A method for preparing a gamma-aminobutyric acid-embedded particle

Abstract

The application discloses a preparation method of gamma-aminobutyric acid particle embedding and health-care product application thereof, and first prepares a double bond starch through lactate dehydrogenase catalysis, obtains an intermediate product by combining allyl glycidyl ether modified cyclodextrin, then carries out cross-linking reaction with cinnamic acid ester to obtain multifunctional modified starch, finally, compounding gelatinized starch and cyclodextrin to construct a composite embedding system, and loading gamma-aminobutyric acid to obtain a finished product. The application introduces active double bond, cyclodextrin cavity and cross-linking structure into the starch skeleton, effectively improves the interface compatibility of each raw material component, improves the defects of uneven dispersion, easy agglomeration and loose structure of the traditional embedding system, and constructs a compact and stable three-dimensional network structure. The structure can significantly improve the encapsulation rate and loading capacity of gamma-aminobutyric acid, effectively resist external environmental interference, inhibit the degradation, loss and burst release of active ingredients, and greatly improve the product stability and bioavailability.

Description

Technical Field

[0001] This invention relates to the field of health product technology, and in particular to a γ-aminobutyric acid (GABA) encapsulated granules and its preparation method. Background Technology

[0002] Gamma-aminobutyric acid (GABA) is a naturally occurring functional non-protein amino acid with a variety of excellent physiological activities, including calming the nerves, improving sleep, regulating blood pressure, and relieving fatigue. It is a core functional ingredient widely used in the health supplement industry, with broad market prospects. However, naturally occurring free GABA has significant limitations. It is highly water-soluble and structurally unstable, making it prone to decomposition and degradation during high-temperature processing, acidic or alkaline environments, light exposure, and long-term storage. Furthermore, free GABA is rapidly metabolized and degraded after entering the human body, significantly reducing its bioavailability and severely limiting its effectiveness and product quality in high-end health supplements.

[0003] To overcome the shortcomings of γ-aminobutyric acid (GABA), such as poor stability, easy loss, and low utilization rate, existing technologies generally employ microencapsulation technology to modify GABA. This involves constructing a protective structure using carrier materials to protect the active ingredient and achieve a sustained-release effect. Currently, the mainstream encapsulation carriers in the industry mostly use single ordinary starch, single cyclodextrin, or conventionally modified starch systems. Among these, ordinary corn starch and tapioca starch are safe, non-toxic, and inexpensive, making them commonly used encapsulation substrates in the food and health product fields. However, ordinary starch has a simple structure, lacks active groups, and has poor porosity and encapsulation properties, resulting in low GABA encapsulation loading and insufficient encapsulation density, failing to effectively lock in the active ingredient. Single cyclodextrin possesses a special cavity structure, enabling the encapsulation and fixation of small molecules; however, using cyclodextrin alone is costly, and its film-forming properties and structural stability are poor, making it difficult to form a complete and stable particulate structure with weak resistance to external environmental interference.

[0004] Existing technologies also include a small number of modified starch-cyclodextrin encapsulation systems, but the modified starches used are mostly simple esterification or etherification products with single functional groups, which cannot form an effective synergistic effect with the matrix starch and cyclodextrin. The chemical compatibility between the raw material components is poor, and the mixed system is prone to problems such as uneven dispersion, particle agglomeration, and component separation. Ultimately, this results in uneven particle size and poor encapsulation stability of the prepared encapsulated particles. During storage, leakage, degradation, and burst release of γ-aminobutyric acid are likely to occur, causing unstable efficacy and shortened shelf life of the finished health products. At the same time, the encapsulation efficiency and loading capacity of conventional systems are limited, making it difficult to meet the needs of high-quality, large-scale health product production and application. Summary of the Invention

[0005] This invention discloses a method for preparing γ-aminobutyric acid (GABA) encapsulated particles and its application in health products. First, double-bonded starch is prepared via lactate dehydrogenase catalysis. Then, cyclodextrin modified with allyl glycidyl ether is used to obtain an intermediate product. This intermediate product is then cross-linked with cinnamic acid ester to prepare a multifunctional modified starch. Finally, a composite encapsulation system is constructed by combining gelatinized starch and cyclodextrin, and GABA is loaded onto the starch to obtain the final product. This invention introduces active double bonds, cyclodextrin cavities, and cross-linking structures into the starch backbone, effectively improving the interfacial compatibility of the raw material components and overcoming the defects of uneven dispersion, easy aggregation, and loose structure in traditional encapsulation systems, thus constructing a dense and stable three-dimensional network structure. This structure can significantly improve the encapsulation efficiency and loading of GABA, effectively resist external environmental interference, inhibit the degradation, loss, and burst release of active ingredients, and greatly improve product stability and bioavailability. This process is mild, safe, and highly controllable, suitable for industrial mass production, and applicable to the preparation of various health products.

[0006] Therefore, it is necessary to provide a method for preparing γ-aminobutyric acid (GABA) particles, comprising the following steps: S1. Gelatinize the starch to obtain gelatinized starch; S2. Mix the gelatinized starch with the modified starch to obtain a suspension, and then mix it with cyclodextrin to obtain encapsulated particles; S3. Mix the encapsulated particles with γ-aminobutyric acid to obtain γ-aminobutyric acid encapsulated particles; The modified starch is obtained by reacting double-bonded starch, allyloxycyclodextrin, and cinnamic acid ester.

[0007] Furthermore, the starch is selected from one or more of corn starch, potato starch, wheat starch, and tapioca starch.

[0008] Furthermore, in step S1, the gelatinization temperature is 90-95℃.

[0009] Furthermore, the mass ratio of the gelatinized starch, modified starch, and cyclodextrin is 10-12:0.5-1:1-2.

[0010] Furthermore, the mass ratio of the encapsulated particles to γ-aminobutyric acid is 5-10:1-2.

[0011] Furthermore, the modified starch preparation method includes the following steps: A1-1. Mix water, gelatinized corn starch, and emulsifier to obtain a starch emulsion; A1-2. Mix starch emulsion, lactate dehydrogenase, and water, and heat to react to obtain double-bonded starch; A2. Allyl glycidyl ether was mixed with cyclodextrin, the pH was adjusted to alkaline, and the reaction was carried out at room temperature to obtain the intermediate product; A3. The intermediate product, double-bonded starch, and cinnamic acid ester are mixed and heated to react under the action of an emulsifier and an initiator to obtain the modified starch.

[0012] Furthermore, in steps A1-2, the temperature of the heating reaction is 75-85°C; In step A3, the temperature of the heating reaction is 60-80℃.

[0013] Further, in step A2, the mass ratio of the cyclodextrin to allyl glycidyl ether is 1:2-4.

[0014] Further, in step A3, the mass ratio of the intermediate product, double-bonded starch, and cinnamic acid ester is 1-2:1-2:0.5-1.5.

[0015] The present invention also provides the application of the γ-aminobutyric acid (GABA) particles prepared by the above-described method in the field of health products.

[0016] Meanwhile, this invention uses common food-grade raw materials such as starch, combined with mild modification reactions and encapsulation processes. The overall preparation process is simple, the conditions are controllable, and it is green and safe, without the introduction of any toxic or harmful substances. It is compatible with the safety production standards in the health product field, and can achieve large-scale industrial production, effectively reducing production costs. It has extremely high industrialization promotion value and market application prospects.

[0017] The present invention has the following beneficial effects: This invention prepares double-bonded starch by lactate dehydrogenase catalysis, and then reacts it with allyloxycyclodextrin and cinnamic acid ester to obtain regular and dense modified starch. At the same time, the introduction of cinnamic acid ester can introduce hydrophobic groups, UV-resistant active structures and cyclodextrin cavity functional structures into the polymerization system. By introducing allyloxycyclodextrin, the problems of free cyclodextrin precipitation and uneven dispersion can be avoided. At the same time, a large number of cavities for adsorbing γ-aminobutyric acid (GABA) can be retained for system stability, which greatly improves the encapsulation loading and encapsulation efficiency of GABA. It effectively enhances the binding and protection of GABA, and can block the destruction of GABA activity by external environmental factors such as temperature, humidity, acid and alkali. It avoids the degradation, loss and burst release of active ingredients during processing and storage, significantly improves the storage stability and bioavailability of encapsulated particles, and ensures the stability of the efficacy of health care products.

[0018] Meanwhile, cinnamic acid esters have a hydrophobic shielding structure and UV blocking properties, which effectively compensate for the poor light resistance, heat resistance and oxidation resistance of traditional starch carriers. They form multiple protective barriers for γ-aminobutyric acid, which can effectively block the interference of harsh external environments such as temperature, humidity, light, acid and alkali, and effectively avoid problems such as oxidative degradation, leakage and loss of active ingredients during the processing and storage of γ-aminobutyric acid. Detailed Implementation

[0019] To more clearly illustrate the technical solution of the present invention, the following embodiments are provided. Unless otherwise stated, the raw materials, reactions, and post-processing methods appearing in the embodiments are all commercially available raw materials and technical methods well known to those skilled in the art.

[0020] The terms "preferred," "more preferably," and "more suitable" used in this invention refer to embodiments of the invention that provide certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of this invention.

[0021] It should be understood that, except in any operational instance or otherwise indicated, all figures representing the amounts of ingredients used, for example, in the specification and claims, should be understood to be modified in all cases by the term "about". Therefore, unless otherwise stated, the numerical parameters set forth in the following specification and appended claims are approximations varying with the desired performance to be obtained according to the invention.

[0022] Cyclodextrin: β-Cyclodextrin: Content: ≥98.0% Hebei Baiyou Biotechnology Co., Ltd.

[0023] γ-Aminobutyric acid: Content: ≥98.0% Bloomage Biotechnology Co., Ltd.

[0024] Cinnamyl ester: Cinnamyl cinnamate.

[0025] Corn starch: High amylose HI-55, purchased from Quanyinxiangyu (Beijing) Biotechnology Co., Ltd.

[0026] Lactate dehydrogenase: D65820-5KU, purchased from Acmec.

[0027] Emulsifier: Triton X-100.

[0028] Initiator: Ammonium persulfate. Example 1

[0029] A method for preparing γ-aminobutyric acid (GABA) particles includes the following steps: S1. Mix corn starch with water and heat to 95°C for 35 minutes to gelatinize the corn starch. The mass ratio of corn starch to water is 1:20; S2. Using anhydrous ethanol as a solvent, the gelatinized starch and modified starch are mixed and ultrasonically treated for 30 minutes to obtain a suspension. Then, the suspension is mixed with cyclodextrin until homogeneous, the solvent is removed, and the mixture is freeze-dried to obtain the encapsulated particles. The mass ratio of the gelatinized starch, modified starch, and cyclodextrin is 10:0.8:1; S3. Using water as a solvent, the embedded particles are mixed with γ-aminobutyric acid, the mixture is thoroughly mixed, the solvent is removed, and the mixture is freeze-dried to obtain embedded γ-aminobutyric acid particles. The mass ratio of the encapsulated particles to γ-aminobutyric acid is 8:1; The modified starch preparation method includes the following steps: A1-1. Mix water, gelatinized corn starch, and emulsifier to obtain a starch emulsion; The mass ratio of water, gelatinized corn starch, and emulsifier is 1:10:0.05; A1-2. Mix starch emulsion, lactate dehydrogenase and water, heat to 80℃ and react for 30 min, centrifuge, take the precipitate, freeze dry to obtain double bond starch; The mass ratio of the starch emulsion, lactate dehydrogenase, and water is 10:0.02:1. A2. Using water as a solvent, allyl glycidyl ether and cyclodextrin were mixed, and 10 wt% NaOH was added to adjust the pH to 10. The mixture was reacted at room temperature for 10 h, the solution was neutralized with hydrochloric acid, filtered, washed, and the solvent was removed to obtain the intermediate product. The mass ratio of allyl glycidyl ether to cyclodextrin is 2:1; A3. Using water as a solvent, the intermediate product, double-bonded starch and cinnamic acid ester are mixed and reacted at 65°C for 4 hours under the action of emulsifier and initiator and nitrogen. Anhydrous ethanol is added to break the emulsion, the precipitate is washed with anhydrous ethanol and dried to obtain the modified starch. The mass ratio of water, intermediate product, double-bonded starch, cinnamic acid ester, emulsifier and initiator is 20:2:2:1.5:0.5:0.05. Example 2

[0030] A method for preparing γ-aminobutyric acid (GABA) particles includes the following steps: S1. Mix corn starch with water and heat to 92℃ for 35 minutes to gelatinize, thus obtaining gelatinized corn starch; The mass ratio of corn starch to water is 1:20; S2. Using anhydrous ethanol as a solvent, the gelatinized starch and modified starch are mixed and ultrasonically treated for 30 minutes to obtain a suspension. Then, the suspension is mixed with cyclodextrin until homogeneous, the solvent is removed, and the mixture is freeze-dried to obtain the encapsulated particles. The mass ratio of the gelatinized starch, modified starch, and cyclodextrin is 10:1:2; S3. Using water as a solvent, the embedded particles are mixed with γ-aminobutyric acid, the mixture is thoroughly mixed, the solvent is removed, and the mixture is freeze-dried to obtain embedded γ-aminobutyric acid particles. The mass ratio of the encapsulated particles to γ-aminobutyric acid is 10:1.5; The modified starch preparation method includes the following steps: A1-1. Mix water, gelatinized corn starch, and emulsifier to obtain a starch emulsion; The mass ratio of water, gelatinized corn starch, and emulsifier is 1:10:0.05; A1-2. Mix starch emulsion, lactate dehydrogenase and water, heat to 80℃ and react for 30 min, centrifuge, take the precipitate, freeze dry to obtain double bond starch; The mass ratio of the starch emulsion, lactate dehydrogenase, and water is 10:0.02:1. A2. Using water as a solvent, allyl glycidyl ether and cyclodextrin were mixed, and 10 wt% NaOH was added to adjust the pH to 10. The mixture was reacted at room temperature for 10 h, the solution was neutralized with hydrochloric acid, filtered, washed, and the solvent was removed to obtain the intermediate product. The mass ratio of allyl glycidyl ether to cyclodextrin is 2:1; A3. Using water as a solvent, the intermediate product, double-bonded starch and cinnamic acid ester are mixed and reacted at 65°C for 4 hours under the action of emulsifier and initiator and nitrogen. Anhydrous ethanol is added to break the emulsion, the precipitate is washed with anhydrous ethanol and dried to obtain the modified starch. The mass ratio of water, intermediate product, double-bonded starch, cinnamic acid ester, emulsifier and initiator is 20:2:2:1.5:0.5:0.05. Example 3

[0031] A method for preparing γ-aminobutyric acid (GABA) particles includes the following steps: S1. Mix corn starch with water and heat to 95°C for 35 minutes to gelatinize the corn starch. The mass ratio of corn starch to water is 1:20; S2. Using anhydrous ethanol as a solvent, the gelatinized starch and modified starch are mixed and ultrasonically treated for 30 minutes to obtain a suspension. Then, the suspension is mixed with cyclodextrin until homogeneous, the solvent is removed, and the mixture is freeze-dried to obtain the encapsulated particles. The mass ratio of the gelatinized starch, modified starch, and cyclodextrin is 12:1:1.5; S3. Using water as a solvent, the embedded particles are mixed with γ-aminobutyric acid, the mixture is thoroughly mixed, the solvent is removed, and the mixture is freeze-dried to obtain embedded γ-aminobutyric acid particles. The mass ratio of the encapsulated particles to γ-aminobutyric acid is 10:2; The modified starch preparation method includes the following steps: A1-1. Mix water, gelatinized corn starch, and emulsifier to obtain a starch emulsion; The mass ratio of water, gelatinized corn starch, and emulsifier is 1:10:0.05; A1-2. Mix starch emulsion, lactate dehydrogenase and water, heat to 80℃ and react for 30 min, centrifuge, take the precipitate, freeze dry to obtain double bond starch; The mass ratio of the starch emulsion, lactate dehydrogenase, and water is 10:0.02:1. A2. Using water as a solvent, allyl glycidyl ether and cyclodextrin were mixed, and 10 wt% NaOH was added to adjust the pH to 10. The mixture was reacted at room temperature for 10 h, the solution was neutralized with hydrochloric acid, filtered, washed, and the solvent was removed to obtain the intermediate product. The mass ratio of allyl glycidyl ether to cyclodextrin is 2:1; A3. Using water as a solvent, the intermediate product, double-bonded starch and cinnamic acid ester are mixed and reacted at 65°C for 4 hours under the action of emulsifier and initiator and nitrogen. Anhydrous ethanol is added to break the emulsion, the precipitate is washed with anhydrous ethanol and dried to obtain the modified starch. The mass ratio of water, intermediate product, double-bonded starch, cinnamic acid ester, emulsifier and initiator is 20:2:2:1.5:0.5:0.05.

[0032] Comparative Example 1 A comparative example of γ-aminobutyric acid (GABA) particles differs from Example 1 in that: step A2 is removed, and in step A3, double-bonded starch is used to replace an equal mass of intermediate product; other components and preparation methods are the same.

[0033] Comparative Example 2 A comparative example of γ-aminobutyric acid (GABA) particles differs from Example 1 in that, in step A3, ethyl acrylate is used to replace an equal mass of cinnamate, while the other components and preparation methods remain the same.

[0034] Test case The performance of the γ-aminobutyric acid-encapsulated particles prepared in Example 1 and Comparative Examples 1-2 was tested.

[0035] Encapsulation efficiency test: The encapsulated γ-aminobutyric acid particles from Examples 1 and Comparative Examples 1-2 were mixed with water to obtain a suspension of 10 mg / mL. After centrifugation for 30 min, the precipitate was washed, and all supernatant was collected. After filtration through a 0.22 μm filter membrane, the supernatant was added to an inner-lined tube and placed in a liquid chromatography vial for analysis. The encapsulation efficiency was calculated as follows:

[0036] In the formula: M1 represents the total amount of added γ-aminobutyric acid (mg), and M2 represents the amount of free (unencapsulated) γ-aminobutyric acid (mg). Stability Test: Constant Temperature and Humidity Stability: The γ-aminobutyric acid particles of Example 1 and Comparative Examples 1-2 were placed in an environment with a temperature of 37±2℃ and a humidity of 75% for 2 months. After being kept at this temperature, they were immediately removed and observed. If there was no obvious discoloration or clumping, the constant temperature and humidity stability was deemed to be qualified.

[0037] Room temperature stability: The γ-aminobutyric acid particles of Example 1 and Comparative Examples 1-2 were placed in an environment at 25±2℃ and kept at this temperature for 2 months. After that, they were taken out and observed immediately. If there was no obvious discoloration or clumping, the room temperature stability was deemed to be qualified.

[0038] Sleep test: Quarantined male ICR mice were randomly divided into three groups of 10 mice each. Each group received 50 mg / kg of the γ-aminobutyric acid (GABA) granules from Example 1 and Comparative Examples 1-2, respectively. After a week of acclimatization, the mice were administered the drugs via gavage for seven consecutive days, once daily. All tests were conducted between 9:00 AM and 11:00 AM. On day 7, all mice were injected intraperitoneally with an above-threshold dose of sodium pentobarbital (50 mg / kg), and the mice were placed on a warm mat (37°C) with their abdomens facing upwards. The duration of sleep in the mice was recorded (the time from the disappearance of the righting reflex to the recovery of the righting reflex).

[0039] The test results are shown in Table 1.

[0040] Table 1. Performance test results of the γ-aminobutyric acid particles prepared in Example 1 and Comparative Examples 1-2

[0041] According to Table 1, the embedding rate of the sample prepared by the present invention is significantly higher than that of comparative examples 1-2. After being placed in constant temperature and humidity environment and normal temperature environment for 2 months, the sample of the present invention did not show discoloration or clumping, demonstrating excellent stability, and the sleep time was longer after taking it.

[0042] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

[0043] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for preparing γ-aminobutyric acid (GABA) particles, characterized in that, Includes the following steps: S1. Gelatinize the starch to obtain gelatinized starch; S2. Mix the gelatinized starch with the modified starch to obtain a suspension, and then mix it with cyclodextrin to obtain encapsulated particles; S3. Mix the encapsulated particles with γ-aminobutyric acid to obtain γ-aminobutyric acid encapsulated particles; The modified starch is obtained by reacting double-bonded starch, allyloxycyclodextrin, and cinnamic acid ester.

2. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 1, characterized in that, The starch is selected from one or more of corn starch, potato starch, wheat starch, and cassava starch.

3. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 1, characterized in that, In step S1, the gelatinization temperature is 90-95℃.

4. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 1, characterized in that, The mass ratio of the gelatinized starch, modified starch, and cyclodextrin is 10-12:0.5-1:1-2.

5. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 1, characterized in that, The mass ratio of the encapsulated particles to γ-aminobutyric acid is 5-10:1-2.

6. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 1, characterized in that, The modified starch preparation method includes the following steps: A1-1. Mix water, gelatinized corn starch, and emulsifier to obtain a starch emulsion; A1-2. Mix starch emulsion, lactate dehydrogenase, and water, and heat to react to obtain double-bonded starch; A2. Allyl glycidyl ether was mixed with cyclodextrin, the pH was adjusted to alkaline, and the reaction was carried out at room temperature to obtain the intermediate product; A3. The intermediate product, double-bonded starch, and cinnamic acid ester are mixed and heated to react under the action of an emulsifier and an initiator to obtain the modified starch.

7. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 6, characterized in that, In steps A1-2, the temperature of the heating reaction is 75-85℃; In step A3, the temperature of the heating reaction is 60-80℃.

8. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 6, characterized in that, In step A2, the mass ratio of the cyclodextrin to allyl glycidyl ether is 1:2-4.

9. The method for preparing γ-aminobutyric acid (GABA) particles according to claim 6, characterized in that, In step A3, the mass ratio of the intermediate product, double-bonded starch, and cinnamic acid ester is 1-2:1-2:0.5-1.

5.

10. The application of the γ-aminobutyric acid (GABA) particles prepared by the method of any one of claims 1-9 in the field of health products.

Images