Red ginseng nutrient solution containing wood frog enzymolysis peptide and preparation method of red ginseng nutrient solution

Abstract

The invention belongs to the field of health-care drinks, and particularly relates to a red ginseng nutrient solution containing wood frog enzymatic hydrolysis peptide and a preparation method of the red ginseng nutrient solution. Raw materials of the wood frog enzymatic hydrolysis peptide beverage comprise the following components in percentage by mass: 10%-15% of wood frog enzymatic hydrolysis peptide, 2%-5% of a red ginseng extract, 2%-5% of an agaric extract, 3%-5% of soybean peptide, 0.2%-0.5% of erythritol, 0.1%-0.3% of a pH regulator, 0.05%-0.1% of a preservative, 0.1%-0.15% of hydroxypropyl-beta-cyclodextrin, 0.1%-0.15% of sulfobutyl-beta-cyclodextrin and the balance of water. The wood frog enzymolysis peptide, the red ginseng extract, the agaric extract, the soybean peptide and other main components are rich in various bioactive substances, the stability of the nutrient solution is improved under the synergistic effect of hydroxypropyl-beta-cyclodextrin and sulfobutyl-beta-cyclodextrin, and the free radical scavenging efficiency of the nutrient solution is improved.

Description

Technical Field

[0001] The present invention belongs to the field of health care beverages, and particularly relates to a red ginseng nutrient solution containing Rana chensinensis enzymolysis peptides and a preparation method thereof. Background Art

[0002] Oviductus Ranae (OR) is the dried product of the oviduct of female Rana temporaria chensinesis David, also known as Rana chensinensis oil. Its economic value mainly comes from the dried oviduct, and it is a traditional Chinese medicine with a history of application in China for hundreds of years. It was probably first recorded in Compendium of Materia Medica in the Ming Dynasty and was included in the Chinese Pharmacopoeia in 1985. Oviductus Ranae has medicinal effects such as benefiting the stomach and spleen, tonifying the kidney and essence, nourishing yin and tonifying the kidney, moistening the lungs and promoting fluid production. Clinically, it is used to treat symptoms such as weakness due to yin deficiency, fatigue, cough with hemoptysis, etc. Rana temporaria chensinesis mainly grows in Northeast China, and Changbai Mountain in Jilin Province and its surrounding areas are known as the "Hometown of Rana temporaria chensinesis in China". In recent years, with the improvement of people's requirements for the quality of food and medicine, more and more people have begun to favor natural medicines to achieve health care and treatment goals, and Oviductus Ranae, as a natural animal traditional Chinese medicine, has also received extensive attention.

[0003] Oviductus Ranae contains various active ingredients such as proteins, sterols, fatty acids, and hormones, among which the protein content is the highest. As early as in the 1930s, Japanese scholars measured its protein content to be 56.3%, and the protein contents of different types of Oviductus Ranae also vary. Oviductus Ranae is not only rich in proteins, but also contains various nutrients required for human growth and development, and has pharmacological effects such as enhancing the body's resistance, delaying aging, relieving fatigue, and antioxidant effects. As a new type of health care product, it has been widely welcomed and trusted by people.

[0004] Red Ginseng is the dried root and stem of Panax ginseng C. A. Meyer after high-temperature steaming and processing. Compared with ordinary ginseng, the tissue structure of Red Ginseng is more compact, which makes it more convenient for long-term storage. The main active ingredients of Red Ginseng are saponins and polysaccharides. Auricularia auricula is a fungal plant with both medicinal and edible values. In China, artificial cultivation began in the Sui and Tang dynasties, and it was listed as a medicinal plant. It has extremely high nutritional and medicinal values and is praised by nutritionists as "the meat among vegetables" and "the king among vegetables". The polysaccharides in Red Ginseng and Auricularia auricula have effects such as reducing blood lipid, anti-thrombosis, anti-coagulation, promoting immunity, and anti-tumor.

[0005] However, there are few studies on preparing a nutrient solution by combining Rana temporaria chensinesis, Red Ginseng, and Auricularia auricula at present. Summary of the Invention

[0006] To solve the problems existing in the prior art, the present invention provides a red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate and a preparation method thereof.

[0007] The present invention is achieved by the following technical solutions: A red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate, the raw materials of which are composed of the following components by mass percentage: 10% - 15% of Rana temporaria enzyme hydrolysate, 2% - 5% of red ginseng extract, 2% - 5% of Auricularia auricula extract, 3% - 5% of soybean peptide, 0.2% - 0.5% of erythritol, 0.1% - 0.3% of pH regulator, 0.05% - 0.1% of preservative, 0.1% - 0.15% of hydroxypropyl-β-cyclodextrin and 0.1% - 0.15% of sulfobutyl-β-cyclodextrin, and the balance is water.

[0008] Further, the raw materials of which are composed of the following components by mass percentage: 10% of Rana temporaria enzyme hydrolysate, 2% of red ginseng extract, 2% of Auricularia auricula extract, 3% of soybean peptide, 0.2% of erythritol, 0.1% of pH regulator, 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.

[0009] Further, the preparation method of the Rana temporaria enzyme hydrolysate is as follows: Take 10.00 g of Rana temporaria oil, dissolve it in 800 mL of deionized water, adjust the pH of the solution to 1.5 with 0.1 M HCl, add 0.08 g of pepsin, then carry out constant-temperature enzymatic hydrolysis 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 carry out constant-temperature enzymatic hydrolysis 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 vacuum freeze-dry to obtain.

[0010] Further, the preparation method of the red ginseng extract is as follows: After the red ginseng is pulverized and passed through an 80-mesh sieve, accurately weigh 10.0 g of red ginseng powder, add it to 400 mL of 70% (v / v) ethanol water, add 0.1 g of cellulase and 0.3 g of pectinase, carry out constant-temperature enzymatic hydrolysis at 50°C for 4 h, then inactivate the enzyme in a water bath at 100°C for 10 min, centrifuge to obtain the supernatant, and finally concentrate and dry to obtain the red ginseng extract.

[0011] Further, the preparation method of the Auricularia auricula extract is as follows: After the Auricularia auricula is pulverized and passed through an 80-mesh sieve, accurately weigh 10.0 g of Auricularia auricula powder, add it to 400 mL of 70% (v / v) ethanol water, add 0.1 g of cellulase and 0.3 g of pectinase, carry out constant-temperature enzymatic hydrolysis at 50°C for 4 h, then inactivate the enzyme in a water bath at 100°C for 10 min, centrifuge to obtain the supernatant, and finally concentrate and dry to obtain the Auricularia auricula extract.

[0012] Further, the preservative is selected from ethyl p - hydroxybenzoate. The pH regulator is selected from citric acid.

[0013] Further, the mass percentages of hydroxypropyl - β - cyclodextrin and sulfobutyl - β - cyclodextrin are both 0.15%.

[0014] The present invention also provides a preparation method of red ginseng nutrient solution containing Rana temporaria enzyme - hydrolyzed peptides, comprising the following steps: Add Rana temporaria enzyme - hydrolyzed peptides, red ginseng extract, auricularia auricula extract, soybean peptides, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p - hydroxybenzoate, hydroxypropyl - β - cyclodextrin, and sulfobutyl - β - cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature to obtain the red ginseng nutrient solution containing Rana temporaria enzyme - hydrolyzed peptides. The conditions for high - temperature sterilization are sterilization at 120 °C for 20 min.

[0015] Compared with the prior art, the present invention has the following advantages and beneficial effects: In the present invention, the main components such as Rana temporaria enzyme - hydrolyzed peptides, red ginseng extract, auricularia auricula extract, and soybean peptides 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 its free - radical scavenging efficiency is enhanced. Description of the Drawings

[0016] The drawings described herein are used to provide a further understanding of the embodiments of the present invention, form a part of this application, and do not constitute a limitation to the embodiments of the present invention.

[0017] Figure 1 It is for the DPPH free - radical scavenging activity test. Detailed Embodiments

[0018] To make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention will be further described in detail below in conjunction with the embodiments and the drawings. The illustrative embodiments and descriptions of the present invention are only used to explain the present invention and do not constitute a limitation to the present invention.

[0019] In the present invention, hydroxypropyl - β - cyclodextrin, CAS: 128446 - 35 - 5; sulfobutyl - β - cyclodextrin, CAS: 182410 - 00 - 0.

[0020] The preparation method of Rana temporaria enzyme - hydrolyzed peptides is as follows: Take 10.00 g of Rana temporaria oil, dissolve it in 800 mL of deionized water, and adjust the pH of the solution to 1.5, add 0.08 g of pepsin, then enzymatically hydrolyze at a constant temperature of 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 enzymatically hydrolyze at a constant temperature of 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 vacuum freeze-dry to obtain.

[0021] Preparation method of red ginseng extract: After red ginseng is crushed, it is sieved through a 80-mesh sieve. Accurately weigh 10.0 g of red ginseng powder, add it to 400 mL of 70% (v / v) ethanol water, then add 0.1 g of cellulase and 0.3 g of pectinase, enzymatically hydrolyze at a constant temperature of 50 °C for 4 h, then inactivate the enzyme in a water bath at 100 °C for 10 min, centrifuge (4000 r / min, 10 min) to obtain the supernatant, and finally concentrate and dry to obtain red ginseng extract.

[0022] The preparation method of auricularia auricula extract is similar to that of red ginseng. Specifically: After auricularia auricula is crushed, it is sieved through a 80-mesh sieve. Accurately weigh 10.0 g of auricularia auricula powder, add it to 400 mL of 70% (v / v) ethanol water, then add 0.1 g of cellulase and 0.3 g of pectinase, enzymatically hydrolyze at a constant temperature of 50 °C for 4 h, then inactivate the enzyme in a water bath at 100 °C for 10 min, centrifuge (4000 r / min, 10 min) to obtain the supernatant, and finally concentrate and dry to obtain auricularia auricula extract.

[0023] Example 1 A red ginseng nutrient solution containing Rana temporaria enzymatic hydrolysate, the raw materials of which are composed of the following components by mass percentage: 10% Rana temporaria enzymatic hydrolysate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.15% hydroxypropyl-β-cyclodextrin and 0.15% sulfobutyl-β-cyclodextrin, and the balance is water.

[0024] Preparation method: Add Rana temporaria enzymatic hydrolysate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymatic hydrolysate.

[0025] Example 2 A red ginseng nutrient solution containing Rana temporaria enzymatic hydrolysate, the raw materials of which are composed of the following components by mass percentage: 10% Rana temporaria enzymatic hydrolysate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.05% hydroxypropyl-β-cyclodextrin and 0.05% sulfobutyl-β-cyclodextrin, and the balance is water.

[0026] Preparation method: Add Rana temporaria enzyme hydrolysate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate.

[0027] Example 3 A red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate, and its raw materials are composed of the following components in mass percentage: 10% Rana temporaria enzyme hydrolysate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin, and 0.2% sulfobutyl-β-cyclodextrin, and the balance is water.

[0028] Preparation method: Add Rana temporaria enzyme hydrolysate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate.

[0029] Example 4 A red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate, and its raw materials are composed of the following components in mass percentage: 10% Rana temporaria enzyme hydrolysate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.2% hydroxypropyl-β-cyclodextrin, and 0.2% sulfobutyl-β-cyclodextrin, and the balance is water.

[0030] Preparation method: Add Rana temporaria enzyme hydrolysate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzyme hydrolysate.

[0031] Example 5 A red ginseng nutrient solution containing Rana temporaria enzyme hydrolyzate, the raw materials of which are composed of the following components by mass percentage: 10% Rana temporaria enzyme hydrolyzate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.2% hydroxypropyl-β-cyclodextrin, and 0.1% sulfobutyl-β-cyclodextrin, with the balance being water.

[0032] Preparation method: Add Rana temporaria enzyme hydrolyzate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzyme hydrolyzate.

[0033] Example 6 A red ginseng nutrient solution containing Rana temporaria enzyme hydrolyzate, the raw materials of which are composed of the following components by mass percentage: 10% Rana temporaria enzyme hydrolyzate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin, and 0.2% sulfobutyl-β-cyclodextrin, with the balance being water.

[0034] Preparation method: Add Rana temporaria enzyme hydrolyzate, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid to pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, hydroxypropyl-β-cyclodextrin, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization conditions are sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzyme hydrolyzate.

[0035] Comparative Example 1 A red ginseng nutrient solution containing Rana temporaria enzyme hydrolyzate, the raw materials of which are composed of the following components by mass percentage: 10% Rana temporaria enzyme hydrolyzate, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.1% hydroxypropyl-β-cyclodextrin, with the balance being water.

[0036] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and hydroxypropyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at a high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0037] Comparative Example 2 A red ginseng nutrient solution containing Rana temporaria enzymolysis peptide, and its raw materials are composed of the following components in mass percentage: 10% Rana temporaria enzymolysis peptide, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.15% hydroxypropyl-β-cyclodextrin, and the balance is water.

[0038] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and hydroxypropyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at a high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0039] Comparative Example 3 A red ginseng nutrient solution containing Rana temporaria enzymolysis peptide, and its raw materials are composed of the following components in mass percentage: 10% Rana temporaria enzymolysis peptide, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.3% hydroxypropyl-β-cyclodextrin, and the balance is water.

[0040] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and hydroxypropyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, and then cool and fill; sterilize the filled nutrient solution at a high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0041] Comparative Example 4 A red ginseng nutrient solution containing Rana temporaria enzymolysis peptide, and its raw materials are composed of the following components in mass percentage: 10% Rana temporaria enzymolysis peptide, 2% red ginseng extract, 2% auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.1% sulfobutyl-β-cyclodextrin, and the balance is water.

[0042] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, Auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0043] Comparative Example 5 A red ginseng nutrient solution containing Rana temporaria enzymolysis peptide, and its raw materials are composed of the following components by mass percentage: 10% Rana temporaria enzymolysis peptide, 2% red ginseng extract, 2% Auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.15% sulfobutyl-β-cyclodextrin, and the balance is water.

[0044] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, Auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0045] Comparative Example 6 A red ginseng nutrient solution containing Rana temporaria enzymolysis peptide, and its raw materials are composed of the following components by mass percentage: 10% Rana temporaria enzymolysis peptide, 2% red ginseng extract, 2% Auricularia auricula extract, 3% soybean peptide, 0.2% erythritol, 0.1% citric acid, 0.05% ethyl p-hydroxybenzoate, 0.15% sulfobutyl-β-cyclodextrin, and the balance is water.

[0046] Preparation method: Add Rana temporaria enzymolysis peptide, red ginseng extract, Auricularia auricula extract, soybean peptide, and citric acid into pure water, and stir the mixed solution at 50 °C for 2 h; then add erythritol, citric acid, ethyl p-hydroxybenzoate, and sulfobutyl-β-cyclodextrin and continue to stir at 50 °C for 2 h, then cool and fill; sterilize the filled nutrient solution at high temperature, and the sterilization condition is sterilization at 120 °C for 20 min to obtain a red ginseng nutrient solution containing Rana temporaria enzymolysis peptide.

[0047] Test Example 1 For aerobic bacteria detection, after storing the red ginseng nutrient solution containing Rana temporaria chensinensis enzymolysis peptides prepared in each example and comparative example for different times, dilute it 10 times, take 200 μL and inoculate it onto the tryptic soy agar medium. Use physiological saline as the negative control, culture at 30 - 35 °C for 24 h, observe and record the number of colonies, repeat the experiment in three groups, and calculate the number of aerobic bacteria.

[0048] Table 1 Aerobic bacteria detection of Example 1.

[0049]

[0050] The red ginseng nutrient solutions containing Rana temporaria chensinensis enzymolysis peptides prepared in Examples 2 - 6 and Comparative Examples 1 - 6 were tested under the same conditions as in Example 1, and no aerobic bacteria were detected either.

[0051] Test Example 2 For mold and yeast detection, after storing the red ginseng nutrient solution containing Rana temporaria chensinensis enzymolysis peptides prepared in each example and comparative example for different times, dilute it 10 times, take 200 μL and inoculate it onto the sabouraud dextrose agar medium. Use physiological saline as the negative control, culture at 20 - 25 °C for 24 h, observe and record the number of colonies, repeat the experiment in three groups, and calculate the number of molds and yeasts.

[0052] Table 2 Number of molds and yeasts in Example 1.

[0053]

[0054] The red ginseng nutrient solutions containing Rana temporaria chensinensis enzymolysis peptides prepared in Examples 2 - 6 and Comparative Examples 1 - 6 were tested under the same conditions as in Example 1, and no molds and yeasts were detected either.

[0055] Test Example 3 Escherichia coli detection. Take 1 mL of the nutrient solution prepared in the examples and comparative examples and inoculate it into 30 mL of tryptic soy broth, culture in a shaker at 30 - 35 °C for 24 h. Take 1 mL of the above preculture and inoculate it into 100 mL of MacConkey broth, culture in a shaker at 42 - 44 °C for 24 h. Take 200 μL of the MacConkey broth culture and inoculate it onto the MacConkey agar medium plate, culture at 30 - 35 °C for 72 h, and set up three replicates.

[0056] No Escherichia coli was detected in each example and comparative example at the 0th month, 1st month, 2nd month, and 3rd month.

[0057] Test Example 4 Place three nutrient solutions of the same example or comparative example in an incubator at a constant temperature of 60 °C at the same time. Take two nutrient solutions at the same time on the 5th day and 10th day to measure the polysaccharide retention rate. The phenol-sulfuric acid method is used to measure the polysaccharide content. The method for measuring the polysaccharide retention rate is as follows: Take the solutions before and after the test, measure the polysaccharide content by the phenol-sulfuric acid method, and calculate the polysaccharide retention rate. The calculation formula is as follows: Polysaccharide retention rate (%) = polysaccharide content after the test (g) / polysaccharide content before the test (g) × 100% Table 3 High-temperature polysaccharide retention rate. (82.5%)

[0058] Test Example 5 DPPH free radical scavenging activity: Dissolve DPPH in 95% ethanol solution to make the final concentration of the DPPH solution reach 0.2 mmol / L (7.8 mg / 100 mL), and store it in the dark at low temperature. Take 1 mL of each example and comparative example and put it into a test tube, and sequentially add 2 mL of DPPH solution and 2 mL of 95% ethanol solution, mix well, react in the dark for 30 min, and measure the absorbance A2 at 517 nm with a UV spectrophotometer. At the same time, measure the absorbance value A0 of the DPPH solution without adding the sample solution (2 mL DPPH + 3 mL 95% ethanol) and the absorbance value A1 of the blank solution without adding the DPPH solution (1 mL sample + 4 mL 95% ethanol). Calculation formula: E(%) = 1 - (A2 - A1) / A0 × 100. Each example and comparative example are measured 3 times, and the test results are as Figure 1 shown.

[0059] The specific embodiments described above further elaborate on the purpose, technical solutions, and beneficial effects of the present invention. It should be understood that the above description is only the specific embodiments of the present invention and is not used to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A red ginseng nutrient solution containing Rana sylvaticum hydrolyzed peptides, characterized in that: The raw materials are composed of the following ingredients 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 sylvaticum hydrolyzed peptides according to claim 1, characterized in that: The raw materials are composed of the following ingredients 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 sylvaticum hydrolyzed peptides according to claim 2, characterized in that: The preparation method of the wood frog enzymatic peptide is as follows: Take 10.00g of Rana affine 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 constant temperature 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 constant temperature enzymolysis at 37°C for 5h; then inactivate the enzyme in a boiling water bath for 10min, freeze at -80°C for 12h, and obtain the product by vacuum freeze-drying.

4. The red ginseng nutrient solution containing Rana sylvaticum hydrolyzed peptides according to claim 2, characterized in that: The preparation method of the red ginseng extract is: 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. Then, 0.1 g of cellulase and 0.3 g of pectinase are added. The mixture is enzymolyzed at a constant temperature of 50°C for 4 h, and 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 red ginseng extract.

5. The red ginseng nutrient solution containing Rana sylvaticum hydrolyzed peptides 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 enzymolyzed at a constant temperature of 50 ° C for 4 h, and 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 sylvaticum hydrolyzed peptides according to claim 2, characterized in that: The preservative is selected from ethyl paraben.

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

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

9. The method for preparing the red ginseng nutrient solution containing Rana sylvaticum hydrolyzed peptides according to any one of claims 1 to 8, characterized in that: The following steps are involved: Add frog enzymatic peptides, red ginseng extract, wood ear extract, soybean peptides and citric acid into pure water, and stir the mixed solution at 50 degrees Celsius for 2 hours; then add erythritol, citric acid, ethyl paraben, 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 to obtain red ginseng nutrient solution containing frog enzymatic peptides.

10. The method for preparing the red ginseng nutrient solution containing Rana sylvaticum hydrolyzed 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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