CN105853350A - Genistein solution, preparation method and application - Google Patents

Abstract

The invention discloses a genistein solution, a preparation method and an application thereof, which are prepared from arginine, genistein, aspartame and baking soda raw materials in a certain proportion. The steps are: A. Weigh a certain amount of arginine and add 3-10 times of water, stir and dissolve at 30-58°C, and set aside; B. Add genistein to the solution of step A, and continue stirring at 30-58°C for 5h, Filter and use the filtrate for later use; C. Weigh aspartame and sodium bicarbonate and add them to the solution in step B, stir at room temperature to dissolve, filter, 100ml/bottle, and divide to obtain. It solves the problem that genistein is insoluble in water, and supplemented with lysine, etc., can increase the SOD activity of smooth muscle cells and reduce the content of lipid peroxide MDA, etc., can resist oxidation, improve human immunity, and delay the speed of body degradation , Prevent skin aging and prolong life.

Description

A kind of genistein solution and its preparation method and application

technical field

The invention belongs to the fields of biomedicine and health care products, and more specifically relates to a genistein solution, a method for preparing the genistein solution, and an application of the genistein solution.

Background technique

Antioxidant refers to the abbreviation of antioxidant free radicals, English Anti-Oxidant. The human body continuously generates free radicals in the human body due to continuous contact with the outside world, including breathing (oxidation reaction), external pollution, radiation exposure and other factors. Scientific studies have shown that cancer, aging or other diseases are mostly associated with the production of excess free radicals. Research on anti-oxidation can effectively overcome the harm it brings, so anti-oxidation is listed as one of the main research and development directions by health care products and cosmetics companies, and it is also one of the most important functional demands of the market.

Antioxidant is any substance that can effectively inhibit the oxidation reaction of free radicals at low concentrations. Its mechanism of action can be to directly act on free radicals, or indirectly consume substances that are easy to generate free radicals to prevent further reactions. While the human body inevitably produces free radicals, it also naturally produces antioxidant substances that resist free radicals to counteract the oxidative attacks of free radicals on human cells. Studies have proved that the antioxidant system of the human body is a system with complete and complex functions comparable to the immune system. The stronger the body's antioxidant capacity, the healthier it is and the longer its life.

The body's antioxidant substances are either synthesized by itself or supplied by food. Enzymatic and non-enzymatic antioxidant substances play a crucial role in protecting against peroxidative damage caused by exercise. Supplementing plant active selenium to enhance anti-oxidation is beneficial to the sports body to reduce the production of free radicals or accelerate their removal, so as to combat the side effects of free radicals, so it is beneficial to the health of ordinary people and athletes, and may delay the occurrence of exercise-induced fatigue and accelerate physical recovery, age Older physically active people do better than younger people taking antioxidants.

More and more studies have shown that anti-oxidation is an important step in preventing aging, because free radicals or oxidants can decompose cells and tissues, affect metabolic functions, and cause different health problems. If excessive oxidative free radicals can be eliminated, many free radical-induced and aging-related diseases can be prevented. For example, common cancers, arteriosclerosis, diabetes, cataracts, cardiovascular diseases, senile dementia, arthritis, etc., are all considered to be related to free radicals.

Sufficient intake of antioxidants should be taken to delay the rate of body degeneration, prevent skin aging, and maintain a youthful look at all times. Antioxidants can be found in the natural diet, such as bioflavonoids called antioxidant substances, vitamin E, vitamin C, and beta-carotene. They can use the characteristics of their own structure to stabilize the excess electrons of free radicals and prevent aging of cells. But in the busy city, you can also choose natural nutritional supplements to get enough antioxidants.

Bioflavonoids can effectively remove free radicals (Free Radical) and toxins in the body, prevent and reduce the occurrence of diseases; anti-inflammatory, anti-allergic, broad-spectrum antibacterial, and antiviral effects. Eliminate alcohol poisoning, protect liver and liver. Clearing heat and detoxifying, dispelling rheumatism, strengthening muscles and bones, anti-menopausal syndrome and other effects.

SOD is the abbreviation of Super Oxide Dimutese, and the Chinese name is superoxide dismutase. It is an important antioxidant enzyme in organisms and is widely distributed in various organisms, such as animals, plants, and microorganisms. SOD has special physiological activity and is the primary substance for scavenging free radicals in organisms. The level of SOD in the living body means an intuitive indicator of aging and death; it has been confirmed that there are as many as 60 kinds of diseases caused by oxygen free radicals. It can resist the damage to cells caused by oxygen free radicals, repair damaged cells in time, and restore the damage to cells caused by free radicals.

Genistein, also known as genistein, genistein, 5,7,4′-trihydroxy isoflavone, is mainly obtained from soybean, clover, kudzu root, Sophora japonica, Sophora japonica, Genistein (Genistein Soy isoflavones are called phytohormones, which are similar in molecular structure to endogenous estrogen, and genistein is one of them. Relevant studies have shown that ^[1] genistein has good effects in protecting cardiovascular and cerebrovascular diseases and reducing the occurrence of cardiovascular diseases. Proliferation, widely used in pharmaceuticals, health care products and other fields.

However, due to the strong hydrophobicity of genistein, it is almost insoluble in water, dilute hydrochloric acid (0.1mol L-1), sodium dodecyl sulfate aqueous solution (sodium dodecyl sulfate aqueous solution), artificial intestinal juice pH: 6.8) , The solubility in general organic solvents is poor, but it is easily soluble in dimethyl sulfoxide, and it also has a certain solubility in methanol and ethanol. The structure contains phenolic hydroxyl groups and is weakly acidic, and it is soluble in dilute alkali. Due to the weak lipophilicity and hydrophilicity of genistein, its absorption in the body is affected, the direct oral bioavailability is low, and there is a strong first-pass effect, which makes it difficult to achieve the purpose of clinical treatment of diseases and limits its use. Clinical application.

Arginine is a nutritional supplement and a component in the ornithine cycle with extremely important physiological functions. Arginine is a basic amino acid that has important physiological and biochemical functions in animals. It is not only cytoplasmic and nucleic acid proteins The main component is also an intermediate for converting aspartic acid, glutamic acid, proline, hydroxyproline, polyamines (putrescine, spermidine, spermine) into high-energy phosphate compounds creatine phosphate. The only ammonia source of creatinine; it also plays an important role in the homogeneous metabolism of the body and can be used in a variety of metabolic pathways, including arginase, nitric oxide synthase, arginine/glycine guanidinotransferase ( AGAT), etc., are also important raw materials for protein synthesis, and are also precursors for the synthesis of creatine, polyamines, and nitric oxide (NO) in the body. They play an important role in the nutritional metabolism and regulation of animals. The essential amino acid of mammals is also the conditional essential amino acid of adult mammals, which can improve libido, immune regulation, promote intestinal development, anti-tumor and anti-obesity, etc.

Contents of the invention

The object of the present invention is to provide a genistein solution, which has a reasonable formula and is easy to use, solves the problem that genistein is insoluble in water, prepares it into an oral aqueous solution, avoids the first-pass effect, and improves bioavailability degree; as a soybean extract, genistein has a wide range of raw materials and low price.

Another object of the present invention is to provide a preparation method of genistein solution, which is easy to implement and easy to operate. The reaction is carried out under normal pressure, and the reaction temperature is 20-60°C, so the energy consumption is low, the equipment is easy to operate, and the investment is low.

Another object of the present invention is to provide a kind of application of genistein solution in anti-oxidation (aspect). The solution prepared by the present invention has the functions of improving smooth muscle cell SOD activity and reducing lipid peroxide MDA content after oral administration etc., can resist oxidation, improve human immunity, delay the speed of body degeneration, prevent skin aging, achieve longevity, and improve quality of life.

In order to achieve the above object, the present invention adopts the following technical measures:

The concept is: using genistein as the main raw material, supplemented with arginine, adding an appropriate amount of flavoring agent aspartame, stabilizing agent baking soda, etc., to develop a drug that can improve the SOD activity of smooth muscle cells and reduce lipid peroxidation. MDA content, etc., can resist oxidation, improve human immunity, and serve people's health.

A kind of genistein solution, it is made from the raw material of following weight part:

A genistein solution, which is made from the raw materials of the following parts by weight (preferred range):

A genistein solution, which is made from the raw materials of the following parts by weight (preferable range):

A genistein solution, which is made from the following raw materials in parts by weight (good range):

A kind of genistein solution, it is made by the raw material of following weight parts (best range):

A kind of genistein solution, it is made (best value) by the raw material of following weight part:

A kind of genistein solution preparation and its application, its preparation steps are:

A. Weigh a certain proportion of arginine and add 3-10 times of water, stir and dissolve at 30-58°C, set aside;

B. Weigh a certain proportion of genistein and add it to the solution of the above step A, continue to stir at 30-58°C for 5-7h, filter, and use the filtrate for later use;

C. Weigh a certain proportion of aspartame and baking soda into the solution of the above step B, stir and dissolve at room temperature (20-25°C), filter, pack in 100ml/bottle, and obtain a genistein solution.

The invention provides a solution of genistein, which is prepared into a solution with arginine and taken, which solves the problem of its insolubility in water, and then supplemented with aspartame, baking soda, etc., has the function of improving smooth muscle Cellular SOD activity and reducing lipid peroxide MDA content, etc., can resist oxidation, improve human immunity, delay body degeneration, prevent skin aging, achieve longevity, and improve quality of life.

A kind of application of genistein solution in antioxidant, its step is:

A. By selecting a certain amount of mice, they are randomly divided into a control group, a low-dose genistein solution group, a middle-dose genistein solution group and a high-dose genistein solution group, and observe the effect of genistein on mice in different dose groups. Effects of SOD activity and MDA content in myocardial tissue, brain tissue and liver tissue of rats.

B, genistein solution low-dose group, middle-dose group and high-dose group all can improve SOD activity in mouse myocardial tissue, brain tissue and liver tissue, reduce MDA content in myocardial tissue, brain tissue and liver tissue; The difference was statistically significant in pairwise comparison, and its dose dependence was present.

C. The invented genistein solution has a good antioxidant effect on mice, can significantly increase the SOD activity in the mouse cardiac muscle tissue, brain tissue and liver tissue, and reduce the MDA content.

Compared with the prior art, the present invention has the following advantages and effects:

The problem that genistein is insoluble in water is solved, and it is prepared into an oral aqueous solution, which avoids the first-pass effect and improves bioavailability; as a soybean extract, genistein has wide sources of raw materials and low price. The method is easy to operate and the operation is simple and convenient. The reaction is carried out under normal pressure, and the reaction temperature is 20-60°C, so the energy consumption is low, the equipment is simple, and the investment is low. The solution prepared by the present invention has the functions of improving the SOD activity of smooth muscle cells and reducing the MDA content of lipid peroxides after oral administration, and can resist oxidation, improve human immunity, delay body degeneration, prevent skin aging, prolong life, and improve quality of life etc. purpose.

The invention provides an aqueous solution of genistein, which solves the problem of its insolubility in water, and is supplemented with arginine, etc., which can improve the SOD activity of smooth muscle cells and reduce the content of lipid peroxide MDA, etc., and can resist oxidation. , Improving human immunity, delaying the speed of body degeneration, preventing skin aging, achieving longevity and improving quality of life.

detailed description

Example 1:

A kind of genistein solution, it is made from the raw material of following weight part:

A kind of genistein solution, its preparation steps are:

A. Weigh a certain proportion of arginine and add 3 or 5 or 7 or 9 or 10 times of water, stir to dissolve at 30 or 35 or 38 or 42 or 48 or 52 or 56 or 58°C, and set aside;

B. Add a certain proportion of genistein to the above solution of A, continue to stir at 30 or 36 or 40 or 44 or 47 or 51 or 55 or 58°C for 4 or 5 or 6 hours, filter, and use the filtrate for later use;

C. Weigh a certain proportion of aspartame and baking soda into the solution of step B above, stir and dissolve at room temperature (20 or 22 or 24 or 25°C), filter, and pack in 100ml/bottle.

Example 2:

A kind of genistein solution, it is made from the raw material of following weight part:

Its preparation steps are identical with embodiment 1.

Example 3:

A kind of genistein solution, it is made from the raw material of following weight part:

Its preparation steps are identical with embodiment 1.

Example 4:

A kind of genistein solution, it is made from the raw material of following weight part:

Its preparation steps are identical with embodiment 1.

Example 5:

A. By selecting 80 mice, they were randomly divided into a control group, a low-dose genistein solution group, a middle-dose genistein solution group, and a high-dose genistein solution group, and the effects of genistein on mice in different dose groups were observed. Effects of SOD activity and MDA content in myocardial tissue, brain tissue and liver tissue of rats.

B, genistein solution low-dose group, middle-dose group and high-dose group all can improve the SOD activity in the mouse myocardial tissue, brain tissue and liver tissue, reduce the MDA content in the mouse myocardial tissue, brain tissue and liver tissue; Compared with the control group, the difference was statistically significant, and the dose-dependent.

C. The invented genistein solution has a good antioxidant effect on mice, can significantly increase the SOD activity in the mouse cardiac muscle tissue, brain tissue and liver tissue, and reduce the MDA content.

Antioxidant experiment of genistein solution of the present invention:

1.1 Test animals

A total of 80 clean-grade mice were selected as the research objects, half male and half male, with an average body weight of (25.3±2.26) g. 80 mice were randomly divided into control group, low dose genistein solution group, medium dose genistein solution group and high dose genistein solution group. There was no difference in the ratio of male to female and average body weight among the four groups of mice, which met the requirements of the experiment.

1.2 Test materials

Genistein solution (made by our unit), superoxide dismutase (SOD) (purchased from Suihe Bioengineering (Xiamen) Co., Ltd.) and malondialdehyde (MDA) kit (purchased from Shanghai Enzyme Biotechnology Co., Ltd. ).

1.3 Preparation of samples for experiments

Except that mice in the control group were given 0.9% sodium chloride solution, mice in all other groups were administered by intraperitoneal injection (30.0 mg/kg for the high-dose group containing genistein, 10.0 mg/kg for the middle-dose group, and 10.0 mg/kg for the low-dose group). Dosage group is 5.0mg/kg, 1 time/d, continuous administration 10 days); each group puts to death mice respectively after last administration, gets its myocardial tissue, brain and liver tissue, puts it in water bath and makes it respectively 10 days. % homogenate and store at -80°C for biological indicators to be tested.

1.4 Determination of biochemical indicators

Take the homogenate for the above experiments, use superoxide dismutase (SOD) and malondialdehyde (MDA) kits, operate according to the kit instructions, and measure the SOD activity and MDA content.

The statistical software Prism 4.0 was used to conduct one-way analysis of variance between each dose group and the control group, and the data in each group were expressed as mean ± standard deviation. Indicates; the q test is used for pairwise comparison between groups, and P<0.05 means that the difference is statistically significant.

2 test results

2.1 Effect of genistein on SOD and MDA content in mouse myocardial tissue

The test results show that the genistein solutions of different dose groups (low dose group, middle dose group and high dose group) can increase the SOD activity in the myocardial tissue of mice and can reduce the MDA content. Compared with the control group, the difference between the different dose groups There was statistical significance (P<0.05), and it was dose-dependent. The results are shown in Table 1 below:

Table 1 Different doses of genistein solution to the measurement results of SOD activity and MDA content in mouse myocardial tissue

Note: Compared with the control group ( ^#1 P<0.05, ^#2 P<0.01).

2.2 Effect of genistein solution on SOD and MDA content in mouse brain tissue

The test results show that the genistein solutions of different dose groups (low dose group, middle dose group and high dose group) can increase the activity of SOD in the brain tissue of mice, and can reduce the content of MDA. There was statistical significance (P<0.05), and it was dose-dependent. The effects of different dosage groups of genistein solution on SOD activity and MDA content in mouse brain tissue are shown in the following table 2:

Table 2 Different dose groups of genistein solution to the measurement results of SOD activity and MDA content in mouse brain tissue

Note: Compared with the control group ( ^#1 P<0.05, ^#2 P<0.01).

2.3 Effect of genistein solution on SOD and MDA content in mouse liver tissue

The test results showed that the genistein solutions of different dose groups (low dose group, middle dose group and high dose group) could increase the activity of SOD in mouse liver tissue and reduce the content of MDA. There was statistical significance (P<0.05), and it was dose-dependent. The effects of different dosage groups of genistein solution on SOD activity and MDA content in mouse liver tissue are shown in Table 3 below:

Table 3 Different dose groups of genistein solution to the measurement results of SOD activity and MDA content in mouse liver tissue

Note: Compared with the control group ( ^#1 P<0.05, ^#2 P<0.01).

The test results show that the genistein solution has a good antioxidant protective effect, and it improves the SOD activity of mouse smooth muscle cells and reduces the MDA content through direct or indirect effects.

Claims (10)

1. a genistein solution, it is made up of the raw material of following weight portion:

Parts by weight of raw materials

Genistein 2-22

Arginine 7-45

Aspartame 0.02-0.08

Sodium bicarbonate 0.02-0.09.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 5-20

Arginine 8-40

Aspartame 0.03-0.07

Sodium bicarbonate 0.03-0.08.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 6-18

Arginine 18-38

Aspartame 0.025-0.065

Sodium bicarbonate 0.04-0.07.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 7-15

Arginase 12 5-35

Aspartame 0.03-0.06

Sodium bicarbonate 0.04-0.065.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 8-12

Arginase 12 0-30

Aspartame 0.04-0.05

Sodium bicarbonate 0.05-0.06.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 9

Arginase 12 5

Aspartame 0.05

Sodium bicarbonate 0.06.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 8

Arginine 30

Aspartame 0.05

Sodium bicarbonate 0.06.

A kind of genistein solution the most according to claim 1, it is characterised in that:

Parts by weight of raw materials

Genistein 10

Arginine 35

Aspartame 0.05

Sodium bicarbonate 0.06.

9. the preparation method of a kind of genistein solution described in claim 1, the steps include:

A, weigh a certain proportion of arginine add 3-10 times of water, 30-58 DEG C of stirring and dissolving,

Standby；

B, weigh a certain proportion of genistein and add in step solution A, continue stirring 5-7h at 30-58 DEG C, filter, filtrate Standby；

C, weigh a certain proportion of Aspartame, sodium bicarbonate adds in step B solution, and dissolving is stirred at room temperature, filter, packing, A kind of genistein solution.

10. the application in anti-oxidant of a kind of genistein solution described in claim 1.