CN108201624A - The preparation and application of isoflavones-chitosan nano compound - Google Patents

Abstract

The present invention proposes a preparation method and application of soybean isoflavone-chitosan nanocomposite, a kind of preparation method of soybean isoflavone chitosan nanocomposite, comprising the following steps: (1) acidification of soybean isoflavone ; (2) self-assembly of soybean isoflavones and chitosan. The gall, cistanche motherwort and salvia miltiorrhiza are pulverized, mixed, decocted, filtered, and concentrated to obtain a water extract of the composition, added with soybean isoflavone-chitosan nanocomposite and resveratrol, and spray-dried to obtain a solid medicinal powder. The soybean isoflavone-chitosan nanocomposite and the prepared composition of the invention have obvious radiation protection effect.

Description

Preparation and application of soybean isoflavone-chitosan nanocomposite

technical field

The invention relates to the technical field of medicine, in particular to the preparation and application of a nanocomposite.

Background technique

my country is a big country in the uranium industry, with millions of people engaged in uranium operations. If related industries are added, there will be even more employees. The health follow-up study of uranium workers showed that long-term exposure to uranium dust was associated with an increased incidence of silicosis, lung cancer and other diseases. Guangxi is rich in uranium resources, and occupational hazards in uranium mining operations are also very common. Therefore, effective medical protective measures to prevent uranium dust damage have broad application prospects.

Current studies have shown that soybean isoflavones have a variety of physiological functions such as protective effects on blood vessels, female estrogen-like effects, anti-hormonal effects, antibacterial activity, and prevention of osteoporosis. Natural antioxidants, and many natural antioxidants have anti-radiation and anti-tumor effects. The anti-radiation effect of soybean isoflavones is manifested as the ability to scavenge free radicals in the body to protect cell membrane lipids and inhibit the damage of free radicals induced by radiation to biological macromolecules. Therefore, the research on the antioxidant and antitumor activities of soybean isoflavones aims to provide a theoretical basis for further development of new antioxidants and antitumor drugs with high efficiency and no toxic side effects.

The absorption of soy isoflavones is mainly carried out in the small intestine, and there are two ways: direct absorption and active transport. Most of the sustained-release dosage forms of soybean isoflavones that are still in the laboratory research stage adopt the cyclodextrin embedding method. higher requirements.

Through nanotechnology, some active substances that are easy to oxidize and decompose (such as polyhydric acid, unsaturated fatty acids, carotenoids) can be protected, and the active substances can be released at the correct time and place to achieve site-specific delivery, and can Effectively absorbed by cells. In the selection of nanomaterials, chitosan is the deacetylation product of chitin. The molecular weight of chitosan is usually around tens of thousands of daltons. The presence of amino active groups in the molecular structure is easy to dissociate into positively charged The cations (-NH ³⁺ ) of acidified soy isoflavones can attract each other with the negatively charged anions (-O ^- and -COO ^- ) dissociated in a large number in acidified soybean isoflavones, and self-assemble into stable nanocomposites. In chitosan Under the link and loading, the protection and slow release of soy isoflavones are realized. The emergence of controlled-release polymeric nanoparticles represents a new step in the development of nanocarriers in the field of medicine.

Gallaceae are the dry insect waists on the twigs of the Fagaceae plant Gallia, formed by the parasitic surface of the Gallaceae larvae, which are called "Meha" by Uighur medicine. Gallatum has the functions of solidifying astringent and astringent, benefiting blood and producing essence, hemostasis and anti-inflammation, etc. It is mainly used clinically for the treatment of large intestine slippery, nonstop, habitual enteritis, hemorrhage and other diseases." Gallatum has a long history of medicine in my country, "Tang Materia Medica" , "Haiyao Materia Medica", "Kaibao Materia Medica", etc., are recorded in Uyghur medicine. It is the most popular and traditional medicinal material in Uyghur medicine. A recent study on gallatum found that it also has a good effect on anti-radiation Effect.

Contents of the invention

In order to solve the above-mentioned technical problems, the present invention provides a kind of preparation method of soybean isoflavone-chitosan nanocomposite, and provides a kind of application of this nanocomposite for radiation protection, this nanocomposite and other traditional Chinese medicine composition The water extract is used together, and the test proves that the composition of the present invention has obvious anti-radiation effect.

The invention provides a kind of preparation method of soybean isoflavone-chitosan nanocomposite, comprising the following steps:

(1) Acidification of soybean isoflavones: dissolve the extracted and purified soybean isoflavones powder in dimethyl sulfoxide, add nitric acid, reflux for 8-12 hours, cool to stop the reaction, and obtain acidified soybean isoflavones;

(2) Self-assembly of soybean isoflavones and chitosan: add chitosan aqueous solution to the acidified soybean isoflavone solution prepared in step (1), reflux for 12-24 hours, cool to stop the reaction, centrifuge the reaction solution, remove the upper clear liquid and deionized water to wash the nanoparticles repeatedly to obtain the soybean isoflavone-chitosan nanocomposite.

As a further improvement of the present invention, the reflux time of the acidification reaction of soybean isoflavones is 10 hours.

As a further improvement of the present invention, the reflux time of the self-assembly reaction of soybean isoflavones and chitosan is 18 hours.

As a further improvement of the present invention, the centrifugation speed is 5000 rpm and the time is 20 min.

As a further improvement of the present invention, every 1 g of soybean isoflavones is dissolved with 5 mL of dimethyl sulfoxide.

As a further improvement of the present invention, the chitosan aqueous solution is prepared in a weight ratio of 1:10.

The present invention further protects the application of the soybean isoflavone-chitosan nanocomposite prepared by the above method for radiation protection.

The present invention further protects a radiation protection composition, which is prepared from the following raw materials in parts by weight: 15-20 parts of soybean isoflavone-chitosan nanocomposite, 15-20 parts of resveratrol, gall 10-15 parts, 5-10 parts of Cistanche, 5-7 parts of Motherwort, 2-5 parts of Salvia.

As a further improvement of the present invention, the composition is prepared from the following raw materials in parts by weight: 17 parts of soybean isoflavone-chitosan nanocomposite, 17 parts of resveratrol, 15 parts of gall, 8 parts of cistanche, 6 parts of motherwort , Salvia 3 parts.

The present invention has following beneficial effect:

1. The present invention introduces nano-chitosan linking and loading technology, and prepares soybean isoflavone-chitosan nanocomposite preparation;

2. The present invention focuses on the preparation of the soybean isoflavone-chitosan compound and the compatibility with traditional Chinese medicine and Uyghur medicine. It is proved by experiments that the soybean isoflavone-chitosan compound prevents the damage of uranium mine dust radiation to cells and experimental animals. Anti-radiation has obvious effect;

3. The present invention applies the soybean isoflavone-chitosan compound to the preparation of anti-radiation medicines, which can promote the anti-radiation of modern medicine to a certain extent.

Description of drawings

Fig. 1 is the preparation process flowchart of soybean isoflavone-chitosan nanocomposite.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some representative embodiments of the present invention, rather than all embodiments. All other embodiments obtained by persons of ordinary skill without making creative efforts belong to the protection scope of the present invention.

The preparation of embodiment 1 soybean isoflavones-chitosan nanocomposite

Follow these steps:

(1) Acidification of soybean isoflavones: dissolve 20 g of extracted and purified soybean isoflavones powder in 50 mL of dimethyl sulfoxide, add nitric acid, reflux for 8 hours, cool to stop the reaction, and obtain acidified soybean isoflavones;

(2) Self-assembly of soybean isoflavones and chitosan: add 0.1g/mL chitosan aqueous solution to the acidified soybean isoflavone solution prepared in step (1), reflux for 12h, cool to stop the reaction, and centrifuge the reaction solution at 3000rpm After 10 minutes, the supernatant was removed, and the nanoparticles were repeatedly washed with deionized water to obtain a soybean isoflavone-chitosan nanocomposite with a yield of 67%.

The preparation of embodiment 2 soybean isoflavones-chitosan nanocomposite

Follow these steps:

(1) Acidification of soybean isoflavones: dissolve 20 g of extracted and purified soybean isoflavones powder in 100 mL of dimethyl sulfoxide, add nitric acid, reflux for 10 h, cool to stop the reaction, and obtain acidified soybean isoflavones;

(2) Self-assembly of soybean isoflavones and chitosan: add 0.2g/mL chitosan aqueous solution to the acidified soybean isoflavone solution prepared in step (1), reflux for 18h, cool to stop the reaction, and centrifuge the reaction solution at 5000rpm After 20 minutes, the supernatant was removed, and the nanoparticles were repeatedly washed with deionized water to obtain a soybean isoflavone-chitosan nanocomposite with a yield of 92%.

The preparation of embodiment 3 soybean isoflavones-chitosan nanocomposite

Follow these steps:

(1) Acidification of soybean isoflavones: dissolve 20 g of extracted and purified soybean isoflavones powder in 150 mL of dimethyl sulfoxide, add nitric acid, reflux for 12 hours, cool to stop the reaction, and obtain acidified soybean isoflavones;

(2) Self-assembly of soybean isoflavones and chitosan: add 0.5g/mL chitosan aqueous solution to the acidified soybean isoflavone solution prepared in step (1), reflux for 24h, cool to stop the reaction, and centrifuge the reaction solution at 8000rpm After 30 minutes, the supernatant was removed, and the nanoparticles were repeatedly washed with deionized water to obtain a soybean isoflavone-chitosan nanocomposite with a yield of 79%.

The preparation of embodiment 4 radiation protection composition

Composition of raw materials: 20 parts of soybean isoflavone-chitosan nanocomposite, 20 parts of resveratrol, 15 parts of gall, 10 parts of cistanche, 7 parts of motherwort, 5 parts of salvia miltiorrhiza.

Preparation steps:

a. Take gallaceae, cistanche, motherwort, and salvia miltiorrhiza, mix them, add 10 times the volume of water, soak for 40 minutes, decoct for 2 hours, filter under reduced pressure, and set aside the filtrate;

b. Add 5 times the volume of water to the dregs obtained in step a, decoct for 1.5 hours, filter under reduced pressure, and the filtrate is set aside;

c. Combine the filtrates obtained in step a and step b, pass through a 400-mesh sieve, and concentrate the filtrate to a relative density of 1.0-1.3 to obtain the aqueous extract of the Chinese medicine composition.

d. Dissolving the soybean isoflavone-chitosan nanocomposite and resveratrol with ethanol solution, adding it to the water extract of the traditional Chinese medicine composition, and spraying and drying to obtain a solid medicine powder.

The preparation of embodiment 5 radiation protection composition

Composition of raw materials: 17 parts of soybean isoflavone-chitosan nanocomposite, 17 parts of resveratrol, 15 parts of gall, 8 parts of cistanche, 6 parts of motherwort, 3 parts of salvia miltiorrhiza.

a. Take gallaceae, cistanche, motherwort, and salvia miltiorrhiza, mix them, add 10 times the volume of water, soak for 40 minutes, decoct for 2 hours, filter under reduced pressure, and set aside the filtrate;

b. Add 5 times the volume of water to the dregs obtained in step a, decoct for 1.5 hours, filter under reduced pressure, and the filtrate is set aside;

c. Combine the filtrates obtained in step a and step b, pass through a 400-mesh sieve, and concentrate the filtrate to a relative density of 1.0-1.3 to obtain the aqueous extract of the Chinese medicine composition.

d. Dissolving the soybean isoflavone-chitosan nanocomposite and resveratrol with ethanol solution, adding it to the water extract of the traditional Chinese medicine composition, and spraying and drying to obtain a solid medicine powder.

The preparation of embodiment 6 radiation protection composition

Composition of raw materials: 15 parts of soybean isoflavone-chitosan nanocomposite, 15 parts of resveratrol, 10 parts of gall, 5 parts of cistanche, 5 parts of motherwort, 2 parts of salvia miltiorrhiza.

a. Take gallaceae, cistanche, motherwort, and salvia miltiorrhiza, mix them, add 10 times the volume of water, soak for 40 minutes, decoct for 2 hours, filter under reduced pressure, and set aside the filtrate;

b. Add 5 times the volume of water to the dregs obtained in step a, decoct for 1.5 hours, filter under reduced pressure, and the filtrate is set aside;

c. Combine the filtrates obtained in step a and step b, pass through a 400-mesh sieve, and concentrate the filtrate to a relative density of 1.0-1.3 to obtain the aqueous extract of the Chinese medicine composition.

d. Dissolving the soybean isoflavone-chitosan nanocomposite and resveratrol with ethanol solution, adding it to the water extract of the traditional Chinese medicine composition, and spraying and drying to obtain a solid medicine powder.

Embodiment 7 does not contain the preparation of the radiation protection composition of soybean isoflavone-chitosan nanocomposite

Composition of raw materials: 17 parts of resveratrol, 15 parts of gall, 8 parts of cistanche, 6 parts of motherwort, 3 parts of salvia miltiorrhiza.

a. Take gallaceae, cistanche, motherwort, and salvia miltiorrhiza, mix them, add 10 times the volume of water, soak for 40 minutes, decoct for 2 hours, filter under reduced pressure, and set aside the filtrate;

b. Add 5 times the volume of water to the dregs obtained in step a, decoct for 1.5 hours, filter under reduced pressure, and the filtrate is set aside;

c. Combine the filtrates obtained in step a and step b, pass through a 400-mesh sieve, and concentrate the filtrate to a relative density of 1.0-1.3 to obtain the aqueous extract of the Chinese medicine composition.

d. Dissolving resveratrol with ethanol solution, adding it to the water extract of the traditional Chinese medicine composition, and spray drying to obtain solid medicinal powder.

Test Example 1 Mouse Test

1. Experimental group. Prepare 140 adult mice, be divided into 7 groups, every group of 20, be respectively the soybean isoflavone powder group (1 group), the soybean isoflavone-chitosan nanocomposite group (2 groups) prepared in embodiment 2, The radiation protection composition (3 groups) that does not contain soybean isoflavone-chitosan nanocomposite that embodiment 7 prepares, the radiation protection composition (4 groups) that embodiment 5 prepares, resveratrol group, control group and blank group.

2. Experimental method. Groups 1 to 5 were intragastrically administered every morning: group 1 administered soybean isoflavone powder 100 mg/kg, group 2 administered soybean isoflavone-chitosan nanocomposite 100 mg/kg prepared in Example 2, group 3 administered The radiation protection composition 100mg/kg that does not contain the soybean isoflavone-chitosan nanocomposite prepared in Example 7, 4 groups of administration The radiation protection composition 100mg/kg prepared in Example 5, 5 groups of administration of resveratrol Alcohol 100mg/kg, control group and blank group gavage distilled water 100mg/kg. The medicinal powders of each group were dissolved in water and administered into the stomach. After feeding for 14 days, the rats in groups 1-5 and the control group were irradiated with ⁶⁰ Co radiation, the irradiation distance was 75cm, the irradiation field of view was 20cm×20cm, and the irradiation time was 8min. Blood was collected on the 1st, 7th, 15th, 22nd, and 30th days of irradiation, and blood physiological indicators were measured with an automatic biochemical analyzer.

The experimental results are shown in Table 1-Table 3.

Table 1 Peripheral red blood cell count of mice (×10 ¹² /L)

group 1d 7d 15d 22d 30d Group 1 5.14 5.02 4.92 5.07 5.15 2 teams 5.22 5.29 5.32 5.34 5.39 3 groups 5.25 5.14 5.02 5.23 5.32 4 groups 5.40 5.37 5.45 5.48 5.56 5 groups 5.19 5.04 4.89 5.19 5.25 control group 5.04 4.75 4.59 4.21 4.62 blank group 5.46 5.43 5.47 5.50 5.49

After radiation, the level of red blood cells in the peripheral blood of the mice decreased, and dropped to the lowest point at 22 days, and then the number of red blood cells began to increase slowly. From the above results, it can be seen that the basic change rules of groups 1 to 5 and the control group are consistent, and both decrease first and then increase , but the radiation protection composition group (group 4) of the present invention had significantly higher levels of peripheral red blood cells after radiation than other groups, and it was relatively stable and recovered quickly. The resveratrol group (group 5), without soybean isoflavone- Although the radiation protection composition of chitosan nanocomposite (group 3) and soybean isoflavone-chitosan nanocomposite group (group 2) have a higher level of peripheral blood erythrocytes than the control group, there is a certain radiation protection effect, but The effect is far lower than that of group 4. It can be seen that the combination of resveratrol, soy isoflavone-chitosan nanocomposite and other traditional Chinese medicine compositions in the radiation protection composition of the present invention has obvious synergistic effect.

Table 2 Mouse peripheral platelet count (×10 ⁹ /L)

group 1d 7d 15d 22d 30d Group 1 477.92 453.21 423.79 442.80 476.92 2 teams 483.15 479.02 468.33 490.21 497.35 3 groups 496.74 485.24 480.31 496.07 510.22 4 groups 512.19 503.82 510.23 517.69 523.78 5 groups 474.52 445.39 410.26 431.24 443.17 control group 470.92 421.57 358.95 332.77 389.70 blank group 515.38 520.61 518.32 522.10 519.73

After receiving radiation, the number of platelets in the mice decreased significantly, dropped to the lowest point on the 22nd day, and then began to rise slowly. From the above results, it can be known that groups 1 to 5 and the control group have basically the same changing rules, both of which decrease first and then increase, but the number of platelets in the radiation protection composition group of the present invention (group 4) is significantly higher than that of other groups after radiation, and is relatively Stable and fast recovery, resveratrol group (5 groups), radiation protection composition without soybean isoflavone-chitosan nanocomposite (3 groups) and soybean isoflavone-chitosan nanocomposite group ( Although the number of platelets in 2 groups) is higher than that of the matched group, there is a certain radiation protection effect, but the effect is far lower than that of 4 groups. It can be seen that the radiation protection composition of the present invention combines resveratrol, soybean isoflavone-chitosan nano It has obvious synergistic effect after being combined with other traditional Chinese medicine compositions.

Table 3 Mouse lymphocyte count (×10 ⁹ /L)

The number of lymphocytes in the peripheral blood of mice can reflect the effect of radiation on immune function to a certain extent. The killing effect of radiation on lymphocytes is obvious. increased, but did not return to normal levels by the end of the experiment. From the above results, it can be known that groups 1 to 5 are basically consistent with the control group, but the level of lymphocytes in the radiation protection composition group of the present invention (group 4) is significantly better than other groups after radiation, and the resveratrol group (group 5) , the radiation protection composition (3 groups) without soybean isoflavone-chitosan nanocomposite and the lymphocyte level of soybean isoflavone-chitosan nanocomposite group (2 groups) though higher than the control group, there is a certain Radiation protection effect, but the effect is far lower than 4 groups, it can be seen that the radiation protection composition of the present invention has obvious synergistic effect after combining resveratrol, soybean isoflavone-chitosan nanocomposite and other traditional Chinese medicine compositions effect.

Various modifications can be made to the above contents by those skilled in the art without departing from the spirit and scope of the present invention defined by the claims. Therefore, the scope of the present invention is not limited to the above description, but is determined by the scope of the claims.

Claims (9)

1. the preparation method of a kind of isoflavones-chitosan nano compound, which is characterized in that the method includes following steps Suddenly：

(1) acidification of isoflavones：The isoflavones powder of extraction purification is dissolved in dimethyl sulfoxide, adds in nitric acid, reflux 8-12h, cooling stop reaction, the isoflavones being acidified；

(2) self assembly of isoflavones and chitosan：Shell is added in the isoflavones solution of the acidification prepared to step (1) Water solution, flow back 12-24h, and cooling stops reaction, reaction solution is centrifuged, and removes supernatant, and deionized water is washed and received repeatedly Rice grain is to get isoflavones-chitosan nano compound.

2. preparation method according to claim 1, which is characterized in that the return time of the acidification reaction of the isoflavones For 10h.

3. preparation method according to claim 1, which is characterized in that the self-assembling reaction of the isoflavones and chitosan Return time be 18h.

4. preparation method according to claim 1, which is characterized in that the centrifugal rotational speed is 5000rpm, time 20min.

5. preparation method according to claim 1, which is characterized in that dissolved per 1g isoflavones with 5mL dimethyl sulfoxides.

6. preparation method according to claim 1, which is characterized in that the chitosan aqueous solution is by weight 1:10 prepare.

7. a kind of isoflavones-chitosan nano compound for preparing preparation method according to claim 1 is used to prevent The application of radiation.

8. a kind of radiation-preventing composition, which is characterized in that composition is prepared by the raw material of following parts by weight：The different Huang of soybean Ketone -15-20 parts of chitosan nano compound, 15-20 parts of resveratrol, 10-15 parts of nutgall, 5-10 parts of Herba Cistanches, motherwort 5-7 parts, 2-5 parts of Radix Salviae Miltiorrhizae.

9. composition according to claim 8, which is characterized in that the composition by following parts by weight raw material prepare and Into：Isoflavones -17 parts of chitosan nano compound, 17 parts of resveratrol, 15 parts of nutgall, 8 parts of Herba Cistanches, motherwort 6 Part, 3 parts of Radix Salviae Miltiorrhizae.