Astaxanthin composition and preparation method thereof

Abstract

The present invention relates to a composition comprising astaxanthin-encapsulated microparticles and glycosaminoglycans. The structure of the astaxanthin-encapsulated microparticles comprises an inner core and an outer shell, wherein the inner core comprises astaxanthin, and the outer shell comprises a skeleton component, a thickener, and an emulsifier. The astaxanthin-encapsulated microparticles of the present invention utilize dextrin as a skeleton component, and are combined with a thickener and an emulsifier to construct the astaxanthin-encapsulated outer shell. The astaxanthin-encapsulated microparticles have a stable structure, are compounded with the glycosaminoglycan component, are uniformly dispersed in water without demulsification or stratification, and remain stable under high temperature conditions.

Description

Technical Field

[0001] The present invention belongs to the technical field of embedded preparations, and in particular relates to an astaxanthin-embedded microparticle composition and a preparation method thereof. Background Art

[0002] Astaxanthin, chemically known as 3,3′-dihydroxy-4,4′-diketo-β-carotene, is a keto-carotenoid. A red solid powder, it exhibits excellent coloring properties and can be used as a natural colorant. Furthermore, astaxanthin possesses strong antioxidant properties, 10 times that of β-carotene and 500 times that of vitamin E, earning it the nickname "Super Vitamin E." Due to its excellent antioxidant and coloring properties, it is now widely used in functional foods, cosmetics, and other fields.

[0003] Astaxanthin is an oily substance with excellent fat solubility but is insoluble in water. Since a large number of foods and cosmetics are water-soluble, astaxanthin's solubility greatly limits its industrial applications. A common solution is to increase its hydrophilicity through encapsulation technology, which can also improve the tolerance of astaxanthin preparations to temperature, light, and pH.

[0004] CN108013441A discloses an astaxanthin-encapsulated complex and its preparation method. It uses sodium carboxymethylcellulose (CMC-Na) and microcrystalline cellulose (MCC) to encapsulate astaxanthin, improving its hydrophilicity. The encapsulated astaxanthin can then be used in the development of yogurt products. CN111358762A discloses an astaxanthin microcapsule that uses β-cyclodextrin, oligomaltodextrin, and modified starch to encapsulate astaxanthin, significantly improving its water solubility and enhancing its bioavailability.

[0005] While conventional encapsulation can effectively address astaxanthin solubility issues, the applicant has discovered that many existing astaxanthin encapsulation systems, when used in food and cosmetics, can experience demulsification and stratification when the product contains glycosaminoglycans. Furthermore, this demulsification and stratification is exacerbated during high-temperature sterilization, further leading to astaxanthin degradation and reduced astaxanthin content, affecting the efficacy of astaxanthin in the product. This significantly limits the application of astaxanthin in combination with glycosaminoglycans. Summary of the Invention

[0006] In view of the problem that the glycosaminoglycan components of the existing astaxanthin embedding system are easily demulsified and stratified, the present invention provides an astaxanthin-embedded microparticle composition and a preparation method thereof.

[0007] In a first aspect, the present invention provides a composition comprising astaxanthin-embedded microparticles and glycosaminoglycans; the structure of the astaxanthin-embedded microparticles comprises an inner core and an outer shell; the inner core component comprises astaxanthin, and the outer shell component comprises a skeleton component, a thickener and an emulsifier.

[0008] In certain embodiments, the matrix component is selected from one or more of resistant dextrin, maltodextrin, and β-cyclodextrin.

[0009] In certain embodiments, the thickener is selected from one or more of gum arabic, xanthan gum, pectin, and guar gum.

[0010] In certain embodiments, the emulsifier is selected from medium chain triglycerides, and / or glyceryl succinate.

[0011] In certain embodiments, the astaxanthin is astaxanthin oil.

[0012] In certain embodiments, the composition is uniformly dispersed in water and is in the form of an emulsion.

[0013] In certain embodiments, the mass ratio of the inner core to the outer shell is 1:10-100.

[0014] In certain embodiments, the mass ratio of the inner core to the outer shell is 1:20-50.

[0015] In certain embodiments, the mass ratio of the skeleton component, thickener, and emulsifier is 1-2:2-6:1-2.

[0016] In certain embodiments, the mass ratio of the skeleton component, thickener, and emulsifier is 1:3:1.

[0017] In certain embodiments, the glycosaminoglycan is selected from one or more of hyaluronic acid, chondroitin sulfate, dermatan sulfate, and keratan sulfate.

[0018] In certain embodiments, the composition further comprises collagen.

[0019] In certain embodiments, the glycosaminoglycan molecules are polymerized onto collagen molecules.

[0020] In certain embodiments, the collagen is glycosaminoglycan-rich collagen.

[0021] In certain embodiments, the mass ratio of astaxanthin to glycosaminoglycan is 1:2-7.

[0022] In certain embodiments, the composition further comprises a pH adjuster.

[0023] In certain embodiments, the pH adjuster is selected from one or more of citric acid, malic acid, and tartaric acid.

[0024] In a second aspect, the present invention provides a method for preparing a composition, the preparation method comprising the following steps:

[0025] (1) Add the inner core and outer shell components described in the first aspect to an appropriate amount of water and stir until the components are fully dissolved to prepare a solution for use;

[0026] (2) The solution in step (1) is homogenized using a homogenizer to obtain astaxanthin-embedded microparticles, wherein the astaxanthin-embedded microparticles are uniformly dispersed in water to form an emulsion;

[0027] (3) Glycosaminoglycan is added to the emulsion and stirred until fully dissolved, and then sterilized in an autoclave to obtain a composition.

[0028] In certain embodiments, the stirring time in step (1) is 10-40 min.

[0029] In certain embodiments, the homogenization speed in step (2) is 5000-25000 rpm.

[0030] In certain embodiments, the homogenization speed in step (2) is 5-15 minutes.

[0031] In certain embodiments, the stirring time in step (3) is 15-30 min.

[0032] In certain embodiments, the sterilization temperature in step (3) is 105-121° C., and the sterilization time is 15-30 min.

[0033] In a third aspect, the present invention provides the use of the composition described in the above aspects in the preparation of medicines, health foods, and foods.

[0034] Definition of terms

[0035] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. At the same time, in order to better understand the present invention, the definitions and explanations of relevant terms are provided below.

[0036] As used herein, the term "encapsulated microparticles," also known as "microcapsules," refers to particles with tiny enclosed spaces, ranging in size from micrometers to nanometers. Encapsulated microparticles typically consist of a core and an outer shell. The outer shell, also known as the wall, can be made from a variety of materials, including natural or synthetic polymers. Encapsulated microparticle technology allows the core substance to be protected while allowing for controlled release of specific substances through the wall.

[0037] As used herein, the term "glycosaminoglycan" refers to repeating disaccharide units composed of penturonic acid or hexuronic acid linked to N-acetylglucosamine or N-acetylgalactosamine, some of which have sulfate groups substituted at positions 1, 4, or 6 on the glycosaminoglycans. Glycosaminoglycans have a wide molecular weight range, ranging from several thousand daltons to over one million daltons. "Glycosaminoglycans" herein include, but are not limited to, hyaluronic acid, chondroitin sulfate, dermatan sulfate, and keratin sulfate. Glycosaminoglycans can exist solely as glycosaminoglycan structures or be polymerized onto collagen molecules.

[0038] Advantageous Effects of the Invention

[0039] Compared with the astaxanthin-embedded microparticles in the prior art, the astaxanthin-embedded microparticles of the present invention have at least the following beneficial effects: the present invention uses dextrin as a skeleton component, and uses a thickener and an emulsifier to construct an astaxanthin-embedded shell, so that the astaxanthin-embedded microparticles have a stable structure. When compounded with glycosaminoglycan components, they can be evenly dispersed in water without demulsification and stratification. The astaxanthin-embedded microparticle structure remains stable under high temperature conditions, further preventing the degradation of the astaxanthin component. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 is the chromatogram of the astaxanthin standard in Example 6;

[0041] Figure 2 , chromatogram of the test sample of composition 4 in Example 6;

[0042] Figure 3 , chromatogram of the test sample of composition 36 in Example 6. DETAILED DESCRIPTION

[0043] The invention will now be described with reference to the following examples which are intended to illustrate the invention but not to limit it.

[0044] Unless otherwise indicated, the experiments and procedures described in the examples were performed essentially according to conventional methods well known in the art and described in various references.

[0045] In addition, if specific conditions are not specified in the examples, the experiments were performed under conventional conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be obtained commercially. It is understood that the examples describe the present invention by way of example and are not intended to limit the scope of the present invention. All publications and other references mentioned herein are incorporated herein by reference in their entirety.

[0046] Example 1. Preparation of astaxanthin-encapsulated microparticle composition

[0047] The formula of astaxanthin-embedded microparticles 1-24 in this embodiment is shown in Table 1-3 by weight:

[0048] Table 1 Astaxanthin embedded microparticles formula

[0049]

[0050]

[0051] Table 2 Astaxanthin embedded microparticles formula

[0052]

[0053] Table 3 Astaxanthin embedded microparticles formula

[0054]

[0055] Table 4 Astaxanthin embedded microparticles formula

[0056]

[0057] The preparation process of the astaxanthin-embedded microparticles 1-32 of this embodiment is as follows:

[0058] (1) Mix the components of the astaxanthin-encapsulated microparticles and stir for 10-40 minutes until the components are fully dissolved to prepare a solution for use;

[0059] (2) The solution obtained in step (1) is homogenized with a homogenizer at a speed of 5000-25000 rpm for 5-15 minutes to obtain astaxanthin-embedded microparticles, wherein the astaxanthin-embedded microparticles are uniformly dispersed in water to form an emulsion.

[0060] Example 2 Preparation of Astaxanthin-Embedded Microparticle Composition

[0061] Take the astaxanthin-embedded microparticles 1-32 prepared in Example 1, add 70 parts by weight (based on the weight fraction of astaxanthin oil as 1) of glycosaminoglycan-rich collagen peptide (for its preparation method, see CN116410302A, the glycosaminoglycan-rich collagen peptide contains 63 parts of collagen peptide and 7 parts of glycosaminoglycan, glycosaminoglycans including chondroitin sulfate, sodium hyaluronate, dermatan sulfate, etc., wherein the glycosaminoglycans exist in the form of glycosaminoglycan structure alone and polymerized on collagen molecules), then stir with a blender for 15-30 minutes until fully dissolved, and place in an autoclave for sterilization (sterilization conditions: temperature 105-121 ° C, time 15-30 minutes) to obtain a composition 1-32 containing astaxanthin-embedded microparticles.

[0062] Take astaxanthin particles 1, 12, 27, and 31 prepared in Example 1, add 2 parts of sodium hyaluronate respectively, and then stir with a blender for 15-30 minutes until fully dissolved, and place in a high pressure sterilizer for sterilization (sterilization conditions: temperature 105-121 ° C, time 15-30 minutes) to obtain compositions 33-36 of astaxanthin-encapsulated particles.

[0063] Take the astaxanthin particles 3, 7, 28, and 32 prepared in Example 1, add 5 parts of chondroitin sulfate respectively, and then stir with a blender for 15-30 minutes until fully dissolved, and place in a high pressure sterilizer for sterilization (sterilization conditions: temperature 105-121 ° C, time 15-30 minutes) to obtain compositions 37-40 of astaxanthin-encapsulated particles.

[0064] Example 3 Sensory testing of astaxanthin-encapsulated microparticle compositions

[0065] 3.1 Test method

[0066] Take a certain amount of the evenly mixed sample to be tested and place it in a 50mL colorless transparent beaker. Observe the color under natural light to check whether there is any foreign matter.

[0067] 3.2 Test results

[0068] Compositions 1-26, 33-34, 37-38 appeared as orange to orange-red liquids, were uniform and had no stratification and no oily floating matter; Compositions 27-32, 35-36, 39-40 solutions were stratified, with oily matter floating on the upper layer, which was orange-red to light yellow, and the lower layer was a yellow transparent liquid.

[0069] Example 4 Test of Soluble Solids Content in Astaxanthin-Embedded Microparticle Composition 5.1 Test Method

[0070] Separate the two prisms of the refractometer and clean them with absorbent cotton dipped in ether or ethanol. Use a glass rod with a fused end to pick up 2-3 drops of each composition solution and drop them on the center of the prism surface of the refractometer (Note: 1. When dipping the glass rod into the sample, first remove the upper oily substance of the composition that has stratified, and use the glass rod to remove the lower layer of solution; 2. Do not let the glass rod touch the mirror surface when adding the sample). Quickly close the prism and let it stand for 1 minute to allow the test solution to be uniform and free of bubbles and fill the field of view. Aim at the light source and observe the objective lens through the eyepiece. Adjust the indicator gauge to divide the field of view into light and dark parts, then rotate the fine-tuning screw to make the boundary between light and dark clear, and make the dividing line exactly at the cross point of the objective lens. Read the percentage or refractive index in the field of view of the eyepiece and record the prism temperature.

[0071] If the eyepiece reading scale is in percentage, it is the soluble solid content (%). If the eyepiece reading scale is in refractive index, it can be converted to soluble solid content (%) according to Appendix A of GB / T12143. The above percentage content is converted to soluble solid content (%) at 20°C according to Appendix B of GB / T12143.

[0072] 5.2 Test results

[0073] Table 5 Soluble solids test results

[0074]

[0075]

[0076] The emulsions of compositions 1-26, 33-34, 37-38 were stable, without demulsification or separation, and the components did not precipitate. Combinations 27-32, 35-36, 39-40 separated, with oily substances precipitating, resulting in a significantly lower solid content in the emulsion system.

[0077] Example 6 Astaxanthin content test

[0078] 6.1 Experimental methods

[0079] The astaxanthin content was detected by high performance liquid chromatography. The specific detection method is as follows:

[0080] 6.1.1 Chromatographic Reference Conditions Chromatographic column: YMC Carotenoid, C30, 250 × 4.6 mm, 5 μm or equivalent performance column;

[0081] Mobile phase: A: methanol, B: methyl tert-butyl ether, C: 1% phosphoric acid solution;

[0082] Table 6 Gradient elution

[0083] Time (min) Mobile phase A (%) Mobile phase B (%) Mobile phase C (%) 0 81 15 4 15 66 30 4 23 16 80 4 27 16 80 4 30 81 15 4 35 81 15 4

[0084] Flow rate: 1.0 mL / min; detection wavelength: 474 nm; column temperature: 30°C;

[0085] 6.1.2 Solution preparation

[0086] 6.1.2.1 Preparation of reference solution:

[0087] Accurately weigh about 10 mg of all-trans-astaxanthin reference substance and place it in a 100 mL brown volumetric flask. Add 20 mL of chloroform solution to dissolve it, dilute to the mark with acetone solution, and shake well to obtain the reference stock solution (about 100 μg / mL).

[0088] Accurately pipette 2 mL of the control stock solution into a 10 mL brown volumetric flask, dilute to the mark with acetone solution, and shake well to obtain the control intermediate solution (about 20 μg / mL).

[0089] Take an appropriate amount of the control intermediate solution and add acetone solution to prepare a standard series solution of 0.4μg / mL to 10μg / mL, and draw a standard working curve. (The concentration range of the standard curve can be adjusted appropriately according to the sensitivity of the instrument and different instrument models)

[0090] 6.1.2.2 Preparation of test solution

[0091] Using compositions 1-40 as the tested samples, shake each composition sample thoroughly, accurately pipette an appropriate amount of each mixed sample, place it in a 50 mL centrifuge tube, add 10 mL of water and vortex mix for 1 minute, then add 10 mL of anhydrous ethanol and vortex mix for 2 minutes, shake and extract with petroleum ether I twice, 20 mL each time, centrifuge at 8000 r / min for 1 minute, collect the two petroleum ether I extracts, evaporate to dryness under reduced pressure at 40°C, and accurately add 20 mL of acetone solution to dissolve to obtain a sample extract.

[0092] Accurately transfer 3 mL of each sample extract to a 15 mL centrifuge tube. Add 2 mL of 0.05 mol / L Tris (hydroxymethylaminomethane) solution and shake well. Then, add 0.3 mL of cholesterol esterase solution (10 U / mL) and shake well. Incubate in a 37°C water bath for 120 min (shake well every 30 min). Remove the sample and add 0.4 g of anhydrous sodium sulfate and 4 mL of petroleum ether I to the centrifuge tube. Vortex mix for 30 s and allow the solution to separate (if emulsification occurs, centrifuge at 8000 rpm for 1 min). Transfer the petroleum ether layer to a 100 mL flat-bottom flask. Add 4 mL of petroleum ether I to the centrifuge tube and repeat the extraction once. Combine the two petroleum ether extracts and evaporate to dryness under reduced pressure at 40°C. Accurately add 3 mL of acetone solution to dissolve the sample. Filter the organic phase through a 0.45 μm filter membrane to obtain the test solution.

[0093] 6.1.3 Detection

[0094] Inject 20 μL of the standard curve working solution and the test sample solution into the liquid chromatograph to establish the standard curve equation. The test sample should show a peak with the same retention time as the reference peak. (Note: Qualitative confirmation was performed using 9-cis-astaxanthin and 13-cis-astaxanthin as controls. The relative retention times of cis-astaxanthin and all-trans-astaxanthin are shown in Table 1. The injection volume can be adjusted appropriately based on instrument sensitivity and model.)

[0095] Table 7

[0096] Components to be tested Relative retention time ratio to all-trans astaxanthin 13-cis-astaxanthin 0.93 All-trans astaxanthin 1 9-cis-astaxanthin 1.29

[0097] 6.2 Test results

[0098] 6.2.1 Result calculation

[0099]

[0100] Linear equation: Atrans=aCtrans+b

[0101]

[0102] Xtotal=Xtrans+X9-cis+X13-cis

[0103] Where:

[0104] X—content of all-trans astaxanthin / 9-cis astaxanthin / 13-cis astaxanthin in the sample, mg / 100g or mg / 100mL;

[0105] Xtotal—total astaxanthin content in the sample, mg / 100g or mg / 100mL;

[0106] M—mass of sample or sampling volume, g or mL;

[0107] V—dilution volume of the sample, mL;

[0108] C—concentration of all-trans astaxanthin / 9-cis astaxanthin / 13-cis astaxanthin in the sample solution, μg / mL;

[0109] C trans—concentration of all-trans astaxanthin in the sample solution, μg / mL;

[0110] C9-cis—concentration of 9-cis astaxanthin in the sample solution, μg / mL;

[0111] C13-cis—concentration of 13-cis astaxanthin in the sample solution, μg / mL;

[0112] Atrans—peak area of ​​all-trans astaxanthin in the sample;

[0113] A9-cis—peak area of ​​9-cis astaxanthin in the sample;

[0114] A13-cis—peak area of ​​13-cis astaxanthin in the sample;

[0115] a—slope of the linear equation;

[0116] b—intercept of the linear equation;

[0117] 1.1—corrected value for the response factor of 9-cis-astaxanthin;

[0118] 1.3—corrected response factor for 13-cis-astaxanthin;

[0119] K—unit conversion coefficient (K=0.1);

[0120] 6.2.2 Calculation results

[0121]

[0122] Compositions 1-26, 33-34, and 37-38 did not demulsify, nor did the oil precipitate. The system was not disrupted, and astaxanthin remained stable in the solution system. Compositions 27-32, 35-36, and 39-40 exhibited demixing, indicating that the solution system had been disrupted and stratified, with oil precipitating and causing the precipitation of astaxanthin. This degraded astaxanthin due to factors such as light, oxygen, and high temperature, resulting in a decrease in its content.

[0123] Although the specific embodiments of the present invention have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details based on all the teachings published, and these changes are all within the scope of protection of the present invention. The entire invention is given by the appended claims and any equivalents thereof.

Claims

1. A composition comprising astaxanthin-embedded microparticles and glycosaminoglycans; the structure of the astaxanthin-embedded microparticles consists of an inner core and an outer shell; the inner core component is astaxanthin, and the outer shell component is composed of a skeleton component, a thickener, and an emulsifier; the mass ratio of the inner core to the outer shell is 1:10-100; the mass ratio of the skeleton component, the thickener, and the emulsifier is 1-2:2-6:1-2; the skeleton component is one or two of resistant dextrin, maltodextrin, and β-cyclodextrin; the thickener is one or more of gum arabic, xanthan gum, pectin, and guar gum; and the emulsifier is medium-chain triglycerides and / or succinic acid glyceride.

2. The composition according to claim 1, characterized in that The composition is uniformly dispersed in water and is in an emulsion state.

3. The composition according to claim 1, characterized in that The mass ratio of the inner core to the outer shell is 1:20-50.

4. The composition according to claim 1, characterized in that The mass ratio of the skeleton component, the thickener and the emulsifier is 1:3:

1.

5. The composition according to claim 1, characterized in that The glycosaminoglycan is selected from one or more of hyaluronic acid, chondroitin sulfate, dermatan sulfate, and keratin sulfate.

6. The composition according to claim 1, characterized in that The composition also includes collagen.

7. The composition according to claim 6, characterized in that The glycosaminoglycans are polymerized onto the collagen molecules.

8. The composition according to claim 1, characterized in that The mass ratio of the astaxanthin-embedded microparticles to glycosaminoglycan is 1:1-10.

9. The method for preparing the composition according to any one of claims 1 to 5 and claim 8, characterized in that: The preparation method comprises the following steps: (1) Add the inner core and outer shell components according to any one of claims 1 to 5 and claim 8 to an appropriate amount of water and stir until the components are fully dissolved to prepare a solution for use; (2) The solution in step (1) is homogenized using a homogenizer to obtain astaxanthin-embedded microparticles, wherein the astaxanthin-embedded microparticles are uniformly dispersed in water to form an emulsion; (3) Glycosaminoglycan is added to the emulsion and stirred until fully dissolved, and then sterilized in an autoclave to obtain a composition.

10. The preparation method according to claim 9, characterized in that: The stirring time in step (1) is 10-40 min, the homogenizing speed in step (2) is 5000-25000 rpm, and the homogenizing speed time is 5-15 min.

11. The preparation method according to claim 9, characterized in that: The stirring time in step (3) is 15-30 minutes; the sterilization temperature in step (3) is 105-121° C., and the sterilization time is 15-30 minutes.

12. Use of the composition according to any one of claims 1 to 8 in the preparation of medicines and foods.

13. Use of the composition according to any one of claims 1 to 8 in the preparation of health food.

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