Ganoderma triterpenoid sustained-release microcapsule and preparation method thereof

Abstract

The invention belongs to the technical field of sustained-release microcapsules, and particularly discloses a ganoderma triterpenoid sustained-release microcapsule and a preparation method thereof.Bovine serum albumin and octenyl succinic anhydride modified starch are selected to prepare a Maillard product through a carbonyl ammonia condensation method to serve as a wall material, and a core material ganoderma triterpenoid is well wrapped in the wall material; the Maillard reaction product and the ganoderma triterpenoid are crosslinked, the structure of the freeze-dried microcapsule is not damaged, and the prepared granular microcapsule is complete in structure, high in embedding rate and good in stability. The sustained release performance of the ganoderma triterpenoids as an embedding material is improved, the water solubility of the ganoderma triterpenoids is improved, the dissolution speed and the intestinal mucosa permeability of the ganoderma triterpenoids are improved through conjugation change of polymer characteristics, the retention time is prolonged through biological adhesion, the oral bioavailability of the ganoderma triterpenoids is improved, and the bioavailability of the ganoderma triterpenoids is improved. The human body absorption and utilization degree of the ganoderma triterpenoids can be obviously improved.

Description

Technical Field

[0001] The invention relates to the technical field of sustained-release microcapsules, in particular to a ganoderma triterpenoid compound sustained-release microcapsule and a preparation method thereof. Background Art

[0002] The chemical composition of Ganoderma lucidum is very complex, containing a variety of chemical components. These components together give it a variety of biological activities and pharmacological effects. Among them, Ganoderma lucidum triterpenoids are important active ingredients in Ganoderma lucidum, with anti-inflammatory, anti-tumor, antiviral, and lipid-lowering effects. Therefore, they have attracted much attention in the fields of food and medicine. However, due to the shortcomings of Ganoderma lucidum triterpenoids themselves, such as low solubility, poor biomembrane permeability, and poor absorption effect, their wide application is limited.

[0003] Therefore, how to improve the water solubility, absorption rate and stability of Ganoderma triterpenoids, and thus improve their bioaccessibility, has become a key focus that needs urgent attention in the application of related products. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a ganoderma triterpenoid compound sustained-release microcapsule and a preparation method thereof.

[0005] To achieve the above object, the present invention is implemented according to the following technical solutions:

[0006] One of the purposes of the present invention is to provide a method for preparing a ganoderma triterpenoid sustained-release microcapsule, comprising the following steps:

[0007] S1, dissolving 30 g / L bovine serum albumin (BSA) and 30 g / L octenyl succinate starch ester (OSAS) in ultrapure water respectively, and stirring at room temperature overnight to obtain BSA mother solution and OSAS mother solution;

[0008] S2. Mix BSA mother solution and OSAS mother solution in a mass ratio of 1:1, adjust the solution pH to 7, place the mixed solution at 37°C for reaction for 12 hours, freeze-dry the sample for 48 hours, grind and spread it in a culture dish, and react it in a constant temperature incubator at 60°C for 48 hours. After taking out the sample, remove the unreacted octenyl succinate starch ester OSAS to obtain a BSA-OSAS glycosylation product solution;

[0009] S3. Take the ethanol solution of triterpenoids of Ganoderma lucidum and add it to the BSA-OSAS glycosylation product solution to make the mass ratio of the BSA-OSAS glycosylation product solution to the ethanol solution of triterpenoids of Ganoderma lucidum be 50:1. After magnetic stirring for 60 minutes, ultrasonicate the solution at 180W for 6 minutes to obtain the sustained-release microcapsule solution of triterpenoids of Ganoderma lucidum. After freeze-drying, obtain the sustained-release microcapsules of triterpenoids of Ganoderma lucidum.

[0010] Furthermore, in step S2, 1 mol / L NaOH and HCl are used to adjust the pH of the solution.

[0011] Furthermore, in step S3, the concentration of the Ganoderma triterpenoid compound ethanol solution is 10 g / L.

[0012] The second object of the present invention is to provide a Ganoderma triterpenoid compound sustained-release microcapsule prepared by the above method.

[0013] Compared with the prior art, the present invention uses bovine serum albumin and octenyl succinate starch ester to prepare Maillard products by carbonyl-amino condensation as wall materials, so that the core material Ganoderma lucidum triterpenoid compounds are well wrapped in the wall materials, the Maillard reaction products and Ganoderma lucidum triterpenoid compounds are cross-linked, the structure of the microcapsules is not destroyed after freeze-drying, and the granular microcapsules prepared by the present invention have a complete structure, high embedding rate and good stability. The sustained-release performance of the embedded Ganoderma lucidum triterpenoid compounds is improved, the polymer properties are changed by conjugation, thereby increasing its water solubility, improving its dissolution rate and intestinal mucosal permeability, extending the residence time by bioadhesion, and improving its oral bioavailability, which has a relatively obvious improvement and enhancement effect on the human body absorption and utilization of Ganoderma lucidum triterpenoid compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is the degradation rate of Ganoderma triterpenoids and the Ganoderma triterpenoids sustained-release microcapsules prepared in this example in simulated gastric juice.

[0015] Figure 2 It is the degradation rate of Ganoderma triterpenoids and the Ganoderma triterpenoids sustained-release microcapsules prepared in this example in simulated intestinal fluid.

[0016] Figure 3 These are the test results of the antioxidant technology effects of Ganoderma lucidum triterpenoid sustained-release microcapsules and different samples.

[0017] Figure 4 The bioaccessibility effect of Ganoderma triterpenoids in sustained-release microcapsules and Ganoderma triterpenoids treated with different solvents. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The specific embodiments described herein are only used to explain the present invention and are not used to limit the invention.

[0019] Unless otherwise specified, the raw materials and reagents used in the following examples were commercially available.

[0020] This example exemplifies a method for preparing a sustained-release microcapsule of a triterpenoid compound of Ganoderma lucidum. The specific steps are as follows:

[0021] 1) Dissolve 30 g / L of bovine serum albumin (BSA) and 30 g / L of octenyl succinate starch (OSAS) in ultrapure water, respectively, and stir overnight at room temperature to obtain BSA mother solution and OSAS mother solution;

[0022] 2) 20 g of BSA mother solution and 20 g of OSAS mother solution were mixed, and the pH of the solution was adjusted to 7 with 1 mol / L NaOH and HCl. The mixed solution was placed at 37°C for reaction for 12 h. After the sample was freeze-dried for 48 h, it was ground and spread in a culture dish and reacted in a constant temperature incubator at 60°C for 48 h. After taking out the sample, unreacted octenyl succinate starch ester OSAS was removed to obtain a BSA-OSAS glycosylation product solution;

[0023] 3) 1 g of 10 g / L ethanol solution of triterpenoids of Ganoderma lucidum was added to 50 g of BSA-OSAS glycosylation product solution, and the solution was ultrasonicated at 180 W for 6 min after magnetic stirring for 60 min to obtain a sustained-release microcapsule solution of triterpenoids of Ganoderma lucidum, and then freeze-dried to obtain sustained-release microcapsules of triterpenoids of Ganoderma lucidum.

[0024] In order to verify the performance of the sustained-release microcapsules of triterpenoids of Ganoderma lucidum prepared in this example, an in vitro digestion simulation experiment was conducted, with triterpenoids of Ganoderma lucidum alone as a comparison. According to the data, the decomposition of triterpenoids of Ganoderma lucidum in human oral saliva is very limited. This is mainly because the enzymes in saliva do not have the ability to decompose triterpenoids, the physical and chemical environment in saliva also limits the possibility of decomposition, and their residence time in the oral cavity is short. Therefore, the decomposition percentage of triterpenoids of Ganoderma lucidum in oral saliva is very low. The degradation rates of triterpenoids of Ganoderma lucidum and the sustained-release microcapsules of triterpenoids of Ganoderma lucidum prepared in this example in simulated gastric fluid are shown in Figure 2. Figure 1 As shown by Figure 1 It can be seen that at this stage, the digestibility of microcapsules and pure Ganoderma triterpenes is not much different, but there is also a slight protective effect.

[0025] The degree of decomposition of Ganoderma triterpenoids in gastric juice mainly depends on their chemical properties as well as the acidity of gastric juice and the action of enzymes. Gastric juice contains hydrochloric acid, and the pH value is between 1.5 and 3.5, which is strongly acidic. In addition, the main enzyme contained in gastric juice is pepsin, which mainly acts on the decomposition of protein. Ganoderma triterpenoids usually have a complex three-ring or five-ring structure, have a certain chemical stability, and have good tolerance to acidic environments. And most of them are hydrophobic molecules that are not easily soluble in water, which may reduce their chances of contact with acid and enzymes in gastric juice. Figure 1It can be seen that the decomposition ratio of Ganoderma triterpenoids in simulated gastric juice is low, and the microcapsules can significantly improve the stability of Ganoderma triterpenoids in simulated gastric juice by 30%, significantly avoiding their degradation in the gastric acid environment, thereby allowing more active ingredients to enter the intestine.

[0026] Intestinal fluid contains a variety of enzymes, including pancreatic enzymes, enteropeptidases, lipases, etc., and the pH value is approximately between 7.5-8.5. In intestinal fluid, the absorption and metabolism of Ganoderma triterpenoids is the most critical stage. Enzymes in the intestine and higher pH values ​​can promote the dissolution and absorption of triterpenoids. Some triterpenoids can enter the blood circulation through passive diffusion or active transport in the intestine, thereby exerting their biological activity. Triterpenoids generally have low water solubility, which limits their absorption in the digestive tract. The degradation rates of Ganoderma triterpenoids and the Ganoderma triterpenoid sustained-release microcapsules prepared in this embodiment in simulated intestinal fluid are shown in Figure 2. Figure 2 As shown by Figure 2 It can be seen that the reaction products generated by the Maillard reaction can increase the water solubility of triterpenoids, thereby improving their solubility in gastrointestinal fluids; embedding into microcapsules can also help protect triterpenoids from degradation by gastric acid and enzymes, thereby improving their stability. Figure 2 It can be seen that the microcapsules enhance the digestibility of Ganoderma triterpenes, with the rate of improvement being 40% in the three-hour time interval.

[0027] Furthermore, the antioxidant effect of the ganoderma triterpenoid sustained-release microcapsules prepared in this example was tested, and the test results are as follows: Figure 3 As shown by Figure 3 It can be seen that the ABTS cation free radical active scavenging rate (86.12%) and DPPH free radical scavenging rate (84.79%) of the microcapsules are significantly higher than those of other samples. The results show that the microcapsules prepared by the present invention significantly improve the antioxidant activity of Ganoderma lucidum triterpenoid compounds compared with microcapsules prepared by other methods.

[0028] The bioaccessibility effect analysis of the Ganoderma triterpenoid sustained-release microcapsules prepared in this example shows that the bioaccessibility of the unencapsulated Ganoderma triterpenoids after intestinal digestion is (9.94±0.59)%. The bioaccessibility of the Ganoderma triterpenoids after encapsulation with the BSA-OSAS Maillard reaction product is (43.24±2.04)%, which is significantly higher than the microcapsules (26.99±1.01)% prepared by other methods. It can be seen that BSA-OSAS-Ganoderma triterpenoid microcapsules can significantly improve the bioaccessibility of Ganoderma triterpenoids.

[0029] Finally, this embodiment also analyzes the storage stability and stability effect of the prepared Ganoderma triterpenoid sustained-release microcapsules under different temperature, acid-base and ion concentration conditions. It can be seen that the microcapsules prepared by the present invention have significantly improved storage stability and different temperature, acid-base and ion concentration conditions compared with other methods; pH stability effect analysis is performed. As the pH increases from 2.0 to 9.0, the particle size of the microcapsules shows a decreasing trend, and all values ​​are less than 300nm, indicating that the microcapsules show better anti-aggregation stability in a higher pH range. The trend of zeta potential is similar to that of particle size, indicating that the microcapsules have good stability. The particle size of the present microcapsules treated in the range of 20 to 90°C fluctuates around 254nm. The PDI change is similar to the particle size. At different treatment temperatures, the PDI ranges from 0.21 to 0.32, and the PDI of microcapsules with larger particle sizes is higher. In addition, the zeta potential changes slightly, and the potential values ​​are all about -11mV at different treatment temperatures, indicating that the microcapsule solution has good stability.

[0030] In summary, the present invention uses bovine serum albumin and octenyl succinate starch ester to prepare Maillard products by carbonyl-amino condensation as wall materials, so that the core material Ganoderma lucidum triterpenoid compounds are well wrapped in the wall materials, the Maillard reaction products and Ganoderma lucidum triterpenoid compounds are cross-linked, the structure of the microcapsules is not destroyed after freeze-drying, and the granular microcapsules prepared by the present invention have a complete structure, high embedding rate and good stability. The sustained-release performance of the embedded Ganoderma lucidum triterpenoid compounds is improved, the polymer properties are changed by conjugation, thereby increasing its water solubility, improving its dissolution rate and intestinal mucosal permeability, extending the residence time by bioadhesion, and improving its oral bioavailability, which has a relatively obvious improvement and enhancement effect on the human body absorption and utilization of Ganoderma lucidum triterpenoid compounds.

[0031] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims

1. A method for preparing a sustained-release microcapsule of triterpenoid compounds of Ganoderma lucidum, characterized in that: The following steps are involved: S1, dissolving 30 g / L bovine serum albumin (BSA) and 30 g / L octenyl succinate starch ester (OSAS) in ultrapure water respectively, and stirring at room temperature overnight to obtain BSA mother solution and OSAS mother solution; S2. Mix BSA mother solution and OSAS mother solution in a mass ratio of 1:1, adjust the solution pH to 7, place the mixed solution at 37°C for reaction for 12 hours, freeze-dry the sample for 48 hours, grind and spread it in a culture dish, and react it in a constant temperature incubator at 60°C for 48 hours. After taking out the sample, remove the unreacted octenyl succinate starch ester OSAS to obtain a BSA-OSAS glycosylation product solution; S3. Take the ethanol solution of triterpenoids of Ganoderma lucidum and add it to the BSA-OSAS glycosylation product solution to make the mass ratio of the BSA-OSAS glycosylation product solution to the ethanol solution of triterpenoids of Ganoderma lucidum be 50:

1. After magnetic stirring for 60 minutes, ultrasonicate the solution at 180W for 6 minutes to obtain the sustained-release microcapsule solution of triterpenoids of Ganoderma lucidum. After freeze-drying, obtain the sustained-release microcapsules of triterpenoids of Ganoderma lucidum.

2. The method for preparing the ganoderma triterpenoid sustained-release microcapsules according to claim 1, characterized in that: In the step S2, 1 mol / L NaOH and HCl are used to adjust the pH of the solution.

3. The method for preparing the ganoderma triterpenoid sustained-release microcapsules according to claim 1, characterized in that: In the step S3, the concentration of the Ganoderma triterpenoid compound ethanol solution is 10 g / L.

4. A sustained-release microcapsule of triterpenoid compounds of Ganoderma lucidum prepared by the method according to any one of claims 1 to 3.

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