Fermented beverage for alleviating fatigue and method for preparing the same

Abstract

The application provides a fermented beverage for relieving fatigue and a preparation method thereof, and relates to the technical field of functional beverages, and belongs to the technical field of A23L2. The fermented beverage comprises a radix astragali fermentation extract, a platycodon grandiflorum extract, L-arginine, taurine, inositol, caffeine, nicotinamide, vitamin B6, L-lysine hydrochloride, pyrroloquinoline quinone and excipients. The radix astragali fermentation extract can improve ATP generation efficiency, reduce post-exercise lactic acid accumulation, reduce the damage of oxidative stress to muscle and nerve cells, and reduce immune suppression caused by fatigue; the platycodon grandiflorum extract can refresh the mind, improve the cognitive function decline caused by central fatigue, enhance capillary permeability, and promote the removal of metabolic waste. The radix astragali fermentation extract and the platycodon grandiflorum extract have excellent anti-fatigue effect. Under the combined action of various components, fatigue is relieved, energy is quickly supplemented, and physical strength is recovered.

Description

Technical Field

[0001] This invention relates to the field of functional beverage technology, with IPC classification number A23L2, and specifically to a fermented beverage for relieving fatigue and its preparation method. Background Technology

[0002] In today's fast-paced lifestyle, people face increasing work pressure and complex life burdens, making fatigue an extremely common physical condition. Whether it's prolonged, high-intensity mental labor, such as office workers focusing on their work for several hours a day, students dealing with heavy academic workloads, or manual laborers performing long hours of high-intensity work, fatigue seriously affects people's quality of life and work efficiency.

[0003] Beverages, as common drinks in people's daily lives, have a huge potential market for relieving fatigue. However, existing anti-fatigue beverages still have many shortcomings. For example, some beverages have relatively simple ingredients, relying on only one or a few ingredients to exert their anti-fatigue effect, making it difficult to comprehensively address the complex mechanisms of physical fatigue, affecting the consumer's drinking experience and reducing the product's market competitiveness.

[0004] Therefore, developing a beverage with scientifically sound ingredients and significant anti-fatigue effects has important practical significance and broad market prospects. Summary of the Invention

[0005] The purpose of this invention is to provide a fermented beverage that relieves fatigue and its preparation method. The fermented beverage provided by this invention can significantly relieve fatigue and improve immunity.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a fermented beverage for relieving fatigue, comprising the following raw materials in parts by weight: 8-12 parts of Astragalus membranaceus fermented extract, 5-10 parts of Platycodon grandiflorus extract, 0.01-0.02 parts of L-arginine, 0.1-0.4 parts of taurine, 0.06-0.1 parts of inositol, 0.05-0.1 parts of caffeine, 0.005-0.01 parts of nicotinamide, 0.001-0.0015 parts of vitamin B6, 0.3-0.8 parts of L-lysine hydrochloride, 0.01-0.03 parts of pyrrolquinone, and 50-60 parts of excipients.

[0008] Preferably, the preparation method of the Astragalus fermented extract includes: pulverizing Astragalus, mixing it with water, sequentially subjecting it to enzymatic hydrolysis by cellulase and nattokinase via ultrasonication, then sequentially fermenting it with Pediococcus pentosus and Bifidobacterium infantis, filtering it, concentrating the filtrate under reduced pressure, and freeze-drying it to obtain the Astragalus fermented extract.

[0009] More preferably, the cellulase has an enzyme activity of 50,000-150,000 U / g, is added at 2%-5% of the weight of Astragalus membranaceus, has an enzymatic hydrolysis temperature of 45-55℃, a pH value of 4.5-5.5, an ultrasonic power of 350-450W, and an enzymatic hydrolysis time of 30-60 min; the nattokinase has an enzyme activity of 20,000-60,000 FU / g, is added at 1%-3% of the weight of Astragalus membranaceus, has an enzymatic hydrolysis temperature of 37-42℃, a pH value of 7-9, an ultrasonic power of 300-400W, and an enzymatic hydrolysis time of 20-30 min.

[0010] More preferably, the viability of the *Pediococcus pentosaceus* is 10-20 billion CFU / g, the addition amount is 1‰-3‰ of the weight of the *Astragalus membranaceus*, the fermentation temperature is 42-48℃, the pH value is 5.5-6.5, the rotation speed is 150-250 rpm, and the time is 24-36 h; the viability of the fermented *Bifidobacterium infantis* is 20-30 billion CFU / g, the addition amount is 5‰-8‰ of the weight of the *Astragalus membranaceus*, the fermentation temperature is 35-40℃, the pH value is 6.5-7, and the time is 24-72 h.

[0011] Preferably, the preparation method of the Platycodon grandiflorus extract includes: pulverizing Platycodon grandiflorus, mixing it with an ethanol solution of 65%-75% by volume for ultrasonic extraction, filtering, eluting the filtrate through a macroporous adsorption resin column, concentrating and drying the eluent to obtain the Platycodon grandiflorus extract.

[0012] More preferably, the mass-to-volume ratio of Platycodon grandiflorus to ethanol solution is 1:15-20 g / mL, the ultrasonic extraction temperature is 50-60℃, the power is 100-200W, the number of extractions is 1-3, and each extraction lasts 20-30 minutes.

[0013] More preferably, the elution includes: eluting sequentially with an ethanol solution of 40%-45% by volume and an ethanol solution of 55%-60% by volume, and collecting the eluent of the ethanol solution of 55%-60% by volume.

[0014] Preferably, the excipients include sweeteners, acidity regulators, preservatives, and stabilizers; the sweetener is one or more of sucrose, erythritol, xylitol, and sucralose; the acidity regulator is one or more of citric acid, malic acid, and sodium citrate; the preservative is one or more of sodium benzoate, potassium sorbate, and disodium EDTA; and the stabilizer is one or more of sodium carboxymethyl cellulose, xanthan gum, or microcrystalline cellulose.

[0015] The present invention also provides a method for preparing the above-mentioned fermented beverage, comprising: mixing Astragalus membranaceus fermented extract and Platycodon grandiflorus extract with water at 50-60°C for 10-15 min, then sequentially adding L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients, cooling to 30-40°C and then adding caffeine and pyrroloquinoline quinone, adjusting the pH value, sterilizing and then filling to obtain the fermented beverage.

[0016] Preferably, the weight ratio of the Astragalus fermentation extract to water is 8-12:1000.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] This invention provides a fermented beverage for relieving fatigue, comprising: Astragalus fermented extract, Platycodon grandiflorus extract, L-arginine, taurine, inositol, caffeine, nicotinamide, vitamin B6, L-lysine hydrochloride, pyrrolidinequinoline quinone, and excipients. The Astragalus fermented extract of this invention can improve ATP production efficiency, reduce lactic acid accumulation after exercise, alleviate oxidative stress damage to muscle and nerve cells, enhance macrophage activity, and reduce fatigue-induced immunosuppression. The Platycodon grandiflorus extract refreshes the mind, improves cognitive decline caused by central nervous system fatigue, enhances capillary permeability, and promotes the clearance of metabolic waste. The Astragalus fermented extract and Platycodon grandiflorus extract have a synergistic effect, significantly relieving both physical and mental fatigue. All components of this fermented beverage possess excellent anti-fatigue effects. The combined action of these components relieves fatigue, rapidly replenishes energy, and restores physical strength. Detailed Implementation

[0019] This invention provides a fermented beverage for relieving fatigue, comprising the following raw materials in parts by weight: 8-12 parts of Astragalus membranaceus fermented extract, 5-10 parts of Platycodon grandiflorus extract, 0.01-0.02 parts of L-arginine, 0.1-0.4 parts of taurine, 0.06-0.1 parts of inositol, 0.05-0.1 parts of caffeine, 0.005-0.01 parts of nicotinamide, 0.001-0.0015 parts of vitamin B6, 0.3-0.8 parts of L-lysine hydrochloride, 0.01-0.03 parts of pyrrolquinone, and 50-60 parts of excipients.

[0020] The active ingredients polysaccharides and saponins in the Astragalus extract of this invention can enhance the activity of immune cells, strengthen the body's anti-fatigue ability, reduce free radicals, protect cells, delay fatigue, increase ATP production efficiency, reduce lactic acid accumulation after exercise, reduce oxidative stress damage to muscle and nerve cells, enhance macrophage activity, and reduce fatigue-induced immunosuppression.

[0021] The Platycodon grandiflorus extract of this invention has the effects of refreshing the mind and has strong antioxidant properties. It can eliminate free radicals, protect brain cells, maintain a healthy microenvironment in the brain, improve cognitive function decline caused by central fatigue, enhance capillary permeability, and promote the removal of metabolic waste, thereby relieving mental fatigue.

[0022] The present invention demonstrates a synergistic effect between Astragalus membranaceus extract and Platycodon grandiflorus extract, which alleviates fatigue from different aspects and enhances the body's anti-fatigue ability and overall health level.

[0023] The L-arginine described in this invention can improve blood circulation, increase oxygen and nutrient supply, and relieve physical fatigue; it also participates in the urea cycle, removes metabolic waste, and enhances anti-fatigue ability.

[0024] The taurine described in this invention can reduce oxidative stress, promote fat metabolism, provide energy, regulate neurotransmitters, improve brain function, and thus relieve fatigue.

[0025] The inositol described in this invention participates in nerve signal transduction, relieves mental fatigue, helps with energy supply, and alleviates physical fatigue.

[0026] The caffeine in this invention can stimulate the central nervous system, refresh the mind, relieve mental fatigue, promote fat breakdown, provide energy, improve cognitive function, and enhance work efficiency.

[0027] The nicotinamide of this invention, as a precursor of a coenzyme, promotes ATP production, improves blood circulation, increases oxygen and nutrient supply, reduces free radical damage, and protects cells.

[0028] The vitamin B6 of this invention can help in the synthesis and breakdown of proteins and amino acids, promote the synthesis of neurotransmitters, and relieve nerve fatigue.

[0029] The L-lysine hydrochloride of this invention promotes protein synthesis, nourishes nerves, enhances immunity, participates in fatty acid metabolism, provides energy, and relieves fatigue.

[0030] The pyrroloquinoline quinone of this invention can effectively scavenge free radicals, protect cells, enhance cellular energy metabolism, relieve fatigue, support neuronal health, and alleviate mental fatigue.

[0031] The fermented beverage of this invention not only relieves fatigue but also enhances the body's immunity through the combined effects of its various components.

[0032] The preferred method for preparing the Astragalus fermented extract of the present invention includes: pulverizing Astragalus, mixing it with water, sequentially subjecting it to enzymatic hydrolysis by cellulase and nattokinase via ultrasonication, followed by sequential fermentation by Pediococcus pentosus and Bifidobacterium infantis, filtering, concentrating the filtrate under reduced pressure, and freeze-drying to obtain the Astragalus fermented extract.

[0033] The cellulase of the present invention preferably has an enzyme activity of 50,000-150,000 U / g, more preferably 100,000 U / g, and the addition amount is preferably 2%-5% of the weight of Astragalus membranaceus, more preferably 3%. The enzymatic hydrolysis temperature is preferably 45-55℃, more preferably 50℃, the pH value is preferably 4.5-5.5, more preferably 5, the ultrasonic power is preferably 350-450W, more preferably 400W, and the enzymatic hydrolysis time is preferably 30-60min, more preferably 45min. The nattokinase of the present invention preferably has an enzyme activity of 20,000-60,000 U / g, more preferably 40,000 U / g, and the addition amount is preferably 1%-3% of the weight of Astragalus membranaceus, more preferably 2%. The enzymatic hydrolysis temperature is preferably 37-42℃, more preferably 40℃, the pH value is preferably 7-9, more preferably 8, the ultrasonic power is preferably 300-400W, more preferably 350W, and the enzymatic hydrolysis time is preferably 20-30min, more preferably 25min.

[0034] Cellulase can break down the cell wall structure of Astragalus membranaceus, making it easier for the active ingredients inside the cells to be released. Nattokinase further enzymatically breaks down the proteins and other components in Astragalus membranaceus, allowing the polypeptides, amino acids and other substances in Astragalus membranaceus to be released more fully. These small molecules are more easily absorbed by the human body, providing a more sufficient material basis for relieving fatigue.

[0035] The *Pediococcus pentosaceus* described in this invention is *Pediococcus pentosaceus*. Pediococcus pentosaceus CICC ® 22253, the viability of *Pediococcus pentosaceus* is preferably 10-20 billion / g, more preferably 15 billion / g, the addition amount is preferably 1‰-3‰ of the weight of *Astragalus membranaceus*, more preferably 2‰, the fermentation temperature is preferably 42-48℃, more preferably 45℃, the pH value is preferably 5.5-6.5, more preferably 6, the rotation speed is preferably 150-250 rpm, more preferably 200 rpm, and the fermentation time is preferably 24-36 h, more preferably 30 h.

[0036] The fermented Bifidobacterium infantis described in this invention is the Bifidobacterium infantis strain IFA09, purchased from the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: M201131, and disclosed in invention patent CN103131647A. The preferred viability of the fermented Bifidobacterium infantis is 20-30 billion CFU / g, more preferably 25 billion CFU / g. The preferred amount added is 5‰-8‰ of the weight of Astragalus membranaceus, more preferably 6‰. The preferred fermentation temperature is 35-40℃, more preferably 37℃. The preferred pH value is 6.5-7, more preferably 6.8. The preferred fermentation time is 24-72h, more preferably 36h.

[0037] This invention first uses Pediococcus pentosaceus to aerobically ferment the enzymatic hydrolysate, and then uses Bifidobacterium infantis for anaerobic fermentation. This can decompose the active ingredients in Astragalus membranaceus into small molecules with special biological activities (antioxidant, antibacterial, and anti-inflammatory), which are more easily absorbed by the human body. It may also change the chemical structure of the active ingredients in Astragalus membranaceus, producing derivatives with stronger active effects, thereby improving the fatigue relief effect.

[0038] The preferred method for preparing the Platycodon grandiflorus extract of this invention includes: pulverizing Platycodon grandiflorus, mixing it with an ethanol solution of 65%-75% by volume for ultrasonic extraction, filtering, eluting the filtrate through a macroporous adsorption resin column, concentrating and drying the eluent to obtain the Platycodon grandiflorus extract. The preferred mass-to-volume ratio of Platycodon grandiflorus to ethanol solution is 1:15-20 g / mL, more preferably 1:18 g / mL; the preferred ultrasonic extraction temperature is 50-60℃, more preferably 55℃; the preferred power is 100-200W, more preferably 150W; the preferred number of extractions is 1-3 times, more preferably 2 times; and each extraction is preferably 20-30 min, more preferably 25 min. This invention uses a combination of ethanol and ultrasound to extract the effective components of Platycodon grandiflorus, which, compared to water extraction, better enhances the anti-fatigue effect of Platycodon grandiflorus.

[0039] The macroporous adsorption resin column used in this invention is preferably an AB-8 macroporous adsorption resin column or a D101 macroporous adsorption resin column. The elution preferably includes: sequentially eluting with a 40%-45% (v / v) ethanol solution and then with a 55%-60% (v / v) ethanol solution, collecting the eluent with a 55%-60% (v / v) ethanol solution; more preferably, it includes: sequentially eluting with a 43% (v / v) ethanol solution and then with a 58% (v / v) ethanol solution, collecting the eluent with a 55% (v / v) ethanol solution. Purification of Platycodon grandiflorus filtrate using a macroporous adsorption resin column can significantly enhance the anti-fatigue effect of Platycodon grandiflorus extract.

[0040] The excipients of this invention include sweeteners, acidity regulators, preservatives, and stabilizers; the sweeteners are one or more of sucrose, erythritol, xylitol, and sucralose; the acidity regulators are one or more of citric acid, malic acid, and sodium citrate; the preservatives are one or more of sodium benzoate, potassium sorbate, and disodium EDTA; and the stabilizers are one or more of sodium carboxymethyl cellulose, xanthan gum, or microcrystalline cellulose.

[0041] The present invention also provides a method for preparing the above-mentioned fermented beverage, comprising: mixing Astragalus membranaceus fermented extract and Platycodon grandiflorus extract with water at 50-60°C for 10-15 min, then sequentially adding L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients, cooling to 30-40°C and then adding caffeine and pyrroloquinoline quinone, adjusting the pH value, sterilizing and then filling to obtain the fermented beverage.

[0042] The weight ratio of Astragalus fermentation extract to water in this invention is 8-12:1000, more preferably 10:1000.

[0043] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0044] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0045] Unless otherwise specified, the following embodiments are all conventional methods.

[0046] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0047] Example 1

[0048] Preparation of Fermented Beverages for Relieving Fatigue

[0049] (1) Astragalus fermentation extract

[0050] After drying, Astragalus membranaceus is pulverized and passed through a 200-mesh sieve to obtain Astragalus membranaceus powder.

[0051] Astragalus powder was mixed with deionized water at a ratio of 1:15 g / mL. Then, 3% (by weight of Astragalus powder) of cellulase (enzyme activity of 100,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 45 min at 50℃, pH 5, and 400W to inactivate the enzyme. Next, 2% (by weight of Astragalus powder) of nattokinase (enzyme activity of 40,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 25 min at 40℃, pH 8, and 350W to inactivate the enzyme, thus obtaining the enzymatic hydrolysis product.

[0052] The enzymatic hydrolysate was inoculated with 2‰ of Astragalus powder containing Pediococcus pentosaceus (15 billion CFU / g) and fermented aerobically at 45℃, pH 6, and 200 rpm for 30 h. After sterilization, it was inoculated with 5‰ of Astragalus powder containing Bifidobacterium infantis (25 billion CFU / g) and fermented anaerobically at 37℃ and pH 6.8 for 36 h. After sterilization and filtration, the fermentation broth was concentrated under reduced pressure to 1 / 4 of the fermentation broth volume and then freeze-dried under vacuum to obtain the Astragalus fermentation extract.

[0053] (2) Platycodon grandiflorus extract

[0054] After drying, the platycodon root is pulverized and passed through a 200-mesh sieve to obtain platycodon root powder;

[0055] Platycodon grandiflorus powder was mixed with a 70% ethanol solution at a ratio of 1:18 g / mL and then ultrasonically extracted at 55℃ and 150W for 30 min. After filtration, the residue was mixed with a 70% ethanol solution at a ratio of 1:18 g / mL and then ultrasonically extracted at 55℃ and 150W for 30 min. After filtration, the filtrates were combined to obtain the ethanol extract.

[0056] After the ethanol extract was adsorbed onto an AB-8 macroporous adsorption resin column, it was eluted sequentially with 5 column volumes of 43% ethanol solution and 10 column volumes of 58% ethanol solution. The eluent of the 58% ethanol solution was collected, concentrated under reduced pressure to 1 / 4 of the fermentation broth volume, and then freeze-dried under vacuum to obtain the Platycodon grandiflorus extract.

[0057] (3) Preparation of fermented beverages

[0058] Weigh out 10 parts by weight of Astragalus membranaceus fermented extract, 8 parts by weight of Platycodon grandiflorus extract, 0.015 parts by weight of L-arginine, 0.3 parts by weight of taurine, 0.08 parts by weight of inositol, 0.07 parts by weight of caffeine, 0.008 parts by weight of nicotinamide, 0.007 parts by weight of vitamin B6, 0.5 parts by weight of L-lysine hydrochloride, 0.02 parts by weight of pyrrolidone, 56.05 parts by weight of excipients, and 1000 parts by weight of purified water. The excipients consist of the following raw materials in parts by weight: 53 parts by weight of sucrose, 0.03 parts by weight of sucralose, 1 part by weight of sodium citrate, 0.02 parts by weight of sodium benzoate, and 2 parts by weight of sodium carboxymethyl cellulose.

[0059] Astragalus fermentation extract and Platycodon grandiflorus extract were mixed with purified water at 55℃ and 200 rpm for 13 minutes. Then, L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients were added in sequence. After cooling to 35℃, caffeine and pyrroloquinoline quinone were added. The pH was adjusted to 5.5, sterilized and bottled to obtain a fermented beverage.

[0060] Example 2

[0061] Preparation of Fermented Beverages for Relieving Fatigue

[0062] (1) Astragalus fermentation extract

[0063] After drying, Astragalus membranaceus was pulverized and passed through a 150-mesh sieve to obtain Astragalus membranaceus powder.

[0064] Astragalus powder and deionized water were mixed at a ratio of 1:10 g / mL. Then, 5% (by weight of Astragalus powder) of cellulase (enzyme activity of 50,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 60 min at 45℃, pH 4.5, and power 350W to inactivate the enzyme. Then, 3% (by weight of Astragalus powder) of nattokinase (enzyme activity of 20,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 30 min at 37℃, pH 7, and power 300W to inactivate the enzyme and obtain the enzymatic hydrolysis product.

[0065] The enzymatic hydrolysate was inoculated with 3‰ of Astragalus powder containing Pediococcus pentosaceus (10 billion CFU / g) and fermented aerobicly at 42℃, pH 5.5, and 150 rpm for 24 h. After sterilization, it was inoculated with 8‰ of Astragalus powder containing Bifidobacterium infantis (20 billion CFU / g) and fermented anaerobicly at 35℃ and pH 6.5 for 72 h. After sterilization and filtration, the fermentation broth was concentrated under reduced pressure to 1 / 3 of the fermentation broth volume and then freeze-dried under vacuum to obtain the Astragalus fermentation extract.

[0066] (2) Platycodon grandiflorus extract

[0067] After drying, the platycodon root is pulverized and passed through a 150-mesh sieve to obtain platycodon root powder.

[0068] Platycodon grandiflorus powder was mixed with 65% ethanol solution at a ratio of 1:15 g / mL and then ultrasonically extracted at 50℃ and 100W for 20 min. After filtration, the residue was extracted twice more by ultrasonic extraction as described above, filtered, and the filtrates were combined to obtain the ethanol extract.

[0069] After the ethanol extract was adsorbed onto a D101 macroporous adsorption resin column, it was eluted sequentially with 6 column volumes of 40% ethanol solution and 12 column volumes of 55% ethanol solution. The eluent of the 55% ethanol solution was collected, concentrated under reduced pressure to 1 / 3 of the fermentation broth volume, and then freeze-dried under vacuum to obtain the Platycodon grandiflorus extract.

[0070] (3) Preparation of fermented beverages

[0071] Weigh out 8 parts by weight of Astragalus membranaceus fermented extract, 10 parts by weight of Platycodon grandiflorus extract, 0.01 parts by weight of L-arginine, 0.4 parts by weight of taurine, 0.06 parts by weight of inositol, 0.1 parts by weight of caffeine, 0.005 parts by weight of nicotinamide, 0.0015 parts by weight of vitamin B6, 0.3 parts by weight of L-lysine hydrochloride, 0.01 parts by weight of pyrrolidone, 51.6 parts by weight of excipients, and 1000 parts by weight of purified water. The excipients consist of the following raw materials in parts by weight: 45 parts by weight of sucrose, 5 parts by weight of erythritol, 0.5 parts by weight of citric acid, 0.1 parts by weight of potassium sorbate, and 1 part by weight of xanthan gum.

[0072] Astragalus fermentation extract and Platycodon grandiflorus extract were mixed with purified water at 50℃ and 150rpm for 15 minutes. Then, L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients were added in sequence. After cooling to 30℃, caffeine and pyrroloquinoline quinone were added. The pH was adjusted to 5, sterilized and then bottled to obtain a fermented beverage.

[0073] Example 3

[0074] Preparation of Fermented Beverages for Relieving Fatigue

[0075] (1) Astragalus fermentation extract

[0076] After drying, Astragalus membranaceus was pulverized and passed through a 250-mesh sieve to obtain Astragalus membranaceus powder.

[0077] Astragalus powder was mixed with deionized water at a ratio of 1:18 g / mL. Then, 2% (by weight of the astragalus powder) of cellulase (enzyme activity of 150,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 30 min at 55℃, pH 5.5, and power 450W to inactivate the enzyme. Next, 1% (by weight of the astragalus powder) of nattokinase (enzyme activity of 60,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 20 min at 42℃, pH 9, and power 400W to inactivate the enzyme and obtain the enzymatic hydrolysis product.

[0078] The enzymatic hydrolysate was inoculated with 1‰ of Astragalus powder containing Pediococcus pentosaceus (20 billion CFU / g) and fermented aerobicly at 48℃, pH 6.5, and 250 rpm for 36 h. After sterilization, it was inoculated with 5‰ of Astragalus powder containing Bifidobacterium infantis (30 billion CFU / g) and fermented anaerobicly at 40℃ and pH 7 for 24 h. After sterilization and filtration, the fermentation broth was concentrated under reduced pressure to 1 / 5 of the fermentation broth volume and then freeze-dried under vacuum to obtain the Astragalus fermentation extract.

[0079] (2) Platycodon grandiflorus extract

[0080] After drying, the platycodon root is pulverized and passed through a 250-mesh sieve to obtain platycodon root powder;

[0081] Platycodon grandiflorus powder was mixed with 75% ethanol solution at a ratio of 1:20 g / mL, and then ultrasonically extracted at 50℃ and 200W for 30 min. The mixture was then filtered, and the filtrate was the ethanol extract.

[0082] After the ethanol extract was adsorbed onto a D101 macroporous adsorption resin column, it was eluted sequentially with 4 column volumes of 45% ethanol solution and 8 column volumes of 60% ethanol solution. The eluent of the 60% ethanol solution was collected, concentrated under reduced pressure to 1 / 5 of the fermentation broth volume, and then freeze-dried under vacuum to obtain the Platycodon grandiflorus extract.

[0083] (3) Preparation of fermented beverages

[0084] Weigh out 12 parts by weight of Astragalus membranaceus fermentation extract, 5 parts by weight of Platycodon grandiflorus extract, 0.02 parts by weight of L-arginine, 0.1 parts by weight of taurine, 0.1 parts by weight of inositol, 0.05 parts by weight of caffeine, 0.01 parts by weight of nicotinamide, 0.001 parts by weight of vitamin B6, 0.8 parts by weight of L-lysine hydrochloride, 0.01 parts by weight of pyrrolidone, 60.25 parts by weight of excipients, and 1000 parts by weight of purified water. The excipients consist of the following raw materials in parts by weight: 56 parts by weight of sucrose, 3 parts by weight of xylitol, 0.5 parts by weight of malic acid, 0.05 parts by weight of disodium EDTA, and 0.7 parts by weight of microcrystalline cellulose.

[0085] Astragalus fermentation extract and Platycodon grandiflorus extract were mixed with purified water at 60℃ and 250rpm for 15 minutes. Then, L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients were added in sequence. After cooling to 30℃, caffeine and pyrroloquinoline quinone were added. The pH was adjusted to 5, sterilized and then bottled to obtain a fermented beverage.

[0086] Comparative Example 1

[0087] The specific implementation method is the same as in Example 1, except that cellulase in step (1) is replaced with ligninase, nattokinase is replaced with elastase, and the ultrasonic enzymatic hydrolysis of Astragalus membranaceus is as follows:

[0088] Astragalus powder was mixed with deionized water at a ratio of 1:15 g / mL. Then, 3% (by weight of the astragalus powder) of ligninase (enzyme activity of 100,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 45 min at 40℃, pH 4, and power of 400W to inactivate the enzyme. Then, 2% (by weight of the astragalus powder) of elastase (enzyme activity of 40,000 U / g) was added. The mixture was ultrasonically hydrolyzed for 25 min at 50℃, pH 8, and power of 350W to inactivate the enzyme and obtain the enzymatic hydrolysis product.

[0089] Comparative Example 2

[0090] The specific implementation method is the same as in Example 1, except that the Pediococcus pentosaceus in step (1) is replaced with Lactococcus lactis subsp. lactis. Lactococcus lactis subsp. lactis CICC ® 6031 (purchased from the China Industrial Microbial Culture Collection Center), replacing Bifidobacterium infantis with Bifidobacterium longum. Bifidobacterium longum CICC ® 6195 (purchased from the China Industrial Microbial Culture Collection Center), fermentation steps are as follows:

[0091] The enzymatic hydrolysate was inoculated with 2‰ of Astragalus membranaceus powder containing *Lactococcus lactis* subsp. lactis (15 billion live bacteria / g) and fermented aerobically at 37℃, pH 6.5, and 200 rpm for 30 h. After sterilization, the product was inoculated with 5‰ of Astragalus membranaceus powder containing *Bifidobacterium longum* (25 billion live bacteria / g) and fermented anaerobically at 37℃ and pH 6.8 for 36 h. After sterilization and filtration, the fermentation broth was concentrated under reduced pressure to 1 / 4 of its volume and then freeze-dried under vacuum to obtain the Astragalus membranaceus fermentation extract.

[0092] Comparative Example 3

[0093] The specific implementation method is the same as that in Example 1, except that the 70% ethanol solution in step (2) is replaced with pure water.

[0094] Comparative Example 4

[0095] The specific implementation method is the same as that in Example 1, except that "13 parts by weight of Astragalus fermented extract and 10 parts by weight of Platycodon extract" in step (3) are replaced with 23 parts by weight of Astragalus fermented extract.

[0096] Comparative Example 5

[0097] The specific implementation method is the same as that in Example 1, except that "13 parts by weight of Astragalus fermentation extract and 10 parts by weight of Platycodon extract" in step (3) are replaced with 23 parts by weight of Platycodon extract.

[0098] Experimental Example 1

[0099] Anti-fatigue animal experiments

[0100] One hundred and eighty male Kunming mice weighing 25-35g were selected and acclimatized for seven days. They were then divided into nine groups of 20 mice each, with no significant differences between the groups.

[0101] The mice were divided into a treatment group and a control group. The treatment group was administered 0.5 mL / 10 gbw of fermented beverages from Examples 1-3 and Comparative Examples 1-5 by gavage at 10:00 AM every day. The control group was administered an equal volume of physiological saline by gavage. The mice had free access to water during the gavage period. After 30 days of continuous administration, exercise endurance and biochemical indicators were measured.

[0102] (1) Swimming experiment

[0103] On day 27 after gavage, mice were acclimatized to swimming. One hour after gavage, the mice were placed in water at 25±1°C and swam for 10 minutes without any weight. This acclimatization period lasted for 3 days, with the swimming time increasing by 5 minutes each day thereafter. On day 30, one hour after gavage, a swimming exhaustion test was conducted. Mice were fasted for 12 hours before swimming and had a lead weight (6% of their body weight) attached to their tails. The swimming exhaustion time for each group was recorded. The standard was defined as the mouse remaining submerged for 8 seconds without being able to surface for air. The swimming exhaustion time data for each group are shown in Table 1.

[0104] Table 1. Exhaustion Time of Fermented Beverages in Each Group

[0105]

[0106] As shown in Table 1, compared with the control group, the time to exhaustion of mice treated with other fermented beverages via gavage was increased to varying degrees, with the exhaustion time in Examples 1-3 being significantly longer than that in other groups. A comparison of Example 1 and Comparative Examples 1-3 shows that fermented beverages obtained from Astragalus or Platycodon extracts using different enzyme hydrolysis, different microbial fermentation, and different solvents exhibit different anti-fatigue effects. The method used in this invention—sequentially hydrolyzing Astragalus with cellulase and nattokinase, fermenting the Astragalus hydrolysate with Pediococcus pentosaceus and Bifidobacterium infantis, and extracting Platycodon with 70% ethanol—resulted in a better-quality fermented beverage. This indicates that cellulase and nattokinase, and Pediococcus pentosaceus and Bifidobacterium infantis, have synergistic effects in terms of anti-immunity. Examples 1 and Comparative Examples 4-5 show that Astragalus fermented extract and Platycodon extract have a synergistic effect, significantly prolonging exhaustion time and improving anti-fatigue effects.

[0107] (2) Lactic acid content

[0108] Blood lactate is a product of anaerobic metabolism during exercise, and its concentration is closely related to exercise intensity and fatigue level. When exercise intensity exceeds the aerobic metabolic threshold, lactate accumulates in muscles, leading to a decrease in pH and subsequently causing fatigue. Therefore, lactate is an important indicator of exercise-induced fatigue. Before swimming, 5 minutes after exhaustion, and 20 minutes after exhaustion, the tail tip of mice was disinfected with an alcohol swab, and blood was collected by tail cutting. A portable blood lactate analyzer was used to measure the lactate content of each group at each time point. The specific results are shown in Table 2.

[0109] Table 2. Lactic acid content (mmol / L) in each group at different time points.

[0110]

[0111] Table 2 shows that there was no significant difference in lactic acid content among the groups of mice before swimming. Five minutes after swimming exhaustion, lactic acid accumulation was significantly reduced in mice given the fermented beverage via gavage, compared to the control group, indicating that the fermented beverage can alleviate fatigue. Data from Example 1 and Comparative Examples 1-3 show that the fermented beverage prepared using the enzymes and microorganisms of this invention has a superior anti-fatigue effect. Data from Example 1 and Comparative Examples 4-5 show that, at the same concentration of active ingredients, Astragalus fermented extract and Platycodon grandiflorus extract have a synergistic effect, enhancing the anti-fatigue effect.

[0112] (3) Antioxidant stress indicators

[0113] Strenuous exercise can increase the production of reactive oxygen species (ROS) in skeletal muscle. ROS can attack intracellular lipid components, leading to peroxidation, which is one of the causes of muscle fatigue. Changes in serum SOD (superoxide dismutase), MDA (malondialdehyde), LDH (lactate dehydrogenase), and CK (creatine kinase) levels are closely related to the body's anti-fatigue ability. By detecting these indicators, the body's anti-fatigue effect can be assessed.

[0114] Immediately after the mice in each group were exhausted from swimming, blood was collected from their eyes to measure the serum levels of SOD, MDA, LDH, and CK. The specific results are shown in Table 3.

[0115] Table 3. Serum levels of T-SOD, MDA, LDH, and CK in each group

[0116]

[0117] As shown in Table 3, compared to the control group, mice given other fermented beverages via gavage showed increased SOD activity and decreased MDA content, LDH activity, and CK activity. Overall, compared to the fermented beverages in Comparative Examples 1-5, Examples 1-3 of this invention exhibit better antioxidant stress effects and superior anti-fatigue effects.

[0118] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A fermented beverage for relieving fatigue, characterized in that, The ingredients include the following parts by weight: 8-12 parts of Astragalus membranaceus fermented extract, 5-10 parts of Platycodon grandiflorus extract, 0.01-0.02 parts of L-arginine, 0.1-0.4 parts of taurine, 0.06-0.1 parts of inositol, 0.05-0.1 parts of caffeine, 0.005-0.01 parts of nicotinamide, 0.001-0.0015 parts of vitamin B6, 0.3-0.8 parts of L-lysine hydrochloride, 0.01-0.03 parts of pyrroloquinoline quinone, and 50-60 parts of excipients; The preparation method of the Astragalus fermented extract includes: pulverizing Astragalus, mixing it with water, sequentially passing it through cellulase and nattokinase via ultrasonic enzymatic hydrolysis, then sequentially fermenting it with Pediococcus pentosus CICC22253 and Bifidobacterium infantis CCTCC NO:M201131, filtering it, concentrating the filtrate under reduced pressure, and freeze-drying it to obtain the Astragalus fermented extract. The preparation method of the Platycodon grandiflorus extract includes: pulverizing Platycodon grandiflorus, mixing it with an ethanol solution of 65%-75% by volume for ultrasonic extraction, filtering, eluting the filtrate through a macroporous adsorption resin column, concentrating and drying the eluent to obtain the Platycodon grandiflorus extract.

2. The fermented beverage according to claim 1, characterized in that, The cellulase has an enzyme activity of 50,000-150,000 U / g, is added at 2%-5% of the weight of Astragalus membranaceus, has an enzymatic hydrolysis temperature of 45-55℃, a pH of 4.5-5.5, an ultrasonic power of 350-450W, and an enzymatic hydrolysis time of 30-60 min; the nattokinase has an enzyme activity of 20,000-60,000 FU / g, is added at 1%-3% of the weight of Astragalus membranaceus, has an enzymatic hydrolysis temperature of 37-42℃, a pH of 7-9, an ultrasonic power of 300-400W, and an enzymatic hydrolysis time of 20-30 min.

3. The fermented beverage according to claim 1, characterized in that, The viability of the *Pediococcus pentosaceus* is 10-20 billion CFU / g, and the addition amount is 1‰-3‰ of the weight of the *Astragalus membranaceus*. The fermentation temperature is 42-48℃, the pH value is 5.5-6.5, the rotation speed is 150-250 rpm, and the time is 24-36 h. The viability of the fermented *Bifidobacterium infantis* is 20-30 billion CFU / g, and the addition amount is 5‰-8‰ of the weight of the *Astragalus membranaceus*. The fermentation temperature is 35-40℃, the pH value is 6.5-7, and the time is 24-72 h.

4. The fermented beverage according to claim 1, characterized in that, The mass-to-volume ratio of Platycodon grandiflorus to ethanol solution is 1:15-20 g / mL. The ultrasonic extraction temperature is 50-60℃, the power is 100-200W, the number of extractions is 1-3, and each extraction lasts 20-30 minutes.

5. The fermented beverage according to claim 1, characterized in that, The elution process includes: sequentially eluting with an ethanol solution of 40%-45% by volume and an ethanol solution of 55%-60% by volume, and collecting the eluent of the ethanol solution of 55%-60% by volume.

6. The fermented beverage according to claim 1, characterized in that, The excipients include sweeteners, acidity regulators, preservatives, and stabilizers; the sweeteners are one or more of sucrose, erythritol, xylitol, and sucralose; the acidity regulators are one or more of citric acid, malic acid, and sodium citrate; the preservatives are one or more of sodium benzoate, potassium sorbate, and disodium EDTA; and the stabilizers are one or more of sodium carboxymethyl cellulose, xanthan gum, or microcrystalline cellulose.

7. The method for preparing the fermented beverage according to any one of claims 1-6, characterized in that, include: After mixing Astragalus fermentation extract and Platycodon grandiflorus extract with water at 50-60℃ for 10-15 minutes, L-arginine, taurine, inositol, nicotinamide, vitamin B6, L-lysine hydrochloride and excipients are added in sequence. After cooling to 30-40℃, caffeine and pyrroloquinoline quinone are added. After adjusting the pH value and sterilizing, the mixture is bottled to obtain a fermented beverage.

8. The preparation method according to claim 7, characterized in that, The weight ratio of the Astragalus fermentation extract to water is 8-12:1000.

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