Composition of malus toringoides leaf extract, beta-nicotinamide mononucleotide and goose carnosine and application of composition

Abstract

The invention discloses a composition of malus toringoides leaf extract, beta-nicotinamide mononucleotide and anserine and application of the composition. The composition of the malus toringoides leaf extract, the beta-nicotinamide mononucleotide and the goose carnosine is prepared from a plant extract, the beta-nicotinamide mononucleotide and the goose carnosine. The invention also provides a preparation method of the composition. Compared with the prior art, the composition of the malus toringoides leaf extract, the beta-nicotinamide mononucleotide and the goose carnosine prepared by the invention has the advantages of alcoholism-relieving and liver-protecting effects, high storage stability and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceuticals, and in particular to a composition of extract of Rhododendron niveum leaves, β-nicotinamide mononucleotide, and carnosine and its uses. Background Art

[0002] Wine has a long history in China and has formed a wine culture. Traditional Chinese medicine has long recognized wine. Li Shizhen pointed out in "Compendium of Materia Medica" that "wine is the spiritual blessing of heaven. Drinking a little can harmonize the blood, promote qi flow, strengthen the spirit, resist cold, and dispel sorrow. Drinking excessively will damage the spirit, consume blood, harm the stomach, deplete essence, produce phlegm, and stir up fire", believing that moderate drinking is beneficial to health, but excessive drinking will cause damage to the viscera, can also lead to alcohol poisoning, and even death. Acute alcohol poisoning, also known as drunkenness, refers to the damage process of the central nervous system, digestive system, respiratory system, cardiovascular and cerebrovascular systems, etc. caused by excessive drinking at one time.

[0003] CN111632081A discloses a compound blueberry andhovenia tablet with the effect of relieving hangover and protecting the liver and its preparation method. The traditional Chinese medicine composition includes pueraria root extract, flos puerariae extract, hovenia dulcis extract, and blueberry anthocyanin. The mass ratio of pueraria root extract: flos puerariae extract: hovenia dulcis extract: blueberry anthocyanin is (1 - 3):(1 - 2.2):(1 - 1.5):(0.8 - 5), which has the effects of relieving symptoms such as thirst, headache, nausea, and vomiting after drinking, achieving the effects of sobering up and relieving a hangover. However, the properties of the extract are unstable, and the composition may have a low stability phenomenon. CN119258173A discloses a traditional Chinese medicine composition for relieving hangover and protecting the liver and its preparation method, which is composed of 6 - 24 parts of pueraria root, 4 - 16 parts of flos puerariae, 5 - 20 parts of hovenia dulcis, 3 - 15 parts of hawthorn, 3 - 15 parts of smoked plum, and 5 - 20 parts of imperata cylindrica root, and has the effect of relieving hangover and protecting the liver. However, the extract in this composition may be unstable, and the active ingredients may be destroyed by gastric acid after oral administration. Summary of the Invention

[0004] In view of the above - mentioned defects of the prior art, the purpose of the present invention is to provide a composition of extract of Rhododendron niveum leaves, β - nicotinamide mononucleotide, and carnosine, which has the advantages of relieving hangover and protecting the liver, high storage stability, etc.

[0005] In order to achieve the above - mentioned invention purpose, the present invention adopts the following technical scheme:

[0006] A composition of extract of Prunus mongolica Maxim. leaves, β-nicotinamide mononucleotide, and carnosine, which is prepared by mixing extract of Prunus mongolica Maxim. leaves, β-nicotinamide mononucleotide, and carnosine according to a mass ratio of 1-5:1-3:1-3; or, by mixing composite microcapsules of extract of Prunus mongolica Maxim. leaves, β-nicotinamide mononucleotide, and carnosine according to a mass ratio of 1-5:1-3:1-3. "Prunus mongolica Maxim. leaves" is a commonly used medicinal material in folk, which is derived from the dried leaves and leaf buds of Malus toringoides (Rehd.) Hughes or Malus transitoria (Batal.) Schneid. of the Rosaceae family. This medicine is recorded in "Four Medical Canons", "Jingzhu Materia Medica", and "Tibetan Medicine Crystal Mirror Materia Medica", and has the effects of attacking and resolving accumulation, removing greasiness and stagnation, protecting the liver and promoting gallbladder function. Clinically, it is used to treat indigestion, hypertension, hyperglycemia, hyperlipidemia, and liver diseases.

[0007] β-Nicotinamide mononucleotide (NMN), NMN is a nucleotide that naturally exists in all forms of life. It is a biosynthetic precursor of nicotinamide adenine dinucleotide (NAD+). Inside cells, NMN is converted into NAD+ through specific enzymatic reactions (such as NMN adenylyltransferase). It has the effects of anti-aging and promoting DNA repair.

[0008] Carnosine is a dipeptide composed of β-alanine and 1-methylhistidine. It mainly exists in the muscle tissues of animals, etc. It has the effects of anti-fatigue and reducing uric acid.

[0009] Preferably, the preparation method of the extract of Prunus mongolica Maxim. leaves includes the following steps: drying Prunus mongolica Maxim. leaves, crushing them, adding an ethanol aqueous solution with a mass of 15-25 times that of Prunus mongolica Maxim. leaves and a concentration of 70-100wt% for heating under reflux extraction for 2-4 times, filtering, collecting the filtrate, recovering under reduced pressure to obtain an ethanol extract; dissolving the ethanol extract in water to obtain mixture 1, then adding petroleum ether with a volume of 2-4 times that of mixture 1 for extraction 3-5 times, separating the liquid; then adding ethyl acetate with a volume of 2-4 times that of the aqueous phase for repeated extraction 3-5 times, combining the ethyl acetate solutions, and recovering under reduced pressure until there is no ethyl acetate left, thus obtaining the extract of Prunus mongolica Maxim. leaves.

[0010] Preferably, the preparation method of the composite microcapsules of extract of Prunus mongolica Maxim. leaves includes the following steps, calculated by mass parts:

[0011] Step 1: Drying Prunus mongolica Maxim. leaves, crushing them, adding an ethanol aqueous solution with a mass of 15-25 times that of Prunus mongolica Maxim. leaves and a concentration of 70-100wt% for heating under reflux extraction for 2-4 times, filtering, collecting the filtrate, recovering under reduced pressure to obtain an ethanol extract; dissolving the ethanol extract in water to obtain mixture 1, then adding petroleum ether with a volume of 2-4 times that of mixture 1 for extraction 3-5 times, separating the liquid; taking the aqueous phase, and then adding ethyl acetate with a volume of 2-4 times that of the aqueous phase for repeated extraction 3-5 times, combining the ethyl acetate solutions, and recovering under reduced pressure until there is no ethyl acetate left, thus obtaining the extract of Prunus mongolica Maxim. leaves;

[0012] Step 2: Add 1 - 10 parts of animal protein to 500 - 700 parts of water, and stir and homogenize at 2000 - 4000 rpm for 1 - 3 h at room temperature; then add 1 - 5 parts of the extract of Prunus potaninii Batal. leaves and continue to stir for 1 - 3 h to obtain the animal protein - Prunus potaninii Batal. leaf extract composite liquid;

[0013] Step 3: Mix 1 - 5 parts of locust bean gum, 0.1 - 1 part of propylene glycol fatty acid ester, and 40 - 80 parts of water, and disperse at 10000 - 15000 rpm for 3 - 5 min. Repeat the dispersion step 2 - 4 times to obtain a uniform and stable mixed liquid a;

[0014] Step 4: Mix the animal protein - Prunus potaninii Batal. leaf extract composite liquid obtained in Step 2 and the mixed liquid a obtained in Step 3 evenly according to a mass ratio of 1 - 5:1 - 5, and then perform homogenization at a temperature of 58 - 62°C and a pressure of 28 - 32 MPa to obtain a mixed material liquid; then perform spray freeze - drying to obtain the Prunus potaninii Batal. leaf extract composite microcapsules.

[0015] Preferably, the Prunus potaninii Batal. leaves are selected from at least one of the leaf buds of Malus toringoides (Rehd.) Hughes, the leaves of Malus toringoides (Rehd.) Hughes, the leaf buds of Malus transitoria (Batal.) Schneid., and the leaves of Malus transitoria (Batal.) Schneid. Preferably, the animal protein is selected from at least one of squid skin protein, collagen, and whey protein.

[0016] Preferably, the preparation method of the squid skin protein comprises the following steps: Crush the squid skin and add water 3 - 10 times the mass of the squid skin to make a homogenate; adjust the pH value of the homogenate to 8 - 10, add 1 - 5 wt% of alkaline protease based on the mass of the squid skin, and enzymolyze at 35 - 45°C for 30 - 60 min; raise the temperature to inactivate the enzyme, centrifuge and filter, and take the supernatant to obtain the squid skin enzymatic hydrolysate; then adjust the pH value of the squid skin enzymatic hydrolysate to 3 - 5, extract by acid precipitation, discard the supernatant and take the precipitate, wash it to neutral, and freeze - dry to obtain the squid skin protein.

[0017] As a further illustration of the present invention, squid skin protein, collagen, and whey protein in animal proteins have good film-forming properties and emulsifying properties, and can form a thin film on the surface of an object to protect the core material; locust bean gum is a plant galactomannan extracted from the endosperm of carob tree seeds, and has emulsifying and film-forming properties; propylene glycol fatty acid ester is an emulsifier that can reduce the interfacial tension; animal proteins can form a thin film on the surface of an object through the interaction between protein molecules to protect the extract of Russula virescens. After mixing with the emulsion formed by locust bean gum, locust bean gum molecules will aggregate around the emulsion droplets through intermolecular interactions (such as hydrogen bonds, etc.), and they will jointly form a composite wall material on the surface of the emulsion droplets to wrap the emulsion droplets. During the spray freeze-drying process, as the water rapidly evaporates, the concentration of the composite wall material around the emulsion droplets gradually increases; in a low-temperature and low-moisture environment, the intermolecular interaction of the wall material is strengthened, and finally solidifies to form microcapsules; this microcapsule structure can effectively protect the internal active ingredients and at the same time enable the internal ingredients to be released better during reconstitution.

[0018] Advantages of the present invention:

[0019] 1. Compared with the prior art, by reasonably proportioning and utilizing the interaction between various substances and optimizing the preparation process, the present invention obtains a composition of the extract of Russula virescens, β-nicotinamide mononucleotide, and carnosine; the composition of the extract of Russula virescens, β-nicotinamide mononucleotide, and carnosine obtained by the present invention not only has the effects of reducing high blood pressure, high blood sugar, high blood lipid, reducing uric acid, and anti-aging, but also has the advantages of relieving alcohol and protecting the liver, and the composition has high stability.

[0020] 2. Compared with the prior art, the extract of Russula virescens has the effect of relieving alcohol and protecting the liver, and co-using it with β-nicotinamide mononucleotide and carnosine can further improve the effect of relieving alcohol and protecting the liver.

[0021] 3. Compared with the prior art, squid skin protein, collagen, and whey protein in animal proteins have good film-forming properties and emulsifying properties, and can form a thin film on the surface of an object to protect the core material; after mixing with the emulsion formed by locust bean gum, locust bean gum molecules will aggregate around the emulsion droplets through intermolecular interactions (such as hydrogen bonds, etc.), and they will jointly form a composite wall material on the surface of the emulsion droplets to wrap the emulsion droplets. During the spray freeze-drying process, as the water rapidly evaporates, the concentration of the composite wall material around the emulsion droplets gradually increases; in a low-temperature and low-moisture environment, the intermolecular interaction of the wall material is strengthened, and finally solidifies to form microcapsules; this microcapsule structure can protect the extract of Russula virescens and increase the stability. Specific embodiments

[0022] The parameters and sources of some substances in the examples are as follows:

[0023] The leaf buds of Malus transitoria (Batalin) C. K. Schneid., Malus transitoria (Batalin) C. K. Schneid. is a plant of the genus Malus in the Rosaceae family, origin: Gansu;

[0024] Alkaline protease, food grade, product specification: 100,000 - 400,000 U / g, sourced from Nanning Pangbo Bioengineering Co., Ltd.;

[0025] Whey protein, purity: 99%, food grade, sourced from Jiangsu Duoyang Bioengineering Technology Co., Ltd.;

[0026] Collagen, purity: 99%, food grade, sourced from Jiangsu Duoyang Bioengineering Technology Co., Ltd.

[0027] Example 1

[0028] A composition of extract of Acer truncatum Bunge leaves, β - nicotinamide mononucleotide and carnosine, which is composed of 4 g of extract of Acer truncatum Bunge leaves, 3 g of β - nicotinamide mononucleotide and 3 g of carnosine mixed together.

[0029] The preparation method of the extract of Acer truncatum Bunge leaves includes the following steps: drying the leaf buds of Malus transitoria, pulverizing them, adding an aqueous ethanol solution of 75 wt% which is 20 times the mass of the leaf buds of Malus transitoria for heating under reflux extraction for 3 times, filtering, collecting the filtrate, recovering under reduced pressure to obtain a crude product without ethanol; dissolving the crude product without ethanol in water to obtain mixture 1, then adding petroleum ether which is 3 times the volume of mixture 1 for extraction 4 times, separating the liquid; taking the aqueous phase, and then adding ethyl acetate which is 3 times the volume of the aqueous phase for repeated extraction 4 times, combining the ethyl acetate solutions, and recovering under reduced pressure until there is no ethyl acetate left, thus obtaining the extract of Acer truncatum Bunge leaves.

[0030] Example 2

[0031] A composition of extract of Acer truncatum Bunge leaves composite microcapsules, β - nicotinamide mononucleotide and carnosine, which is composed of 4 g of extract of Acer truncatum Bunge leaves composite microcapsules, 3 g of β - nicotinamide mononucleotide and 3 g of carnosine mixed together.

[0032] The preparation method of the extract of Acer truncatum Bunge leaves composite microcapsules includes the following steps:

[0033] Step 1: drying the leaf buds of Malus transitoria, pulverizing them, adding an aqueous ethanol solution of 75 wt% which is 20 times the mass of the leaf buds of Malus transitoria for heating under reflux extraction for 3 times, filtering, collecting the filtrate, recovering under reduced pressure to obtain a crude product without ethanol; dissolving the crude product without ethanol in water to obtain mixture 1, then adding petroleum ether which is 3 times the volume of mixture 1 for extraction 4 times, separating the liquid; taking the aqueous phase, and then adding ethyl acetate which is 3 times the volume of the aqueous phase for repeated extraction 4 times, combining the ethyl acetate solutions, and recovering under reduced pressure until there is no ethyl acetate left, thus obtaining the extract of Acer truncatum Bunge leaves;

[0034] Step 2: Add 6 g of squid skin protein to 600 g of water, and stir and homogenize the mixture at 3000 rpm for 2 h at room temperature; then add 3 g of Russula extract and continue stirring for 2 h to obtain a squid skin protein-Russula extract composite solution.

[0035] Step 3: Mix 3 g of locust bean gum, 0.5 g of propylene glycol fatty acid ester, and 60 g of water, disperse the mixture at 13000 rpm for 4 min, and repeat the dispersion step 3 times to obtain a uniform and stable mixture a.

[0036] Step 4: Mix the squid skin protein-Russula extract composite solution obtained in Step 2 and the mixture a obtained in Step 3 evenly according to a mass ratio of 3:2, then perform homogenization at a temperature of 60 °C and a pressure of 30 MPa to obtain a mixed material liquid; then perform spray freeze-drying to obtain Russula extract composite microcapsules.

[0037] The preparation method of the squid skin protein includes the following steps:

[0038] Crush squid skin and add 7 times the mass of water of squid skin to make a homogenate; adjust the pH value of the homogenate to 9, add 3 wt% of alkaline protease based on the mass of squid skin, and enzymatically hydrolyze at 40 °C for 45 min; heat up to inactivate the enzyme, centrifuge and filter, and take the supernatant to obtain a squid skin enzymatic hydrolysate; then adjust the pH value of the squid skin enzymatic hydrolysate to 4, extract by acid precipitation, discard the supernatant and take the precipitate, wash it with water until neutral, and freeze-dry to obtain squid skin protein.

[0039] Example 3

[0040] A composition of Russula extract, β-nicotinamide mononucleotide, and carnosine, which is composed of 4 g of Russula extract composite microcapsules, 3 g of β-nicotinamide mononucleotide, and 3 g of carnosine.

[0041] The preparation method of the Russula extract composite microcapsules includes the following steps:

[0042] Step 1: Dry and crush the leaf buds of Malus transitoria, add a 75 wt% ethanol aqueous solution 20 times the mass of the leaf buds of Malus transitoria, heat under reflux for extraction for 3 times, filter, collect the filtrate, and recover under reduced pressure to obtain a crude product without ethanol; dissolve the crude product without ethanol in water to obtain a mixed solution 1, then add petroleum ether 3 times the volume of the mixed solution 1 and extract 4 times, separate the liquid; take the aqueous phase, and then add ethyl acetate 3 times the volume of the aqueous phase and extract repeatedly 4 times, combine the ethyl acetate solutions, and recover under reduced pressure until there is no ethyl acetate to obtain Russula extract.

[0043] Step 2: Add 6 g of collagen to 600 g of water, and stir and homogenize the mixture at 3000 rpm for 2 h at room temperature; then add 3 g of Russula extract and continue stirring for 2 h to obtain a squid skin protein-Russula extract composite solution.

[0044] Step 3: Mix 3 g of locust bean gum, 0.5 g of propylene glycol fatty acid ester, and 60 g of water, disperse at 13,000 rpm for 4 min, and repeat the dispersion step 3 times to obtain a uniform and stable mixture a;

[0045] Step 4: Mix the collagen - Russula extract composite solution obtained in Step 2 and the mixture a obtained in Step 3 evenly according to a mass ratio of 3:2, then perform homogenization at a homogenization temperature of 60 °C and a pressure of 30 MPa to obtain a mixed material liquid; then perform spray freeze - drying to obtain the Russula extract composite microcapsules.

[0046] Example 4

[0047] A composition of Russula extract, β - nicotinamide mononucleotide, and carnosine, which is composed of 4 g of Russula extract composite microcapsules, 3 g of β - nicotinamide mononucleotide, and 3 g of carnosine mixed together.

[0048] The preparation method of the Russula extract composite microcapsules includes the following steps:

[0049] Step 1: Dry and crush the leaf buds of Malus transitoria, add 20 times the mass of the leaf buds of Malus transitoria of 75 wt% ethanol aqueous solution for heating under reflux extraction for 3 times, filter, collect the filtrate, and recover under reduced pressure to obtain a crude product without ethanol; dissolve the crude product without ethanol in water to obtain a mixed solution 1, then add petroleum ether 3 times the volume of the mixed solution 1 for extraction 4 times, separate the liquid; take the aqueous phase, and then add ethyl acetate 3 times the volume of the aqueous phase for repeated extraction 4 times, combine the ethyl acetate solutions, and recover under reduced pressure until there is no ethyl acetate to obtain the Russula extract;

[0050] Step 2: Add 6 g of whey protein to 600 g of water, stir and homogenize at 3000 rpm for 2 h at room temperature; then add 3 g of Russula extract and continue to stir for 2 h to obtain a whey protein - Russula extract composite solution;

[0051] Step 3: Mix 3 g of locust bean gum, 0.5 g of propylene glycol fatty acid ester, and 60 g of water, disperse at 13,000 rpm for 4 min, and repeat the dispersion step 3 times to obtain a uniform and stable mixture a;

[0052] Step 4: Mix the whey protein - Russula extract composite solution obtained in Step 2 and the mixture a obtained in Step 3 evenly according to a mass ratio of 3:2, then perform homogenization at a homogenization temperature of 60 °C and a pressure of 30 MPa to obtain a mixed material liquid; then perform spray freeze - drying to obtain the Russula extract composite microcapsules.

[0053] Comparative Example 1

[0054] An anti - hangover and liver - protecting composition, which is composed of 6 g of Russula extract and 4 g of β - nicotinamide mononucleotide mixed together.

[0055] The preparation method of the extract of Phyllanthus emblica L. var. robinsonii (Rehd.) Hand.-Mazz. is the same as that in Example 1.

[0056] Comparative Example 2

[0057] An anti-alcoholism and liver-protecting composition is composed of 6 g of the extract of Phyllanthus emblica L. var. robinsonii (Rehd.) Hand.-Mazz. and 4 g of carnosine, which are mixed together.

[0058] The preparation method of the extract of Phyllanthus emblica L. var. robinsonii (Rehd.) Hand.-Mazz. is the same as that in Example 1.

[0059] Comparative Example 3

[0060] An anti-alcoholism and liver-protecting composition is composed of 5 g of β-nicotinamide mononucleotide and 5 g of carnosine, which are mixed together.

[0061] Test Example 1

[0062] Test on the effects of the administered drugs on serum ALT and AST in mice

[0063] 100 healthy mice with a body weight of 18 - 22 g, half male and half female, were randomly divided into 10 groups, namely Example 1 - 4 groups, Comparative Example 1 - 3, blank control group, model group and the control group of commercially available Haiwang Jinzun tablets (Approval number: Weishi Jianzi (2002) No. 0396). After one week of adaptive feeding, the experiment was carried out.

[0064] In the experiment, in the groups of Example 1 - 4 and Comparative Example 1 - 3, intragastric administration was carried out regularly every day according to the alcohol dose of 0.25 ml / 20 g of 56% ethanol and the drug dose of 0.2 g / 10 g. In the commercially available Haiwang Jinzun control group, intragastric administration was carried out regularly every day according to the alcohol dose of 0.25 ml / 20 g of 56% ethanol and the drug dose of 0.4 g / 10 g. The blank control group and the model control group were intragastrically administered with normal saline. After the administration ended on the 44th day, all animals were fasted for one day. One hour after the administration on the 45th day, blood was taken from the orbital artery and orbital vein of the mice, the serum was separated and stored at -4°C at low temperature for measuring the serum biochemical indexes (ALT, AST); ALT and AST mainly exist in the hepatocyte cytoplasm. When the hepatocytes are damaged, the intracellular transaminases can enter the blood, thus causing an increase in ALT and AST in the blood. The test results are shown in Table 1.

[0065] Table 1 Test on the effects of the administered drugs on serum ALT and AST in mice

[0066]

[0067] Note: When comparing Example 1 - 4 and Comparative Example 1 - 2 with the blank control group, model group and the commercially available Haiwang Jinzun control group, the differences were statistically significant (P < 0.05); when comparing Example 1 and Example 2, the difference was statistically significant (P < 0.05); when comparing Example 2 with Example 3 - 4, the difference was statistically significant (P < 0.05).

[0068] By comparing Examples 1-4, Comparative Examples 1-3, the blank control group, the model group, and the commercially available Haiwang Jinzun control group, it can be found that the anti-alcoholism and liver-protecting effects of Examples 1-4 are better than those of Comparative Examples 1-3. By comparing Examples 1-4, it can be found that the anti-alcoholism and liver-protecting effect of Example 2 is better. The reason may be that although Comparative Examples 1-2 contain extracts of Russula leaves, they lack the synergistic effect of other components. This shows that although the extracts of Russula leaves have certain anti-alcoholism and liver-protecting functions, their effects are limited when used alone. Comparative Example 3 is a composition of β-nicotinamide mononucleotide and carnosine, and it can be found that the anti-alcoholism and liver-protecting effects of these two are weak. Example 1 is a composition of extracts of Russula leaves, β-nicotinamide mononucleotide, and carnosine. The better anti-alcoholism and liver-protecting effect than that of Comparative Examples 1-3 may be because the extracts of Russula leaves act together with β-nicotinamide mononucleotide and carnosine to form a synergistic effect; the flavonoid compounds in the extracts of Russula leaves and β-nicotinamide mononucleotide both have antioxidant capabilities, can scavenge free radicals, reduce the oxidative stress damage caused by alcohol, and protect liver cells; NMN promotes cell energy metabolism and DNA repair by supplementing NAD⁺, enhancing the anti-damage ability of liver cells; carnosine can relieve the fatigue caused by alcohol and reduce the metabolic burden on the liver, thus indirectly protecting the liver. The better anti-alcoholism and liver-protecting effect of Example 2 than that of Example 1 may be because the active ingredients of the extracts of Russula leaves are unstable under acidic conditions, while the composite microcapsules formed by squid skin protein and locust bean gum can protect the extracts of Russula leaves from being structurally damaged under acidic conditions, thus having a better anti-alcoholism and liver-protecting effect. The reason why the anti-alcoholism and liver-protecting effect of Example 2 is better than that of Examples 3-4 may be that the structural stability of the composite microcapsules formed by squid skin protein and locust bean gum in Example 2 is better than that of collagen and whey protein, with better protection effects. Squid skin protein contains rich amino acids and peptide segments, which may be able to scavenge free radicals and reduce the oxidation reaction caused by alcohol.

[0069] Test Example 2

[0070] Anti-alcoholism effect test

[0071] Method of drunkenness experiment: 80 mice (half male and half female) were pre-raised for 3 days and divided into a model group, Examples 1-4, and Comparative Examples 1-3 groups, with 10 mice in each group. Each group of mice was first fasted and water-deprived for 6 hours. The groups of Examples 1-4 and Comparative Examples 1-3 were gavaged according to the dosage of 0.2 g / 10 g, and the model group was gavaged with an equal volume of 9.0 g / L sodium chloride injection. 30 minutes later, each group of mice was gavaged with white liquor at 18 ml / kg, and the time required for the disappearance (drunkenness) to recovery (sobering up) of the righting reflex of the mice was recorded. The experimental results are shown in Table 2.

[0072] Table 2 Mouse drunkenness experiment

[0073]

[0074] Note: Compared with the model group, there were statistically significant differences in Examples 1-4 and Comparative Examples 1-2 (P < 0.05); there were statistically significant differences between Example 1 and Example 2 (P < 0.05); there were statistically significant differences between Example 2 and Examples 3-4 (P < 0.05).

[0075] By comparing Examples 1-4, Comparative Examples 1-3 and the model group, it can be found that the anti-alcohol effects of Examples 1-4 are better than those of Comparative Examples 1-3. By comparing Examples 1-4, it can be found that the anti-alcohol effect of Example 2 is the best. The reason may be that there is a synergistic effect when the extract of Rhododendron hypoglaucum, β-nicotinamide mononucleotide and carnosine are used together in Example 1, which can enhance the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in the liver, improve cell energy metabolism, and enable alcohol to be metabolized more quickly. The reason why Example 2 is better than Examples 1 and 3-4 may be that squid skin protein contains rich amino acids and peptide segments, which may be able to scavenge free radicals and reduce the oxidation reaction caused by alcohol. At the same time, this microcapsule structure can protect the extract of Rhododendron hypoglaucum from being structurally damaged under acidic conditions, and its wall material structure stability is better than that of collagen and whey protein microcapsules in the comparative examples, with better protection effect, enabling the active ingredients to play a better role, so the anti-alcohol effect is better.

[0076] Test Example 3

[0077] Stability test

[0078] Determination of total flavonoid content: It was determined using a UV-2550 double-beam ultraviolet spectrophotometer, Shimadzu Corporation, Japan.

[0079] Method for making the standard curve: Weigh 7.2 g of rutin, dissolve it with methanol and make the volume up to 50 mL to obtain a rutin standard product of 0.144 g / L. Accurately measure 0, 1, 2, 3, 4, 5 mL of the standard product into 25 mL volumetric flasks, add 3 mL of 1% AlCl 3 solution, make the volume up with 75% ethanol, shake well and let it stand for 30 min, and label them as 0#, 1#, 2#, 3#, 4#, 5# respectively. Using 0# as the control, scan the 5# sample between 800 - 190 nm, detect that the maximum absorption peak of rutin is at 412 nm, measure the absorbance values of 1#, 2#, 3#, 4#, 5# samples at 412 nm, and draw a standard curve with the absorbance value as the ordinate and the mass concentration mg / L as the abscissa.

[0080] Preparation method of test solution: Respectively take the samples prepared in Examples 1-4 of the present invention, grind them finely, accurately weigh 1.0 g, place them in a 100 ml stoppered conical flask, ultrasonically extract with 50 mL of methanol (power 120 W, frequency 40 kHz) for 30 min, filter, concentrate the filtrate to dryness, dissolve with methanol and make up the volume to 50 mL, shake well, filter with a 0.45 μm filter membrane, and calculate the content of total flavonoids in the sample by the standard curve method.

[0081] Place the samples of Examples 1-4 under the stability test conditions of temperature 45±2°C and humidity 75%±5%, and detect the retention rate of total flavonoids content at the test cycles of 1, 2, 3, and 6 months. The higher the retention rate, the higher the stability.

[0082] The calculation formula for the retention rate of flavonoid content is as follows:

[0083] Retention rate of flavonoid content % = amount of flavonoids in each experimental cycle ÷ initial amount of flavonoids × 100%

[0084] The experimental results are shown in Table 3.

[0085] Table 3 Stability test

[0086]

[0087] The main flavonoid compounds in E. utilis leaves include quercetin and its glycosides and other components. These flavonoid compounds have multiple phenolic hydroxyl structures and have certain chemical activities. An increase in temperature may promote their oxidation reaction; high humidity may cause hydrolysis reaction.

[0088] Through the comparison of Examples 1-4, it can be found that the retention rate of flavonoid content in Examples 2-4 is better than that in Example 1. By comparing Examples 2-4, it can be found that the retention rate of flavonoid content in Example 2 is the highest, and the retention rate for six months reaches 80.3%. The reason may be that Examples 2-4 have a microcapsule structure, which can protect the flavonoid components in the E. utilis leaf extract. The retention rate of flavonoid content in Example 2 is better than that in Examples 3-4, probably because squid skin protein contains rich specific amino acids that can crosslink with locust bean gum to make the wall material structure of the microcapsule more compact.

Claims

1. A composition of an extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine, characterized in that: The extract of Osmanthus fragrans leaves, β-nicotinamide mononucleotide and anserine are mixed in a mass ratio of 1-5:1-3:1-3; Or, the composite microcapsules of the extract of the Russian leaf, β-nicotinamide mononucleotide and anserine are mixed in a mass ratio of 1-5:1-3:1-3.

2. The composition of the extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine according to claim 1, characterized in that: The preparation method of the extract of Osmanthus fragrans leaves comprises the following steps: The leaves of the Russian Se are dried and crushed, and a 70-100wt% ethanol aqueous solution in an amount of 15-25 times the mass of the Russian Se leaves is added, heated and refluxed for extraction, the extraction is performed 2-4 times, the filtrate is filtered, the filtrate is collected, and the extract is recovered under reduced pressure to obtain an ethanol extract; the extract of the ethanol extract is dissolved in water to obtain a mixed solution 1, and then petroleum ether in an amount of 2-4 times the volume of the mixed solution is added, extracted 3-5 times, and the liquids are separated; the aqueous phase is taken, and then ethyl acetate in an amount of 2-4 times the volume of the aqueous phase is added, and extracted repeatedly 3-5 times, the ethyl acetate solutions are combined, and the reduced pressure is recovered until there is no ethyl acetate, thereby obtaining an Russian Se leaf extract.

3. The composition of the extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine according to claim 1, characterized in that: The preparation method of the composite microcapsules of the extract of the Russian leaf comprises the following steps, calculated by weight: Step 1, drying and crushing the leaves of the Russian color, adding 70-100wt% ethanol aqueous solution in an amount of 15-25 times the mass of the Russian color leaves, heating and refluxing for extraction, extracting 2-4 times, filtering, collecting the filtrate, and recovering under reduced pressure to obtain an ethanol extract; dissolving the ethanol extract in the ethanol part in water to obtain a mixed solution 1, and then adding 2-4 times the volume of petroleum ether of the mixed solution 1 to extract 3-5 times, and separating the liquids; taking the aqueous phase, and then adding 2-4 times the volume of the aqueous phase of ethyl acetate to repeatedly extract 3-5 times, combining the ethyl acetate solutions, and recovering under reduced pressure until there is no ethyl acetate, thereby obtaining an Russian color leaf extract; Step 2: Add 1-10 parts of animal protein to 500-700 parts of water, stir at room temperature 2000-4000 rpm for 1-3 hours to homogenize; then add 1-5 parts of Russian leaf extract and continue stirring for 1-3 hours to obtain an animal protein-Russian leaf extract composite liquid; Step 3: Mix 1-5 parts of locust bean gum, 0.1-1 parts of propylene glycol fatty acid ester and 40-80 parts of water, and disperse at 10000-15000 rpm for 3-5 minutes. Repeat the dispersion step 2-4 times to obtain a uniform and stable mixed solution a. Step 4: Mix the animal protein-Oise leaf extract composite liquid obtained in step 2 and the mixed liquid a obtained in step 3 in a mass ratio of 1-5:1-5, and then homogenize at a homogenization temperature of 58-62°C and a pressure of 28-32MPa to obtain a mixed liquid; then spray freeze-drying is performed to obtain Oise leaf extract composite microcapsules.

4. The composition of the extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine according to claim 3, characterized in that: The animal protein is selected from at least one of squid skin protein, collagen and whey protein.

5. The composition of the extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine according to claim 4, characterized in that: The preparation method of squid skin protein comprises the following steps: The squid skin is crushed, and then mixed with water 3-10 times the mass of the squid skin to prepare a homogenate; the pH value of the homogenate is adjusted to 8-10, 1-5wt% of alkaline protease of the squid skin is added, and enzymolysis is carried out at 35-45°C for 30-60min; the enzyme is inactivated by heating, centrifugation is performed, and the clear liquid is obtained to obtain a squid skin enzymatic hydrolysate; then the pH value of the squid skin enzymatic hydrolysate is adjusted to 3-5, acid precipitation is performed, the supernatant is discarded, the precipitate is taken, the precipitate is washed with water until neutral, and freeze-dried to obtain squid skin protein.

6. The composition of the extract of Euphorbia obesa leaf, β-nicotinamide mononucleotide and anserine according to claim 2 or 3, characterized in that: The Russian leaves are selected from at least one of the leaf buds of the Rosaceae plant Malus variegata, the leaves of the Rosaceae plant Malus variegata, the leaf buds of Malus variegata, and the leaves of Malus variegata.

7. Use of the composition of the extract of Osmanthus fragrans leaves, β-nicotinamide mononucleotide and anserine as claimed in any one of claims 1 to 6 in alcohol sobering and liver protecting drugs.

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