A hyaluronic acid-astaxanthin complex and its preparation method and application

Abstract

The present invention belongs to the field of material technology, and specifically relates to a hyaluronic acid-astaxanthin complex and a preparation method and application thereof. Through the technical solution provided by the present invention, the shortcomings of the astaxanthin oil mixture that it is not easy to disperse, easy to hang on the wall and easy to agglomerate during use are improved. At the same time, through the improvement of the system, the astaxanthin oil is successfully encapsulated to form a colloidal solution, the stability of the astaxanthin oil is improved, and the synergistic effect of sodium hyaluronate and astaxanthin oil is played.

Description

Technical Field

[0001] The invention belongs to the field of material technology, and specifically relates to a hyaluronic acid-astaxanthin complex and a preparation method and application thereof. Background Art

[0002] Astaxanthin is a red carotenoid found in the shells of river crabs and shrimps, oysters and salmon. It has antioxidant, anti-aging, anti-tumor, and cardiovascular and cerebrovascular prevention effects. Natural astaxanthin is the strongest antioxidant discovered by humans in nature so far, and its antioxidant activity far exceeds that of existing antioxidants. However, astaxanthin has a long conjugated unsaturated double bond structure system, is unstable in nature, is easy to isomerize and degrade, and is sensitive to heat, oxygen, natural light, and ultraviolet light, which causes a lot of inconvenience to the promotion and use of astaxanthin.

[0003] In order to solve the above problems, the prior art usually prepares astaxanthin carriers, for example, by dissolving them using biomacromolecule chelating agents or organic solvents. However, the complex structure of biomacromolecule chelating agents does not have an ideal protective effect on astaxanthin, and organic solvents are often volatile, which can cause material instability.

[0004] Currently, there is an urgent need to develop new preparation processes to improve the stability of astaxanthin. Summary of the invention

[0005] In order to overcome the shortcomings of the prior art, the present invention discloses a method for preparing a hyaluronic acid-astaxanthin complex, which is characterized by comprising: dissolving astaxanthin in an aqueous solution system containing chitosan, hydroxytyrosol, and butanediol and heating the solution, and then dropping the solution into a hyaluronic acid solution containing a surfactant to obtain the hyaluronic acid-astaxanthin complex. The technical problem to be solved is that the appearance of the astaxanthin microcapsule solution does not have obvious black particles.

[0006] Preferably, the preparation of the hyaluronic acid solution containing a surfactant comprises adjusting the pH of the hyaluronic acid solution to 3-4, heating the solution, and then adding the surfactant. The technical problem to be solved is to reduce the white floccules in the astaxanthin microcapsule solution.

[0007] Preferably, the molecular weight of the hyaluronic acid is 200000-400000. The technical problem to be solved is that the astaxanthin microcapsule solution has no obvious particles and no obvious precipitation.

[0008] Preferably, the surfactant is Tween 80 or Span 80.

[0009] Preferably, the heating temperature is 60-80°C and the heating time is 1-2 hours.

[0010] Preferably, the aqueous solution system comprising chitosan, hydroxytyrosol and butanediol further comprises acetic acid.

[0011] Preferably, after astaxanthin is dissolved in the aqueous solution system comprising chitosan, hydroxytyrosol and butanediol, the heating temperature is 50-60° C. and the time is 1-2 hours.

[0012] According to another aspect of the present invention, there is provided a hyaluronic acid-astaxanthin complex prepared according to the above preparation method for use in health care products, medicines and cosmetics.

[0013] Compared with the prior art, the compounds disclosed in the present invention have the following beneficial effects: through the technical scheme provided by the present invention, the shortcomings of the astaxanthin oil mixture that it is not easy to disperse, and it is easy to stick to the wall and agglomerate during use are improved. At the same time, through the improvement of the system, the astaxanthin oil is successfully encapsulated to form a colloidal solution, which improves the stability of the astaxanthin oil and plays a synergistic role between sodium hyaluronate and astaxanthin oil. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0015] Figure 1 A schematic diagram showing the appearance of an astaxanthin microcapsule solution provided according to one embodiment of the present invention is shown.

[0016] Figure 2 A schematic diagram showing the appearance of an astaxanthin microcapsule solution provided according to one embodiment of the present invention is shown.

[0017] Figure 3 A schematic diagram showing the appearance of an astaxanthin microcapsule solution provided according to one embodiment of the present invention is shown.

[0018] Figure 4 A structural diagram of a sodium hyaluronate molecule provided according to one embodiment of the present invention is shown.

[0019] Figure 5 A microscopic picture showing modified astaxanthin oil encapsulated by modified sodium hyaluronate and chitosan provided according to one embodiment of the present invention.

[0020] Figure 6 A comparison diagram of samples obtained according to Example 2 and Example 3 of the present invention is shown.

[0021] Figure 7 A rendering showing the phenomenon that the unmodified astaxanthin oil provided according to one embodiment of the present invention is not easy to disperse and is easy to stick to the wall. DETAILED DESCRIPTION

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0024] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0025] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0026] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0027] The reagents used in the examples of the present invention are all commercially available analytically pure or chemically pure.

[0028] The technical solution of the present invention is further illustrated by the following embodiments.

[0029] Example 1

[0030] Sodium hyaluronate and chitosan as encapsulation materials for unmodified astaxanthin oil

[0031] 1. Pre-dissolve 0.3-0.5 g of sodium hyaluronate in 80 g of water to form solution A;

[0032] 2. Add 0.05-0.1 g of astaxanthin oil, 0.5-1 g of Tween-80, and 0.5-1 g of Span-80 to solution A, and disperse them uniformly in water to form solution B;

[0033] 3. Add 1% acetic acid to 20 g of water and add 0.1-0.5 g of chitosan to form solution C;

[0034] 4. Add solution C into solution B at a dropping speed of 0.05 g / s, homogenizing while adding at a homogenizing speed of 8000-9000 rpm to form an astaxanthin microcapsule solution.

[0035] like Figure 1 As shown, the final astaxanthin microcapsule solution has obvious black particles on the appearance, and the inner edge of the beaker is adhered with obvious astaxanthin oil. After being left overnight, there is obvious flocculent precipitation at the bottom of the beaker.

[0036] Example 2

[0037] Sodium hyaluronate (molecular weight: 200000-400000, unless otherwise specified below) and chitosan are used as capsules to encapsulate the modified astaxanthin oil

[0038] 1. Pre-dissolve 0.3-0.5 g of sodium hyaluronate in 80 g of water to form solution A;

[0039] 2. Add 0.5-1 g of Tween-80 and 0.5-1 g of Span-80 to solution A and disperse them evenly in water to form solution B;

[0040] 3. Add 1% acetic acid to 20 g of water, add 0.1-0.5 g of chitosan, 0.05-0.1 g of astaxanthin oil, 0.1-0.5 g of hydroxytyrosol, and 0.5 g of butanediol, and heat at 50-60°C for 1-2 h to form solution C;

[0041] 4. Add solution C into solution B at a dropping speed of 0.05 g / s, homogenizing while adding at a homogenizing speed of 8000-9000 rpm to form an astaxanthin microcapsule solution.

[0042] like Figure 2 As shown, the final astaxanthin microcapsule solution has no obvious particles on the outside, but there are many white flocs in the solution. After standing for a while, there are obvious white flocs precipitated at the bottom of the solution, and the precipitates are in clumps.

[0043] Example 3

[0044] Modified sodium hyaluronate and chitosan as capsules to encapsulate modified astaxanthin oil

[0045] 1. Pre-dissolve 0.3-0.5 g of sodium hyaluronate in 80 g of water, adjust the pH value of the system to 3-4, and heat at 60-80°C for 1-2 hours to form solution A;

[0046] 2. Add 0.5-1 g of Tween-80 and 0.5-1 g of Span-80 to solution A and disperse them evenly in water to form solution B;

[0047] 3. Add 1% acetic acid to 20 g of water, add 0.1-0.5 g of chitosan, 0.05-0.1 g of astaxanthin oil, 0.1-0.5 g of hydroxytyrosol, and 0.5 g of butanediol, and heat at 50-60°C for 1-2 h to form solution C;

[0048] 4. Add solution C into solution B at a dropping speed of 0.05 g / s, homogenizing while adding at a homogenizing speed of 8000-9000 rpm to form an astaxanthin microcapsule solution.

[0049] like Figure 3 As shown, the final astaxanthin microcapsule solution has no obvious particles on the outside and no obvious precipitation at the bottom of the solution. Figure 6 The comparison diagram of the samples obtained in Example 2 and Example 3 shows that there are obvious flocs at the bottom of the sample in Example 2 (black circle standard). Under the conditions of Example 3, the high molecular weight sodium hyaluronate is cross-linked to increase its molecular chain, increase its average molecular weight, and modify its mechanical properties and rheological characteristics. Figure 4 It can be seen that the three sites in the sodium hyaluronate molecule that can be chemically modified are hydroxyl, N-acetyl, and carboxyl. The functional groups that can participate in the cross-linking reaction are mainly hydroxyl and carboxyl. Figure 5 It can be seen that the cross-linked sodium hyaluronate molecules expand from the original linear to a three-dimensional network, and then can be attracted by charge with chitosan to form a package. In the process of cross-linking reaction, more negative charges are stimulated in the sodium hyaluronate macromolecules themselves, making the positive and negative charges of the sodium hyaluronate molecules with chitosan more firmly attracted, and the sodium hyaluronate molecules are wrapped around the astaxanthin oil to form a stable rod-shaped colloidal solution.

[0050] In addition, the conditions of Example 3 allow hydroxytyrosol to be embedded in astaxanthin, and at the same time, hydroxytyrosol and olive oil themselves have a good mutual solubility effect. The addition of hydroxytyrosol improves the disadvantages of astaxanthin, olive oil and water being extremely repellent, and improves the disadvantages of astaxanthin oil being difficult to disperse and easy to adhere to the wall during the emulsification process.

[0051] Example 4

[0052] Study on the stability of astaxanthin oil colloidal solution

[0053] The performance test results of the samples obtained in Examples 1, 2 and 3 are shown in the following table.

[0054]

[0055] It can be seen from the data in the above table that the microcapsules of Examples 1 and 2 show different degrees of aggregation, wall adhesion and precipitation at room temperature, and the aggregation, wall adhesion and precipitation of the microcapsules are not improved after being placed in an environment of 45°C and -15°C for 24 hours; while Example 3 is clear and transparent at room temperature, and is still clear and transparent after being placed in an environment of 45°C and -15°C for 24 hours, without obvious aggregation, wall adhesion and precipitation.

[0056] Example 5

[0057] Modified sodium hyaluronate and chitosan with different molecular weights as capsules to encapsulate modified astaxanthin oil

[0058] 1. Pre-dissolve 0.3-0.5 g of sodium hyaluronate (molecular weight: 5000-10000 or 30000-50000) in 80 g of water, adjust the pH value of the system to 3-4, and heat at 60-80°C for 1-2 hours to form solution A;

[0059] 2. Add 0.5-1 g of Tween-80 and 0.5-1 g of Span-80 to solution A and disperse them evenly in water to form solution B;

[0060] 3. Add 1% acetic acid to 20 g of water, add 0.1-0.5 g of chitosan, 0.05-0.1 g of astaxanthin oil, 0.1-0.5 g of hydroxytyrosol, and 0.5 g of butanediol, and heat at 50-60°C for 1-2 h to form solution C;

[0061] 4. Add solution C into solution B at a dropping speed of 0.05 g / s, homogenizing while adding at a homogenizing speed of 8000-9000 rpm to form an astaxanthin microcapsule solution.

[0062] Finally, there were obvious clumping precipitations at the bottom of the astaxanthin colloidal solution.

[0063] Example 6

[0064] Improve the dispersibility of astaxanthin oil

[0065] Commercially available astaxanthin-related raw materials are generally astaxanthin microcapsule powder and astaxanthin oil. Since astaxanthin microcapsule powder may contain a variety of capsule materials, its astaxanthin content is not high. At the same time, subsequent operations such as spray drying may cause astaxanthin loss. Therefore, this experiment selected a mixture of astaxanthin and olive oil, hereinafter referred to as astaxanthin oil, in which the astaxanthin content is 10%.

[0066] Preparation of emulsion containing astaxanthin oil

[0067]

[0068] Under the conditions of Example 6, hydroxytyrosol can be embedded in astaxanthin. The addition of hydroxytyrosol improves the disadvantages of astaxanthin being extremely repellent to olive oil and water, and improves the disadvantages of astaxanthin oil being difficult to disperse and easy to adhere to the wall during the emulsification process. Figure 7 This is a rendering of the phenomenon that the unmodified astaxanthin oil is difficult to disperse and easy to stick to the wall. The modified astaxanthin oil will not stick to the wall, so the rendering is not provided.

[0069] Example 7

[0070] Advantages of sodium hyaluronate replacing sodium alginate

[0071] Conventional hydrogel encapsulation is generally carried out with sodium alginate and chitosan. This patent innovatively uses modified sodium hyaluronate instead of sodium alginate for encapsulation, which can give full play to the excellent moisturizing properties of sodium alginate and form a synergistic effect with the antioxidant properties of astaxanthin oil.

[0072] 1. Pre-dissolve 0.3-0.5 g of sodium alginate in 80 g of water to form solution A;

[0073] 2. Add 0.5-1 g of Tween-80 and 0.5-1 g of Span-80 to solution A and disperse them evenly in water to form solution B;

[0074] 3. Add 1% acetic acid to 20 g of water, add 0.1-0.5 g of chitosan, 0.05-0.1 g of astaxanthin oil, 0.1-0.5 g of hydroxytyrosol, and 0.5 g of butanediol, and heat at 50-60°C for 1-2 h to form solution C;

[0075] 4. Add solution C into solution B at a dropping speed of 0.05 g / s, homogenizing while adding at a homogenizing speed of 8000-9000 rpm to form an astaxanthin microcapsule solution.

[0076] 5. The obtained microcapsule solution of astaxanthin oil wrapped in sodium alginate and chitosan is adjusted to a pH value of 5-6 using triethanolamine. The standby number is "microcapsule essence 1"

[0077] 6. The sodium hyaluronate and chitosan microcapsule solution of astaxanthin oil in Example 3 was adjusted to a pH value of 5-6 using triethanolamine, and the standby number was "microcapsule essence 2"

[0078] Example 8

[0079] Moisturizing efficacy test

[0080] 1. Instrument name: Skin moisture tester Corneometer CM825

[0081] 2. Test site: inner side of forearm

[0082] 3. Test time: 0h, 6h

[0083] 4. Test method:

[0084] 4.1 Preparation before the test: The subjects cleaned the inner forearms of both hands and sat quietly in the efficacy evaluation room (temperature: 20℃22℃, humidity: 40%60%) for 1 hour. They were not allowed to drink water or eat during this period. The test site was exposed, and they were relaxed and avoided contact with the test site. The test area was at least 3cm*3cm, with a spacing of 1cm. The smeared area and the blank control area were randomly distributed.

[0085] 4.2 Establishment of skin barrier damage: The laboratory uses a special tape tearing method to damage the skin barrier of the test area and the control area.

[0086] 4.3 Determine the dosage: Test sample at (2.0±0.1) mg / cm 2 Apply a single application of 100% ethanol.

[0087] 5. Subject Information

[0088] Gender of subjects: 10 females.

[0089] Age of subjects: 28-45 years old

[0090] Skin condition requirements for the subjects: fair forearm skin color and no scars.

[0091] Those who have the following conditions cannot be selected as subjects

[0092] (1) Those who have used antihistamines in the past week or immunosuppressants in the past month.

[0093] (2) Those who have used any anti-inflammatory drugs in the test area within the past two months.

[0094] (3) Subjects with inflammatory skin disease that has not been clinically cured.

[0095] (4) Patients with insulin-dependent diabetes mellitus.

[0096] (5) Patients who are receiving treatment for asthma or other chronic respiratory diseases.

[0097] (6) Patients who have received anticancer chemotherapy in the past 6 months.

[0098] (7) Patients with immunodeficiency or autoimmune diseases.

[0099] (8) Women who are breastfeeding or pregnant.

[0100] (9) Bilateral mastectomy and bilateral axillary lymph node removal.

[0101] (10) The judge shall be aware that scars, pigmentation, atrophy, port-wine stains, uneven skin tone, folliculitis or other blemishes on the skin to be tested may affect the test results.

[0102] (11) Participating in other clinical trial researchers.

[0103] (12) Participated in other clinical trials including patch test researchers within 2 months.

[0104] (13) People with highly sensitive constitutions.

[0105] (14) Those who are unwilling to participate or are unable to complete the required content according to the experimental requirements.

[0106] (15) Those who are deemed by the researcher to be unsuitable for participation in this study.

[0107] 6. Calculation of results

[0108] Table 6-1 Descriptive statistics of moisture content in the stratum corneum of the skin (n=33)

[0109] Microcapsule Essence 1 Microcapsule Essence 2 Time h0 h6 h6 Mean 26.32 32.55 38.01 Standard Deviation 8.33 9.44 8.56 Maximum 48.67 56.33 60.4 Minimum 17.32 14.01 15.05 Normality test P value 0.241 0.128 0.226

[0110] Note: Normality test P value>0.050, that is, the data obeys normal distribution, and paired T test is used;

[0111] The moisture content of the stratum corneum of the subjects was tested 6 hours before and after the use of the samples. From the data in the table, it can be seen that the moisturizing effect of microcapsule essence 2 is better than that of microcapsule essence 1, indicating that sodium hyaluronate, as the encapsulating material, can improve the moisturizing effect of astaxanthin microcapsules themselves.

[0112] The above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Those skilled in the art may make various modifications and variations to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing a hyaluronic acid-astaxanthin complex, characterized in that: include, Pre-dissolve sodium hyaluronate, adjust the pH value of the system to 3-4, and then add a surfactant after heating to obtain a hyaluronic acid solution containing a surfactant; The aqueous solution system containing astaxanthin, acetic acid, chitosan, hydroxytyrosol and butanediol is heated, and then added dropwise to the hyaluronic acid solution containing the surfactant and homogenized to obtain the hyaluronic acid-astaxanthin complex.

2. The preparation method according to claim 1, characterized in that: The molecular weight of the hyaluronic acid is 200,000-400,000.

3. The preparation method according to claim 1, characterized in that: The surfactants are Tween 80 and Span 80.

4. The preparation method according to claim 1, characterized in that: Sodium hyaluronate is pre-dissolved, and the system pH is adjusted to 3-4, and then the heating temperature is 60-80°C for 1-2 hours.

5. The preparation method according to claim 1, characterized in that: After astaxanthin is dissolved in an aqueous solution system containing acetic acid, chitosan, hydroxytyrosol and butanediol, the heating temperature is 50-60° C. and the time is 1-2 hours.

6. The preparation method according to claim 1, characterized in that: The homogenizing speed during the homogenizing process is 8000-9000 rpm.

7. Application of the hyaluronic acid-astaxanthin complex prepared by the preparation method according to any one of claims 1 to 6 in health products, medicines and cosmetics.

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