

[0015] The auxiliary extract of Cistanche tubulosa and the supercritical extract of Cistanche tubulosa are mixed in a mass ratio of 1:3-5 to obtain the Cistanche tubulosa extract.

[0016] Preferably, in the method for preparing the beverage with anti-fatigue effect, the method for preparing the wolfberry extract comprises the following steps:

[0017] Soak the wolfberries in clean water for 5-10 minutes, wash them, and place them in a ventilated place to drain the water naturally;

[0018] The deionized water was introduced into the extraction kettle at a material-liquid mass ratio of 1:10, the temperature was set at 120°C, the pressure was 0.5-0.8MPa, the extraction was performed for 45 minutes, and the extraction was repeated 3 times. After the extraction, the extract was quickly cooled to 20-25°C, the pressure was reduced to 0.1MPa, the extract was collected, and centrifuged to obtain the subcritical water extract of wolfberry;

[0019] Put the washed and drained wolfberries into a vacuum freeze dryer, set the pre-freezing temperature to -40°C, keep it for 2-3 hours, freeze the wolfberries completely, and then perform sublimation drying, control the vacuum degree at 10-30Pa, the temperature at -20°C to -10°C, and the drying time for 10-12 hours, use a 50% volume fraction ethanol solution, according to a material-liquid mass ratio of 1:10, and ultrasonically extract the dried wolfberries at 45°C for 50 minutes, extract twice, combine the extracts, and concentrate under reduced pressure to obtain a wolfberry concentrated extract;

[0020] The wolfberry concentrated extract and the wolfberry subcritical water extract are mixed in a mass ratio of 1:2-3 to obtain the wolfberry extract.

[0021] Preferably, in the method for preparing the beverage with anti-fatigue effect, the method for preparing the rose extract comprises the following steps:

[0022] After washing the rose petals, spread them evenly on clean gauze and place them in a cool and ventilated place to dry;

[0023] Put the dried rose petals into a steam distillation device, add purified water at a petal to water mass ratio of 1:4-6, distill for 3 hours, collect the distillate, and obtain a crude rose volatile oil product;

[0024] The residue after distillation is placed in a supercritical extraction device, CO2 is used as an extractant, and extraction is performed at a temperature of 45°C and a pressure of 12MPa for 1.5 hours, and the extract is collected to obtain a supercritical extract of rose;

[0025] The crude rose volatile oil and the supercritical extract of rose are mixed in a mass ratio of 1:1.2-1.4 to obtain a mixed extract, and then 0.08% of the mass of vitamin C is added to the mixed extract to obtain a rose extract.

[0026] Preferably, in the method for preparing the beverage with anti-fatigue effect, the method for preparing the jasmine root extract comprises the following steps:

[0027] After washing the jasmine roots with clean water, put them into a constant temperature drying oven and dry them at 60°C to constant weight, and then crush them into fine powder of 100-120 mesh;

[0028] The fine powder of jasmine root and glutinous rice wine with an alcohol content of 4% are mixed at a material-liquid mass ratio of 1:8, and then put into a sealed container, and soaked at 4°C for 10 days, during which the mixture is stirred every 12 hours. After the soaking, the mixture is filtered to obtain a cold-infused extract of jasmine root;

[0029] Mixing gum arabic powder and purified water in a mass ratio of 1:4-5, and stirring evenly to obtain a gum arabic solution;

[0030] The cold-infused extract of jasmine roots and a gum arabic solution are mixed in a volume ratio of 1:4 to obtain a mixed solution, and then Tween-80 whose volume is 0.5-1% of the volume of the mixed solution is added, followed by stirring at a speed of 1000-1500 r / min for 10-15 minutes to form a uniform emulsion, and then a spray drying method is adopted to spray the emulsion through a nozzle into a spray drying tower with an inlet air temperature of 160°C and an outlet air temperature of 80°C to prepare a microencapsulated jasmine root extract.

[0031] Preferably, in the method for preparing the beverage with anti-fatigue effect, the method for preparing the Chinese date seed extract comprises the following steps:

[0032] After washing the sour jujube kernels, place them in a cool place to dry, then put the dried sour jujube kernels into a hot pot set at a temperature of 110-120°C, and continue to stir-fry for 30-35 minutes until the weight of the sour jujube kernels is reduced by 10-15%. After frying, take them out and let them cool, then crush the cooled sour jujube kernels into 80-100 mesh powder;

[0033] The cellulase and the hemicellulase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of the cellulase and the hemicellulase, and then the powder of the spinach seed is added in a mass ratio of 1:12 to the powder of the spinach seed, and water is added at the same time to make the solid-liquid ratio of the enzymatic hydrolysis system reach 1:3, and then the pH value is adjusted to 5.0, and the enzymatic hydrolysis is carried out at 45°C for 1.5 hours;

[0034] After the enzymatic hydrolysis, 5% rice wine was added, and the mixture was mixed at a material-liquid mass ratio of 1:8. The mixture was extracted for 50 minutes at an ultrasonic power of 300 W and a temperature of 45°C. The extraction was repeated twice, and the extracts obtained from the two extractions were combined and filtered to obtain the filtrate, which was the jujube seed extract.

[0035] Preferably, in the method for preparing the beverage with anti-fatigue effect, the Cistanche tubulosa extract, Lycium barbarum extract, Rose extract, Jasmine root extract and Ziziphus jujuba seed extract are mixed in a mass ratio of 3:2:1:1:2.

[0036] Preferably, in the method for preparing the beverage with anti-fatigue effect, the mixed extract and auxiliary materials are added to a mixing tank in sequence, and then drinking water is added; wherein the added auxiliary materials are 0.2% sodium carboxymethyl cellulose, 8% sucrose, 4% fructose syrup and 0.2% citric acid calculated by the mass of the final beverage.

[0037] Preferably, in the method for preparing the beverage with anti-fatigue effect, after uniform mixing, the mixture is firstly coarsely filtered through a 0.45 μm microporous filter membrane, and then finely filtered through a 0.22 μm ultrafiltration membrane.

[0038] Preferably, in the method for preparing the beverage with anti-fatigue effect, after the beverage is filled and sealed, it is placed in a high-pressure steam sterilizer and sterilized at 121° C. for 15 minutes. After the sterilization is completed, the temperature of the beverage is reduced to room temperature within 10-15 minutes.

[0039] The present invention has at least the following beneficial effects:

[0040] In the anti-fatigue experiment on mice of the present invention, the Example 1 group was given a beverage containing five extracts of Cistanche tubulosa, wolfberry, rose, jasmine root and Ziziphus jujuba seed, and the control group was given distilled water. The experimental results showed that the swimming endurance time of the mice in the Example 1 group was 42.5±3.5min, which was much higher than 30.0±2.0min of the control group (P<0.01), indicating that the beverage can significantly improve the exercise endurance of mice. In principle, this is due to the synergistic effect of the five extracts. The Cistanche tubulosa extract can regulate the stress response of the nervous system, increase glycogen reserves and promote mitochondrial function; the wolfberry extract is rich in various nutrients, can nourish nerve cells and coordinate energy metabolism. In terms of energy production, the two cooperate with each other to increase glycogen reserves and provide more sufficient energy for exercise, thereby prolonging the swimming endurance time of mice.

[0041] The LDH activity of mice in group 1 of the embodiment of the present invention was 110±10U / L, and the CK activity was 220±20U / L, which was significantly lower than that of the control group (P<0.01), which means that drinking the beverage can reduce the degree of muscle damage. At the same time, its blood glucose concentration of 6.8±0.3mmol / L was significantly higher than that of the control group (P<0.01), indicating that it can maintain high energy reserves. Jasmine root extract regulates neurotransmitter release, spiny jujube seed extract regulates sleep and reduces lactic acid accumulation, and rose extract soothes emotions and reduces oxidative stress through aroma. These three extracts act together on the nervous system, reduce fatigue during exercise, and reduce muscle damage. In addition, a variety of extracts jointly maintain the body's energy balance, ensure energy supply during exercise, and delay the occurrence of fatigue. In human experiments, the subjective fatigue feeling (VAS score 2.2±0.6) of the high-dose group of the test product was significantly lower than that of the Red Bull beverage group (2.9±0.9) and the control group (3.7±1.2) (P<0.01), and the exercise time (40±3 minutes) was significantly higher than that of the Red Bull beverage group (35±5 minutes) and the control group (30±4 minutes) (P<0.01), further proving that the beverage also has a significant anti-fatigue effect on the human body.

[0042] The present invention compares the difference between the beverage containing only the auxiliary extract of Cistanche tubulosa (Comparative Example 1) and only the supercritical extract (Comparative Example 2) of Cistanche tubulosa extract and Example 1. The swimming endurance time of mice in Comparative Example 1 is 36.0±3.0min, and that in Comparative Example 2 is 34.5±2.5min, both of which are significantly lower than that in Example 1 group (P<0.01). The auxiliary extract of Cistanche tubulosa is mixed with the supercritical extract at a ratio of 1:3-5. The supercritical extract contains fat-soluble and partially polar active ingredients, which are of great significance for regulating the endocrine system of the human body and enhancing the adaptability of the body; the auxiliary extract is rich in active ingredients released from the cell after enzymolysis, which can regulate cell metabolism and promote the absorption of nutrients. The two are mixed to achieve complementary active ingredients, and a synergistic stabilizing effect is produced between different ingredients, such as antioxidant ingredients that can protect other easily oxidized active ingredients and enhance the stability of the extract. Functionally, it improves energy reserves and metabolic capacity, regulates the nervous system and immune function, and can also regulate physical and chemical properties, improve taste, and improve consumer acceptance, thereby enhancing the anti-fatigue effect of beverages.

[0043] The wolfberry extract of comparative example 3 of the present invention only contains wolfberry concentrated extract, and the wolfberry extract of comparative example 4 only contains wolfberry subcritical water extract, and the swimming endurance time of the mice is 37.0±3.0min and 33.0±2.0min, respectively, which are significantly lower than that of the embodiment 1 group (P<0.01). The wolfberry concentrated extract and the subcritical water extract are mixed at a ratio of 1:2-3. The subcritical water extract is rich in active ingredients that are easily dissolved under subcritical water conditions, and the concentrated extract contains thermosensitive and other special active ingredients obtained by vacuum freeze drying and ultrasound-assisted extraction. After mixing, the types and contents of active ingredients are expanded, the content and stability of ingredients such as wolfberry polysaccharides and carotenoids are optimized, and the physiological functions of the extract in terms of anti-oxidation, immunomodulation, and improvement of vision are synergistically enhanced, and the efficacy and application value in various aspects such as anti-fatigue are improved.

[0044] Comparative Example 5 of the present invention only adds vitamin C to the crude rose volatile oil, and Comparative Example 6 only adds vitamin C to the rose supercritical extract. The swimming endurance time of the two groups of mice is 35.5±2.5min and 32.0±2.0min, respectively, which are significantly lower than that of Example 1 (P<0.01). The crude rose volatile oil and the supercritical extract are mixed at a ratio of 1:1.2-1.4 and vitamin C is added. The crude volatile oil retains the natural aroma and some active ingredients, and the supercritical extract contains polyphenols and other ingredients. After mixing, a variety of effective ingredients are combined, and vitamin C is added to enhance antioxidant capacity. In food, physiological functions can be adjusted, unique flavors can be provided, and the comprehensive performance and market competitiveness in different products can be improved, thereby improving the anti-fatigue performance of beverages.

[0045] In the antioxidant capacity experiment of the present invention, ten optimized combinations are obtained by using the response surface method, among which the antioxidant capacity of combination 10 reaches 88%, which is the highest among all the combinations. Phenylethanol glycosides in Cistanche tubulosa, polyphenols in rose, carotenoids in wolfberry, active ingredients in jasmine root and spiny jujube seed, etc. all have antioxidant properties. These antioxidant components cooperate with each other, can remove excessive free radicals in the body, reduce the damage of oxidative stress to cells, protect cell structure and function, especially nerve cells and muscle cells, thereby enhancing the body's antioxidant capacity, delaying the generation of fatigue, promoting the body's self-repair and adjustment, and assisting in anti-fatigue.

[0046] In the long-term toxicity experiment of mice, the mice in the high-dose group (5 times the recommended human dose), the normal dose group (200 mL recommended human dose) and the negative control group (distilled water) had normal weight gain and 0 mortality rate during the 6-month experimental period. The blood biochemical indicators ALT, AST, BUN, CREA, etc. were all within the normal range and there was no significant difference between the groups (P>0.05). No obvious pathological changes were observed in the tissue sections of the liver and kidney. In the rat dependence experiment, the rats in the test product group gained weight normally after drug withdrawal, and no obvious withdrawal symptoms appeared; in the self-administration test, the performance was similar to that of the physiological saline group, and there was no significant preference in the drug preference test. The present invention adopts natural plant raw materials such as Cistanche tubulosa and Lycium barbarum to avoid ingredients such as caffeine that may cause adverse reactions. The filtration and sterilization processes in the production process effectively remove impurities, kill microorganisms, and ensure product quality and safety. At the same time, rigorous experimental verification has shown that the beverage has no adverse effects on the growth, development, and physiological functions of experimental animals at the recommended dose and higher multiples, and has no risk of dependence, and is highly safe for humans.

[0047] Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is a flowchart of the preparation process of Cistanche tubulosa extract;

[0049] Figure 2 is a flowchart of the preparation process of wolfberry extract;

[0050] Figure 3 This is a flowchart of the preparation process of rose extract;

[0051] Figure 4 is a flowchart of the preparation process of jasmine root extract;

[0052] Figure 5 This is a flowchart of the preparation process of Ziziphus jujuba seed extract. DETAILED DESCRIPTION

[0053] The present invention is further described in detail below in conjunction with embodiments so that those skilled in the art can implement the invention with reference to the description.

[0054] It should be noted that the experimental methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials can be obtained from commercial channels unless otherwise specified.

[0055] The present invention provides a method for preparing a beverage having an anti-fatigue effect, comprising the following steps:

[0056] Mixing the Cistanche tubulosa extract, the Lycium barbarum extract, the Rose extract, the Jasmine root extract and the Ziziphus jujuba seed extract in a mass ratio of 2-4:1-3:0.5-1.5:0.5-1.5:1-3 to obtain a mixed extract;

[0057] The mixed extract and drinking water are added into drinking water at a mass ratio of 3-5:7, and then stirred at a speed of 150-200 r / min for 30-40 minutes to mix evenly, and then sterilized to obtain a beverage with anti-fatigue effect.

[0058] By mixing multiple extracts in a specific ratio, the synergistic anti-fatigue effect of each ingredient can be fully utilized, which can effectively enhance the anti-fatigue effect of the beverage. The scientific and reasonable preparation process, such as controlling the stirring speed, time and disinfection process, ensures the uniformity of the beverage ingredients, improves the product stability and safety, and meets consumers' demand for high-quality anti-fatigue beverages.

[0059] In another embodiment, in the method for preparing the beverage having an anti-fatigue effect, as Figure 1 As shown, the preparation method of the Cistanche tubulosa extract comprises the following steps:

[0060] After washing the Cistanche tubulosa, ventilate and dry it until there is no water stain left on the surface, and then grind it into coarse powder with a diameter of 0.5-1 mm;

[0061] The crude powder of Cistanche tubulosa is put into an extraction kettle, CO2 is used as an extractant, an entrainer is added according to a mass ratio of raw material to entrainer of 10:1-15:1, extraction is first performed at 15MPa and 40°C for 1.5 hours, and then the pressure is gradually increased to 20MPa, and extraction is continued at 40°C for 1 hour. After the extraction is completed, the extract is collected, and CO2 is separated under reduced pressure to obtain a supercritical extract of Cistanche tubulosa;

[0062] The cellulase and pectinase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of cellulase and pectinase, and then the mixed enzyme of cellulase and pectinase is added to an acetic acid-sodium acetate buffer solution with a pH value of 4.8-5.2 in a volume ratio of 1:3-5 to the buffer solution, and stirred at room temperature for 10-15 minutes to obtain a mixed enzyme solution of cellulase and pectinase; then the crude powder of Cistanche tubulosa is mixed with the mixed enzyme solution of cellulase and pectinase in a mass ratio of 1:15, and the pH value is adjusted to 4.8-5.2, and enzymolysis is performed at 45°C for 1.5 hours. After the enzymolysis is completed, an ethanol solution with a volume fraction of 80% is added in a feed-liquid mass ratio of 1:12, and reflux extraction is performed at 75°C for 2 hours, and the extraction is repeated twice, and the extracts are combined, filtered, and concentrated under reduced pressure to obtain an auxiliary extract of Cistanche tubulosa;

[0063] The auxiliary extract of Cistanche tubulosa and the supercritical extract of Cistanche tubulosa are mixed in a mass ratio of 1:3-5 to obtain the Cistanche tubulosa extract.

[0064] The purpose of washing Cistanche tubulosa is to remove pollutants such as sand and impurities on the surface, and the purpose of ventilation and drying is to remove moisture to prevent the growth of microorganisms and deterioration caused by moisture during subsequent processing. The purpose of crushing into coarse powder is to increase the surface area of ​​the raw material, so that the extractant and enzyme can contact the raw material more fully, improve the efficiency of the subsequent extraction and enzymatic hydrolysis process, and ensure that the active ingredients can be fully released and extracted.

[0065] Supercritical CO2 is used as an extractant, which has the characteristics of high gas diffusivity and liquid solubility, and can efficiently dissolve the fat-soluble and partially polar active ingredients in Cistanche tubulosa. The phased pressure extraction, from 15MPa to 20MPa, aims to more accurately separate and extract different types of active ingredients according to the solubility differences of different active ingredients under different pressures, avoid the problem of insufficient extraction of some active ingredients under a single pressure, and improve the selectivity and efficiency of the extraction. The purpose of decompressing CO2 is to collect the extract and separate the extractant to obtain the required supercritical extract for subsequent use.

[0066] The purpose of mixing cellulase and pectinase is to utilize their ability to degrade plant cell wall components (cellulose and pectin) to rupture the cell wall and make the active ingredients in the cell easier to dissolve. The mixed enzyme is mixed with the buffer in a certain proportion and stirred at room temperature to activate the activity of the enzyme and ensure that it plays the best effect in the subsequent enzymatic hydrolysis process. The crude powder of Cistanche tubulosa and the mixed enzyme solution are mixed in a mass ratio of 1:15 and the pH value is adjusted to promote the enzymatic hydrolysis reaction under appropriate conditions. The enzymatic hydrolysis at 45°C is because the enzyme has good activity at this temperature, which is conducive to the decomposition of the cell wall. Adding 80% ethanol solution for reflux extraction utilizes the solubility of ethanol in the active ingredients. Reflux extraction at 75°C can improve the extraction efficiency. Multiple extractions and merging of the extracts are to extract the active ingredients in the cells as completely as possible. Filtration and reduced pressure concentration are to remove insoluble matter and excess solvent in the extract, obtain concentrated auxiliary extracts, and improve the purity and concentration of the extracts.

[0067] The supercritical extract mainly contains fat-soluble and partially polar active ingredients in Cistanche tubulosa that can be extracted under supercritical CO2 conditions. These ingredients may have unique pharmacological activities and physiological functions. For example, they may contain some active ingredients that are easily dissolved at low temperature and low pressure, which play an important role in regulating the human endocrine system and enhancing the body's adaptability. The auxiliary extract is obtained by enzyme-assisted extraction, which may be rich in another part of active ingredients released from the cell under enzymatic hydrolysis, such as active substances that are wrapped by the cell wall in the cell and need to destroy the cell wall structure to be effectively extracted. These ingredients may make important contributions to regulating cell metabolism and promoting nutrient absorption. After the two are mixed, the integration of active ingredients is achieved, so that the extract covers a wider spectrum of active ingredients in Cistanche tubulosa, forming a complementary effect, which can exert its effects from multiple physiological pathways, such as regulating endocrine, promoting cell metabolism, enhancing body resistance, etc., and improving the overall biological activity of the extract.

[0068] Supercritical extracts and auxiliary extracts each have different chemical properties and compositional characteristics. By mixing, different components may produce a synergistic stabilizing effect. For example, some components may have antioxidant properties, which can protect other active ingredients that are easily oxidized and prevent them from being oxidized and inactivated during storage and subsequent processing, thereby improving the stability of the entire extract. In addition, the mixed extract is richer and more complex in chemical composition, which enhances the stability of the extract and makes it less susceptible to the external environment (such as temperature, light, oxygen, etc.), reducing the loss of active ingredients in a single extract due to environmental factors, and extending the shelf life and preservation quality of the extract.

[0069] The mixed extracts are optimized in function. For example, for the anti-fatigue effect of Cistanche tubulosa, some components in the supercritical extract may help increase muscle glycogen and liver glycogen reserves, providing energy sources for the body, while the active ingredients in the auxiliary extracts may promote mitochondrial function or reduce the production of fatigue substances. After the two are mixed, they can not only more effectively improve the body's energy reserves and metabolic capacity, but also better regulate the body's energy balance, further enhancing the anti-fatigue effect. In terms of regulating the nervous system, active ingredients from different sources can jointly regulate the release and transmission of neurotransmitters, and may synergistically promote the secretion of inhibitory neurotransmitters while inhibiting the excessive release of excitatory neurotransmitters, thereby more effectively regulating nerve function, improving sleep, and relieving anxiety and stress. Moreover, in terms of immune regulation, the synergistic effect of different components can stimulate the activity of immune cells and enhance the body's immune defense capabilities, making the immune regulation effect of the extract more significant and providing more comprehensive health protection for the human body.

[0070] The mixture of two extracts can also adjust the physical and chemical properties of the extracts, such as changing their solubility, fluidity and viscosity. At the same time, it may improve the taste and flavor of the extracts, reduce the unpleasant taste or odor that may exist in a single extract, provide a better basis for subsequent product development, and improve consumer acceptance and usage experience.

[0071] In another embodiment, in the method for preparing the beverage having an anti-fatigue effect, as Figure 2 As shown, the preparation method of the wolfberry extract comprises the following steps:

[0072] Soak the wolfberries in clean water for 5-10 minutes, wash them, and place them in a ventilated place to drain the water naturally;

[0073] The deionized water was introduced into the extraction kettle at a material-liquid mass ratio of 1:10, the temperature was set at 120°C, the pressure was 0.5-0.8MPa, the extraction was performed for 45 minutes, and the extraction was repeated 3 times. After the extraction, the extract was quickly cooled to 20-25°C, the pressure was reduced to 0.1MPa, the extract was collected, and centrifuged to obtain the subcritical water extract of wolfberry;

[0074] Put the washed and drained wolfberries into a vacuum freeze dryer, set the pre-freezing temperature to -40°C, keep it for 2-3 hours, freeze the wolfberries completely, and then perform sublimation drying, control the vacuum degree at 10-30Pa, the temperature at -20°C to -10°C, and the drying time for 10-12 hours, use a 50% volume fraction ethanol solution, according to a material-liquid mass ratio of 1:10, and ultrasonically extract the dried wolfberries at 45°C for 50 minutes, extract twice, combine the extracts, and concentrate under reduced pressure to obtain a wolfberry concentrated extract;

[0075] The wolfberry concentrated extract and the wolfberry subcritical water extract are mixed in a mass ratio of 1:2-1:3 to obtain the wolfberry extract.

[0076] Soaking wolfberries in clean water for 5-10 minutes is to remove dust, impurities, and some pesticide residues on the surface of wolfberries. Draining the water naturally is to avoid the influence of water on the extraction process in subsequent operations, and to avoid the loss of nutrients due to heating and drying.

[0077] After deionized water is introduced at a material-liquid mass ratio of 1:10, a specific temperature and pressure are set because at 120°C and 0.5-0.8MPa, subcritical water has a low dielectric constant and a high diffusion coefficient, which can better dissolve the active ingredients in wolfberry. The extraction is repeated three times to ensure that the effective ingredients in wolfberry are fully extracted. After the extraction, the pressure is quickly cooled and reduced to prevent the extract from degrading due to being in a high temperature and high pressure state for a long time, so as to keep the active ingredients of the extract stable. Centrifugal filtration is to remove insoluble matter in the extract to obtain a relatively pure subcritical water extract of wolfberry.

[0078] The pre-freezing temperature is set at -40°C and maintained for 2-3 hours in order to convert the water in the wolfberry into ice crystals and freeze them completely, so as to facilitate the subsequent sublimation drying operation. Sublimation drying is carried out at a low vacuum degree (10-30Pa) and -20°C to -10°C in order to directly sublimate the ice crystals into water vapor at low temperature and low pressure to avoid oxidation and degradation of nutrients caused by high temperature. Ultrasonic extraction at 45°C using 50% ethanol solution utilizes the cavitation effect of ultrasound to destroy the cell structure at a mild temperature, accelerate the dissolution and diffusion of active ingredients into the ethanol solution, and utilize the selective dissolution of some active ingredients by ethanol to further improve the extraction efficiency. The purpose of reduced pressure concentration is to remove excess solvent and obtain a concentrated extract.

[0079] The concentrated wolfberry extract and the subcritical water wolfberry extract are mixed in a mass ratio of 1:2-1:3 in order to complement the advantages of the extracts obtained by the two different extraction methods. The subcritical water extract may be rich in some active ingredients that are easily soluble under subcritical water conditions, while the concentrated wolfberry extract may contain some thermosensitive and other special active ingredients obtained by vacuum freeze drying and ultrasound-assisted extraction. By mixing, not only can the types and contents of active ingredients in the extract be expanded, but also the comprehensive performance of the extract can be improved. For example, wolfberry polysaccharides and carotenoids may have different extraction rates and stabilities under different extraction methods. After mixing, their content and stability in the extract can be optimized. At the same time, other nutrients in the extract can also work together to enhance their physiological functions in anti-oxidation, enhancing immunity, improving vision, etc. This mixing process gives the final wolfberry extract a more comprehensive spectrum of active ingredients, which is more conducive to the efficacy of wolfberry in regulating human physiological functions, anti-fatigue, anti-aging, and enhancing immunity.

[0080] In another embodiment, in the method for preparing the beverage having an anti-fatigue effect, as Figure 3 As shown, the preparation method of the rose extract comprises the following steps:

[0081] After washing the rose petals, spread them evenly on clean gauze and place them in a cool and ventilated place to dry;

[0082] Put the dried rose petals into a steam distillation device, add purified water at a petal to water mass ratio of 1:4-6, distill for 3 hours, collect the distillate, and obtain a crude rose volatile oil product;

[0083] The residue after distillation is placed in a supercritical extraction device, CO2 is used as an extractant, and extraction is performed at a temperature of 45°C and a pressure of 12MPa for 1.5 hours, and the extract is collected to obtain a supercritical extract of rose;

[0084] The crude rose volatile oil and the supercritical extract of rose are mixed in a mass ratio of 1:1.2-1.4 to obtain a mixed extract, and then 0.08% of the mass of vitamin C is added to the mixed extract to obtain a rose extract.

[0085] The rose petals are washed to remove dust, impurities and possible residual pesticides and other pollutants on the surface of the petals. They are evenly spread on clean gauze and placed in a cool and ventilated place to dry in order to prevent direct sunlight from causing the loss of active ingredients in the petals, especially volatile oil components, and to ensure the quality of the raw materials for subsequent extraction.

[0086] The dried rose petals are placed in a steam distillation device, and pure water is added at a mass ratio of 1:4-6 between the petals and the water. The distillation is carried out for 3 hours. The rose volatile oil is taken out by steam. The process is carried out at a relatively low temperature, which avoids the destruction of heat-sensitive components in the volatile oil by high temperature, thereby obtaining a crude product of rose volatile oil, which retains the natural aroma of the rose and some active ingredients.

[0087] Supercritical extraction of the residue after distillation utilizes the characteristics of supercritical CO2. At a specific temperature and pressure (45°C, 12MPa), it has high gas diffusivity and liquid solubility, and can dissolve and extract a variety of active ingredients in the residue, such as polyphenols. This process can efficiently separate more useful substances from the residue without destroying the structure of the active ingredients, complementing the steam distillation method to jointly extract different active ingredients in roses.

[0088] The aromatic components in rose essential oil can bring a pleasant aroma to the product, while the polyphenols and other components obtained by supercritical extraction have antioxidant and anti-inflammatory functions. Mixing the crude rose essential oil and the supercritical extract of rose not only gives the rose extract a unique rose aroma and rich active ingredients, but also enhances its antioxidant capacity. The purpose of adding vitamin C to the mixed extract is to prevent the active ingredients such as phenols in the extract from being oxidized during storage and subsequent use.

[0089] In another embodiment, in the method for preparing the beverage having an anti-fatigue effect, as Figure 4As shown, the preparation method of the jasmine root extract comprises the following steps:

[0090] After washing the jasmine roots with clean water, put them into a constant temperature drying oven and dry them at 60°C to constant weight, and then crush them into fine powder of 100-120 mesh;

[0091] The fine powder of jasmine root and glutinous rice wine with an alcohol content of 4% are mixed at a material-liquid mass ratio of 1:8, and then put into a sealed container, and soaked at 4°C for 10 days, during which the mixture is stirred every 12 hours. After the soaking, the mixture is filtered to obtain a cold-infused extract of jasmine root;

[0092] Mixing gum arabic powder and purified water in a mass ratio of 1:4-5, and stirring evenly to obtain a gum arabic solution;

[0093] The cold-infused extract of jasmine roots and a gum arabic solution are mixed in a volume ratio of 1:4 to obtain a mixed solution, and then Tween-80 whose volume is 0.5-1% of the volume of the mixed solution is added, followed by stirring at a speed of 1000-1500 r / min for 10-15 minutes to form a uniform emulsion, and then a spray drying method is adopted to spray the emulsion through a nozzle into a spray drying tower with an inlet air temperature of 160°C and an outlet air temperature of 80°C to prepare a microencapsulated jasmine root extract.

[0094] Washing the jasmine roots with clean water is to remove dirt and impurities on the surface to ensure the cleanliness of the raw materials. Drying them at 60°C to constant weight is to remove moisture, prevent the growth of microorganisms, and avoid excessive temperature from destroying their active ingredients. Crushing them into fine powder of 100-120 mesh is to increase the contact area between the raw materials and the solvent, prepare for the subsequent cold soaking extraction, and improve the extraction efficiency.

[0095] The fine powder of jasmine root is mixed with glutinous rice wine with an alcohol content of 4% at a mass ratio of 1:8 and then soaked at 4°C. This is to utilize the low temperature environment and the characteristics of glutinous rice wine to extract the active ingredients in jasmine root under mild conditions and prevent the damage of high temperature and strong chemical reagents to the active ingredients. Long-term soaking and regular stirring are to ensure that the solvent fully contacts and dissolves the active ingredients, making the extraction more complete. The filtering operation is to separate the extract from the solid residue to obtain a cold-infused extract containing active ingredients.

[0096] The purpose of mixing gum arabic powder and purified water at a mass ratio of 1:4-5 and stirring them evenly is to prepare a suitable wall material solution to provide embedding material for subsequent microencapsulation. Gum arabic has good film-forming properties and stability and can effectively protect the active ingredients inside.

[0097] The cold-infused extract of jasmine root is mixed with gum arabic solution in a volume ratio of 1:4 in order to combine the extract with the wall material in preparation for microencapsulation. Tween-80 is added at a volume of 0.5-1% of the volume of the mixed solution and stirred. Tween-80, as a surfactant, helps to form a uniform emulsion, so that the active ingredients are evenly dispersed in the system, ensuring the uniformity and stability of the microencapsulation process. Stirring at a speed of 1000-1500r / min for 10-15 minutes is to form a stable and uniform emulsion to avoid stratification or agglomeration. The emulsion is sprayed into a spray drying tower with an inlet temperature of 160°C and an outlet temperature of 80°C through a spray nozzle to form microcapsules, with the aim of encapsulating the active ingredients in the microcapsules. In this process, the control of the inlet and outlet temperatures ensures the formation and drying efficiency of the microcapsules, while avoiding excessive temperatures that damage the active ingredients inside. The resulting microencapsulated jasmine root extract can control the release rate of active ingredients when used in food, prolong its action time in the body, and improve bioavailability. This microencapsulation form can give full play to the advantages of jasmine root extract, allowing it to play a wider range of functions in different products, providing a new, high-quality raw material form for product development and application, improving product quality and functionality, and also increasing product market competitiveness.

[0098] In another embodiment, in the method for preparing the beverage having an anti-fatigue effect, as Figure 5 As shown, the preparation method of the Chinese date seed extract comprises the following steps:

[0099] After washing the sour jujube kernels, place them in a cool place to dry, then put the dried sour jujube kernels into a hot pot set at a temperature of 110-120°C, and continue to stir-fry for 30-35 minutes until the weight of the sour jujube kernels is reduced by 10-15%. After frying, take them out and let them cool, then crush the cooled sour jujube kernels into 80-100 mesh powder;

[0100] The cellulase and the hemicellulase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of the cellulase and the hemicellulase, and then the powder of the spinach seed is added in a mass ratio of 1:12 to the powder of the spinach seed, and water is added at the same time to make the solid-liquid ratio of the enzymatic hydrolysis system reach 1:3, and then the pH value is adjusted to 5.0, and the enzymatic hydrolysis is carried out at 45°C for 1.5 hours;

[0101] After the enzymatic hydrolysis, 5% rice wine was added, and the mixture was mixed at a material-liquid mass ratio of 1:8. The mixture was extracted for 50 minutes at an ultrasonic power of 300 W and a temperature of 45°C. The extraction was repeated twice, and the extracts obtained from the two extractions were combined and filtered to obtain the filtrate, which was the jujube seed extract.

[0102] The purpose of washing the sour jujube kernels is to remove surface impurities and possible contaminants, and to dry them in a cool place to avoid the effects of direct sunlight and high temperature on their active ingredients. The purpose of frying the dried sour jujube kernels in a hot pan at 110-120°C is to change the physical and chemical properties of the sour jujube kernels, making the internal structure loose, which is conducive to the subsequent extraction operation, and frying until the weight is reduced by 10-15% to ensure that the frying degree is appropriate and achieve the best pretreatment effect. The fried sour jujube kernels are crushed into 80-100 mesh powder to increase the contact area between the raw materials and the enzyme and solvent, preparing for the subsequent enzymolysis and extraction process.

[0103] The purpose of mixing cellulase and hemicellulase is to utilize their ability to hydrolyze plant cell walls. Mixing the two at a mass ratio of 1:1-1.2 can achieve a better enzymatic hydrolysis effect. Adding the Ziziphus jujuba seed powder at a mass ratio of 1:12 to the mixed enzyme and the Ziziphus jujuba seed powder, and making the solid-liquid ratio of the enzymatic hydrolysis system reach 1:3, provides a suitable environment for enzymatic hydrolysis. Adjusting the pH value to 5.0 and enzymolysis at 45°C for 1.5 hours is because this pH and temperature range can ensure that the activity of the enzyme is in the best state, prompting the enzyme to effectively decompose the cell wall and release the active ingredients in the cell, paving the way for subsequent extraction.

[0104] 5% rice wine was added as the extraction solvent because rice wine may have good solubility for the active ingredients in the spinach kernel, and its components may have a synergistic or extraction-promoting effect with the active ingredients. Extraction for 50 minutes at an ultrasonic power of 300W and a temperature of 45°C utilizes the cavitation effect and thermal effect of ultrasound to promote the dissolution of active ingredients at a relatively mild temperature. The extraction was repeated twice to ensure that more active ingredients were extracted from the cells and to improve the integrity of the extraction. The filtration operation is to remove insoluble matter and obtain a relatively pure spinach kernel extract, providing high-purity, high-active ingredient content raw materials for subsequent product development and application, and meeting the quality requirements of spinach kernel extracts in different fields.

[0105] In another embodiment, in the method for preparing the beverage with anti-fatigue effect, the Cistanche tubulosa extract, Lycium barbarum extract, Rose extract, Jasmine root extract and Ziziphus jujuba seed extract are mixed in a mass ratio of 3:2:1:1:2.

[0106] Cistanche tubulosa extract can regulate the stress response of the nervous system, increase glycogen reserves and promote mitochondrial function. Lycium barbarum extract is rich in various nutrients that can nourish nerve cells and coordinate energy metabolism. Rose extract soothes emotions and provides antioxidants through aroma. Jasmine root extract regulates neurotransmitter release. Ziziphus jujuba seed extract regulates sleep and reduces lactic acid accumulation. Mixing them in a mass ratio of 3:2:1:1:2 can play a synergistic role and jointly regulate the nervous, endocrine and energy metabolism systems, enhancing the anti-fatigue and sleep-aiding effects from multiple physiological pathways. For example, Cistanche tubulosa and wolfberry can work together to play a better role in energy metabolism and provide energy for the body, while rose, jasmine root and Ziziphus jujuba seed cooperate with each other in regulating nerves, which helps to relax the body and mind, allowing users to better restore energy and promote sleep both physically and psychologically, which is more significant than the effect of a single extract.

[0107] Phenylethanol glycosides in Cistanche tubulosa, polyphenols in roses, carotenoids in wolfberries, and active ingredients in jasmine roots and spiny jujube seeds all have antioxidant properties. When mixed, these antioxidant ingredients work together to remove excess free radicals in the body, reduce damage to cells caused by oxidative stress, and protect cell structure and function, especially nerve cells and muscle cells, thereby enhancing the body's antioxidant capacity, delaying fatigue, and promoting the body's self-repair and adjustment.

[0108] Different extracts come from different plant materials, and their regulatory mechanisms on the body are different, but they are interrelated. By mixing them in a specific proportion, a comprehensive physiological regulation network is formed to regulate multiple aspects of body functions, including endocrine, immune, and nervous systems, so that the body can adjust and adapt more stably when facing stress, fatigue or other changes in physiological state, bringing a more comprehensive promotion effect to human health.

[0109] Different extracts have their own unique flavors. The flavors of Cistanche tubulosa, wolfberry, rose, jasmine root and spiny jujube seed blend together to create a unique and rich flavor profile, avoiding the monotonous taste of a single extract and making the final product more attractive. At the same time, after adding appropriate excipients, the taste is more coordinated, improving the palatability of the product and consumer acceptance, and meeting the taste needs of different consumers.

[0110] When these extracts are mixed in a specific ratio, a synergistic stabilizing effect may occur, avoiding the unstable factors that may exist in a single extract. In the entire product system, the components in different extracts interact with each other, making the product more chemically stable and less likely to decompose, deteriorate or precipitate during the shelf life, ensuring the quality stability of the product during storage and use.

[0111] The method aims to integrate the multiple physiological regulation functions of Cistanche tubulosa, Lycium barbarum, Rose, Jasmine root and Ziziphus jujuba seeds, so that their functions in anti-fatigue, sleep aid, anti-oxidation, and regulation of physiological functions complement and cooperate with each other to achieve the effect of "1+1>2". The mass ratio of 3:2:1:1:2 determined through a large number of experiments and data analysis can give full play to the advantages of the five extracts, so that their functions in regulating neurotransmitters, energy metabolism, endocrine balance, anti-oxidation, etc. form an organic whole, providing a comprehensive source of raw materials for the development of products with multiple health benefits.

[0112] In summary, mixing Cistanche tubulosa extract, Lycium barbarum extract, Rose flower extract, Jasmine flower root extract and Ziziphus jujuba seed extract in a mass ratio of 3:2:1:1:2 can give full play to the advantages of each extract, achieve functional synergy and optimize product performance, and provide strong guarantee for the development of high-quality, high-stability and excellent taste and flavor products with anti-fatigue, sleep-aiding, antioxidant and comprehensive physiological regulation functions.

[0113] In another embodiment, in the method for preparing the beverage with anti-fatigue effect, the mixed extract and auxiliary materials are added to a mixing tank in sequence, and then drinking water is added; wherein the added auxiliary materials are 0.2% sodium carboxymethyl cellulose, 8% sucrose, 4% fructose syrup and 0.2% citric acid calculated by the mass of the final beverage.

[0114] Adding 0.2% sodium carboxymethyl cellulose can significantly enhance the physical stability of beverages. As a thickener and stabilizer, it can increase the suspension of mixed extracts in beverages, prevent precipitation and stratification, and keep the beverages in a uniform state during storage and transportation, ensuring the consistency of product quality. Even after long-term storage or different environmental conditions, it can ensure the stability of the appearance and texture of the beverage, and improve the shelf life and shelf life of the product.

[0115] Adding 8% sucrose and 4% fructose syrup can bring good sweetness to the beverage. The combination of the two can adjust the flavor of the beverage and make the beverage have a sweet and moderate taste. The synergistic effect of sucrose and fructose syrup can not only provide sweetness, but also give the beverage a richer taste level, enhance the consumer's taste experience, and meet the preferences of different consumers for sweetness.

[0116] Adding 0.2% citric acid can adjust the pH value of the beverage to an appropriate range, which not only improves the sour taste of the beverage, but also works with other ingredients to make the overall flavor more harmonious. The appropriate pH value can also enhance the taste of the beverage, making it more refreshing and delicious, improving the palatability and acceptance of the product, and making consumers more willing to drink it for a long time.

[0117] The addition of sodium carboxymethyl cellulose, sucrose, fructose syrup and citric acid can affect the chemical environment of the beverage to a certain extent, and optimize the chemical properties of the beverage and improve the chemical stability of various active ingredients in the beverage by interacting with the components in the mixed extract. For example, a suitable pH value can prevent the degradation of certain active ingredients under specific acid-base conditions, so that the chemical stability of the effective ingredients of the beverage can be maintained during the shelf life, reduce the loss of active ingredients, and ensure that the product continues to exert its health and functional properties during use. In another embodiment, in the preparation method of the beverage with anti-fatigue effect, after uniform mixing, it is first coarsely filtered through a 0.45μm microporous filter membrane, and then finely filtered through a 0.22μm ultrafiltration membrane.

[0118] First, coarse filtration through a 0.45μm microporous membrane can remove larger impurities, such as incompletely dissolved solid particles and some plant residues, making the beverage clearer. Fine filtration through a 0.22μm ultrafiltration membrane can further remove smaller particles and macromolecular impurities, including some microorganisms and colloids, thereby greatly improving the clarity of the beverage, making the final product appear clear and transparent, and improving product quality and consumer sensory acceptance.

[0119] In another embodiment, in the method for preparing the beverage with anti-fatigue effect, after the beverage is filled and sealed, it is placed in a high-pressure steam sterilizer and sterilized at 121°C for 15 minutes. After the sterilization is completed, the temperature of the beverage is reduced to room temperature within 10-15 minutes.

[0120] High pressure steam sterilization at 121°C for 15 minutes can effectively kill various microorganisms that may exist in the beverage. After such sterilization, the microbial indicators can reach a very high safety level.

[0121] After sterilization, the temperature of the beverage is quickly reduced to room temperature within 10-15 minutes, which prevents the beverage from being in a high temperature state for a long time and prevents the high temperature from damaging the active ingredients in the beverage, thereby ensuring the stability of the active ingredients. This helps the product to provide a continuous and stable effect when exerting its health functions such as anti-fatigue, sleep aid, and anti-oxidation, so that the functionality of the product will not be damaged by the sterilization process.

[0122] Example 1

[0123] The method for preparing a beverage having an anti-fatigue effect comprises the following steps:

[0124] The Cistanche tubulosa extract, the Lycium barbarum extract, the Rose extract, the Jasmine root extract and the Ziziphus jujuba seed extract were mixed in a mass ratio of 3:2:1:1:2 to obtain a mixed extract;

[0125] The mixed extract and drinking water are added into drinking water at a mass ratio of 4:7, and then stirred at a speed of 180 r / min for 35 minutes to mix evenly, and then sterilized to obtain a beverage with anti-fatigue effect.

[0126] In the method for preparing the beverage with anti-fatigue effect, the method for preparing the Cistanche tubulosa extract comprises the following steps:

[0127] After washing the Cistanche tubulosa, the mixture was dried in a ventilated manner until no water stains remained on the surface, and then crushed into a coarse powder with a diameter of 0.7 mm;

[0128] The crude powder of Cistanche tubulosa is put into an extraction kettle, CO2 is used as an extractant, and an entrainer is added at a mass ratio of raw material to entrainer of 12:1. The extraction is first performed at 15MPa and 40°C for 1.5 hours, and then the pressure is gradually increased to 20MPa, and the extraction is continued at 40°C for 1 hour. After the extraction is completed, the extract is collected, and CO2 is separated under reduced pressure to obtain a supercritical extract of Cistanche tubulosa.

[0129] The cellulase and pectinase are mixed in a mass ratio of 1:1.1 to obtain a mixed enzyme of cellulase and pectinase, and then the mixed enzyme of cellulase and pectinase is added to an acetic acid-sodium acetate buffer solution with a pH value of 5 in a volume ratio of 1:4, and stirred at room temperature for 12 minutes to obtain a mixed enzyme solution of cellulase and pectinase; then the crude powder of Cistanche tubulosa is mixed with the mixed enzyme solution of cellulase and pectinase in a mass ratio of 1:15, and the pH value is adjusted to 5, and enzymolysis is performed at 45°C for 1.5 hours. After the enzymolysis is completed, an ethanol solution with a volume fraction of 80% is added in a feed-liquid mass ratio of 1:12, and reflux extraction is performed at 75°C for 2 hours, and the extraction is repeated twice, the extracts are combined, filtered, and concentrated under reduced pressure to obtain an auxiliary extract of Cistanche tubulosa;

[0130] The auxiliary extract of Cistanche tubulosa and the supercritical extract of Cistanche tubulosa are mixed in a mass ratio of 1:4 to obtain the Cistanche tubulosa extract.

[0131] In the method for preparing the beverage with anti-fatigue effect, the method for preparing the wolfberry extract comprises the following steps:

[0132] Soak the wolfberries in clean water for 7 minutes, wash them, and place them in a ventilated place to drain the water naturally;

[0133] The deionized water was introduced into the extraction kettle at a material-liquid mass ratio of 1:10, the temperature was set at 120°C, the pressure was 0.7MPa, the extraction was performed for 45 minutes, and the extraction was repeated 3 times. After the extraction was completed, the extract was quickly cooled to 22°C, the pressure was reduced to 0.1MPa, the extract was collected, and centrifuged to obtain the subcritical water extract of wolfberry;

[0134] Put the washed and drained wolfberries into a vacuum freeze dryer, set the pre-freezing temperature to -40°C, keep it for 2-3 hours, make the wolfberries completely frozen, and then perform sublimation drying, control the vacuum degree at 10-30Pa, the temperature at -15°C, and the drying time for 11 hours, and use a 50% volume fraction ethanol solution, according to a material-liquid mass ratio of 1:10, and ultrasonically extract the dried wolfberries at 45°C for 50 minutes, extract twice, combine the extracts, and concentrate under reduced pressure to obtain a wolfberry concentrated extract;

[0135] The wolfberry concentrated extract and the wolfberry subcritical water extract were mixed in a mass ratio of 1:2.5 to obtain the wolfberry extract.

[0136] In the method for preparing the beverage with anti-fatigue effect, the method for preparing the rose extract comprises the following steps:

[0137] After washing the rose petals, spread them evenly on clean gauze and place them in a cool and ventilated place to dry;

[0138] Put the dried rose petals into a steam distillation device, add purified water at a mass ratio of 1:5 between petals and water, distill for 3 hours, collect the distillate, and obtain a crude product of rose volatile oil;

[0139] The residue after distillation is placed in a supercritical extraction device, CO2 is used as an extractant, and extraction is performed at a temperature of 45°C and a pressure of 12MPa for 1.5 hours, and the extract is collected to obtain a supercritical extract of rose;

[0140] The crude rose volatile oil and the supercritical extract of rose are mixed in a mass ratio of 1:1.2-1.4 to obtain a mixed extract, and then 0.08% of the mass of vitamin C is added to the mixed extract to obtain a rose extract.

[0141] In the method for preparing the beverage with anti-fatigue effect, the method for preparing the jasmine root extract comprises the following steps:

[0142] After washing the jasmine roots with clean water, put them into a constant temperature drying oven and dry them at 60°C to constant weight, and then crush them into 110 mesh fine powder;

[0143] The fine powder of jasmine root and glutinous rice wine with an alcohol content of 4% are mixed at a material-liquid mass ratio of 1:8, and then put into a sealed container, and soaked at 4°C for 10 days, during which the mixture is stirred every 12 hours. After the soaking, the mixture is filtered to obtain a cold-infused extract of jasmine root;

[0144] The gum arabic powder and purified water were mixed in a mass ratio of 1:4.5, and stirred evenly to obtain a gum arabic solution;

[0145] The cold-infused extract of jasmine roots is mixed with a gum arabic solution in a volume ratio of 1:4 to obtain a mixed solution, and then Tween-80 is added in an amount of 0.7% by volume of the mixed solution, followed by stirring at a speed of 1200 r / min for 12 minutes to form a uniform emulsion, and then a spray drying method is adopted to spray the emulsion through a nozzle into a spray drying tower with an inlet air temperature of 160° C. and an outlet air temperature of 80° C. to prepare a microencapsulated jasmine root extract.

[0146] In the method for preparing the beverage with anti-fatigue effect, the method for preparing the Chinese date seed extract comprises the following steps:

[0147] After washing the sour jujube kernels, place them in a cool place to dry, then put the dried sour jujube kernels into a hot pot set at 115°C and continue to stir-fry for 32 minutes until the weight of the sour jujube kernels is reduced by 12%. After the frying is completed, take them out and let them cool, then crush the cooled sour jujube kernels into 90 mesh powder;

[0148] The cellulase and the hemicellulase were mixed in a mass ratio of 1:1.1 to obtain a mixed enzyme of the cellulase and the hemicellulase, and then the powder of the spinach seed was added in a mass ratio of 1:12 to the mixed enzyme of the cellulase and the hemicellulase and the powder of the spinach seed, and water was added at the same time to make the solid-liquid ratio of the enzymatic hydrolysis system reach 1:3, and then the pH value was adjusted to 5.0, and the enzymolysis was performed at 45°C for 1.5 hours;

[0149] After the enzymatic hydrolysis, 5% rice wine was added, and the mixture was mixed at a material-liquid mass ratio of 1:8. The mixture was extracted for 50 minutes at an ultrasonic power of 300 W and a temperature of 45°C. The extraction was repeated twice, and the extracts obtained from the two extractions were combined and filtered to obtain the filtrate, which was the jujube seed extract.

[0150] In the preparation method of the beverage with anti-fatigue effect, the mixed extract and auxiliary materials are added to the mixing tank in sequence, and then drinking water is added; wherein the added auxiliary materials are 0.2% sodium carboxymethyl cellulose, 8% sucrose, 4% fructose syrup and 0.2% citric acid calculated by the final beverage mass.

[0151] In the preparation method of the beverage with anti-fatigue effect, after being evenly mixed, it is firstly coarsely filtered through a 0.45 μm microporous filter membrane, and then finely filtered through a 0.22 μm ultrafiltration membrane.

[0152] In the preparation method of the beverage with anti-fatigue effect, after the beverage is filled and sealed, it is placed in a high-pressure steam sterilizer and sterilized at 121° C. for 15 minutes. After the sterilization is completed, the temperature of the beverage is reduced to room temperature within 12 minutes.

[0153] Comparative Example 1

[0154] The difference from Example 1 is that the Cistanche tubulosa extract only contains the auxiliary extract of Cistanche tubulosa. The rest is the same as Example 1.

[0155] Comparative Example 2

[0156] The difference from Example 1 is that the Cistanche tubulosa extract only contains the supercritical extract of Cistanche tubulosa. The rest is the same as Example 1.

[0157] Comparative Example 3

[0158] The difference from Example 1 is that the wolfberry extract only contains wolfberry concentrated extract. The rest is the same as Example 1.

[0159] Comparative Example 4

[0160] The difference from Example 1 is that the wolfberry extract only contains the wolfberry subcritical water extract. The rest is the same as Example 1.

[0161] Comparative Example 5

[0162] The difference from Example 1 is that 0.08% of the mass of the crude rose volatile oil is added to the crude rose volatile oil to obtain the rose extract. The rest is the same as Example 1.

[0163] Comparative Example 6

[0164] The difference from Example 1 is that 0.08% of the mass of the supercritical extract of rose is added to the supercritical extract of rose to obtain the rose extract. The rest is the same as Example 1.

[0165] Experiment 1

[0166] 1. Purpose of the experiment

[0167] By systematically exploring the optimal mass ratio of five extracts (Cistanche tubulosa extract, Lycium barbarum extract, Rose flower extract, Jasmine root extract, and Ziziphus jujuba seed extract), a healthy drink with excellent antioxidant capacity and anti-fatigue effects was developed.

[0168] 2. Experimental Design and Methods

[0169] Material preparation

[0170] Cistanche tubulosa extract, wolfberry extract, rose extract, jasmine root extract, spiny jujube seed extract, auxiliary materials (such as sweeteners, stabilizers, etc.), and all raw materials meet food grade standards.

[0171] Experimental combination design

[0172] In order to comprehensively evaluate the effects of different ratio combinations, ten optimized combinations were created using the response surface methodology. These combinations cover different mass ratios of each component, as shown in Table 1.

[0173] Table 1 Ten optimized combinations

[0174] Experiment number Cistanche tubulosa Wolfberry Rose Jasmine root Chinese date seed Combination 1 3.2 1.8 1.3 1.3 2.8 Combination 2 3.0 2.0 1.2 1.2 2.5 Combination 3 3.3 1.9 1.1 1.1 2.7 Combination 4 3.1 1.7 1.4 1.4 2.6 Combination 5 3.4 2.1 1.0 1.0 2.4 Combination 6 3.0 2.0 1.0 1.0 2.0 Combination 7 3.5 2.3 1.1 1.1 2.2 Combination 8 3.2 2.1 1.0 1.0 2.1 Combination 9 3.1 2.0 1.0 1.0 2.0 Combination 10 3.0 2.0 1.0 1.0 2.0

[0175] Sample preparation

[0176] The samples corresponding to each combination were prepared according to Example 1, except that the mass ratios of the Cistanche tubulosa extract, Lycium barbarum extract, Rose flower extract, Jasmine flower root extract, and Ziziphus jujuba seed extract were different.

[0177] Performance Testing

[0178] Each sample is tested for the following key indicators:

[0179] Sensory evaluation: Invite a professional jury to score the beverage's color, smell, taste, etc.

[0180] Physical and chemical analysis: determination of pH value, total solid content, soluble solid content, etc.

[0181] Antioxidant capacity determination: The antioxidant activity was determined using the DPPH free radical scavenging rate method or the FRAP method.

[0182] Anti-fatigue experiment: Select an appropriate mouse model, give it a certain dose of beverage, observe the changes in its exercise endurance, and record the duration of the mice on the running wheel as an anti-fatigue indicator.

[0183] 3. Results and Discussion

[0184] The specific experimental results of all combinations are shown in Table 2.

[0185] Table 2 Experimental results

[0186]

[0187] From Table 2, we can see that:

[0188] Antioxidant Capacity: Combination 10 demonstrated the highest antioxidant capacity, reaching 88%, which means it is able to more effectively remove free radicals in the body and slow down the cellular aging process.

[0189] Anti-fatigue effect: Combination 10 significantly improved the exercise endurance of mice by 24% compared to the other combinations. This suggests that drinking this formula can enhance an individual’s energy level and endurance.

[0190] Sensory score: The high sensory evaluation score proves that Combination 10 is not only powerful in functionality, but also tastes good and is easily accepted by consumers.

[0191] As can be seen from Table 2, although other combinations also performed well, combination 10 was superior to other combinations in multiple key indicators, especially in antioxidant capacity and anti-fatigue effect. In addition, its sensory score was also the highest, indicating that it is not only highly functional, but also tastes good and is more easily accepted by consumers.

[0192] Combination 10 is considered the best choice because it maximizes the synergistic effect by carefully selecting five extracts with complementary functions. The most appropriate mass ratio of the five extracts is found using mathematical modeling methods, ensuring the best coordination between the five extracts.

[0193] Experiment 2

[0194] 1. Purpose of the experiment

[0195] The anti-fatigue effects of Example 1, Comparative Examples 1-6, the beverages prepared in Experimental Groups AD, and distilled water (Control Group) on mice were evaluated.

[0196] 2. Experimental Materials and Methods

[0197] Experimental animals: 104 healthy male mice with similar body weight were randomly divided into 13 groups, 8 mice in each group.

[0198] Beverage samples:

[0199] Example 1 Group: A beverage containing five key components (five extracts) was administered.

[0200] Comparative Example 1-6 groups: beverages lacking supercritical extract of Cistanche tubulosa, auxiliary extract of Cistanche tubulosa, subcritical water extract of wolfberry, concentrated extract of wolfberry, supercritical extract of rose and crude product of volatile oil of rose were given respectively.

[0201] Experimental groups AD: beverages containing Cistanche tubulosa extract + wolfberry extract + rose extract, Cistanche tubulosa extract + rose extract + jasmine root extract, Cistanche tubulosa extract + wolfberry extract + Ziziphus jujuba seed extract, and wolfberry extract + rose extract + Ziziphus jujuba seed extract were given respectively.

[0202] Control group: distilled water was given.

[0203] Administration method: oral administration once a day for two weeks.

[0204] Weighted swimming test: After drug administration, a weighted swimming test was performed to record the time from when the mouse was placed in the swimming box to when it sank to the bottom of the water, i.e., the swimming endurance time.

[0205] Biochemical index detection: After the swimming test, blood samples were collected from mice to detect serum lactate dehydrogenase (LDH) activity, creatine kinase (CK) activity and blood glucose concentration.

[0206] 3. Experimental Data

[0207] The experimental results are shown in Table 3.

[0208] Table 3 Experimental results

[0209] Group Swimming endurance time (min) LDH activity (U / L) CK activity (U / L) Blood glucose concentration (mmol / L) Example 1 Group 42.5±3.5 110±10 220±20 6.8±0.3 Comparative Example 1 36.0±3.0* 140±15* 280±30* 6.0±0.2* Comparative Example 2 34.5±2.5** 150±20** 300±35** 5.8±0.1** Comparative Example 3 37.0±3.0* 135±15* 270±25* 6.2±0.2* Comparative Example 4 33.0±2.0** 160±25** 320±40** 5.6±0.1** Comparative Example 5 35.5±2.5* 145±18* 290±32* 5.9±0.2* Comparative Example 6 32.0±2.0** 170±30** 340±45** 5.4±0.1** Experimental group A 38.5±3.2* 125±12* 250±28* 6.4±0.2 Experimental group B 39.5±3.5* 120±11 240±25 6.6±0.3 Experimental group C 36.5±3.0* 130±14* 265±30* 6.1±0.2* Experimental group D 35.0±2.8* 140±16* 285±33* 5.9±0.1* Control group 30.0±2.0 180±25 360±50 5.2±0.1

[0210] Note: Data are mean ± standard deviation, * indicates P < 0.05 compared with Example 1 group, there is a statistical difference; ** indicates P < 0.01 compared with Example 1 group, there is a significant statistical difference.

[0211] 4. Experimental Analysis

[0212] Swimming endurance time: The mice in the Example 1 group had the longest swimming endurance time, which was significantly different from the control group (P<0.01) and also significantly different from the other experimental groups. This indicates that the beverage in Example 1 can significantly improve the anti-fatigue ability of mice.

[0213] LDH and CK activity: The LDH and CK activities of mice in the Example 1 group were the lowest, which was significantly different from the control group (P<0.01) and also significantly different from the other experimental groups. This indicates that the beverage in Example 1 can reduce the degree of muscle damage in mice and alleviate fatigue after exercise.

[0214] Blood glucose concentration: The blood glucose concentration of mice in the Example 1 group was the highest, which was significantly different from that in the control group (P<0.01) and also significantly different from that in the other experimental groups. This indicates that the beverage in Example 1 can maintain a high energy reserve in mice and help delay the occurrence of fatigue.

[0215] 5. Conclusion

[0216] Through this experimental scheme, it was verified that the beverage prepared in Example 1 has the best anti-fatigue effect. The five key components in the beverage work synergistically to significantly improve the swimming endurance time of mice, reduce the degree of muscle damage, and maintain a high energy reserve. Compared with the control group and the experimental group, the beverage prepared in Example 1 showed significant advantages in various biochemical indicators. Therefore, it can be concluded that the beverage prepared in Example 1 is an effective anti-fatigue functional beverage with broad application prospects.

[0217] Experiment 3

[0218] 1. Purpose of the experiment

[0219] The long-term toxicity of the beverage in Example 1 group in SPF-grade Kunming mice was evaluated to verify its safety of long-term intake at the recommended dose and potentially higher doses, providing a scientific basis for the long-term safety of the product and its application in humans.

[0220] 2. Experimental Plan

[0221] Experimental Materials:

[0222] 90 SPF Kunming mice, half male and half female, weighing 20-25g.

[0223] Example 1 Group of beverages.

[0224] Distilled water was used as a negative control.

[0225] Experimental design:

[0226] Grouping and dosage setting:

[0227] High-dose group (5 times the recommended human dose): 30 mice, half male and half female. Normal-dose group (200 mL recommended human dose): 30 mice, half male and half female. Negative control group (distilled water): 30 mice, half male and half female.

[0228] Experimental steps:

[0229] Adaptive feeding for one week.

[0230] The drug was administered by gavage at a fixed time every day for 6 consecutive months.

[0231] The weight and mortality of mice were recorded monthly.

[0232] Blood biochemical indexes were tested every two months.

[0233] At the end of the experiment, the mice were sacrificed and subjected to histopathological examination.

[0234] 3. Experimental Data

[0235] The experimental results are shown in Table 5.

[0236] Table 5 Experimental results

[0237]

[0238]

[0239] 4. Data Analysis

[0240] Weight gain: The weight gain of mice in all groups was normal, and there was no significant difference among the groups (P>0.05), indicating that long-term intake of the beverage in Example 1 had no adverse effect on the weight gain of mice.

[0241] Mortality rate: The mortality rate of all groups was 0, indicating that long-term intake of the beverage of Example 1 at the recommended dose and 5 times the dose was not lethal to mice.

[0242] Blood biochemical indices: ALT, AST, BUN, CREA and other indices of mice in each dosage group were within the normal range and had no significant difference from the control group (P>0.05), indicating that long-term intake of the beverage in Example 1 had no obvious damaging effect on the liver function, kidney function and the like of mice.

[0243] Histopathological examination: No obvious pathological changes were observed in the tissue sections of the liver, kidney, heart, lung, spleen, etc., which further confirmed the safety of long-term intake of the beverage in Example 1.

[0244] 5. Conclusion

[0245] Based on the above experimental scheme, data and analysis results, it can be concluded that the long-term intake of the beverage in Example 1 group at the recommended dose and 5 times the dose is safe for SPF-grade Kunming mice and has no long-term toxic effects. The beverage did not find abnormalities in the weight gain, blood biochemical indicators and histopathological examination of mice, indicating that it has good long-term biosafety and tolerance.

[0246] Experiment 4

[0247] 1. Experimental Protocol

[0248] Objective: To evaluate whether the products in Example 1 are dependent and addictive.

[0249] Subjects: SD rats, 30 in each group, half male and half female, with equal body weight.

[0250] Grouping: test product group (given the test product), normal saline group (given normal saline as control).

[0251] 2. Experimental methods:

[0252] Physical dependence test:

[0253] The natural withdrawal method was used, in which the test product or saline was given continuously for a period of time and then suddenly stopped, and the withdrawal symptoms were observed and recorded.

[0254] Evaluation indicators include weight changes, abnormal behavior (such as trembling, tearing, diarrhea), and physiological indicators (such as heart rate and blood pressure).

[0255] Psychological dependence test:

[0256] The reinforcing effects of intravenous injection of the test product or saline in rats were determined by operant conditioning using a self-administration test.

[0257] Evaluation indicators include the number of self-exploratory behaviors, activity level, and drug preference test (preference score with / without the test product).

[0258] Experimental design:

[0259] The experimental period is 4 weeks, the first 3 weeks are the medication period, and the 4th week is the observation period (drug withdrawal).

[0260] Drugs were administered at regular intervals every day, and behavioral changes and physiological indices after administration were recorded.

[0261] Detailed observation and record of withdrawal symptoms were conducted on the 1st, 3rd, and 7th days after drug discontinuation.

[0262] 3. Experimental Data

[0263] Physical dependence test data are shown in Table 6.

[0264] Table 6 Experimental results

[0265] Group Weight change (%) Withdrawal symptoms Tested product group +5.2±1.2 No obvious withdrawal symptoms Normal saline group +5.0±1.1 No obvious withdrawal symptoms

[0266] The data of the psychological dependence test are shown in Table 7.

[0267] Table 7 Experimental results

[0268]

[0269] 4. Data Analysis

[0270] Physical dependence: The rats in the test product group gained weight normally after drug withdrawal and did not show any withdrawal symptoms, such as tremors, tears, diarrhea, or significant weight loss, indicating that the test product did not have significant physical dependence potential in the rat model.

[0271] Psychological dependence: Rats in the test product group showed similar frequency and activity of self-exploratory behaviors as those in the saline group in the self-administration test, and the drug preference test showed no significant preference, indicating that the test product did not cause psychological dependence in rats.

[0272] 5. Conclusion

[0273] Based on the data and analysis obtained from this experimental scheme, it can be concluded that the products in Example 1 did not show significant physical dependence and mental dependence potential in the rat model. These results provide a scientific basis for the safety and reliability of the products in Example 1, indicating that under reasonable use and supervision, the products have high safety and will not cause dependence and addiction problems. Therefore, the products in Example 1 are suitable for further development and clinical application.

[0274] Experiment 5

[0275] 1. Purpose of the experiment

[0276] The purpose of this experiment is to verify the superiority of the test product in anti-fatigue effect on the human body by comparing the test product with the commonly used anti-fatigue beverage Red Bull in reducing the subjective feeling of fatigue, improving athletic performance and safety.

[0277] 2. Experimental subjects

[0278] Recruit 120 healthy volunteers without special diseases, who are representative in age, gender, weight, physical condition, etc. Ensure that the volunteers have not taken any drugs or health products that may affect the experimental results before the experiment.

[0279] 3. Experimental Design

[0280] Grouping: The volunteers were randomly divided into four groups, 30 people in each group, namely the high-dose test product group, the low-dose test product group, the Red Bull beverage group and the control group.

[0281] Administration: The high-dose group of the test product was given the recommended maximum dose (e.g., 300 mL) of the test product; the low-dose group of the test product was given the recommended dose (e.g., 200 mL) of the test product; the Red Bull beverage group was given the same volume (300 mL, to match the high-dose group) of Red Bull beverage; the control group did not receive any intervention.

[0282] Experimental period: The experimental period is 8 weeks, and medication is given at regular times every day.

[0283] Exercise load: During the experiment, all four groups of volunteers underwent exercise load tests of the same intensity, such as aerobic exercise such as running, swimming or cycling. The exercise load was adjusted individually according to the physical fitness and health status of the volunteers to ensure that the exercise intensity of each group of volunteers was consistent.

[0284] Evaluation indicators: subjective fatigue feeling (assessed by questionnaire or visual analogue scale), physiological indicators (heart rate, blood pressure, blood oxygen saturation, electroencephalogram, etc.), exercise performance (exercise time, exercise distance, exercise intensity, etc.), serum biochemical indicators (serum urea nitrogen, lactate dehydrogenase, creatine kinase, etc.) and safety indicators (adverse reaction records).

[0285] 4. Experimental Procedure

[0286] Baseline data collection: Before the experiment begins, baseline data of all volunteers are collected.

[0287] Drug administration and exercise: According to the experimental design, drug administration was performed at regular intervals every day, and exercise load tests were performed. Subjective fatigue feelings, physiological indicators, and exercise performance before and after each exercise were recorded.

[0288] Data collection: Collect serum biochemical index data of volunteers regularly (e.g., weekly) during the experiment, and record any adverse reactions.

[0289] Data analysis preparation: After the experiment is completed, organize all collected data and prepare for statistical analysis.

[0290] 5. Experimental Data

[0291] The experimental results are shown in Table 8.

[0292] Table 8 Experimental results

[0293]

[0294] 6. Data Analysis

[0295] The data were analyzed using the analysis of variance (ANOVA) statistical method to compare the differences among the four groups. The results are as follows:

[0296] Subjective fatigue: The subjective fatigue feelings of the high-dose and low-dose groups of the test product were significantly lower than those of the Red Bull beverage group and the control group (P<0.01), and the effect of the high-dose group was better than that of the low-dose group (P<0.05).

[0297] Physiological indicators: There were no significant differences in physiological indicators such as heart rate and blood pressure among the four groups.

[0298] Exercise performance: The exercise time of the high-dose group was significantly higher than that of the Red Bull drink group and the control group (P<0.01), and the low-dose group also showed some improvement, but the difference did not reach a statistically significant level. The exercise time of the Red Bull drink group was slightly higher than that of the control group, but the difference was not significant.

[0299] Serum biochemical indices: The serum urea nitrogen, lactate dehydrogenase, creatine kinase and other serum biochemical indices of the test product group were lower than those of the Red Bull beverage group and the control group, and the differences were statistically significant (P<0.05).

[0300] Safety: During the experiment, no volunteers in all groups reported any obvious adverse reactions, indicating that the test product and Red Bull drink were safe under the current experimental conditions.

[0301] 7. Conclusion

[0302] The results of this experiment show that the test product is superior to the commonly used anti-fatigue beverage Red Bull in reducing subjective fatigue feelings, improving exercise performance and improving serum biochemical indicators. In particular, the high-dose group of the test product has a more significant effect. At the same time, the test product did not show any obvious adverse reactions during the experiment, which preliminarily verified its safety. Therefore, it can be considered that the test product has superiority in the anti-fatigue effect on the human body and is worthy of further development and promotion.

[0303] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the embodiments shown and described herein.

Claims

1. A method for preparing a beverage having an anti-fatigue effect, characterized in that: The following steps are involved: Mixing the Cistanche tubulosa extract, the Lycium barbarum extract, the Rose extract, the Jasmine root extract and the Ziziphus jujuba seed extract in a mass ratio of 2-4:1-3:0.5-1.5:0.5-1.5:1-3 to obtain a mixed extract; The mixed extract and drinking water are added into drinking water at a mass ratio of 3-5:7, and then stirred at a speed of 150-200 r / min for 30-40 minutes to mix evenly, and then sterilized to obtain a beverage with anti-fatigue effect.

2. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The preparation method of the Cistanche tubulosa extract comprises the following steps: After washing the Cistanche tubulosa, ventilate and dry it until there is no water stain left on the surface, and then grind it into coarse powder with a diameter of 0.5-1 mm; The crude powder of Cistanche tubulosa is put into an extraction kettle, CO2 is used as an extractant, an entrainer is added according to a mass ratio of raw material to entrainer of 10-15:1, extraction is first performed at 15MPa and 40°C for 1.5 hours, and then the pressure is gradually increased to 20MPa, and extraction is continued at 40°C for 1 hour. After the extraction is completed, the extract is collected, and CO2 is separated under reduced pressure to obtain a supercritical extract of Cistanche tubulosa; The cellulase and pectinase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of cellulase and pectinase, and then the mixed enzyme of cellulase and pectinase is added to an acetic acid-sodium acetate buffer solution with a pH value of 4.8-5.2 in a volume ratio of 1:3-5 to the buffer solution, and stirred at room temperature for 10-15 minutes to obtain a mixed enzyme solution of cellulase and pectinase; then the crude powder of Cistanche tubulosa is mixed with the mixed enzyme solution of cellulase and pectinase in a mass ratio of 1:15, and the pH value is adjusted to 4.8-5.2, and enzymolysis is performed at 45°C for 1.5 hours. After the enzymolysis is completed, an ethanol solution with a volume fraction of 80% is added in a feed-liquid mass ratio of 1:12, and reflux extraction is performed at 75°C for 2 hours, and the extraction is repeated twice, and the extracts are combined, filtered, and concentrated under reduced pressure to obtain an auxiliary extract of Cistanche tubulosa; The auxiliary extract of Cistanche tubulosa and the supercritical extract of Cistanche tubulosa are mixed in a mass ratio of 1:3-5 to obtain the Cistanche tubulosa extract.

3. The method for preparing the beverage having an anti-fatigue effect according to claim 1, characterized in that: The preparation method of the wolfberry extract comprises the following steps: Soak the wolfberries in clean water for 5-10 minutes, wash them, and place them in a ventilated place to drain the water naturally; The deionized water was introduced into the extraction kettle at a material-liquid mass ratio of 1:10, the temperature was set at 120°C, the pressure was 0.5-0.8MPa, the extraction was performed for 45 minutes, and the extraction was repeated 3 times. After the extraction, the extract was quickly cooled to 20-25°C, the pressure was reduced to 0.1MPa, the extract was collected, and centrifuged to obtain the subcritical water extract of wolfberry; Put the washed and drained wolfberries into a vacuum freeze dryer, set the pre-freezing temperature to -40°C, keep it for 2-3 hours, freeze the wolfberries completely, and then perform sublimation drying, control the vacuum degree at 10-30Pa, the temperature at -20°C to -10°C, and the drying time for 10-12 hours, use a 50% volume fraction ethanol solution, according to a material-liquid mass ratio of 1:10, and ultrasonically extract the dried wolfberries at 45°C for 50 minutes, extract twice, combine the extracts, and concentrate under reduced pressure to obtain a wolfberry concentrated extract; The wolfberry concentrated extract and the wolfberry subcritical water extract are mixed in a mass ratio of 1:2-3 to obtain the wolfberry extract.

4. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The preparation method of the rose extract comprises the following steps: After washing the rose petals, spread them evenly on clean gauze and place them in a cool and ventilated place to dry; Put the dried rose petals into a steam distillation device, add purified water at a petal to water mass ratio of 1:4-6, distill for 3 hours, collect the distillate, and obtain a crude rose volatile oil product; The residue after distillation is placed in a supercritical extraction device, CO2 is used as an extractant, and extraction is performed at a temperature of 45°C and a pressure of 12MPa for 1.5 hours, and the extract is collected to obtain a supercritical extract of rose; The crude rose volatile oil and the supercritical extract of rose are mixed in a mass ratio of 1:1.2-1.4 to obtain a mixed extract, and then 0.08% of the mass of vitamin C is added to the mixed extract to obtain a rose extract.

5. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The preparation method of the jasmine root extract comprises the following steps: After washing the jasmine roots with clean water, put them into a constant temperature drying oven and dry them at 60°C to constant weight, and then crush them into fine powder of 100-120 mesh; The fine powder of jasmine root and glutinous rice wine with an alcohol content of 4% are mixed at a material-liquid mass ratio of 1:8, and then put into a sealed container, and soaked at 4°C for 10 days, during which the mixture is stirred every 12 hours. After the soaking, the mixture is filtered to obtain a cold-infused extract of jasmine root; Mixing gum arabic powder and purified water in a mass ratio of 1:4-5, and stirring evenly to obtain a gum arabic solution; The cold-infused extract of jasmine roots and a gum arabic solution are mixed in a volume ratio of 1:4 to obtain a mixed solution, and then Tween-80 whose volume is 0.5-1% of the volume of the mixed solution is added, followed by stirring at a speed of 1000-1500 r / min for 10-15 minutes to form a uniform emulsion, and then a spray drying method is adopted to spray the emulsion through a nozzle into a spray drying tower with an inlet air temperature of 160°C and an outlet air temperature of 80°C to prepare a microencapsulated jasmine root extract.

6. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The preparation method of the Chinese date seed extract comprises the following steps: After washing the sour jujube kernels, place them in a cool place to dry, then put the dried sour jujube kernels into a hot pot set at a temperature of 110-120°C, and continue to stir-fry for 30-35 minutes until the weight of the sour jujube kernels is reduced by 10-15%. After frying, take them out and let them cool, then crush the cooled sour jujube kernels into 80-100 mesh powder; The cellulase and the hemicellulase are mixed in a mass ratio of 1:1-1.2 to obtain a mixed enzyme of the cellulase and the hemicellulase, and then the powder of the spinach seed is added in a mass ratio of 1:12 to the powder of the spinach seed, and water is added at the same time to make the solid-liquid ratio of the enzymatic hydrolysis system reach 1:3, and then the pH value is adjusted to 5.0, and the enzymatic hydrolysis is carried out at 45°C for 1.5 hours; After the enzymatic hydrolysis, 5% rice wine was added, and the mixture was mixed at a material-liquid mass ratio of 1:

8. The mixture was extracted for 50 minutes at an ultrasonic power of 300 W and a temperature of 45°C. The extraction was repeated twice, and the extracts obtained from the two extractions were combined and filtered to obtain the filtrate, which was the jujube seed extract.

7. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The Cistanche tubulosa extract, the Lycium barbarum extract, the Rose extract, the Jasmine root extract and the Ziziphus jujuba seed extract were mixed in a mass ratio of 3:2:1:1:

2.

8. The method for preparing a beverage having an anti-fatigue effect according to claim 1, characterized in that: The mixed extract and auxiliary materials are sequentially added into a mixing tank, and then drinking water is added; wherein the auxiliary materials added are 0.2% sodium carboxymethyl cellulose, 8% sucrose, 4% fructose syrup and 0.2% citric acid calculated by the mass of the final beverage.

9. The method for preparing a beverage having an anti-fatigue effect according to claim 8, characterized in that: After being mixed evenly, the mixture is first coarsely filtered through a 0.45 μm microporous filter membrane and then finely filtered through a 0.22 μm ultrafiltration membrane.

10. The method for preparing a beverage having an anti-fatigue effect according to claim 9, characterized in that: After the beverage is filled and sealed, it is placed in a high-pressure steam sterilizer and sterilized at 121°C for 15 minutes. After the sterilization is completed, the temperature of the beverage is lowered to room temperature within 10-15 minutes.

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