Preparation process of microcapsule composition containing adenosine and application of microcapsule composition in preparation of anti-aging effect product

Abstract

The invention discloses a preparation process of an adenosine-containing microcapsule composition and application of the adenosine-containing microcapsule composition in preparation of an anti-aging effect product in the technical field of microcapsule compositions. The preparation method comprises the following steps: dispersing the succinic acid modified cyclodextrin coated adenosine microcapsules, the polygonatum sibiricum fermented polypeptide, the humectant and the emulsifier into deionized water, stirring, homogenizing and dispersing to obtain a mixed solution, stirring and mixing the mixed solution, the thickener and the antioxidant, homogenizing and dispersing at high pressure, and finally defoaming, slowly stirring, standing and cooling the mixed solution. The stability and effect exertion of adenosine are remarkably improved through the succinic acid modified cyclodextrin coating technology, the succinic acid modified cyclodextrin coated adenosine microcapsules and the polygonatum sibiricum fermented polypeptide are in synergistic compatibility, the composition exerts the excellent anti-aging effect and meanwhile improves the moisturizing capacity, in addition, the polygonatum sibiricum fermented polypeptide can inhibit formation of AGEs, and the anti-aging effect of the composition is improved. A comprehensive anti-aging system is formed through oxidation resistance, saccharification resistance and moisturizing, and the anti-aging composition has a wide application prospect in the field of cosmetics.

Description

Technical Field

[0001] This invention relates to the field of microcapsule composition technology, and more specifically to a manufacturing process for a microcapsule composition containing adenosine and its use in the preparation of anti-aging products. Background Technology

[0002] Adenosine (AD) is a nucleoside composed of adenine molecules and ribose. Studies have found that cordycepin and adenosine are the most important bioactive components of Cordyceps militaris. Adenosine possesses a wide range of pharmacological effects, such as anti-aging, anti-inflammatory, and anti-tumor properties. The physiological functions of adenosine in the human body have also been extensively studied, including regulating cardiovascular function and participating in sleep regulation. Furthermore, adenosine can exert various pharmacological effects through interaction with specific adenosine receptors, such as reducing pathological arrhythmias. Therefore, adenosine, as a bioactive substance, has broad application prospects in the fields of medicine, cosmetics, and food.

[0003] In the prior art, for example, patent publication number CN109363990A describes an anti-wrinkle and anti-aging skincare composition, the raw materials of which include adenosine, chamomile extract, baobab extract, deionized water, sodium hyaluronate solution, and polypeptide complex. The composition is obtained by stirring and dispersing evenly. This composition is effective, gentle, non-irritating, and has no side effects. Patent publication number CN107411982A describes a whitening, moisturizing, and repairing skincare composition and its preparation method and application, the raw materials of which include water, sodium hyaluronate, sodium dihydrogen phosphate dihydrate, potassium chloride, sodium hydroxide, calcium chloride, glutamine, and benzalkonium chloride. This composition contains magnesium sulfate, GABA, sodium ascorbate, alanine, arginine, lysine hydrochloride, valine, histidine, leucine, taurine, coenzyme A, alcohol, polysorbate 80, thiamine, disodium diphosphate, and recombinant human epidermal growth factor. This combination can penetrate the skin's base layer and has whitening, moisturizing, and repairing effects. However, in practical applications, adenosine, as an active ingredient, suffers from poor water solubility and low stability at room temperature, which is detrimental to its dispersion in skincare compositions and limits its application and development in the field of cosmetic skin penetration. Therefore, improving the water solubility and stability of adenosine has become a current research focus.

[0004] Microcapsule technology can effectively protect the core ingredients inside through its shell, while altering the physical properties of the substance, controlling release, reducing volatility and toxic side effects, and achieving the delivery of active ingredients. In addition, with consumers paying more attention to the safety of ingredients, the development of skin care products containing plant extracts has gradually gained attention. Therefore, this invention proposes a manufacturing process for a microcapsule composition containing adenosine and its use in the preparation of anti-aging products. Summary of the Invention

[0005] The purpose of this invention is to solve the above-mentioned technical problems by providing a manufacturing process for a microcapsule composition containing adenosine and its use in the preparation of anti-aging products.

[0006] The present invention achieves the above objectives through the following technical solutions: As a first aspect of the present invention, a process for manufacturing a microcapsule composition containing adenosine is provided, comprising the following steps: (1) Disperse 0.5-5% by mass of succinic acid-modified cyclodextrin-coated adenosine microcapsules, 2-5% by mass of Polygonatum odoratum fermented polypeptide, 1-4% by mass of humectant, and 0.05-0.2% by mass of emulsifier into deionized water and stir to mix. Then, homogenize the mixture at 80℃-85℃ and 10000rpm-15000rpm for 30s-60s to obtain a mixture. (2) After cooling the mixture to 50℃-55℃, add 0.05%-0.1% by mass of thickener and 0.2%-0.5% by mass of antioxidant. After stirring and mixing, homogenize under high pressure at 600bar-800bar and 10000rpm-15000rpm for 60s-90s. Then, after degassing, slow stirring and cooling, the adenosine microcapsule composition can be obtained.

[0007] As a further optimized solution of the present invention, the manufacturing process of the succinic acid modified cyclodextrin-coated adenosine microcapsules is as follows: succinic acid modified cyclodextrin is dispersed in an ethanol aqueous solution with a mass fraction of 50%-80%, stirred and dissolved at 60℃-80℃, and then adenosine is added and stirred and dissolved evenly to obtain a solution. The solution is then vacuum spray-dried to obtain the succinic acid modified cyclodextrin-coated adenosine microcapsules.

[0008] As a further optimization of the present invention, the amount of adenosine added is 10%-30% of the mass of the succinic acid-modified cyclodextrin, and the cyclodextrin is β-cyclodextrin or hydroxypropyl-β-cyclodextrin.

[0009] As a further optimized solution of the present invention, the manufacturing process of the succinic acid modified cyclodextrin is as follows: succinic acid and cyclodextrin are dispersed in N,N-dimethylformamide solvent, heated to 90℃-120℃ and stirred until uniformly dissolved, an acidic catalyst is added and stirred to react, and then the solvent and unreacted raw materials are removed by vacuum evaporation to obtain succinic acid modified cyclodextrin.

[0010] As a further optimized solution of the present invention, the manufacturing process of the Polygonatum fermentation polypeptide is as follows: first, the crushed Polygonatum tuber is added to the fermentation culture medium to obtain the fermentation substrate, then activated brewing yeast is inoculated into the fermentation substrate for liquid fermentation, after liquid fermentation, the fermentation liquid is obtained by centrifugation, and then the fermentation liquid is obtained by protein extraction and enzymatic hydrolysis with compound enzymes.

[0011] As a further optimization of the present invention, the inoculation amount of the brewing yeast is 5%-10% of the fermentation substrate volume.

[0012] As a further optimization of the present invention, the complex enzyme is a mixture of equal masses of neutral protease and alkaline protease.

[0013] As a further optimization of the present invention, the moisturizer is hyaluronic acid or glycerin, the emulsifier is sucrose palmitate or glyceryl linoleate, the thickener is carbomer, and the antioxidant is tocopheryl acetate.

[0014] As a second aspect of the invention, an adenosine microcapsule composition obtained by any of the manufacturing processes described above is also provided.

[0015] As a third aspect of the invention, the use of the adenosine microcapsule composition as described in any of the above descriptions in the field of cosmetics is also provided.

[0016] The beneficial effects of this invention are as follows: This invention first involves dispersing succinic acid-modified cyclodextrin-coated adenosine microcapsules, Polygonatum sibiricum fermented polypeptides, humectants, and emulsifiers in deionized water, stirring, and homogenizing to obtain a mixture. Then, the mixture is mixed with a thickener and an antioxidant, stirred, and homogenized under high pressure. Finally, the mixture is defoamed, slowly stirred, and cooled to obtain an adenosine microcapsule composition. The succinic acid-modified cyclodextrin encapsulation technology significantly improved the stability and efficacy of adenosine. Furthermore, the combination of succinic acid-modified cyclodextrin-encapsulated adenosine microcapsules with Polygonatum sibiricum fermented peptides can effectively neutralize environmental oxidative stress, reduce oxidative damage, and thus delay the skin photoaging process, enhance the antioxidant capacity of the composition, and obtain a more superior anti-aging effect. At the same time, it also significantly improves the moisturizing effect of the composition, which can replace the use of some traditional moisturizers and is conducive to simplifying the formula. By fermenting Polygonatum sibiricum with Saccharomyces cerevisiae, followed by protein extraction and enzymatic hydrolysis with compound enzymes, Polygonatum sibiricum fermented polypeptides were obtained. The anti-glycation efficacy test results showed that it has a positive effect on inhibiting the formation of advanced glycation end products. Further compounding with succinic acid modified cyclodextrin-coated adenosine microcapsules can make the composition exert a more excellent anti-glycation effect. The synergistic combination of succinic acid-modified cyclodextrin-encapsulated adenosine microcapsules and Polygonatum sibiricum fermented peptides has created a comprehensive anti-aging system with antioxidant, anti-glycation, and moisturizing effects. Patch test results show that the composition did not cause skin irritation at any observation time point. The formula is simple and safe, and has broad application prospects in the cosmetic field. Detailed Implementation

[0017] The present application will now be described in further detail. It should be noted that the following specific embodiments are only used to further illustrate the present application and should not be construed as limiting the scope of protection of the present application. Those skilled in the art can make some non-essential improvements and adjustments to the present application based on the above application content.

[0018] Unless otherwise specified, all reagents and materials used below are commercially available products. Unless otherwise specified, all methods used below are conventional methods known to those skilled in the art.

[0019] 1. Obtaining adenosine Adenosine can be obtained commercially (CAS: 58-61-7) or through the following biosynthetic pathway. The biosynthetic process of adenosine includes the following steps: (1.1) Using an inoculation loop, pick 3-5 colonies of Cordyceps militaris (Cordyceps militaris strain ACCC50623) growing on agar plates and inoculate them into the culture medium. Incubate on slant at 25°C for 7 days, then transfer to a 200L seed tank and incubate at 25°C for 3 days. Then transfer to a 1000L seed tank and incubate at 25°C for 3-4 days to obtain the seed culture solution. The composition of the agar plate culture medium is as follows: potato extract powder 20.0 g / L, peptone 30.0 g / L, yeast extract powder 3.0 g / L, glucose 40.0 g / L, K2HPO4·3H2O 2.0 g / L, MgSO4·7H2O 0.20 g / L. (1.2) Fermentation tank culture: The seed culture obtained in the above steps was inoculated into a fermentation tank containing fermentation medium at an inoculation rate of 2%. The fermentation process lasted for 8 days. The first 3 days were for dark culture, and the 4th day was for light culture (light intensity: 360 lux). During the fermentation culture, the carbon source (glucose) concentration in the culture system was monitored. A 70% (mass percentage) glucose aqueous solution was used as feed to control the carbon source concentration in the culture system at 5-10 g / L. The cultivation parameters were as follows: fermenter aeration rate of 20 L / min, stirring speed of 200 rpm, tank pressure of 0.8 MPa, temperature of 25℃, and dissolved oxygen of 35%. After fermentation, the entire culture system was collected, and mycelium was collected using a plate centrifuge (centrifuged at 3000 rpm for 30 min), and then dried at 60℃ until the moisture content was 7%. (1.3) Adenosine extraction: The mycelium was ground, and 0.5 g of powder was taken. Water was added to a final volume of 10 mL, and the mixture was extracted with ultrasound (40 kHz, 225 W) for 60 min. After centrifugation at 5000 rpm for 15 min, the supernatant was collected and filtered through a 0.22 μm microporous membrane to obtain the adenosine test solution. The test solution was transferred to a vacuum drying oven, and the temperature was controlled at 60 °C and the vacuum degree at 0.092 MPa for 10 h to obtain dried adenosine crystals. (1.4) HPLC detection: The chromatographic column was Sunfire RC18 (4.6 mm × 250 mm, 5 μm), the mobile phase was methanol-water (20:80), the flow rate was 1 mL / min, the detection wavelength was 260 nm, the column temperature was 25 ± 5 ℃, and the injection volume was 10 μL. The product was confirmed to be adenosine by HPLC detection.

[0020] 2. Preparation of succinic acid-modified cyclodextrin-coated adenosine microcapsules First, succinic acid and cyclodextrin are added to N,N-dimethylformamide solvent at a mass ratio of 30:100 and heated to 90℃-120℃ (preferably 100℃ in this embodiment) and stirred until uniformly dissolved. After adding the acidic catalyst sodium dihydrogen phosphate and stirring for 6 hours, the solvent and unreacted raw materials are removed by vacuum evaporation at 120℃ to obtain succinic acid modified cyclodextrin. The cyclodextrin can be β-cyclodextrin or hydroxypropyl-β-cyclodextrin, and in this embodiment, hydroxypropyl-β-cyclodextrin is preferred. Then, succinic acid modified cyclodextrin is added to an ethanol aqueous solution with a mass fraction of 50%-80% (preferably 75% in this embodiment). After stirring and dissolving evenly at 80°C, adenosine (preferably 30% in this embodiment) is added at a mass fraction of 10%-30% of succinic acid modified cyclodextrin. The mixture is stirred and dissolved evenly to obtain a mixture. The mixture is then vacuum spray-dried to obtain succinic acid modified cyclodextrin-coated adenosine microcapsules.

[0021] 3. Preparation of Polygonatum odoratum fermented polypeptides First, crush the rhizome of Polygonatum sibiricum and then add it to the fermentation medium at a rate of 15g / 100ml to obtain the fermentation substrate. The fermentation medium consists of 15g / L glucose, 10g / L peptone, 2g / L KH2PO4, 1g / L MgSO4, 0.05g / L manganese sulfate and 1.0g / L Tween 80, with the pH adjusted to 6.8. The activated brewing yeast is then inoculated into the fermentation substrate at an inoculation rate of 5%-10% of the fermentation substrate volume (preferably 8% in this embodiment) for liquid fermentation. The fermentation temperature is 20℃ and the culture time is 7 days. After liquid fermentation, the mixture is centrifuged at 5000 rpm / min for 10 min and the upper fermentation liquid is collected. Add ammonium sulfate to the fermentation broth at a rate of 25g per 100ml of fermentation broth, refrigerate overnight, then centrifuge at 5000rpm / min for 10min in a high-speed refrigerated centrifuge, discard the supernatant and collect the protein precipitate at the bottom; Finally, using a complex enzyme (neutral protease with an enzyme activity of 10 × 10⁻⁶) 4 (U / g) and alkaline protease (enzyme activity 5×10) 4The protein precipitate was enzymatically hydrolyzed by an equal mass mixture of protein precipitate and complex enzyme (U / g), with a mass ratio of protein precipitate to complex enzyme of 1:30 and a hydrolysis time of 12 h. After filtration, the mixture was vacuum spray-dried to obtain the fermented polypeptide of Polygonatum sibiricum.

[0022] 4. Preparation of adenosine microcapsule composition The formulations of the adenosine microcapsule compositions are shown in Table 1.

[0023] Table 1. Formulation (mass percentage) of adenosine microcapsule composition AG Note: "-" indicates that it has not been added.

[0024] Adenosine microcapsule composition AG was prepared according to the formulation given in Table 1 and the following process: The preparation method of adenosine microcapsule composition A is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition A can be obtained. The preparation method of adenosine microcapsule composition B is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition B can be obtained. The preparation method of adenosine microcapsule composition C is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition C can be obtained. The preparation method of adenosine microcapsule composition D is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition D can be obtained. The preparation method of adenosine microcapsule composition E is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition E can be obtained. The preparation method of adenosine microcapsule composition F is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed first, and homogenized and dispersed at 85°C and 12000rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800bar and 12000rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition F can be obtained. The preparation method of adenosine microcapsule composition G is as follows: According to the formulation dosage given in Table 1, the raw materials other than thickener and antioxidant are mixed and homogenized at 85°C and 12000 rpm for 60s to obtain a mixture. After cooling the mixture to 55°C, thickener and antioxidant are added according to the formulation dosage given in Table 1. The mixture is then homogenized under high pressure at 800 bar and 12000 rpm for 60s. After degassing, slow stirring, and cooling, adenosine microcapsule composition G can be obtained.

[0025] Furthermore, to investigate the effect of the composition of the adenosine microcapsule composition on its efficacy, the formulation of adenosine microcapsule composition G in Table 1 was adjusted, and the adenosine microcapsule composition AG was obtained according to its manufacturing process: The difference between adenosine microcapsule composition a and adenosine microcapsule composition G is that, based on the actual adenosine content in the adenosine microcapsules coated with succinic acid-modified cyclodextrin, adenosine is used instead of succinic acid-modified cyclodextrin to coat the adenosine microcapsules.

[0026] The difference between adenosine microcapsule composition b and adenosine microcapsule composition G lies in the fact that cyclodextrin-coated adenosine microcapsules are used instead of succinic acid-modified cyclodextrin-coated adenosine microcapsules with the same mass percentage. The preparation method of cyclodextrin-coated adenosine microcapsules is as follows: they are added to an ethanol aqueous solution with a mass fraction of 75%, stirred and dissolved evenly at 80°C, and then adenosine accounting for 30% of the mass of succinic acid-modified cyclodextrin is added. The mixture is stirred and dissolved evenly to obtain a mixed solution. The mixed solution is then vacuum spray-dried to obtain succinic acid-modified cyclodextrin-coated adenosine microcapsules.

[0027] The difference between the formulations of adenosine microcapsule composition C and adenosine microcapsule composition G is that: the ethanol extract of Polygonatum rhizome is used instead of the fermented polypeptide of Polygonatum rhizome. The preparation method of the ethanol extract of Polygonatum rhizome is as follows: after pulverizing Polygonatum rhizome, it is mixed with an ethanol solution with a volume concentration not exceeding 40% at a ratio of 1:10, and then heated and refluxed in an 85℃ water bath for 1 hour. After cooling and filtration, the filtrate and the residue are obtained. Ten times the amount of ethanol solution with a volume concentration not exceeding 40% is added to the residue, and the process is repeated twice. The filtrates are combined and then evaporated using a vacuum rotary evaporator at a water bath of 40℃, a rotation speed of 20 r / min, and a pressure of 0.1 MPa to remove the solvent. The ethanol extract of Polygonatum rhizome is then obtained by vacuum spray drying.

[0028] The difference between the formulation of adenosine microcapsule composition d and adenosine microcapsule composition G is that the former does not contain Polygonatum odoratum fermented polypeptide.

[0029] In addition, a blank control group was set up. The difference between the blank control group and the adenosine microcapsule composition G was that the adenosine microcapsules were not coated with succinic acid-modified cyclodextrin.

[0030] 5. Efficacy Test (1) Moisturizing efficacy test The moisturizing efficacy of adenosine microcapsule composition AG, adenosine microcapsule composition ad, and a blank control group were tested. The specific steps are as follows: Take 3 mL of each sample solution and evenly coat it onto a glass plate covered with 3M tape. Place it in an environment with a relative humidity of RH=43% for 72 hours to maintain a constant weight. Then transfer it to a desiccator containing 200 g of dry silica gel and place it for 4, 8, and 12 hours respectively. Weigh it after each time. The moisture retention rate is calculated according to the following formula: ; Where m0 is the moisture mass before placement; m1 is the moisture mass after placement for a period of time.

[0031] The results are shown in Table 2.

[0032] Table 2 Results of Moisturizing Efficacy Test As shown in Table 2, the formulation of the composition affects its moisturizing effect. Compared with the adenosine microcapsule composition AD and the blank control group, the adenosine microcapsule composition AG has a better moisturizing effect. The addition of succinic acid modified cyclodextrin-coated adenosine microcapsules and Polygonatum fermented peptides to the composition has a positive promoting effect on the moisturizing effect of the composition. Succinic acid modified cyclodextrin-coated adenosine microcapsules can replace part of the moisturizing agent in the composition, so that the composition can exert a better moisturizing performance.

[0033] (2) In vitro antioxidant test (DPPH free radical method) In vitro antioxidant tests were performed on adenosine microcapsule compositions AG and AD, as well as a blank control group. DPPH· is a nitrogen-centered stable free radical, and its anhydrous ethanol solution exhibits maximum absorption at a wavelength of 517 nm. In the presence of free radical scavengers, DPPH· can bind to or replace DPPH·, reducing the number of free radicals and thus decreasing absorbance. The specific test steps are as follows: First, weigh DPPH and dilute it to 250 mL in a brown volumetric flask with anhydrous ethanol to prepare 2 × 10⁻⁶ ppm. -4 Store the mol / L DPPH solution in a refrigerator protected from light.

[0034] Then, take 4.0 mL of 2×10 -4 After mixing 1.0 mL of 95% ethanol with 1 mol / L DPPH solution and the reaction stabilized, the absorbance was measured at 515 nm using 95% ethanol as a reference, and recorded as A0. Take 4.0 mL of 95% ethanol and 1.0 mL of the test sample solution (adenosine microcapsule composition AG, adenosine microcapsule composition ad and blank control group), mix well and after the reaction is stable, use 95% ethanol as a reference, and measure the absorbance at 515 nm, which is recorded as A1; Take 4.0 mL of 2×10 -4 After mixing 1.0 mL of mol / L DPPH solution and the test sample and allowing the reaction to stabilize, the absorbance was measured at 515 nm using 95% ethanol as a reference, and recorded as A2. Finally, the DPPH radical scavenging rate was calculated using the following formula: DPPH free radical scavenging rate (%) = [1 - (A2 - A1) / A0] × 100%.

[0035] The results are shown in Table 3.

[0036] Table 3 Results of in vitro antioxidant tests As shown in Table 3, compared with the blank control group, the addition of succinic acid-modified cyclodextrin-coated adenosine microcapsules to the composition can enhance the composition's DPPH free radical scavenging rate, reduce oxidative loss, and thus delay skin aging. Furthermore, the table shows that succinic acid-modified cyclodextrin coating of adenosine allows its anti-aging activity to be better exerted. In addition, comparing adenosine microcapsule composition G and adenosine microcapsule composition cd reveals that the Polygonatum sibiricum fermented polypeptide also has certain antioxidant activity, and it has a synergistic promoting effect with the succinic acid-modified cyclodextrin-coated adenosine microcapsules in enhancing the composition's antioxidant capacity.

[0037] (3) Anti-glycation efficacy test Studies have shown that inhibiting the formation of AGEs can effectively prevent skin aging. Measuring the AGE content can reflect the anti-glycation effect of various compositions. The specific testing steps are as follows: 10 ml of 0.01 mol / L PBS buffer solution (pH ≈ 7.2) was added to test tubes containing adenosine microcapsule composition AG, adenosine microcapsule composition ad, and a blank control group. After sonication, the sample solutions were pipetteed and placed into centrifuge tubes. The tubes were vortexed for 30 min and then centrifuged at 4000 rpm for 10 min. The supernatant was collected, and HRP-labeled standards, adenosine microcapsule compositions AG and ad, and the blank control group were added to microplates coated with specific antibodies and labeled as group A. HRP-labeled standards were added to another... In a microplate coated with a specific antibody and labeled as group B, 100 μL of the antibody was added to each well. After the sample was added, the wells were sealed with adhesive sealant and incubated at 37°C with an ultrasonic shaker for 30 min. The solution in the wells was discarded, and the plates were washed three times with PBS buffer. 100 μL of chemiluminescent solution was added to each well and mixed thoroughly for luminescence development. After 5 min, the wavelength of the multi-mode microplate reader was adjusted to 425 nm and used to collect data. The content of Nε-carboxymethyl lysine (CML) was calculated, and then the AGEs content was calculated in μg / mL. The results are shown in Table 4.

[0038] Table 4. Results of Anti-glycation Efficacy Test Table 4 shows that, compared to adenosine microcapsule composition c with added Polygonatum sibiricum extract and adenosine microcapsule composition d without added Polygonatum sibiricum fermented peptides, the addition of Polygonatum sibiricum fermented peptides has a positive promoting effect on enhancing the anti-glycation efficacy of the compositions. Among them, adenosine microcapsule composition D has the lowest AGEs content, less than 5.0 μg / mL. Furthermore, the table also shows that, under appropriate dosage, combining succinic acid-modified cyclodextrin-coated adenosine microcapsules with Polygonatum sibiricum fermented peptides enables the compositions to exhibit optimal anti-glycation activity.

[0039] (5) Security testing Thirty participants were selected for the experiment, referencing the 2015 edition of the "Cosmetic Safety Technical Specifications". The area selected did not exceed 50mm². 2 A qualified patch test apparatus with a depth of approximately 1 mm is used. The test substance is placed in the patch test apparatus chamber at a volume of approximately 0.020-0.025 mL (liquid). The test substance is an adenosine microcapsule composition AG diluted to a concentration of 1%. The control well uses the same diluent for the adenosine microcapsule composition. The patch test apparatus containing the test substance is applied to the back or flexor side of the forearm of the subject using hypoallergenic adhesive tape. Gently press with the palm of the hand to ensure even application to the skin, and leave for 24 hours. Skin reactions are observed according to the skin reaction grading criteria for closed patch tests at 30 minutes (after the indentation disappears), 24 hours, and 48 hours after removing the patch test apparatus.

[0040] The grading criteria for skin reactions in the closed patch test are shown in Table 5, and the observation results should be recorded.

[0041] Table 5. Grading Criteria for Skin Reactions in Occlusive Patch Tests The results are shown in Table 6. The adenosine microcapsule composition AG was non-irritating to the skin and had a high safety profile.

[0042] Table 6. Results of patch test The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A manufacturing process for a microcapsule composition containing adenosine, characterized in that, Includes the following steps: (1) Disperse 0.5-5% by mass of succinic acid-modified cyclodextrin-coated adenosine microcapsules, 2-5% by mass of Polygonatum odoratum fermented polypeptide, 1-4% by mass of humectant, and 0.05-0.2% by mass of emulsifier into deionized water and stir to mix. Then, homogenize the mixture at 80℃-85℃ and 10000rpm-15000rpm for 30s-60s to obtain a mixture. (2) After cooling the mixture to 50℃-55℃, add 0.05%-0.1% by mass of thickener and 0.2%-0.5% by mass of antioxidant. After stirring and mixing, homogenize under high pressure at 600bar-800bar and 10000rpm-15000rpm for 60s-90s. Then, after degassing, slow stirring and cooling, the adenosine microcapsule composition can be obtained.

2. The manufacturing process of a microcapsule composition containing adenosine according to claim 1, characterized in that, The manufacturing process of the succinic acid modified cyclodextrin-coated adenosine microcapsules is as follows: succinic acid modified cyclodextrin is dispersed in an ethanol aqueous solution with a mass fraction of 50%-80%, and stirred and dissolved at 60℃-80℃. Then, adenosine is added and stirred until dissolved evenly to obtain a solution. The solution is then vacuum spray-dried to obtain the succinic acid modified cyclodextrin-coated adenosine microcapsules.

3. The manufacturing process of a microcapsule composition containing adenosine according to claim 2, characterized in that, The amount of adenosine added is 10%-30% of the mass of the succinic acid-modified cyclodextrin, and the cyclodextrin is β-cyclodextrin or hydroxypropyl-β-cyclodextrin.

4. The manufacturing process of a microcapsule composition containing adenosine according to claim 2, characterized in that, The manufacturing process of the succinic acid modified cyclodextrin is as follows: succinic acid and cyclodextrin are dispersed in N,N-dimethylformamide solvent, heated to 90℃-120℃ and stirred until uniformly dissolved. After adding an acidic catalyst and stirring to react, the solvent and unreacted raw materials are removed by vacuum evaporation to obtain the succinic acid modified cyclodextrin.

5. The manufacturing process of a microcapsule composition containing adenosine according to claim 1, characterized in that, The manufacturing process of the Polygonatum fermentation polypeptide is as follows: first, the crushed Polygonatum tuber is added to the fermentation culture medium to obtain the fermentation substrate, then activated brewing yeast is inoculated into the fermentation substrate for liquid fermentation, after liquid fermentation, the fermentation liquid is obtained by centrifugation, and then the fermentation liquid is obtained by protein extraction and enzymatic hydrolysis with compound enzymes.

6. The manufacturing process of a microcapsule composition containing adenosine according to claim 5, characterized in that, The inoculation amount of the brewing yeast is 5%-10% of the fermentation substrate volume.

7. The manufacturing process of a microcapsule composition containing adenosine according to claim 5, characterized in that, The complex enzyme is a mixture of equal masses of neutral protease and alkaline protease.

8. The manufacturing process of a microcapsule composition containing adenosine according to claim 1, characterized in that, The moisturizer is hyaluronic acid or glycerin, the emulsifier is sucrose palmitate or glyceryl linoleate, the thickener is carbomer, and the antioxidant is tocopheryl acetate.

9. An adenosine microcapsule composition obtained by the manufacturing process according to any one of claims 1-8.

10. The use of the adenosine microcapsule composition as described in claim 9 in the preparation of anti-aging products.

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