CN111635844A - Method for preparing highland barley compound wine with function of improving anoxia tolerance - Google Patents
Method for preparing highland barley compound wine with function of improving anoxia tolerance Download PDFInfo
- Publication number
- CN111635844A CN111635844A CN202010558438.7A CN202010558438A CN111635844A CN 111635844 A CN111635844 A CN 111635844A CN 202010558438 A CN202010558438 A CN 202010558438A CN 111635844 A CN111635844 A CN 111635844A
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- highland barley
- wine
- ginseng
- concentrated solution
- extracting
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- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention discloses a preparation method of highland barley compound wine capable of improving anoxia endurance, which comprises the following steps: selecting highland barley Chinese liquor, aging for more than 5 years, adding the concentrated solution of Ginseng radix and radix Puerariae, stirring, standing for 48 hr, and filtering with active carbon. The invention can improve the drinking experience of drinking people in plateau local areas and can also improve the anoxia endurance of foreign people.
Description
Technical Field
The invention relates to the technical field of wine brewing, in particular to a method for preparing highland barley compound wine capable of improving anoxia tolerance.
Background
The altitude reaction is various pathological reactions generated after the human body is exposed to a low-pressure hypoxic environment in an acute way, and is a unique common symptom in the plateau area, wherein the hypoxia of the body is one of the main causes of the symptom. Many medicines for preventing altitude diseases and improving the anoxia endurance of organisms are researched and used in China, and most of the medicines are mainly Chinese herbal medicines. In recent years, Chinese medicines and Tibetan medicines have made great progress on the prevention and treatment research of acute altitude diseases. Most of the traditional Chinese medicine composition is composed of natural plants, and has the advantages of definite curative effect, small toxic and side effect, economy, practicability and the like.
The active component ginsenoside of ginseng has obvious effect of resisting lipid peroxidation, and has obvious effect of protecting the ultrastructure of cerebral cortical neuron from oxygen deficiency damage under the high oxygen deficiency condition of 7000m altitude. Research of military medical science and academy indicates that the compatibility of the two monomers of the ginsenoside Rg1 and the tanshinone IIA has an obvious protective effect on myocardial cells damaged by hypoxia-reoxygenation, and the mechanism of the anti-oxidative damage of the ginsenoside Rg1 and the tanshinone IIA is probably realized by increasing the activity of SOD enzyme. The combination effect of the ginsenoside Rg160 mu mol/L and the tanshinone IIA 2 mu mol/L is the best. The research of Tianjianming et al shows that: the ginsenoside Rg2 has no influence on the pulse amplitude and survival rate of normal cultured cardiomyocytes at the concentration of 10-40 μmol. L, and has effects of increasing pulse amplitude and survival rate of anoxic and glucose-deficient cardiomyocytes.
Radix Puerariae is dried root of Pueraria lobata Ohwi of Leguminosae, also called Pueraria lobata Ohwi, and contains flavonoids, triterpenoid saponins, puerarin, coumarins, organic acids, etc. Modern scientific researches find that the kudzuvine root has pharmacological functions of protecting the liver, regulating immunity, resisting tumors, protecting the heart and blood vessels, reducing blood sugar and the like. The midday friendship hospital of Jilin university establishes an acute alcoholic liver injury model by establishing blank, model, high (0.8g/kg) and low (0.4g/kg) puerarin dose groups to carry out animal experiments, and observes the prevention effect of puerarin on acute alcoholic liver injury, so that the serum TG, ALT and AST of rats in the high and low dose groups of puerarin are lower than those in the model group (P <0.05), and have no significant difference (P >0.05) compared with the blank group. The liver mass/body mass ratio of the model control group is higher than that of the puerarin high and low dose groups, and the puerarin high and low dose groups have no significant difference (P >0.05) with the blank group, so that the puerarin can prevent acute alcoholic liver injury of rats and has a protective effect on the liver.
Highland barley wine is generally drunk by residents in Tibet plateau well, the highland barley wine is more in amount, and when the highland barley wine is drunk excessively, due to oxygen deficiency and reduction of liver detoxification function, liver cells can be directly damaged by alcohol, so that chronic alcoholism or liver cirrhosis is caused. Therefore, the intensive research is carried out by mainly improving the anoxia endurance and by assisting in reducing liver damage, the great significance is brought to the development of the highland barley compound wine capable of effectively improving the anoxia endurance, and the highland barley wine has strong plateau identification, so that the product attribute for improving the anoxia endurance can enhance the regional characteristics.
Disclosure of Invention
The invention aims to provide highland barley compound wine capable of improving anoxia endurance and a preparation method thereof, which can improve the drinking experience of drinking people in local plateau areas and can also improve the anoxia endurance of foreign people.
In order to achieve the above object, the present invention provides the following technical solutions:
a preparation method of highland barley compound wine for improving anoxia tolerance comprises the following steps:
selecting highland barley Chinese liquor, aging for more than 5 years, adding concentrated solution of Ginseng radix and radix Puerariae, stirring the highland barley Chinese liquor and concentrated solution at a ratio of (100-.
Preferably, the highland barley white spirit and the concentrated solution are fully and uniformly stirred according to the proportion of 100: 1.
Preferably, the concentrated solution of ginseng and kudzuvine root is prepared by mixing ginseng extract and kudzuvine root extract according to the weight ratio of 3: extracting with alcohol of 1, compounding with 42-degree alcohol solution, and filtering with membrane.
Preferably, the ginseng extract is prepared by the following method: taking a proper amount of ginseng medicinal materials, extracting the ginseng medicinal materials for 3 times by using an extraction solvent process with ethanol concentration of 60% voL-65% voL at 60 ℃ for 3h, 1h and 1h, and directly concentrating and drying three extracting solutions.
Preferably, the kudzu root extract is prepared by the following method: taking a proper amount of medicinal materials, and extracting with 60-80% ethanol; the extraction temperature is 80 +/-5 ℃; the extraction times are 3 times; the extraction time is 2.0h for the first time and 2h for the second time and the third time respectively; the solvent times are respectively 7 times, 5 times and 4 times; concentrating the eluate to 1/10 corresponding to the amount of the medicinal materials, adding 0.5 times of water, stirring with stirring paddle at 100rpm, standing, cooling overnight for crystallization; centrifuging the crystallization liquid until no more centrifugate flows out, centrifuging for 10min, and directly concentrating and drying the crystallization mother liquor; dissolving the crystal precipitate in purified water under heating, cooling, recrystallizing, centrifuging, repeating the above steps for 2-3 times, collecting the mother liquor, concentrating, and drying.
The invention also provides the highland barley compound wine with the function of improving anoxia endurance, which is prepared by the method.
Preferably, the puerarin of the highland barley compound wine is more than 155mg/L, and the total ginsenoside is more than 64.1 mg/L.
Compared with the prior art, the invention has the following advantages:
1) the highland barley compound wine with the function of improving and relieving the altitude stress (improving the anoxia endurance) is developed for the first time.
2) The modern biotechnology is used for extracting the functional components free from the damage of oxygen deficiency for the first time to improve the added value of the highland barley wine.
The highland barley wine prepared by the invention has the effect of improving anoxia endurance, and a large number of clinical animal tests and safety evaluation tests prove that the beverage wine is safe and nontoxic and has the function of improving anoxia endurance.
Drawings
FIG. 1 is a process flow diagram of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Examples
As shown in figure 1, a preparation method of highland barley compound wine with improved anoxia endurance comprises selecting highland barley Chinese liquor as base wine, and aging the highland barley Chinese liquor for more than 5 years. The following are the detection components of the highland barley raw materials of different varieties and the detection components of the highland barley raw wine brewed by the highland barley raw materials, and the results are as follows:
TABLE 1 Table of the test ingredients of raw highland barley and raw highland barley wine
| Detecting items | Raw material of Ganlihuang highland barley | Herba Orostachyos highland barley raw material | Highland barley wine base |
| Total Flavonoids | 0.096％ | 0.291％ | Not detected out |
| Aspartic acid | 0.77％ | 0.51％ | Not detected out |
| Serine | 2.08％ | 1.24％ | Not detected out |
| Glutamic acid | 0.38％ | 0.30％ | Not detected out |
| Glycine | 0.071％ | 0.054％ | Not detected out |
| Histidine | 0.36％ | 0.45％ | Not detected out |
| Arginine | 0.18％ | 0.39％ | Not detected out |
| Threonine | 1.47％ | 0.90％ | Not detected out |
| Alanine | 0.24％ | 0.19％ | Not detected out |
| Proline | 0.44％ | 0.31％ | Not detected out |
| Cysteine | 0.061％ | 0.055％ | Not detected out |
| Tyrosine | 0.11％ | 0.09％ | Not detected out |
| Valine | 0.31％ | 0.22％ | Not detected out |
| Methionine | 0.65％ | 0.51％ | Not detected out |
| Lysine | 0.30％ | 0.17％ | Not detected out |
| Isoleucine | 0.41％ | 0.27％ | Not detected out |
| Leucine | 1.23％ | 0.79％ | Not detected out |
As can be seen in the table: the highland barley raw materials all contain a certain amount of flavonoids, but the highland barley wine brewed by the highland barley has no active ingredients through detection. The highland barley wine base is prepared by adding the concentrated solution of ginseng and kudzuvine root into highland barley wine base according to the proportion of 100:1, fully and uniformly stirring, standing for 48 hours and filtering by active carbon.
The ginseng and kudzuvine root concentrated solution is prepared by mixing a ginseng extract and a kudzuvine root extract according to the weight ratio of 3: extracting with alcohol of 1, compounding with 42-degree alcohol solution, and filtering with membrane.
The ginseng extract is prepared by the following method: taking a proper amount of ginseng medicinal materials, extracting the ginseng medicinal materials for 3 times by using an extraction solvent process with ethanol concentration of 60% voL-65% voL at 60 ℃ for 3h, 1h and 1h, and directly concentrating and drying three extracting solutions.
The kudzu root extract is prepared by the following method: taking a proper amount of medicinal materials, and extracting with 60-80% ethanol; the extraction temperature is 80 +/-5 ℃; the extraction times are 3 times; the extraction time is 2.0h for the first time and 2h for the second time and the third time respectively; the solvent times are respectively 7 times, 5 times and 4 times; concentrating the eluate to 1/10 corresponding to the amount of the medicinal materials, adding 0.5 times of water, stirring with stirring paddle at 100rpm, standing, cooling overnight for crystallization; centrifuging the crystallization liquid until no more centrifugate flows out, centrifuging for 10min, and directly concentrating and drying the crystallization mother liquor; dissolving the crystal precipitate in purified water under heating, cooling, recrystallizing, centrifuging, repeating the above steps for 2-3 times, collecting the mother liquor, concentrating, and drying.
The highland barley compound wine prepared in the embodiment is detected, and the result is as follows:
TABLE 2 ingredient table for testing highland barley blended wine of the present invention
| Detecting items | Content, mg/L | Detecting items | Content, mg/L |
| Total saponins of Ginseng radix | 64.10 | Arginine | Not detected out |
| Puerarin and its preparation method | 155 | ||
| Linoleic acid | Not detected out | Threonine | Not detected out |
| Potassium salt | 23 | Alanine | Not detected out |
| Magnesium alloy | 9.8 | Proline | Not detected out |
| Methanol | 30 | Cystine | Not detected out |
| Fusel oil | 260 | Tyrosine | Not detected out |
| Lead (II) | ＜0.02mg/L | Valine | Not detected out |
| Manganese oxide | ＜0.02mg/L | Methionine | Not detected out |
| Aspartic acid | Not detected out | Lysine | Not detected out |
| Serine | Not detected out | Isoleucine | Not detected out |
| Glycine | Not detected out | Leucine | Not detected out |
| Histidine | Not detected out | Phenylalanine | Not detected out |
Verification example
1 laboratory animal
The types are as follows: mouse
Strain: kunming seed
Grade: SPF stage
Quantity: 150, female.
Body weight range: 18g to 22 g.
Adaptive feeding days: 4 days
Animal status: good effect
Animal provision units: research center for experimental animals in Hubei province
Production license number of experimental animal: SCXK (jaw) 2015-0018.
Quality certification number of experimental animal: 42000600034584
Reason for animal selection: the technical Specification for testing and evaluating health food (2003 edition).
Study system labeling program: each group of animals is marked by staining the hairs on different body surface parts of the animals with picric acid solution by a hair dyeing method. The original data was identified using the experimental animal number. The mice are bred in groups in cages, 5 mice are bred in one cage, and cage cards are marked with information such as special number, animal species, sex, cage number, group, animal number, first test date, special responsible person and the like.
Animal feed
Wuhan Wanqianjiaxing Biotechnology, Inc., and the license number is SCXK (Hubei) 2016-.
Feeding environment
The SPF animal laboratory has the temperature of 20-26 ℃ and the humidity of 40-70 percent, and uses the license number of SYXK (jaw) 2017-.
2 Main instruments and reagents
Scale model: PL602-L, numbered: WY0207, Mettler-Tollido instruments (Shanghai) Inc.;
scale model: AR1530, number: WY0216, mettler-toledo instruments (shanghai) ltd;
an electronic stopwatch: TF807 type, produced by Shenzhen Shanghao Industrial and trade Co., Ltd;
sodium nitrite: batch number: 20150518, manufactured by national drug group chemical agents, Inc.;
soda lime, batch number: 20190212, produced by Shanghai Nahui Dry reagents works;
oxyhemoglobin ELISA reagent: batch number: 08/2019, Shanghai enzyme-Linked Bio Inc.;
phosphocreatine kinase: batch number: 19031, tokyo wittman biotechnology (nanjing) ltd;
lactate dehydrogenase: batch number: 19021, Toho Witmann Biotechnology (Nanjing) Ltd;
malondialdehyde (MDA) test kit 100T/96 sample, lot number: 20190717, Nanjing was established as a institute of bioengineering;
SOD test box, 100T/96 samples, batch number: 20190715, Nanjing was established as a institute of bioengineering;
3 dose design and grouping
The recommended amount of finished product population is 100 mL/person/day, namely 1.667mL/kg BW. The recommended daily intake of 20.0mL/60kg BW (0.333 mL/kg BW) of the population of 5-fold concentrate to be inspected is taken as a dosage design basis, 3.33mL/kg BW, 6.67mL/kg BW and 10.00mL/kg BW (respectively corresponding to 10-fold, 20-fold and 30-fold of the recommended population) are taken as the dosages of three dosage groups, and a negative control group and a wine-based control group are simultaneously set. The test subjects were administered by continuous gavage and the test was started 30 days later.
4 preparation and administration of samples
Sample preparation: accurately measuring 33.3mL, 66.7mL and 100.0mL of 5 times of the concentrated solution sample in the embodiment, respectively adding 15 ° wine base to respectively constant volume to 200mL (final alcohol content is 15%), and mixing uniformly to be used for intragastric administration of low, medium and high dose groups; the negative control group was given distilled water and the wine-based control group was given 15% alcohol.
Preparing a wine-based reference substance: 71.4mL of 42-degree wine base is measured, distilled water is added to the wine base to be constant volume to 200mL (the content of alcohol is 15 percent), and the mixture is shaken up and used for intragastric administration of a wine base control group.
The gavage volume of the sample group, the wine-based control group and the negative control group is 20mL/kg BW, and the gavage period is 30 days. The weight of the animals is weighed once a week, and the gavage amount is adjusted according to the weight.
5 atmospheric hypoxia tolerance test
The experimental method comprises the following steps: the mice were continuously gavaged for 30 days, 1 hour after the last administration of the test substance, each group of mice was placed in 250mL ground bottles (1 per bottle) containing 5g soda lime, the bottle mouth was sealed with vaseline, the bottles were tightly closed to prevent air leakage, a stopwatch was immediately used to time, and the time of death of the mice due to oxygen deficiency was observed with the cessation of respiration as an index.
Compared with a negative control group (a negative pair), the normal-pressure hypoxia-resistant survival time of mice in the wine-based control group (the wine-based pair) is not obviously different (P is more than 0.05), the normal-pressure hypoxia-resistant survival time of the mice in each dose group in the embodiment can be prolonged, and the mice in each dose group are obviously different (P is less than 0.01) compared with the negative control group, compared with the wine-based control group, the normal-pressure hypoxia-resistant survival time of the mice in each dose group in the embodiment can be prolonged, and the mice in each dose group in the embodiment can be obviously prolonged (P is less than 0.01) compared with the wine-based control group, and the results show that the normal-pressure hypoxia-resistant survival time of the mice in each dose group in.
TABLE 3 Effect of the invention on survival time of mice in normbaric hypoxia tolerance experiments (mean. + -. standard deviation)
6 acute cerebral ischemic hypoxia experiment
The experimental method comprises the following steps: the mice were continuously gavaged for 30 days, 1h after the test object was last given, each group of animals broke their heads one by one from the neck, and the time from the mouse's head breaking to the time of stopping breathing through the mouth opening was immediately recorded according to a stopwatch.
Compared with a negative control group, the acute cerebral ischemic anoxia-gasping time of the mice of the wine-based control group has no obvious difference (P is more than 0.05), each dosage group in the embodiment can prolong the gasping time of the mice of the acute cerebral ischemic anoxia-experimental group, and the dosage groups have significant difference (P is less than 0.01) compared with the negative control group; compared with a wine-based control group, each dose group in the embodiment can prolong the gasping time of the acute cerebral ischemic anoxia experimental mice, and has significant difference (P is less than 0.01) compared with the wine-based control group; the above results illustrate that: in the embodiment, each dosage group can obviously prolong the gasping time of the acute cerebral ischemic anoxia experimental mice (see table 4).
TABLE 4 influence of this example on gasping time in mouse acute cerebral ischemic anoxia test (mean. + -. standard deviation)
7 biochemical index determination
The experimental method comprises the following steps: the eyes of mice in the atmospheric hypoxia tolerance experimental group are taken 25 days after the test sample is given
Collecting blood 0.5ml, separating serum, measuring content of serum phosphocreatine kinase and lactate dehydrogenase, and measuring antioxidant index (MDA, SOD, etc.) in serum. In 28 days after the test sample is given, the eyes of the mice of the acute cerebral ischemic anoxia experimental group are taken
Blood collection, measurement of the content of oxygenated hemoglobin by whole blood.
Compared with a negative control group, the content of Oxyhemoglobin (OHG) in the blood of mice in the wine-based control group has no obvious difference (P is more than 0.05), each dose group in the embodiment can increase the content of oxyhemoglobin in the blood of the mice, and the content of oxyhemoglobin in the blood of the mice in the wine-based control group has obvious difference (P is less than 0.05) compared with the negative control group and the wine-based control group; the above results illustrate that: in the embodiment, each dose group can obviously increase the content of oxygenated hemoglobin in the blood of the mice.
Compared with a negative control group, the CK of the low-dose group is obviously increased, and the difference is significant (P is less than 0.05); the difference in CK was not significant (P >0.05 for each dose group) compared to the wine-based control group. Compared with the negative control group and the wine-based control group, MDA, SOD and LDH of each dose group have no obvious difference (all are P >0.05), and the result is shown in Table 5.
Table 5 effect of this example on several biochemical parameters of mice (n 10, mean ± sd)
Remarking: compared with the wine-based control group,*p is less than 0.05; compared with the negative control group, the test results show that,#P＜0.05
according to the evaluation program regulation of the function of improving the anoxia endurance of the health food inspection and evaluation technical specification (2003 edition), the animal with the function of improving the anoxia endurance is judged to have positive test results by combining the test results. The wine can prolong the normal-pressure hypoxia-resistant survival time of a mouse, prolong the gasping time of an acute cerebral ischemic hypoxia experiment mouse and increase the content of oxygenated hemoglobin in blood to improve the hypoxia tolerance, and one of the mechanisms of the wine can be to improve the content of the oxygenated hemoglobin in the blood to improve the hypoxia tolerance.
The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (7)
1. The preparation method of the highland barley compound wine with the function of improving anoxia tolerance is characterized by comprising the following steps:
selecting highland barley Chinese liquor, aging for more than 5 years, adding concentrated solution of Ginseng radix and radix Puerariae, stirring the highland barley Chinese liquor and concentrated solution at a ratio of (100-.
2. The preparation method according to claim 1, wherein the highland barley white spirit and the concentrated solution are fully and uniformly stirred according to the proportion of 100: 1.
3. The method of claim 1, wherein the concentrated solution of ginseng and pueraria lobata is prepared from ginseng extract and pueraria lobata extract according to a ratio of 3: extracting with alcohol of 1, compounding with 42-degree alcohol solution, and filtering with membrane.
4. The method according to claim 1, wherein the ginseng extract is prepared by the following method: taking a proper amount of ginseng medicinal materials, extracting the ginseng medicinal materials for 3 times by using an extraction solvent process with ethanol concentration of 60% voL-65% voL at 60 ℃ for 3h, 1h and 1h, and directly concentrating and drying three extracting solutions.
5. The method of claim 1, wherein the kudzu root extract is prepared by the following method: taking a proper amount of medicinal materials, and extracting with 60-80% ethanol; the extraction temperature is 80 +/-5 ℃; the extraction times are 3 times; the extraction time is 2.0h for the first time and 2h for the second time and the third time respectively; the solvent times are respectively 7 times, 5 times and 4 times; concentrating the eluate to 1/10 corresponding to the amount of the medicinal materials, adding 0.5 times of water, stirring with stirring paddle at 100rpm, standing, cooling overnight for crystallization; centrifuging the crystallization liquid until no more centrifugate flows out, centrifuging for 10min, and directly concentrating and drying the crystallization mother liquor; dissolving the crystal precipitate in purified water under heating, cooling, recrystallizing, centrifuging, repeating the above steps for 2-3 times, collecting the mother liquor, concentrating, and drying.
6. The highland barley compound wine with the function of improving anoxia endurance prepared by the preparation method of claims 1-5.
7. The highland barley wine of claim 6, wherein the highland barley wine has puerarin of more than 155mg/L and total ginsenoside of more than 64.1 mg/L.
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| CN113025454A (en) * | 2021-02-07 | 2021-06-25 | 青海景阳川生物科技有限公司 | Raspberry and highland barley wine rich in superoxide dismutase and preparation method thereof |
| CN113956936A (en) * | 2021-08-27 | 2022-01-21 | 西藏达热瓦青稞酒业股份有限公司 | Preparation method of canned highland barley wine |
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| CN113956936A (en) * | 2021-08-27 | 2022-01-21 | 西藏达热瓦青稞酒业股份有限公司 | Preparation method of canned highland barley wine |
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Application publication date: 20200908 |