A red ginseng nutrient solution containing ranae enzyme hydrolyzed peptide and a preparation method thereof

Abstract

The present application belongs to the field of health-care drinks, and particularly relates to a red ginseng nutrient solution containing forest frog enzymatic peptides and a preparation method. The raw materials are composed of the following components in mass percentage: forest frog enzymatic peptides 10-15%, red ginseng extract 2-5%, agaric extract 2-5%, soybean peptides 3-5%, erythritol 0.2-0.5%, pH regulator 0.1-0.3%, preservative 0.05-0.1%, hydroxypropyl-beta-cyclodextrin 0.1-0.15% and sulfobutyl-beta-cyclodextrin 0.1-0.15%, and the balance is water. The forest frog enzymatic peptides, red ginseng extract, agaric extract and soybean peptides and other main components used in the present application are rich in various bioactive substances, and the stability of the nutrient solution is improved under the synergistic action of hydroxypropyl-beta-cyclodextrin and sulfobutyl-beta-cyclodextrin, and the free radical scavenging efficiency is improved.

Description

Technical Field

[0001] The invention belongs to the field of health drinks, and in particular relates to a red ginseng nutrient solution containing rana enzymatic peptides and a preparation method thereof. Background Art

[0002] Toad oil (Oviductus Ranae, OR) is a dried product from the oviduct of the female Chinese wood frog (Rana temporaria chensinesis David), also known as wood frog oil. Its economic value primarily stems from the dried oviduct, and it is a traditional Chinese medicine with a centuries-old history in China. It may have been first recorded in the Ming Dynasty's Compendium of Materia Medica and was included in the Chinese Pharmacopoeia in 1985. Wood frog oil boasts medicinal benefits such as nourishing the stomach and spleen, tonifying the kidneys and replenishing essence, nourishing yin and kidneys, and moistening the lungs and promoting fluid production. Clinically, it is used to treat symptoms such as yin deficiency, fatigue, and coughing up blood. Wood frogs primarily grow in Northeast China, with Changbai Mountain and its surrounding areas in Jilin Province known as the "Homeland of the Wood Frog." In recent years, with rising demands for food and drug quality, more and more people are turning to natural remedies for health and therapeutic purposes. As a natural animal-based traditional Chinese medicine, wood frog oil has garnered widespread attention.

[0003] Rana oil contains a variety of active ingredients, including protein, sterols, fatty acids, and hormones, with protein being the highest. As early as the 1930s, Japanese researchers measured its protein content at 56.3%, and protein content varies among different types of Rana oil. Rana oil is not only rich in protein but also contains a variety of nutrients essential for human growth and development. It has pharmacological effects such as boosting immunity, delaying aging, relieving fatigue, and providing antioxidant benefits. As a new health product, it has gained widespread popularity and trust.

[0004] Red ginseng is the dried root and stem of Panax ginseng (C.A. Meyer) that has been steamed at high temperatures. Compared to regular ginseng, red ginseng has a denser tissue structure, making it easier to preserve for long periods of time. The main active ingredients in red ginseng are saponins and polysaccharides. Wood ear mushrooms are a fungus with both medicinal and edible properties. They were cultivated in my country during the Sui and Tang dynasties and listed as a medicinal plant. They possess extremely high nutritional and medicinal value, being hailed by nutritionists as the "vegetarian equivalent" and the "king of vegetarian foods." The polysaccharides in red ginseng and wood ear mushrooms have been shown to lower blood lipids, fight thrombosis, promote blood coagulation, boost immunity, and fight tumors.

[0005] However, there are few studies on preparing nutrient solution from Rana sylvatica, red ginseng and Auricularia auricula together. Summary of the Invention

[0006] In order to solve the problems existing in the prior art, the present invention provides a red ginseng nutrient solution containing rana enzymatic peptides and a preparation method thereof.

[0007] The present invention is achieved through the following technical solutions:

[0008] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10%-15% frog enzymatic peptides, 2%-5% red ginseng extract, 2%-5% wood ear extract, 3%-5% soybean peptide, 0.2%-0.5% erythritol, 0.1%-0.3% pH regulator, 0.05%-0.1% preservative, 0.1%-0.15% hydroxypropyl-β-cyclodextrin and 0.1%-0.15% sulfobutyl-β-cyclodextrin, and the balance is water.

[0009] Furthermore, its raw materials are composed of the following ingredients in the following mass percentages: 10% of wood frog enzymatic peptide, 2% of red ginseng extract, 2% of wood ear extract, 3% of soybean peptide, 0.2% of erythritol, 0.1% of pH adjuster, 0.05% of preservative, 0.1%~0.15% of hydroxypropyl-β-cyclodextrin and 0.1%~0.15% of sulfobutyl-β-cyclodextrin, and the balance is water.

[0010] Furthermore, the preparation method of the wood frog enzymatic peptide is:

[0011] Take 10.00g of Rana sylvatica oil, dissolve it in 800mL of deionized water, adjust the pH of the solution to 1.5 with 0.1M HCl, add 0.08g of pepsin, and then perform enzymolysis at 37°C for 2h. Then, adjust the pH to 8.0 with 0.1M NaOH solution, add 0.16g of trypsin, and then perform enzymolysis at 37°C for 5h. Then, inactivate the enzyme in a boiling water bath for 10min, freeze at -80°C for 12h, and freeze-dry in a vacuum.

[0012] Furthermore, the preparation method of the red ginseng extract is:

[0013] After red ginseng was crushed, it was passed through an 80-mesh sieve. 10.0 g of red ginseng powder was accurately weighed and added to 400 mL of 70% (v / v) ethanol water. 0.1 g of cellulase and 0.3 g of pectinase were then added. The mixture was enzymatically hydrolyzed at 50°C for 4 h, then inactivated in a water bath at 100°C for 10 min. The supernatant was centrifuged and finally concentrated and dried to obtain the red ginseng extract.

[0014] Furthermore, the preparation method of the fungus extract is:

[0015] After the fungus is crushed, it is passed through an 80-mesh sieve. 10.0 g of fungus powder is accurately weighed and added to 400 mL of 70% (v / v) ethanol water. Then, 0.1 g of cellulase and 0.3 g of pectinase are added. The mixture is enzymatically hydrolyzed at a constant temperature of 50°C for 4 h, then the enzyme is inactivated in a water bath at 100°C for 10 min. The supernatant is obtained by centrifugation and finally concentrated and dried to obtain the fungus extract.

[0016] Furthermore, the preservative is selected from ethyl p-hydroxybenzoate. The pH regulator is selected from citric acid.

[0017] Furthermore, the mass percentages of hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin are both 0.15%.

[0018] The present invention also provides a method for preparing a red ginseng nutrient solution containing Rana sylvatica enzymatic peptides, comprising the following steps:

[0019] Rana sylvaticum enzymatic peptides, red ginseng extract, wood ear mushroom extract, soybean peptides, and citric acid were added to purified water and stirred at 50°C for 2 hours. Erythritol, citric acid, ethyl paraben, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin were then added and stirred at 50°C for another 2 hours. The mixture was then cooled and bottled. The bottled nutrient solution was then sterilized at high temperature to obtain a red ginseng nutrient solution containing Rana sylvaticum enzymatic peptides. The sterilization conditions were 120°C for 20 minutes.

[0020] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0021] The present invention uses the main ingredients of wood frog enzymatic peptide, red ginseng extract, wood ear extract and soybean peptide, which are rich in various bioactive substances. Under the synergistic effect of hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin, the stability of the nutrient solution is improved and the free radical scavenging efficiency thereof is improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The drawings described herein are used to provide a further understanding of the embodiments of the present invention, constitute a part of this application, and do not constitute a limitation of the embodiments of the present invention.

[0023] Figure 1 This is a test of DPPH free radical scavenging activity. DETAILED DESCRIPTION

[0024] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with examples and drawings. The exemplary embodiments of the present invention and their descriptions are only used to explain the present invention and are not intended to limit the present invention.

[0025] The hydroxypropyl-β-cyclodextrin in the present invention has a CAS number of 128446-35-5 and a sulfobutyl-β-cyclodextrin has a CAS number of 182410-00-0.

[0026] The preparation method of the enzymatic peptide of Rana sylvatica is as follows:

[0027] Take 10.00g of Rana sinensis oil and dissolve it in 800mL of deionized water. Adjust the pH of the solution to

[0028] 1.5, add 0.08 g of pepsin, and then perform enzymolysis at 37°C for 2 h, then adjust the pH to 8.0 with 0.1 M NaOH solution, add 0.16 g of trypsin, and then perform enzymolysis at 37°C for 5 h; then inactivate the enzyme in a boiling water bath for 10 min, freeze at -80°C for 12 h, and freeze-dry in a vacuum.

[0029] Preparation method of red ginseng extract: After red ginseng is crushed, it is passed through an 80-mesh sieve, 10.0 g of red ginseng powder is accurately weighed and added to 400 mL of 70% (v / v) ethanol water, followed by 0.1 g of cellulase and 0.3 g of pectinase. The mixture is enzymatically hydrolyzed at 50°C for 4 hours, then inactivated in a water bath at 100°C for 10 minutes, and the supernatant is obtained by centrifugation (4000 r / min, 10 minutes). Finally, the supernatant is concentrated and dried to obtain the red ginseng extract.

[0030] The preparation method of wood ear extract is similar to that of red ginseng, specifically: after the wood ear is crushed, it is passed through an 80-mesh sieve, 10.0 g of wood ear powder is accurately weighed, added to 400 mL of 70% (v / v) ethanol water, and then 0.1 g of cellulase and 0.3 g of pectinase are added. The mixture is enzymatically hydrolyzed at a constant temperature of 50°C for 4 hours, and then the enzyme is inactivated in a water bath at 100°C for 10 minutes. The supernatant is obtained by centrifugation (4000 r / min, 10 minutes), and finally concentrated and dried to obtain the wood ear extract.

[0031] Example 1

[0032] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.15% hydroxypropyl-β-cyclodextrin and 0.15% sulfobutyl-β-cyclodextrin, and the balance is water.

[0033] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0034] Example 2

[0035] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.05% hydroxypropyl-β-cyclodextrin and 0.05% sulfobutyl-β-cyclodextrin, and the balance is water.

[0036] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0037] Example 3

[0038] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin and 0.2% sulfobutyl-β-cyclodextrin, and the balance is water.

[0039] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0040] Example 4

[0041] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.2% hydroxypropyl-β-cyclodextrin and 0.2% sulfobutyl-β-cyclodextrin, and the balance is water.

[0042] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0043] Example 5

[0044] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.2% hydroxypropyl-β-cyclodextrin and 0.1% sulfobutyl-β-cyclodextrin, and the balance is water.

[0045] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0046] Example 6

[0047] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin and 0.2% sulfobutyl-β-cyclodextrin, and the balance is water.

[0048] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0049] Comparative Example 1

[0050] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following ingredients in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin, and the balance being water.

[0051] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and hydroxypropyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0052] Comparative Example 2

[0053] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.15% hydroxypropyl-β-cyclodextrin, and the balance being water.

[0054] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and hydroxypropyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0055] Comparative Example 3

[0056] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.3% hydroxypropyl-β-cyclodextrin, and the balance being water.

[0057] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and hydroxypropyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0058] Comparative Example 4

[0059] The invention discloses a red ginseng nutrient solution containing frog enzymatic peptides. The raw materials of the red ginseng nutrient solution are composed of the following components in percentage by mass: 10% frog enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.1% sulfobutyl-β-cyclodextrin, and the balance being water.

[0060] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0061] Comparative Example 5

[0062] A red ginseng nutrient solution containing rana enzymatic peptides is prepared from the following ingredients in percentage by mass: 10% rana enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.15% sulfobutyl-β-cyclodextrin, and the balance being water.

[0063] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0064] Comparative Example 6

[0065] A red ginseng nutrient solution containing rana enzymatic peptides is prepared from the following ingredients in percentage by mass: 10% rana enzymatic peptides, 2% red ginseng extract, 2% wood ear extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl parahydroxybenzoate, 0.15% sulfobutyl-β-cyclodextrin, and the balance being water.

[0066] Preparation method: add frog enzymatic peptide, red ginseng extract, wood ear extract, soybean peptide and citric acid to pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate and sulfobutyl-β-cyclodextrin and continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 degrees Celsius for 20 minutes to obtain red ginseng nutrient solution containing frog enzymatic peptide.

[0067] Test Example 1

[0068] For aerobic bacteria testing, the red ginseng nutrient solutions containing rana hydrolyzed peptides prepared in each example and comparative example were stored for different periods of time, diluted 10-fold, and 200 μL was inoculated onto trypticase soy agar. A physiological saline solution was used as a negative control. The culture was incubated at 30–35°C for 24 hours, and the number of colonies was observed and recorded. This experiment was repeated three times, and the number of aerobic bacteria was calculated.

[0069] Table 1 Aerobic bacteria detection of Example 1.

[0070]

[0071] The red ginseng nutrient solutions containing Rana sylvatic peptides prepared in Examples 2 to 6 and Comparative Examples 1 to 6 were tested under the same conditions as in Example 1, and no aerobic bacteria were detected.

[0072] Test Example 2

[0073] For mold and yeast testing, the red ginseng nutrient solution containing rana hydrolyzed peptides prepared in each example and comparative example was stored for different periods of time, diluted 10-fold, and 200 μL was inoculated onto Sabouraud dextrose agar. A physiological saline solution was used as a negative control. The culture was incubated at 20-25°C for 24 hours, and the number of colonies was observed and recorded. This experiment was repeated three times, and the number of mold and yeast was calculated.

[0074] Table 2 The number of molds and yeasts in Example 1.

[0075]

[0076] The red ginseng nutrient solutions containing Rana sylvatic peptides prepared in Examples 2 to 6 and Comparative Examples 1 to 6 were tested under the same conditions as in Example 1, and no mold or yeast was detected.

[0077] Test Example 3

[0078] Escherichia coli detection. Inoculate 1 mL of the nutrient solution prepared in the Examples and Comparative Examples into 30 mL of trypticase soy broth and culture at 30-35°C on a shaker for 24 h. Inoculate 1 mL of the above preculture into 100 mL of MacConkey broth and culture at 42-44°C on a shaker for 24 h. Inoculate 200 μL of the MacConkey broth onto a MacConkey agar plate and culture at 30-35°C for 72 h. Set up three replicates.

[0079] No Escherichia coli was detected in the samples prepared in the Examples and Comparative Examples at the 0th, 1st, 2nd and 3rd months.

[0080] Test Example 4

[0081] Three nutrient solutions from the same example or comparative example were placed simultaneously in a constant temperature incubator at 60°C. Two nutrient solutions were taken at the same time on days 5 and 10 to measure the polysaccharide retention rate. The polysaccharide content was measured using the phenol-concentrated sulfuric acid method. The polysaccharide retention rate was determined as follows: the polysaccharide content was determined using the phenol-concentrated sulfuric acid method in the solutions before and after the test. The polysaccharide retention rate was calculated using the following formula:

[0082] Polysaccharide retention rate (%) = polysaccharide content after the test (g) / polysaccharide content before the test (g) × 100%

[0083] Table 3 High temperature polysaccharide retention rate. (82.5%)

[0084]

[0085] Test Example 5

[0086] DPPH free radical scavenging activity, take DPPH and dissolve it in 95% ethanol solution to make the final concentration of DPPH solution reach 0.2mmol / L (7.8mg / 100mL), and store it at low temperature and away from light. Take 1 mL of each embodiment and comparative example and put it into a test tube, add 2mL DPPH solution and 2mL 95% ethanol solution in turn, mix thoroughly, react in the dark for 30 minutes, and use an ultraviolet spectrophotometer to measure the absorbance A2 at 517nm. At the same time, measure the absorbance value A0 of the DPPH solution when no sample solution is added (2 mL DPPH + 3 mL95% ethanol) and the absorbance value A1 of the blank solution when no DPPH solution is added (1 mL sample + 4 mL 95% ethanol). Calculation formula: E(%) = 1-(A2-A1) / A0×100. Each embodiment and comparative example is measured 3 times. The test results are as follows: Figure 1 shown.

[0087] The specific implementation methods described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A red ginseng nutrient solution containing Rana sylvatica enzymatic peptides, characterized in that: The raw materials are composed of the following components in percentage by mass: 10% to 15% of wood frog enzymatic peptide, 2% to 5% of red ginseng extract, 2% to 5% of wood ear extract, 3% to 5% of soybean peptide, 0.2% to 0.5% of erythritol, 0.1% to 0.3% of pH adjuster, 0.05% to 0.1% of preservative, 0.1% to 0.15% of hydroxypropyl-β-cyclodextrin and 0.1% to 0.15% of sulfobutyl-β-cyclodextrin, and the balance is water.

2. The red ginseng nutrient solution containing rana enzymatic peptides according to claim 1, characterized in that: The raw materials are composed of the following components in percentage by mass: 10% of wood frog enzymatic peptide, 2% of red ginseng extract, 2% of wood ear extract, 3% of soybean peptide, 0.2% of erythritol, 0.1% of pH adjuster, 0.05% of preservative, 0.1%-0.15% of hydroxypropyl-β-cyclodextrin and 0.1%-0.15% of sulfobutyl-β-cyclodextrin, and the balance is water.

3. The red ginseng nutrient solution containing rana enzymatic peptide according to claim 2, characterized in that: The preparation method of the wood frog enzymatic peptide is as follows: Take 10.00g of Rana sylvatica oil, dissolve it in 800mL of deionized water, adjust the pH of the solution to 1.5 with 0.1M HCl, add 0.08g of pepsin, and then perform enzymolysis at 37°C for 2h. Then, adjust the pH to 8.0 with 0.1M NaOH solution, add 0.16g of trypsin, and then perform enzymolysis at 37°C for 5h. Then, inactivate the enzyme in a boiling water bath for 10min, freeze at -80°C for 12h, and freeze-dry in a vacuum.

4. The red ginseng nutrient solution containing rana enzymatic peptides according to claim 2, characterized in that: The preparation method of the red ginseng extract is: After red ginseng was crushed, it was passed through an 80-mesh sieve. 10.0 g of red ginseng powder was accurately weighed and added to 400 mL of 70% (v / v) ethanol water. 0.1 g of cellulase and 0.3 g of pectinase were then added. The mixture was enzymatically hydrolyzed at 50°C for 4 h, then inactivated in a water bath at 100°C for 10 min. The supernatant was centrifuged and finally concentrated and dried to obtain the red ginseng extract.

5. The red ginseng nutrient solution containing rana enzymatic peptide according to claim 2, characterized in that: The preparation method of the fungus extract is: After the fungus is crushed, it is passed through an 80-mesh sieve. 10.0 g of fungus powder is accurately weighed and added to 400 mL of 70% (v / v) ethanol water. Then, 0.1 g of cellulase and 0.3 g of pectinase are added. The mixture is enzymatically hydrolyzed at a constant temperature of 50°C for 4 h, then the enzyme is inactivated in a water bath at 100°C for 10 min. The supernatant is obtained by centrifugation and finally concentrated and dried to obtain the fungus extract.

6. The red ginseng nutrient solution containing rana enzymatic peptides according to claim 2, characterized in that: The preservative is selected from ethyl paraben.

7. The red ginseng nutrient solution containing rana enzymatic peptides according to claim 2, characterized in that: The pH adjuster is selected from citric acid.

8. The red ginseng nutrient solution containing rana enzymatic peptides according to claim 2, characterized in that: The mass percentages of the hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin are both 0.15%.

9. The method for preparing a red ginseng nutrient solution containing Rana sylvatic peptides according to any one of claims 1 to 8, characterized in that: The following steps are involved: Add the enzymatic hydrolyzed peptides of Chinese frog, red ginseng extract, wood ear extract, soybean peptide and citric acid into pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl parahydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin, continue stirring at 50 degrees Celsius for 2 hours, and then cool and fill; sterilize the filled nutrient solution at high temperature to obtain the red ginseng nutrient solution containing the enzymatic hydrolyzed peptides of Chinese frog.

10. The method for preparing the red ginseng nutrient solution containing Rana sylvatica enzymatic peptides according to claim 9, characterized in that: The high temperature sterilization condition is sterilization at 120 degrees Celsius for 20 minutes.

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