Functional composite fruit juice capable of supplementing energy and preparation method of functional composite fruit juice

Abstract

The invention discloses functional composite fruit juice capable of supplementing energy and a preparation method of the functional composite fruit juice, and belongs to the technical field of composite fruit juice. The functional composite fruit juice is prepared from the following components in parts by weight: plant source functional instant powder, an anti-fatigue compound, an antioxidant extract, sodium alginate, taurine, fruit juice stock solution and deionized water. The plant source functional instant powder, the anti-fatigue compound and the anti-oxidation extract are added into the compound fruit juice, so that the immunity of the organism is quickly improved, and energy is recovered.

Description

Technical Field

[0001] The invention belongs to the technical field of compound fruit juices, and specifically refers to a functional compound fruit juice for supplementing energy and a preparation method thereof. Background Art

[0002] The development of functional compound juice is one of the research hotspots in modern food science, especially in meeting the needs of rapid energy replenishment and nutrition. Traditional fruit juice is mainly based on the natural flavor and basic nutrition of fruits. Functional juice further enhances its health value and specific functions through reasonable raw material matching and process optimization, such as energy replenishment, anti-oxidation, and immunity improvement. Functional compound juice usually uses a combination of different fruits, plant extracts and other functional ingredients to achieve the purpose of energy replenishment and health enhancement, such as fruits containing natural fructose and glucose, high-vitamin fruits, and fruits containing minerals. It also includes functional added ingredients such as glucose, maltose, honey, etc. to enhance the direct energy replenishment effect, ginseng, maca, green tea extracts, etc. to enhance vitality and metabolic capacity such as branched-chain amino acids (BCAA), glutamine, etc., to promote energy metabolism and muscle recovery. Functional compound juices have rich varieties of nutrients and sufficient content, so they mainly have the following problems: Nutrient stability: Many functional ingredients (such as vitamin C) are easily decomposed or inactivated during processing, and need to be optimized through process optimization and the addition of antioxidants to maintain their activity; Flavor balance: The combination of multiple raw materials may produce an unpleasant taste, which needs to be solved through formula adjustment and flavor improvement technology (such as blending spices); Absorption and metabolism: How to design energy supplement formulas that are easier for the human body to absorb and utilize is an important direction, such as controlling the proportion of reducing sugars to avoid large fluctuations in blood sugar; Product preservation: The water and nutrients in the juice are easy to breed microorganisms, and the shelf life needs to be extended through low-temperature pasteurization, high-pressure treatment or the addition of natural preservatives. Summary of the invention

[0003] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a functional compound juice for replenishing energy and a preparation method thereof. The present invention quickly improves the body's immunity and restores energy by adding plant-derived functional instant powder, anti-fatigue complex, and antioxidant extract to the compound juice.

[0004] In order to achieve the above-mentioned object, the technical scheme adopted by the present invention is as follows: the present invention proposes a functional compound juice for supplementing energy, the functional compound juice comprises the following components in parts by weight: 5-6 parts of plant-derived functional instant powder, 8-10 parts of anti-fatigue complex, 1.5-2 parts of antioxidant extract, 5-8 parts of sodium alginate, 0.2-0.4 parts of taurine, 20-30 parts of juice stock solution, and 80-100 parts of deionized water; Preferably, the plant-derived functional instant powder comprises the following components in parts by weight: 8-10 parts of purple alfalfa instant powder and 4-5 parts of American ginseng instant powder; Preferably, the anti-fatigue extract comprises the following components in parts by weight: 10-12 parts of acetylated sea cucumber polysaccharide and 6-8 parts of sea cucumber polypeptide; Preferably, the anti-fatigue extract is prepared by a polysaccharide-polypeptide co-production method; Preferably, the juice stock solution comprises the following components in parts by weight: 10-12 parts of freshly squeezed orange stock solution, 3-4 parts of passion fruit juice stock solution and 1-2 parts of lemon juice stock solution; Preferably, the method for preparing the plant-derived functional instant powder specifically comprises the following steps: (1) Wash the plant rhizome tissue and cut it into 5-10 mm stem segments. After longitudinal sectioning, place it in a proliferation medium containing indoleacetic acid and kinetin. Culture it in the dark at 22-28°C for 21-30 days until the cambium shows obvious proliferation of explants. (2) Isolate explant cells and transfer them to an expansion medium containing indoleacetic acid and 6-benzylaminoadenine, and culture them at 28-32°C in a light-dark alternating environment for 21-42 days, with a light period of 8-12 hours and a dark period of 12-16 hours; subculture once every two weeks to obtain plant tissue; (3) freeze-drying the plant tissue obtained in step (2) at -40°C, grinding it into powder in liquid nitrogen, and storing it in a frozen state to obtain a plant-derived functional instant powder; Preferably, in step (1), the proliferation medium is based on MS medium, wherein the concentrations of indoleacetic acid and kinetin are 0.5-1 mg / L and 1-2 mg / L, respectively; Preferably, in step (1), the plant stems include at least one of alfalfa stems and American ginseng roots; Preferably, in step (2), the expansion medium is based on MS medium, wherein the concentrations of indoleacetic acid and 6-benzylaminoadenine are 1-2 mg / L and 1-1.5 mg / L, respectively.

[0005] Preferably, the preparation method of the anti-fatigue extract comprises the following steps: ① Wash the fresh sea cucumber, place it in deionized water at a temperature of 80-100°C, soak it for 10-20 minutes, vacuum dry it, and crush it through an 80-mesh sieve to obtain sea cucumber powder; ② Soak the sea cucumber powder obtained in step ① in water according to a mass ratio of 1:30-40, raise the temperature to 40-50° C. and treat in a constant temperature water bath for 3-5 hours, add a composite enzyme for enzymolysis, inactivate the enzyme in a boiling water bath, obtain a sea cucumber enzymolysis solution, add ammonium sulfate precipitation, centrifuge at a speed of 5000-6000 rpm for 20-30 minutes, and collect the supernatant and precipitate respectively; ③ Add an equal volume of 10% trichloroacetic acid solution to the supernatant obtained in step ②, let stand at 4°C for 12-24h, centrifuge at 5000-6000rpm for 10-20min, take the supernatant, add anhydrous ethanol for precipitation, centrifuge at 2000-3000rpm for 20-30min, take the precipitate, wash it with distilled water 3 times, and freeze-dry it to obtain sea cucumber polysaccharide; ④ The sea cucumber polysaccharide obtained in step ③ is dissolved in deionized water, and ultrasonic treatment is performed for 20-30 minutes, the ultrasonic power is 500-600W, the ultrasonic frequency is 10-20kHz, the pH is adjusted to 8.0-10.0, acetic anhydride is added dropwise according to a solid-liquid ratio of 1:15, and the reaction is stirred at 45-55°C for 2-3 hours, during which the pH is kept stable at 8.0-8.5, and then the pH is adjusted to 7.0. The product is dialyzed with distilled water in a dialysis bag with a molecular weight cutoff of 3500Da for 48 hours, and the dialyzate is concentrated under reduced pressure at 50-60°C, anhydrous ethanol is added, and alcohol precipitation is carried out at 4°C overnight, and freeze-dried to obtain acetylated sea cucumber polysaccharide; ⑤ Redissolving the precipitate obtained in step ② with water, filtering it with an inorganic nanofiltration ceramic membrane with a molecular weight cutoff of 800Da, concentrating the filtrate, and freeze-drying it to obtain a sea cucumber polypeptide; Preferably, in step ②, the compound enzyme is a compound enzyme system of papain and trypsin, the added amount of papain is 2-3% of the mass of the sea cucumber powder, and the added amount of trypsin is 1-1.2% of the mass of the sea cucumber powder; Preferably, in step ②, the enzymatic hydrolysis conditions are pH 8.0-9.0, temperature 30-40°C, and time 3-5h; Preferably, in step ②, the amount of ammonium sulfate precipitate added to the enzymatic hydrolyzate is 1-1.2 mg / mL; Preferably, in step ④, the mass concentration of the sea cucumber polysaccharide in deionized water is 20 mg / mL; Preferably, the method for preparing the antioxidant extract comprises the following steps: The dried cyanobacteria biomass powder is added to a Tris-HCl buffer with a pH of 7.0-7.5 and a concentration of 50 mmol / L, and stirred thoroughly to dissolve it, and a hydrolase is added, and enzymolysis is performed at 10-20°C for 6-10 hours, and the enzyme is inactivated in a boiling water bath, and the mixture is cooled to room temperature, and centrifuged at 8000-12000 rpm for 30-60 minutes, and the supernatant is taken, and a saturated ammonium sulfate solution is added, and the mixture is stirred at 50-100 rpm for 1 hour, and the mixture is cooled to 4°C and centrifuged at 9000-10000 rpm for 8-10 minutes, and the supernatant is collected, and ultrafiltration is performed using a 100 kDa hollow fiber membrane, and the mixture is filtered through a 0.22 μm filter membrane to obtain an antioxidant extract; Preferably, the blue algae biomass includes one of Spirulina, Pueraria lobata or Auricularia auricula; Preferably, the mass volume ratio of the cyanobacteria biomass powder and Tris-HCl buffer is 1-2 g:100 mL; Preferably, the hydrolase is a composite enzyme system of bromelain and papain, the added amount of bromelain is 1-2% of the mass of the cyanobacteria biomass powder, and the added amount of papain is 0.8-1% of the mass of the cyanobacteria biomass powder; Preferably, the volume ratio of the saturated ammonium sulfate solution to the supernatant is 1-2:10.

[0006] The present invention also provides a method for preparing a functional composite juice for supplementing energy, which specifically comprises the following steps: S1. Dissolve the plant-derived functional instant powder and the antioxidant extract in phosphate buffer respectively, stir at 5000-6000 rpm for 5-7 hours to mix the two evenly, freeze-dry, and place the freeze-dried powder in a constant temperature oven at 100° C. for 24-48 hours to obtain a pretreatment complex; S2. Add acetylated sea cucumber polysaccharide and sea cucumber polypeptide into deionized water according to the mass fraction to obtain an anti-fatigue complex solution; stir the pretreated complex prepared in step S1, sodium alginate, taurine and juice stock solution at a speed of 500-700 rpm for 10 minutes under ice bath conditions to obtain a functional composite juice.

[0007] The beneficial effects achieved by the present invention are as follows: The present invention provides a functional composite juice for replenishing energy and a preparation method thereof. The present invention rapidly improves the body's immunity and restores energy by adding plant-derived functional instant powder, anti-fatigue compound and antioxidant extract into the composite juice. The plant-derived functional instant powder is prepared by freeze-drying callus tissue of alfalfa and American ginseng and then grinding into powder. The synergistic effect of the tissues of the cambium of alfalfa and American ginseng can inhibit the generation of enzymes related to inflammation such as cyclooxygenase, help increase the number of lymphocytes, activate immune cells such as macrophages and NK cells, promote the activation of immune cells and exert excellent immune effects. The invention discloses a kind of anti-fatigue compound, which is a kind of acetylated sea cucumber polysaccharide and sea cucumber polypeptide, and is a kind of acetylated sea cucumber polysaccharide and sea cucumber polypeptide. The invention discloses a kind of anti-fatigue compound, which is a kind of acetylated sea cucumber polysaccharide and sea cucumber polypeptide, and ... Acetylation modification is performed, and the hydroxyl groups of sea cucumber polysaccharides are replaced by acetyl groups. Acetylated sea cucumber polysaccharides are generated through a nucleophilic substitution reaction. The introduction of acetyl groups can cause the sea cucumber polysaccharide branches to extend and change direction to form transversely arranged polysaccharide molecules. The spatial arrangement change of the polysaccharide chain will expose the polysaccharide hydroxyl groups, thereby improving its solubility. In addition, the acetyl group can supply hydrogen, which can inhibit the chain reaction of free radicals and convert the free radicals into a more stable form. The acetylated sea cucumber polysaccharide has better antioxidant properties and enhances the scavenging effect on hydroxyl free radicals. In the present invention, the antioxidant extract is extracted from sea cucumbers. Sea cucumber polysaccharides and sea cucumber polypeptides are extracted, and the sea cucumber polysaccharides are acetylated. The hydroxyl groups of the sea cucumber polysaccharides are replaced by acetyl groups, and acetylated sea cucumber polysaccharides are generated through a nucleophilic substitution reaction. The introduction of acetyl groups can cause the sea cucumber polysaccharide branches to extend and change direction to form laterally arranged polysaccharide molecules. The change in the spatial arrangement of the polysaccharide chains will expose the polysaccharide hydroxyl groups, thereby increasing their solubility. In addition, the acetyl group can supply hydrogen, which can inhibit the chain reaction of free radicals and convert the free radicals into a more stable form. The acetylated sea cucumber polysaccharides have better antioxidant properties and enhanced the scavenging effect on hydroxyl free radicals. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 The graph showing the reducing sugar content of the composite juices prepared in Examples 1-3 and Comparative Examples 1-3 of the present invention in simulated gastric juice; Figure 2 The antioxidant performance results of the composite juices prepared in Examples 1-3 and Comparative Examples 1-3 of the present invention are shown in FIG. Figure 3 The graph is a result of the weighted swimming time of mice in the embodiment group of the present invention and the comparative example group.

[0009] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention. DETAILED DESCRIPTION

[0010] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0011] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only and are not intended to limit the content of this application.

[0012] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials and test strains used in the following examples are purchased from commercial channels unless otherwise specified.

[0013] Papain (CasNo: 9001-73-4) was purchased from Hebei Chuangzhiyuan Biotechnology Co., Ltd.; Trypsin (CasNo: 9002-07-7) was purchased from Beijing Puxitang Biotechnology Co., Ltd.; Bromelain (CasNo: 9001-00-7) was purchased from Hunan Century Huaxing Bioengineering Co., Ltd.; Sodium alginate (CasNo: 9005-38-3) was purchased from Hubei Yongkuo Technology Co., Ltd. Example

[0014] A functional compound juice for energy supplementation, comprising the following components in parts by weight: 6 parts of plant-derived functional instant powder, 8 parts of anti-fatigue compound, 1.5 parts of antioxidant extract, 5 parts of sodium alginate, 0.4 parts of taurine, 30 parts of juice stock solution, and 80 parts of deionized water; The plant-derived functional instant powder comprises the following components in parts by weight: 10 parts of purple alfalfa instant powder and 5 parts of American ginseng instant powder; The anti-fatigue extract comprises the following components in parts by weight: 12 parts of acetylated sea cucumber polysaccharide and 8 parts of sea cucumber polypeptide; The juice stock solution includes the following components in parts by weight: 10 parts of freshly squeezed orange stock solution, 4 parts of passion fruit juice stock solution and 1 part of lemon juice stock solution; The preparation method of plant-derived functional instant powder specifically comprises the following steps: (1) Wash 3 g of alfalfa and 3 g of American ginseng roots, cut them into 5 mm stem segments, cut them longitudinally, and place them in a proliferation medium containing 0.5 mg / L indoleacetic acid and 1 mg / L kinetin. Culture them in the dark at 22°C for 21 days until the cambium shows obvious proliferation of explants. (2) Isolate explant cells and transfer them to an expansion medium containing 1 mg / L indoleacetic acid and 1 mg / L 6-benzylaminoadenine, and culture them at 28°C in a light-dark alternating medium for 21 days, with a light period of 8 h and a dark period of 12 h. Subculture the cells every two weeks to obtain alfalfa tissue and American ginseng tissue. (3) freeze-drying the plant tissue obtained in step (2) at -40°C, grinding it into powder in liquid nitrogen, and storing it in a frozen state to obtain a plant-derived functional instant powder; The preparation method of the anti-fatigue extract specifically comprises the following steps: ① Remove the lime from the mouth of the fresh sea cucumber, cut open the abdomen to remove the internal organs and mud, cut off the inner tendons, first use 20℃ water ultrasonic cleaning twice, each time for 10 minutes, ultrasonic power of 100W, ultrasonic frequency of 10kHz, then place in deionized water at 80℃, soak for 10 minutes, vacuum dry, vacuum degree of 80Pa, temperature of 55℃, time for 3 hours, crush through 80 mesh sieve to obtain sea cucumber powder; ② Soak 16g of the sea cucumber powder obtained in step ① with 300mL of water, treat it in a constant temperature water bath at 40°C for 3h, add 0.2g of papain and 0.1g of trypsin for enzymolysis, the enzymolysis pH is 8.0, the temperature is 30°C, the time is 3h, and the enzyme is inactivated in a boiling water bath to obtain a sea cucumber enzymolysis solution, add 0.31g of ammonium sulfate precipitation, centrifuge at 5000rpm for 20min, and collect the supernatant and precipitate respectively; ③ Add an equal volume of 10% trichloroacetic acid solution to the supernatant obtained in step ②, let stand at 4°C for 12h, centrifuge at 5000rpm for 10min, take the supernatant, add 3 times the volume of anhydrous ethanol to the supernatant, let stand at 4°C for 12h, centrifuge at 2000rpm for 20min, take the precipitate, wash it with distilled water 3 times, and freeze-dry it to obtain sea cucumber polysaccharide; ④ Dissolve 10g of the sea cucumber polysaccharide obtained in step ③ in 500mL of deionized water, perform ultrasonic treatment for 20min, the ultrasonic power is 500W, the ultrasonic frequency is 10kHz, adjust the pH to 8.0, add 33.3mL of acetic anhydride dropwise, stir and react at 45°C for 2h, keep the pH stable at 8.0, then adjust the pH to 7.0, dialyze the product with distilled water in a dialysis bag with a molecular weight cutoff of 3500Da for 48h, concentrate the dialysate under reduced pressure at 50°C, add anhydrous ethanol, precipitate overnight at 4°C, and freeze-dry to obtain acetylated sea cucumber polysaccharide; ⑤ Dissolve 6 g of the precipitate obtained in step ② in water, filter it using an inorganic nanofiltration ceramic membrane with a molecular weight cutoff of 800 Da, concentrate the filtrate, and freeze-dry it to obtain sea cucumber polypeptides.

[0015] The preparation method of the antioxidant extract specifically comprises the following steps: 10 g of dry spirulina powder was added to 1 L of Tris-HCl buffer with a pH of 7.0 and a concentration of 50 mmol / L, and stirred thoroughly to dissolve it. 0.1 g of bromelain and 0.08 g of papain were added, and enzymatic hydrolysis was carried out at 10°C for 6 h. The enzyme was inactivated in a boiling water bath, cooled to room temperature, and centrifuged at 8000 rpm for 30 min. The supernatant was taken, 100 mL of saturated ammonium sulfate solution was added, and the mixture was stirred at 50 rpm for 1 h. The mixture was cooled to 4°C and centrifuged at 9000 rpm for 8 min. The supernatant was collected, ultrafiltered with a 100 kDa hollow fiber membrane, and passed through a 0.22 μm filter membrane to obtain an antioxidant extract.

[0016] This embodiment provides a method for preparing a functional composite juice for energy supplementation, which specifically comprises the following steps: S1. Dissolve the plant-derived functional instant powder and the antioxidant extract in phosphate buffer respectively, stir at 5000 rpm for 5 hours to mix the two evenly, freeze-dry, and place the freeze-dried powder in a constant temperature oven at 100° C. for 48 hours to obtain a pretreatment complex; S2. Add acetylated sea cucumber polysaccharide and sea cucumber polypeptide into deionized water according to the mass fraction to obtain an anti-fatigue complex solution; stir the pretreated complex prepared in step S1, sodium alginate, taurine and juice stock solution at 500 rpm for 10 min in an ice bath to obtain a functional composite juice. Example

[0017] A functional compound juice for energy supplementation, comprising the following components in parts by weight: 5 parts of plant-derived functional instant powder, 9 parts of anti-fatigue compound, 2 parts of antioxidant extract, 8 parts of sodium alginate, 0.2 parts of taurine, 20 parts of juice stock solution, and 100 parts of deionized water; The plant-derived functional instant powder comprises the following components in parts by weight: 9 parts of purple alfalfa instant powder and 4 parts of American ginseng instant powder; The anti-fatigue extract comprises the following components in parts by weight: 12 parts of acetylated sea cucumber polysaccharide and 6 parts of sea cucumber polypeptide; The juice stock solution includes the following components in parts by weight: 12 parts of freshly squeezed orange stock solution, 3 parts of passion fruit juice stock solution and 2 parts of lemon juice stock solution; The preparation method of the plant-derived functional instant powder is the same as that of Example 1; The preparation method of the anti-fatigue extract specifically comprises the following steps: ① Remove the lime from the mouth of the fresh sea cucumber, cut open the abdomen to remove the internal organs and mud, cut off the inner tendons, first use 25 ℃ water ultrasonic cleaning twice, each time for 12 minutes, ultrasonic power of 150W, ultrasonic frequency of 15kHz, then place in deionized water at 90 ℃, soak for 15 minutes, vacuum dry, vacuum degree of 90Pa, temperature of 60 ℃, time for 4 hours, crush through 80 mesh sieve to obtain sea cucumber powder; ② Soak 18g of the sea cucumber powder obtained in step ① with 385mL of water, treat it in a constant temperature water bath at 45°C for 4h, add 0.22g of papain and 0.11g of trypsin for enzymolysis, the enzymolysis pH is 8.5, the temperature is 35°C, the time is 4h, and the enzyme is inactivated in a boiling water bath to obtain a sea cucumber enzymolysis solution, add 0.4g of ammonium sulfate precipitation, centrifuge at 5500rpm for 25min, and collect the supernatant and precipitate respectively; ③ Add an equal volume of 10% trichloroacetic acid solution to the supernatant obtained in step ②, let stand at 4°C for 18h, centrifuge at 5500rpm for 15min, take the supernatant, add 3 times the volume of anhydrous ethanol to the supernatant, let stand at 4°C for 18h, centrifuge at 2500rpm for 25min, take the precipitate, wash it 3 times with distilled water, and freeze-dry it to obtain sea cucumber polysaccharide; ④ Dissolve 11g of the sea cucumber polysaccharide obtained in step ③ in 550mL of deionized water, perform ultrasonic treatment for 25min, the ultrasonic power is 550W, the ultrasonic frequency is 15kHz, adjust the pH to 9.0, add 36.6mL of acetic anhydride dropwise, stir and react at 50°C for 2.5h, keep the pH stable at 8.2, then adjust the pH to 7.0, dialyze the product with distilled water in a dialysis bag with a molecular weight cutoff of 3500Da for 48h, concentrate the dialysate under reduced pressure at 55°C, add anhydrous ethanol, precipitate overnight at 4°C, and freeze-dry to obtain acetylated sea cucumber polysaccharide; ⑤ Dissolve 7 g of the precipitate obtained in step ② in water, filter it using an inorganic nanofiltration ceramic membrane with a molecular weight cutoff of 800 Da, concentrate the filtrate, and freeze-dry it to obtain a sea cucumber polypeptide.

[0018] The preparation method of the antioxidant extract is the same as that of Example 1; This embodiment provides a method for preparing a functional composite juice for energy supplementation, which specifically comprises the following steps: S1. Dissolve the plant-derived functional instant powder and the antioxidant extract in phosphate buffer respectively, stir at 7000 rpm for 5 h to mix the two evenly, freeze-dry, and place the freeze-dried powder in a constant temperature oven at 100° C. for 24 h to obtain a pretreatment complex; S2. Add acetylated sea cucumber polysaccharide and sea cucumber polypeptide into deionized water according to the mass fraction to obtain an anti-fatigue complex solution; stir the pretreated complex prepared in step S1, sodium alginate, taurine and juice stock solution at 700 rpm for 10 min under ice bath conditions to obtain a functional composite juice. Example

[0019] A functional compound juice for energy supplementation, comprising the following components in parts by weight: 6 parts of plant-derived functional instant powder, 10 parts of anti-fatigue compound, 1.8 parts of antioxidant extract, 6 parts of sodium alginate, 0.3 parts of taurine, 25 parts of juice stock solution, and 90 parts of deionized water; The plant-derived functional instant powder comprises the following components in parts by weight: 80 parts of purple alfalfa instant powder and 4.5 parts of American ginseng instant powder; The anti-fatigue extract comprises the following components in parts by weight: 10 parts of acetylated sea cucumber polysaccharide and 7 parts of sea cucumber polypeptide; The juice stock solution includes the following components in parts by weight: 11 parts of freshly squeezed orange stock solution, 3 parts of passion fruit juice stock solution and 1 part of lemon juice stock solution; The preparation method of the plant-derived functional instant powder is the same as that of Example 1; The preparation method of the anti-fatigue extract specifically comprises the following steps: The preparation method of the modified sea cucumber extract specifically comprises the following steps: ① Remove the lime from the mouth of the fresh sea cucumber, cut open the abdomen to remove the internal organs and mud, cut off the inner tendons, first use clean water at a temperature of 30°C for ultrasonic cleaning 3 times, each time for 15 minutes, the ultrasonic power is 200W, the ultrasonic frequency is 20kHz, then place it in deionized water at a temperature of 100°C, soak for 20 minutes, vacuum dry, the vacuum degree is 100Pa, the temperature is 65°C, the time is 5 hours, and crush it through an 80-mesh sieve to obtain sea cucumber powder; ② Soak 20g of the sea cucumber powder obtained in step ① with 480mL of water, treat it in a constant temperature water bath at 50°C for 5h, add 0.24g of papain and 0.12g of trypsin for enzymolysis, the enzymolysis pH is 9.0, the temperature is 40°C, the time is 5h, and the enzyme is inactivated in a boiling water bath to obtain a sea cucumber enzymolysis solution, add 0.5g of ammonium sulfate precipitation, centrifuge at 6000rpm for 30min, and collect the supernatant and precipitate respectively; ③ Add an equal volume of 10% trichloroacetic acid solution to the supernatant obtained in step ②, let stand at 4°C for 24 hours, centrifuge at 6000 rpm for 20 minutes, take the supernatant, add 3 times the volume of anhydrous ethanol to the supernatant, let stand at 4°C for 24 hours, centrifuge at 3000 rpm for 30 minutes, take the precipitate, wash it with distilled water 3 times, and freeze-dry it to obtain sea cucumber polysaccharide; ④ Dissolve 12g of the sea cucumber polysaccharide obtained in step ③ in 600mL of deionized water, perform ultrasonic treatment for 30min, the ultrasonic power is 600W, the ultrasonic frequency is 20kHz, adjust the pH to 10.0, add 40mL of acetic anhydride dropwise, stir and react at 55°C for 3h, keep the pH stable at 8.5, then adjust the pH to 7.0, dialyze the product with distilled water in a dialysis bag with a molecular weight cutoff of 3500Da for 48h, concentrate the dialysate under reduced pressure at 60°C, add anhydrous ethanol, precipitate overnight at 4°C, and freeze-dry to obtain acetylated sea cucumber polysaccharide; ⑤ Redissolve 8 g of the precipitate obtained in step ② in water, filter it using an inorganic nanofiltration ceramic membrane with a molecular weight cutoff of 800 Da, concentrate the filtrate, and freeze-dry it to obtain a sea cucumber polypeptide.

[0020] The preparation method of the antioxidant extract is the same as that of Example 1; This embodiment provides a method for preparing a functional composite juice for energy supplementation, which specifically comprises the following steps: S1. Dissolve the plant-derived functional instant powder and the antioxidant extract in phosphate buffer respectively, stir at 6000 rpm for 7 hours to mix the two evenly, freeze-dry, and place the freeze-dried powder in a constant temperature oven at 100° C. for 36 hours to obtain a pretreatment complex; S2. Add acetylated sea cucumber polysaccharide and sea cucumber polypeptide into deionized water according to the mass fraction to obtain an anti-fatigue complex solution; stir the pretreated complex prepared in step S1, sodium alginate, taurine and juice stock solution at 600 rpm for 10 min under ice bath conditions to obtain a functional composite juice.

[0021] Comparative Example 1 This comparative example provides a functional compound juice for supplementing energy and a preparation method thereof. The difference between the comparative example and Example 1 is that the compound juice does not include plant-derived functional instant powder, and the remaining components, component contents, and preparation methods are the same as those in Example 1.

[0022] Comparative Example 2 This comparative example provides a functional composite juice for supplementing energy and a preparation method thereof, which is different from Example 1 only in that the anti-fatigue composite does not include acetylated sea cucumber polysaccharide, and the remaining components, component contents, and preparation methods are the same as those in Example 1.

[0023] Comparative Example 3 This comparative example provides a functional composite juice for supplementing energy and a preparation method thereof, which is different from Example 1 only in that the anti-fatigue composite does not include sea cucumber polypeptides, and the remaining components, component contents, and preparation methods are the same as those in Example 1.

[0024] Experimental Example 1 In this experimental example, the composite juice prepared in Example 1-3 and Comparative Example 1-3 was subjected to an in vitro simulated gastrointestinal digestion experiment. 50 mL of the composite juice of Example 1-3 and Comparative Example 1-3 was added with a 5 mol / L HCl solution, the pH was adjusted to 2.0, pepsin was added at a mass concentration of 10 g / L, and the enzyme activity of pepsin was 3000 U / g. The mixture was placed in a constant temperature shaker at a set temperature of 37° C. Samples were taken at different time points, the sample solutions were placed in a water bath at 95° C. for enzyme inactivation, and the concentrations of various substances in the sample solutions were determined; Figure 1 The reducing sugar content of the composite juices prepared in Examples 1-3 and Comparative Examples 1-3 of the present invention in simulated gastric juice is shown in the figure. As shown in the figure, the reducing sugar content of the composite juices prepared in Examples 1-3 and Comparative Examples 1-3 in simulated gastric juice has a trend of first increasing and then decreasing. The reducing sugar converted from the composite juice prepared in Examples 1-3 in simulated gastric juice is significantly higher than that of Comparative Examples 1-3. At the same time, the decreasing rate of the reducing sugar content is slow. The molecular structure of reducing sugar is simple, and no complicated digestion process is required. It can be directly absorbed into the blood through the small intestine, and can quickly replenish energy.

[0025] Experimental Example 2 In this experimental example, the antioxidant properties of the composite juices prepared in Example 13 and Comparative Examples 1-3 were measured. 2 mL of the composite juices prepared in Example 1-3 and Comparative Examples 1-3 were taken, and fresh DPPH ethanol solution (0.1 mM) was added. After mixing evenly, the mixture was reacted in the dark for 30 minutes, and the absorbance was measured at 517 nm. The DPPH scavenging rate was calculated according to the following formula: DPPH free radical scavenging rate = (A0-A1) / A0×100%; Where A0 is the absorbance of DPPH radical solution; A1 is the absorbance of the solution after adding juice; The ABTS solution and K2S2O8 solution were mixed evenly, and then allowed to stand at room temperature in the dark for 12 hours to obtain the ABTS free radical stock solution. Then, 0.2 mL of the compound juice and 0.8 mL of the diluted ABTS stock solution (A734nm=0.70±0.02) were mixed evenly, and the mixture was reacted for 6 minutes in the dark, and the absorbance at 734 nm was measured. The scavenging rate of ABTS free radicals was calculated using the following formula: The scavenging rate of ABTS free radicals = (A0–A1) / A0×100%, where A0 is the absorbance of the ABTS free radical solution; A1 is the absorbance of the solution after adding the sample.

[0026] Figure 2 The antioxidant performance result diagram of the composite juice prepared by Examples 1-3 of the present invention and Comparative Examples 1-3; as shown in the figure, the free clearance rate of the composite juice prepared by Examples 1-3 is above 50%, and Example 1 lacks the plant-derived functional instant powder, and the content of flavonoids, polyphenols and other substances is greatly reduced, resulting in a decrease in antioxidant performance.

[0027] Experimental Example 3 In this experimental example, the anti-fatigue performance of the composite juice prepared in Examples 1-3 and Comparative Examples 1-3 was measured; 140 ICR mice weighing 20-25g were selected, and the light / dark time was maintained at 12h in a greenhouse at 25±2°C and a relative humidity of 45-50%. The mice were adaptively raised in separate cages for 7 days, and the initial body weight was weighed. The mice were randomly divided into 14 groups according to their body weight, with 10 mice in each group, half of which were male and half were female. The composite juice of Examples 1-3 of the present invention was used in the Example group at a dosage of 10g / Kg and 20g / Kg. The control group was gavaged with the compound juice of the comparative examples 1-3 of the present invention at a dose of 10 g / Kg and 20 g / Kg, and the blank group was gavaged with physiological saline at a dose of 10 g / kg and 20 g / kg, and the above gavages were performed once a day, from 9:00 to 11:00 a.m. every day, for 30 consecutive days; 30 minutes after the last gavage, a lead wire of 5% of the body weight was loaded on the tail root of each group of mice, and the mice were placed in a 30 cm deep water tank at a temperature of 25±2°C, and the time from the mice swimming to death or fainting was observed and recorded. The measurement results are as follows: Figure 3 As shown in the figure, the weighted swimming time of the mice in the Example 1-3 groups in the water was significantly longer than that of the mice in the Comparative Example 1-3 groups and the blank group. It can be seen that the energy supplement functional compound juice prepared in the Example 1-3 groups of the present invention is beneficial to improving the body's anti-fatigue degree. While the embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that many changes, modifications, substitutions and variations can be made to the embodiments without departing from the principles and spirit of the invention.

[0028] The present invention and its implementation methods are described above, which is not restrictive. The drawings are only one of the implementation methods of the present invention, and the actual application is not limited thereto. In short, if ordinary technicians in the field are inspired by it and design methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

Claims

1. A functional compound juice for supplementing energy, characterized by: The functional compound juice comprises the following components in parts by weight: 5-6 parts of plant-derived functional instant powder, 8-10 parts of anti-fatigue complex, 1.5-2 parts of antioxidant extract, 5-8 parts of sodium alginate, 0.2-0.4 parts of taurine, 20-30 parts of juice concentrate, and 80-100 parts of deionized water; the plant-derived functional instant powder comprises the following components in parts by weight: 8-10 parts of alfalfa instant powder and 4-5 parts of American ginseng instant powder.

2. The energy-supplementing functional compound juice according to claim 1, characterized in that: The anti-fatigue extract comprises the following components in parts by weight: 10-12 parts of acetylated sea cucumber polysaccharide and 6-8 parts of sea cucumber polypeptide; the anti-fatigue extract is prepared by a polysaccharide-polypeptide co-production method.

3. The energy-supplementing functional compound juice according to claim 2, characterized in that: The juice stock solution comprises the following components in parts by weight: 10-12 parts of freshly squeezed orange stock solution, 3-4 parts of passion fruit juice stock solution and 1-2 parts of lemon juice stock solution.

4. A method for preparing the energy-supplementing functional composite juice according to any one of claims 1 to 3, characterized in that: The specific steps include: S1. Dissolve the plant-derived functional instant powder and the antioxidant extract in phosphate buffer respectively, stir at 5000-6000 rpm for 5-7 hours to mix the two evenly, freeze-dry, and place the freeze-dried powder in a constant temperature oven at 100° C. for 24-48 hours to obtain a pretreatment complex; S2. Add acetylated sea cucumber polysaccharide and sea cucumber polypeptide into deionized water according to the mass fraction to obtain an anti-fatigue complex solution; stir the pretreated complex prepared in step S1, sodium alginate, taurine and juice stock solution at a speed of 500-700 rpm for 10 minutes under ice bath conditions to obtain a functional composite juice.

5. The method for preparing the energy-supplementing functional composite juice according to claim 4, characterized in that: The method for preparing the plant-derived functional instant powder specifically comprises the following steps: (1) Wash the plant rhizome tissue and cut it into 5-10 mm stem segments. After longitudinal sectioning, place it in a proliferation medium containing indoleacetic acid and kinetin. Culture it in the dark at 22-28°C for 21-30 days until the cambium shows obvious proliferation of explants. (2) Isolate explant cells and transfer them to an expansion medium containing indoleacetic acid and 6-benzylaminoadenine, and culture them at 28-32°C in a light-dark alternating environment for 21-42 days, with a light period of 8-12 hours and a dark period of 12-16 hours; subculture once every two weeks to obtain plant tissue; (3) The plant tissue obtained in step (2) is freeze-dried at -40°C, ground into powder in liquid nitrogen, and stored frozen to obtain a plant-derived functional instant powder.

6. The method for preparing the energy-supplementing functional composite juice according to claim 5, characterized in that: In step (1), the proliferation culture medium is based on MS culture medium, wherein the concentrations of indoleacetic acid and kinetin are 0.5-1 mg / L and 1-2 mg / L respectively; the plant stems include at least one of alfalfa stems and American ginseng roots; in step (2), the expansion culture medium is based on MS culture medium, wherein the concentrations of indoleacetic acid and 6-benzylaminoadenine are 1-2 mg / L and 1-1.5 mg / L respectively.

7. The method for preparing the energy-supplementing functional composite juice according to claim 6, characterized in that: The preparation method of the anti-fatigue extract specifically comprises the following steps: ① Wash the fresh sea cucumber, place it in deionized water at a temperature of 80-100°C, soak it for 10-20 minutes, vacuum dry it, and crush it through an 80-mesh sieve to obtain sea cucumber powder; ② Soak the sea cucumber powder obtained in step ① in water according to a mass ratio of 1:30-40, raise the temperature to 40-50° C. and treat in a constant temperature water bath for 3-5 hours, add a composite enzyme for enzymolysis, inactivate the enzyme in a boiling water bath, obtain a sea cucumber enzymolysis solution, add ammonium sulfate precipitation, centrifuge at a speed of 5000-6000 rpm for 20-30 minutes, and collect the supernatant and precipitate respectively; ③ Add an equal volume of 10% trichloroacetic acid solution to the supernatant obtained in step ②, let stand at 4°C for 12-24h, centrifuge at 5000-6000rpm for 10-20min, take the supernatant, add anhydrous ethanol for precipitation, centrifuge at 2000-3000rpm for 20-30min, take the precipitate, wash it with distilled water 3 times, and freeze-dry it to obtain sea cucumber polysaccharide; ④ The sea cucumber polysaccharide obtained in step ③ is dissolved in deionized water, and ultrasonic treatment is performed for 20-30 minutes, the ultrasonic power is 500-600W, the ultrasonic frequency is 10-20kHz, the pH is adjusted to 8.0-10.0, acetic anhydride is added dropwise according to a solid-liquid ratio of 1:15, and the reaction is stirred at 45-55°C for 2-3 hours, during which the pH is kept stable at 8.0-8.5, and then the pH is adjusted to 7.

0. The product is dialyzed with distilled water in a dialysis bag with a molecular weight cutoff of 3500Da for 48 hours, and the dialyzate is concentrated under reduced pressure at 50-60°C, anhydrous ethanol is added, and alcohol precipitation is carried out at 4°C overnight, and freeze-dried to obtain acetylated sea cucumber polysaccharide; ⑤ The precipitate obtained in step ② is redissolved in water, filtered using an inorganic nanofiltration ceramic membrane with a molecular weight cutoff of 800Da, and the filtrate is concentrated and freeze-dried to obtain sea cucumber polypeptides.

8. The method for preparing the energy-supplementing functional composite juice according to claim 7, characterized in that: In step ②, the complex enzyme is a complex enzyme system of papain and trypsin, the added amount of papain is 2-3% of the mass of the sea cucumber powder, and the added amount of trypsin is 1-1.2% of the mass of the sea cucumber powder; the enzymatic hydrolysis conditions are pH 8.0-9.0, temperature 30-40°C, and time 3-5h; the added amount of ammonium sulfate precipitate in the enzymatic hydrolysis solution is 1-1.2 mg / mL; in step ④, the mass concentration of the sea cucumber polysaccharide in deionized water is 20 mg / mL.

9. The method for preparing the energy-supplementing functional composite juice according to claim 8, characterized in that: The preparation method of the antioxidant extract specifically comprises the following steps: The dried cyanobacteria biomass powder is added to a Tris-HCl buffer with a pH of 7.0-7.5 and a concentration of 50 mmol / L, and stirred thoroughly to dissolve it. A hydrolase is added, and enzymolysis is performed at 10-20°C for 6-10 hours. The enzyme is inactivated in a boiling water bath, cooled to room temperature, and centrifuged at 8000-12000 rpm for 30-60 minutes. The supernatant is taken, a saturated ammonium sulfate solution is added, and the mixture is stirred at 50-100 rpm for 1 hour. The mixture is cooled to 4°C and centrifuged at 9000-10000 rpm for 8-10 minutes. The supernatant is collected, ultrafiltered with a 100 kDa hollow fiber membrane, and passed through a 0.22 μm filter membrane to obtain an antioxidant extract.

10. The method for preparing the energy-supplementing functional compound juice according to claim 9, characterized in that: The cyanobacteria biomass comprises one of spirulina, kudzu vine or ground fungus; the mass volume ratio of the cyanobacteria biomass powder to the Tris-HCl buffer is 1-2 g:100 mL; the hydrolase is a composite enzyme system of bromelain and papain, the added amount of bromelain is 1-2% of the mass of the cyanobacteria biomass powder, and the added amount of papain is 0.8-1% of the mass of the cyanobacteria biomass powder; the volume ratio of the saturated ammonium sulfate solution to the supernatant is 1-2:10.

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