A complex with antioxidant and whitening effects, and its preparation method and application

Abstract

The present invention belongs to the field of biomaterial preparation technology and cosmetics, and specifically relates to a complex with antioxidant whitening effect, a preparation method and application thereof. The preparation method of the complex of the present invention comprises the following steps: (1) mixing water with one or more of organic acid, amino acid and amine reagent to obtain solution A; (2) mixing copper salt with alcohol solution to obtain solution B; (3) adding solution A to solution B, adjusting pH to obtain solution C; (4) adding surfactant to solution C, reacting, washing and drying to obtain the complex. The raw materials used in the complex prepared in the present invention have low toxicity and good biocompatibility, and can be widely used in the field of biomedicine. In addition, the complex of the present invention has the function of removing hydroxyl radicals and superoxide anion radicals, and has certain enzyme-like activity.

Description

Technical Field

[0001] The invention belongs to the field of biomaterial preparation technology and cosmetics, and specifically relates to a complex with antioxidant and whitening effects, a preparation method and application thereof. Background Art

[0002] Metal-organic frameworks (MOFs) are a new type of coordination material. They are composed of metal ions or ion clusters as vertices and organic ligands as connectors, and are assembled through coordination bonds to form a topological structure with a three-dimensional periodic space. Biometal-organic frameworks, as a type of MOFs material, are composed of metal ions and organic ligands with good biocompatibility.

[0003] Copper ions, as one of the essential trace elements for the human body, not only have low toxicity, but also have good biocompatibility and certain antibacterial and anti-inflammatory properties. Using copper ions as the metal ion center gives the invention the potential and development prospects for practical application in the biological field.

[0004] The generation of free radicals and the appearance of active oxygen are one of the important causes of skin aging. Common cosmetics on the market now need to have a certain antioxidant effect, so how to effectively remove oxygen free radicals has become a powerful means to resist aging. In addition, the market demand for whitening and spot-removing cosmetics is increasing.

[0005] Chinese invention patent application CN108704663A discloses a method for preparing a bimetallic carbon nanocomposite electrocatalytic material, comprising the following steps: 1) dissolving copper salt in water and stirring until blue and transparent to obtain solution A; 2) dissolving trimesic acid in a mixed solvent of anhydrous ethanol and DMF and stirring to obtain solution B; 3) uniformly mixing solution A and solution B and ultrasonicating for 1 to 3 hours, washing the precipitate and drying it in an oven at 40 to 80°C for 1 to 3 hours to obtain a Cu3(BTC)2 precursor; 4) dispersing Cu3(BTC)2 in a mixed solvent of deionized water and anhydrous ethanol, adding RuCl3·xH2O and stirring for 4 to 24 hours, centrifuging to obtain a precipitate, washing it with anhydrous ethanol and drying it to obtain a MOFs material containing Cu and Ru bimetallics; 5) placing the MOFs material in a tubular furnace, heating it to 400 to 700°C in an inert atmosphere and keeping it warm for 1 to 24 hours. 5h, and cooled to room temperature to obtain Cu and Ru bimetallic carbon nanocomposite electrocatalytic material CuRu / C. However, the catalytic material prepared by the invention is not suitable for application in the field of cosmetics.

[0006] Chinese invention patent application CN114591509A discloses a metal organic framework material with antioxidant activity, wherein the metal organic framework material uses tri(triphenylphosphine) ruthenium dichloride [Ru(PPh3)3Cl2] as a metal node and HATP·6HCl as an organic ligand, and is formed in water with ruthenium ions as the center and coordinated with the organic ligands; the metal organic framework has a three-dimensional pore structure, with metal ions as connection points, and organic ligands supporting the formation of a 3D extension of the space, with large pores, a very high specific surface area, multiple enzyme activities, and the ability to adsorb and remove multiple active oxygen and active nitrogen free radicals, and can be used for biological antioxidant stress treatment. However, the application effect in the field of cosmetics is not described.

[0007] In the past, metal-organic frameworks were often used in biocatalysis and sensing, medical imaging, bone tissue engineering, drug delivery, etc., but little is known about metal-organic framework materials used in the cosmetics field. Summary of the invention

[0008] In view of the deficiencies of the prior art, the present invention provides a complex with antioxidant and whitening effects, a preparation method and application thereof. In the present invention, an organic ligand with good biocompatibility is used, and a complex with antioxidant and whitening effects is successfully prepared by hydrothermal synthesis. In the past, metal organic frameworks were often used in biocatalysis and sensing, medical imaging, bone tissue engineering, drug delivery, etc., but little is known about metal organic framework materials used in the field of cosmetics. The present invention creatively develops the application of bio-MOFs materials in the field of cosmetics, and has antioxidant and whitening effects, providing a new idea for the application of bio-MOFs materials.

[0009] In order to achieve the purpose of the present invention, the following technical solutions are adopted:

[0010] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0011] (1) mixing water with one or more of an organic acid, an amino acid and an amine reagent to obtain a solution A;

[0012] (2) mixing the copper salt with water or an alcohol solution to obtain a solution B;

[0013] (3) adding solution A to solution B and adjusting the pH to obtain solution C;

[0014] (4) Adding a surfactant to solution C, reacting, washing, and drying to obtain the complex.

[0015] Preferably, in step (1), the organic acid is selected from one or more of terephthalic acid, nicotinic acid and trimesic acid; the amino acid is proline or histidine; and the amine reagent is nicotinamide.

[0016] Preferably, the molar concentration of solution A in step (1) is 0.2-0.6 mol / L.

[0017] Preferably, the copper salt in step (2) is copper sulfate or copper nitrate.

[0018] Preferably, the alcohol solution in step (2) is ethylene glycol, and the molar concentration of solution B is 0.03-0.1 mol / L.

[0019] Preferably, the reagent for adjusting pH in step (3) is sodium hydroxide or anhydrous sodium acetate, and the adjusted pH is 5-7.

[0020] Preferably, the surfactant in step (4) is selected from one or more of sodium dodecyl sulfate, polyvinyl pyrrolidone and hexadecyltrimethylammonium bromide, and the molar ratio of the surfactant to the copper salt is 1:2.5-15.

[0021] Preferably, the reaction temperature in step (4) is 80-140° C., the reaction pressure is 2-3 MPa, and the reaction time is 24-48 h.

[0022] The present invention also relates to a complex with antioxidant and whitening effects prepared by the above preparation method.

[0023] The present invention also relates to the application of the complex prepared by the preparation method in the preparation of cosmetics.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The raw materials of the present invention are easily available, the preparation process is simple, and the morphology and size of the sample can be effectively controlled by controlling the pressure, pH, and addition of surfactants during the preparation process;

[0026] (2) The raw materials used in the complex (metal organic framework) prepared in the present invention have low toxicity and good biocompatibility, and can be widely used in the field of biomedicine;

[0027] (3) The present invention innovatively explores the application potential of a series of complexes (metal organic frameworks) in the field of cosmetics, which is conducive to expanding the application field of metal organic frameworks, especially since there is currently no data or literature showing the application of metal organic frameworks in the field of cosmetics;

[0028] (4) The series of complexes (metal organic frameworks) prepared in the present invention have whitening and anti-ultraviolet effects, have the function of removing hydroxyl radicals and superoxide anion radicals, and have certain enzyme-like activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1is a scanning electron microscope photograph of sample 1 in Example 1;

[0030] Figure 2 is the energy spectrum analysis result diagram of sample 1 in Example 1;

[0031] Figure 3 is the powder diffraction pattern (XRD) of sample 1 in Example 1;

[0032] Figure 4 is a polarizing microscope photograph of sample 2 in Example 2;

[0033] Figure 5 is the infrared spectrum of sample 4 in Example 4;

[0034] Figure 6 It is the ultraviolet absorption spectra of the samples in Example 3 and Example 5 and other control samples, wherein curves 1-5 are 1-sample 5, 2-sample 3, 3-nicotinamide powder, 4-sodium carboxymethyl cellulose powder, and 5-silicon powder, respectively. DETAILED DESCRIPTION

[0035] The technical solutions in the embodiments of the present invention are further clearly described. The described embodiments are only a part of the present invention and are used to explain the present invention but not to limit the present invention. Therefore, other embodiments obtained by other technicians in the field without creative work all belong to the protection scope of the present invention.

[0036] Polyvinyl pyrrolidone, specification: average molecular weight 10000, K13-18; sodium hyaluronate, specification: >99.0% biotechnology grade; carbomer (carbomer 940), product number C832684-25g; xanthan gum, specification USP grade, product number G810381-100g; PEG-40 hydrogenated castor oil, specification HLB value: 13-14, pH: 5-7, product number E875014-25g; all purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0037] Example 1

[0038] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0039] (1) Take 5 ml of water, add 0.5 mmol of proline and 0.5 mmol of nicotinamide, and stir to obtain solution A;

[0040] (2) Take 15 ml of ethylene glycol, add 0.5 mmol of copper sulfate thereto, and stir to obtain solution B;

[0041] (3) adding solution A to solution B and mixing, and then dropping NaOH until the pH is 5.5 to obtain solution C;

[0042] (4) 0.2 mmol sodium dodecyl sulfate (SDS) was added to solution C and mixed evenly to obtain a mixed solution, and then the mixed solution was transferred to a reactor, set to 110° C., adjusted to a pressure of 2.5 MPa, and reacted for 48 hours. After the reaction was completed, the reaction solution was taken out and separated, washed three times with deionized water and three times with anhydrous ethanol, and dried to obtain sample 1.

[0043] Sample 1 was subjected to SEM, energy spectrum analysis and XRD analysis tests: Figure 1 This is the SEM image of sample 1. It can be seen that the particle size of sample 1 is about 10 μm, most of them are square, and there are defects and grooves on the surface; Figure 2 The energy spectrum analysis shown shows that Cwt% = 52.9, N wt% = 10.5, O wt% = 18.73%, Cu wt% = 18.22%; the powder diffraction analysis (XRD) results ( Figure 3 ) shows that the product is pure and uniform with good crystallinity.

[0044] Example 2

[0045] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0046] (1) Take 10 ml of distilled water, add 0.5 mmol of proline and 0.5 mmol of terephthalic acid to the solution to obtain solution A;

[0047] (2) Take 10 ml of ethylene glycol, add 0.5 mmol of copper sulfate thereto, and stir to obtain solution B;

[0048] (3) Solution A was added to solution B and mixed, and NaOH was added dropwise until the pH value was 5.5 to obtain solution C;

[0049] (4) Add 0.2 mmol sodium dodecyl sulfate (SDS) to solution C and mix well. Then transfer the mixed solution to a reactor, set the temperature to 110° C., adjust the pressure to 2 MPa, and react for 48 h. After the reaction is completed, take out the reaction solution and centrifuge it. Wash it three times with deionized water and anhydrous ethanol respectively, and dry it to obtain sample 2.

[0050] The polarizing microscope photo of sample 2 is shown in Figure 4 shown.

[0051] Example 3

[0052] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0053] (1) Take 5 ml of distilled water, add 1 mmol of nicotinamide thereto and mix to obtain solution A;

[0054] (2) Take 15 ml of ethylene glycol, add 0.5 mmol of copper sulfate thereto and mix to obtain solution B;

[0055] (3) adding solution A to solution B and mixing, adding anhydrous sodium acetate to adjust the pH to 6, to obtain solution C;

[0056] (4) Add 0.1 mmol of polyvinyl pyrrolidone (PVP) to solution C and mix well to obtain a mixed solution, then transfer the mixed solution to a reactor, set the temperature to 100° C., adjust the pressure to 3 MPa, and react for 24 hours. After the reaction, take out the reaction solution and separate it, wash it three times with deionized water and anhydrous ethanol respectively, and dry it to obtain sample 3.

[0057] The infrared spectrum of sample 3 is as follows Figure 5 shown.

[0058] Example 4

[0059] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0060] (1) Take 5 ml of distilled water, add 1 mmol of nicotinic acid and 1 mmol of trimesic acid, mix, and obtain solution A;

[0061] (2) Take 15 ml of ethylene glycol, add 1 mmol of copper nitrate thereto, mix, and obtain solution B;

[0062] (3) adding solution A to solution B and mixing, adding anhydrous sodium acetate to adjust the pH to 6, to obtain solution C;

[0063] (4) Add 0.1 mmol of polyvinyl pyrrolidone (PVP) to solution C and mix well to obtain a mixed solution, then transfer the mixed solution to a reactor, set it to 140° C., adjust the pressure to 2.5 MPa, and react for 24 hours. After the reaction, take out the reaction solution and separate it, wash it three times with deionized water and anhydrous ethanol respectively, and dry it to obtain sample 4.

[0064] Example 5

[0065] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0066] (1) Take 5 ml of distilled water, add 1 mmol of nicotinamide and 1 mmol of histidine, mix, and obtain solution A;

[0067] (2) Take 15 ml of ethylene glycol, add 1 mmol of copper nitrate thereto, mix, and obtain solution B;

[0068] (3) Solution A was added to solution B and mixed, and NaOH was added dropwise until the pH value was 6.5 to obtain solution C;

[0069] (4) Add 0.1 mmol of hexadecyltrimethylammonium bromide (CTAB) to solution C and mix well to obtain a mixed solution, then transfer the mixed solution to a reactor, set the temperature to 120° C., adjust the pressure to 2.5 MPa, and react for 48 hours. After the reaction, take out the reaction solution, separate it, wash it three times with deionized water and anhydrous ethanol respectively, and dry it to obtain sample 5.

[0070] Example 6

[0071] A method for preparing a complex with antioxidant and whitening effects, comprising the following steps:

[0072] (1) Take 5 ml of distilled water, add 1 mmol of nicotinic acid, 1 mmol of terephthalic acid and 1 mmol of histidine, mix and obtain solution A;

[0073] (2) Take 15 ml of ethylene glycol, add 1.5 mmol of copper sulfate thereto, mix, and obtain solution B;

[0074] (3) adding solution A to solution B and mixing, and adding anhydrous sodium acetate to adjust the pH to 5 to obtain solution C;

[0075] (4) Add 0.1 mmol sodium dodecyl sulfate (SDS) to solution C and mix well to obtain a mixed solution, then transfer the mixed solution to a reactor, set it to 80° C., adjust the pressure to 2.5 MPa, and react for 36 hours. After the reaction, take out the reaction solution and separate it, wash it three times with deionized water and anhydrous ethanol respectively, and dry it to obtain sample 6.

[0076] Example 7

[0077] A whitening facial cream, using sample 3 prepared in Example 3 as a raw material, the specific raw materials and proportions of the facial cream are shown in Table 1.

[0078] The preparation method of the above-mentioned whitening facial cream is:

[0079] (1) Under room temperature stirring, add deionized water, sample 3, glycerol, 1,3-butylene glycol, betaine and sodium hyaluronate triethanolamine of phase A into the main pot in proportion, start stirring, stir at room temperature for 30 minutes to disperse evenly, and use triethanolamine to adjust the solution pH to 5.5;

[0080] (2) Raise the temperature to 40°C, add carbomer and xanthan gum in phase B, and stir for 1 h until a sky blue gel-like substance is obtained;

[0081] (3) Add the essence, PEG-40 hydrogenated castor oil and potassium sorbate from phase C and stir until uniform;

[0082] (4) After cooling to room temperature, the obtained sky blue gel mask product is placed in a sealed container and stored at a low temperature (2° C.) away from light.

[0083] Table 1 Raw materials and proportions of moisturizing cream

[0084]

[0085]

[0086] Comparative Example 1

[0087] A method for preparing a complex with antioxidant and whitening effects, the difference from Example 5 is that the histidine in step (1) is replaced by 2-methylimidazole.

[0088] Comparative Example 2

[0089] A method for preparing a complex with antioxidant and whitening effects, the difference from Example 5 is that the nicotinamide in step (1) is replaced by 2-amino-terephthalic acid.

[0090] Comparative Example 3

[0091] A method for preparing a complex with antioxidant and whitening effects, the difference from Example 5 is that the hexadecyltrimethylammonium bromide in step (4) is replaced by sodium benzenesulfonate.

[0092] Comparative Example 4

[0093] A method for preparing a complex with antioxidant and whitening effects, the only difference from Example 5 is that the reaction temperature in step (4) is 180°C.

[0094] Comparative Example 5

[0095] A method for preparing a complex with antioxidant and whitening effects, the difference from Example 5 is that the ethylene glycol in step (2) is replaced by propylene glycol.

[0096] Comparative Example 6

[0097] A method for preparing a complex with antioxidant and whitening effects, the only difference from Example 5 is that 1 mmol of copper nitrate in step (2) is replaced by 1 mmol of zinc acetate.

[0098] Comparative Example 7

[0099] A whitening facial cream, which is different from Example 7 only in that Sample 3 in Phase A is replaced with niacinamide.

[0100] Effect test

[0101] 1. Antioxidant performance test

[0102] (1) Test on the effect of removing hydroxyl radicals

[0103] The scavenging test of hydroxyl radicals is usually carried out by competitively capturing hydroxyl radicals with salicylic acid. During the test, hydrogen peroxide generates hydroxyl radicals under the action of ferrous ions. Salicylic acid is added to the system to capture hydroxyl radicals, and purple products are generated at the same time, with characteristic absorption in the visible light region. Subsequently, antioxidants are added to the reaction system to compete with salicylic acid to capture hydroxyl radicals, reduce the generated purple products, change the absorbance of the solution, and compare with the blank group to know the antioxidant activity of the sample.

[0104] The test method is:

[0105] Preparation of sample solution: Before testing, samples 1 and 4-6 prepared in Example 1, Examples 4-6, and samples prepared in Comparative Examples 1-6 were respectively prepared into solutions with a concentration of 1 mg / ml, the solvent was pure water, and the concentration of vitamin C in the control group was 1 mg / ml.

[0106] Add 0.067 mL of 2.2 mmol / L H2O2, 2.25 mmol / L FeSO4 aqueous solution, 2.25 mmol / L salicylic acid and 95% ethanol solution to a 96-well plate, and 0.1 mL of sample solution. After reacting at room temperature and away from light for 5 minutes, detect the absorbance at 510 nm. The blank group replaced the sample solution with 0.1 mL of water, and the control group replaced the sample solution with 0.1 mL of vitamin C. Each group of experiments was repeated 3 times and the average value was taken. The hydroxyl radical scavenging rate was calculated as follows:

[0107] Hydroxyl radical scavenging rate = (A1-A2) / A1×100%

[0108] Where: A1 is the average absorbance of the blank; A2 is the average absorbance of the sample.

[0109] The test results are shown in Table 2 and Table 3.

[0110] Table 2 Test results of the effect of removing hydroxyl radicals

[0111]

[0112] Table 3 Test results of the effect of removing hydroxyl radicals

[0113]

[0114] (2) Superoxide anion removal test

[0115] Pyrogallol undergoes a self-oxidation chain reaction in an alkaline environment and releases a large amount of superoxide anions. The generated superoxide anions further accelerate the oxidation of pyrogallol, thereby generating a series of intermediates with characteristic absorption in the visible light region. The addition of antioxidants can reduce the self-oxidation rate of pyrogallol, remove the generated superoxide anions, inhibit the generation of intermediates, and reduce the absorbance of the reaction solution. By comparing the self-oxidation rate of pyrogallol in the test group and the blank group, the antioxidant capacity of the test substance can be obtained. Using the pyrogallol self-oxidation system to determine the scavenging effect of the test substance on superoxide anion free radicals is a simple and quick method to evaluate the antioxidant activity of natural products.

[0116] The test method is:

[0117] Preparation of sample solution: Before testing, samples 3-6 prepared in Examples 3-6 and samples prepared in Comparative Examples 1-6 were prepared into solutions with a concentration of 1 mg / ml. The solvent was pure water. The concentration of vitamin C in the control group was also 1 mg / ml.

[0118] 0.095mL of Tris-HCl buffer (0.05mol / L) with pH=8.2, 0.005mL of pyrogallol solution (25mmol / L) and 0.2mL of sample solution were added to a 96-well plate in sequence. After reacting at room temperature and in the dark for 5 minutes, the absorbance was detected at 320nm. The blank group was replaced with 0.2mL of water instead of the sample solution. Three replicate wells were performed for each group of experiments and the average value was taken. The clearance rate was calculated as follows:

[0119] Superoxide anion radical scavenging rate = (A1-A2) / A1×100%

[0120] Where: A1 is the average absorbance of the blank; A2 is the average absorbance of the sample.

[0121] The test results of superoxide anion removal are shown in Table 4-5.

[0122] Table 4 Superoxide anion removal test results

[0123]

[0124] Table 5 Superoxide anion removal test results

[0125]

[0126] (3) Total antioxidant capacity test

[0127] The DPPH method is widely used to quantitatively determine the antioxidant capacity of biological samples or foods. The principle of this method is that the DPPH free radical has a single electron, a strong absorption peak at 517nm, and its alcohol solution is purple. When a free radical scavenger is present, it will pair with its single electron and gradually disappear its absorption. The degree of fading is quantitatively related to the number of electrons it accepts. This method has become a common method for evaluating the antioxidant activity of natural products.

[0128] Test method:

[0129] Preparation of sample solution: Before testing, samples 3-6 prepared in Examples 3-6 and samples prepared in Comparative Examples 1-6 were prepared into solutions with a concentration of 1 mg / ml. The solvent was pure water. The concentration of vitamin C in the control group was also 1 mg / ml.

[0130] Prepared at a concentration of 2×10 -4 mol / L DPPH ethanol solution (ethanol volume fraction is 95%), stored in dark. Take 100μL DPPH solution, add 100μL of different sample solutions respectively, mix thoroughly, react at 25℃ in dark for 30min, measure its absorption value at 517nm (maximum absorption wavelength), and calculate DPPH free radical active scavenging rate according to the following formula:

[0131] P=[(1-(A i -A j ) / A c )]×100%

[0132] In the formula, A i Indicates the absorbance of 100 μL DPPH solution + 100 μL sample solution; A j Indicates the absorbance of 100 μL sample solution + 100 μL solvent; A c It represents the absorbance of 100 μL DPPH solution + 100 μL solvent.

[0133] The total antioxidant capacity test results are shown in Tables 6-7.

[0134] Table 6 Total antioxidant capacity test results

[0135]

[0136] Table 7 Total antioxidant capacity test results

[0137]

[0138] From the results in Table 6, it can be seen that although the samples in Examples 3-6 are not as good as the antioxidant performance of vitamin C, these are the results obtained at its lower concentration. Considering that vitamin C is completely soluble in water, but the samples in the examples are all insoluble in water, the antioxidant capacity of the samples has not been fully demonstrated within the experimental conditions, and in this case, the free radical scavenging rate is still close to half of that of the positive control group, so it can be inferred that the samples have relatively good antioxidant performance. Compared with the results in Table 7, the antioxidant capacity of the samples in the examples is also much higher than that of comparative examples 1-6, and the data in the above Tables 2-5 show that the effect of the samples is also significantly better than that of the comparative examples.

[0139] 2. Tyrosinase inhibition rate test

[0140] In the process of melanin production, tyrosine is first converted into dopa, and then dopa is converted into dopaquinone, in which tyrosinase plays a major catalytic role. Dopaquinone is a colored substance with characteristic absorption at a wavelength of 475nm. When a sample solution that inhibits tyrosinase is added, the amount of dopaquinone synthesis decreases, and the amount of decrease has a certain dose-effect relationship with the amount of sample solution added. Therefore, the change in absorbance at 475nm can be used to compare the strength of the sample solution's inhibitory effect on tyrosinase.

[0141] Test method:

[0142] Preparation of sample solution: Before testing, samples 3-6 prepared in Examples 3-6 and samples prepared in Comparative Examples 1-6 were prepared into solutions with a concentration of 1 mg / ml, the solvent was pure water, and the concentrations of vitamin C and niacinamide in the control group were also 1 mg / ml.

[0143] Prepare phosphate buffer solution (PBS) with a pH of 6.83, tyrosinase solution with a mass concentration of 0.07 mg / mL, and L-tyrosine solution with a mass concentration of 1.0 mg / mL. Accurately prepare test solution 1: 2.0 mL PBS + 0.5 mL tyrosinase solution + 0.5 mL L-tyrosine solution; test solution 2: 2.5 mL PBS + 0.5 mL tyrosinase solution; test solution 3: 0.5 mL sample solution + 1.5 mL PBS + 0.5 mL tyrosinase solution + 0.5 mL L-tyrosine solution; test solution 4: 0.5 mL sample solution + 2.0 mL PBS + 0.5 mL tyrosinase solution; test solutions 1-4 were first placed in a 37°C water bath for 30 minutes and then the absorbance was immediately measured at 475 nm. The absorbances were recorded as A1, A2, A3, and A4, respectively. The tyrosinase inhibition rate was calculated according to the following formula.

[0144] I=[(A1-A2)-(A3-A4)] / (A1-A2)×100%

[0145] The tyrosinase inhibition rate test results are shown in Table 8-9.

[0146] Table 8 Tyrosinase inhibition test results

[0147]

[0148] Table 9 Tyrosinase inhibition test results

[0149]

[0150] As shown in Table 8, the samples obtained in Examples 3 and 5 have a good tyrosinase inhibition rate, even higher than the control group, and the tyrosinase inhibition of vitamin C is mainly attributed to its antioxidant properties, and its unstable and easily oxidized generally does not take into account its actual whitening effect. The tyrosinase inhibition rate of samples 4 and 6 is low, but it is more than half overall, and the tyrosinase inhibition rate of nicotinamide as a commonly used whitening ingredient in cosmetics is not obvious in its in vitro test, and is even much lower than that of Examples 4 and 6, and is much lower than that of the samples in Examples 3 and 5, so it can be seen from this result that the samples in Examples 3-6 have a good tyrosinase inhibition rate, and it can be said that they have a good whitening potential. Compared with the results of Table 9, the tyrosinase inhibition rate of each sample is also generally higher than that of each comparative example.

[0151] 3. UV absorption test

[0152] Ultraviolet radiation is considered to be one of the main causes of photoaging damage to the skin, and it can be divided into three bands according to different wavelengths. The wavelength of 200-280nm is called short-wave ultraviolet (ultraviolet C, UVC), the ultraviolet at 280-320nm is called medium-wave ultraviolet (ultraviolet B, UVB), and the wavelength of 320-400nm is called long-wave ultraviolet (ultravioletA, UVA). UVC will be absorbed by the ozone layer in the atmosphere and will not directly cause damage to the human body, while UVB is more phototoxic than UVA and mainly targets the epidermis of the skin. The epidermis is located in the outermost layer of the skin. After long-term exposure to UVB radiation, the skin barrier is destroyed and the skin will show photoaging. The external manifestations of skin photoaging are mostly abnormal thickening, roughness, skin sagging, and local abnormal pigmentation of the dermis and epidermis of the skin. In severe cases, it may even lead to the formation of benign or malignant tumors. Therefore, in order to avoid damage to the skin by ultraviolet rays in the sun, active ingredients with good absorption in the UVA (320-400nm) and UVB (280-320 nm) bands are usually added to skin care products to achieve the effect of preventing tanning and sunburn. The effective absorption and shielding of ultraviolet rays can also have a whitening effect.

[0153] Test method:

[0154] Weigh accurately (35 mg of sample is weighed accurately and filled evenly in a 0.1 mm detachable quartz cuvette, and the reference substance weighs the same amount), and scan its absorption spectrum at 200-600 nm using a UV-visible spectrophotometer. Test the powder samples in Examples 3 and 5 and the nicotinamide powder, one of the raw materials, and compare them with two reference substances: 4-sodium carboxymethylcellulose (CMC) and 5-silicon powder.

[0155] Depend on Figure 6 The results show that the ultraviolet absorption of the reference substances (5-silicon powder and 4-sodium carboxymethyl cellulose powder) at 200-600nm is relatively flat and weak, and the taste absorption of nicotinamide in this wavelength range is strong, especially the absorption at 200-300nm. The ultraviolet absorption of samples 3 and 5 is stronger than that of nicotinamide at 200-600nm, and the absorption peak range is also wider, with strong absorption between 200-380nm and 500-600nm.

[0156] 4. Sun protection factor analysis test

[0157] The whitening creams prepared in Example 7 and Comparative Example 7 were tested for their sun protection performance. The test method was as follows: 0.1 ml of the prepared whitening cream was drawn out using a special flat-head syringe and evenly applied on the 3M tape, then spread evenly, and then analyzed using an SPF sun protection factor analyzer.

[0158] SPF: Sun Protection Factor; UVA / UVB radio: This ratio is used to measure the relative absorbance of sunscreen samples in the UVA and UVB ranges. The UVA wavelength range is 320-400nm, while the UVB wavelength range is 290-320nm; Boots Star Rating (2004): Boots Star Rating Standard given by the British Boots Company, widely used in countries such as the United Kingdom, Ireland and Thailand. This method evaluates the effectiveness of sunscreen by calculating the relative protection factor of UVA to the UVB wavelength range; UVA I / UV Ratio: A new in vitro UVA test standard proposed by the US Food and Drug Administration FDA in 2004. PFA: PFA (Protection Factor of UVA) refers to the ability to defend against UVA. The calculation formula is PFA = MPPD value of skin with sunscreen applied / MPPD value of skin without sunscreen applied. The higher the PFA value, the longer the protection time; PA grade: PA is the abbreviation of Protection Grade of UVA, and "+" indicates the ability of the product to defend against long-wave ultraviolet rays. The PA grade is determined based on the long-wave UV protection index (PFA) of sunscreen cosmetics. It reflects the protective effect against long-wave UV tanning and is a protection indicator for evaluating the ability of sunscreen cosmetics to prevent skin tanning.

[0159] The test results are shown in Table 10.

[0160] Table 10 Sun protection factor analysis test results

[0161] project Example 7 Face Cream Comparative Example 7 Cream SPF 3.05 1.40 UVA / UVB radio 0.586 -0.131 Boots star rating(2004) 2 0.1 UVA I / UV Ratio 0.6 -1.54 PFA 2.06 1.18 PA Level + none

[0162] It can be seen from Table 10 that, compared with the cream of Comparative Example 7, the sun protection performance of the cream of Example 7 is significantly improved, and the SPF value is increased to 3.05, indicating that only about 1 / 3 of the solar radiation can reach the skin, wherein the Boots star rating (2004) is increased from 0 to 2. According to the Boots star rating standard given by the British Boots company, the sun protection level of the cream can be rated as *, that is, it has a sun protection effect; according to the new in vitro UVA test standard proposed by the US Food and Drug Administration FDA, UVA I / UV Ratio=0.6 can reach the "medium" ordinary level; according to the most common PA grade index, after adding the sample, the PFA of the cream is increased to 2.06, reaching the PA+ level, and also has a basic sun protection effect.

[0163] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. A method for preparing a complex with antioxidant and whitening effects, characterized in that: The following steps are involved: (1) mixing water with one or more of an organic acid, an amino acid and an amine reagent to obtain a solution A; (2) mixing the copper salt with the solvent to obtain solution B; (3) Add solution A to solution B and adjust the pH to obtain solution C; (4) adding a surfactant to solution C, reacting, washing, and drying to obtain the complex; One or more of the organic acids, amino acids and amine reagents in step (1) is selected from a combination of proline and nicotinamide, a combination of proline and terephthalic acid, a combination of nicotinic acid and trimesic acid, a combination of nicotinamide and histidine, a combination of nicotinic acid, terephthalic acid and histidine, or nicotinamide; The solvent in step (2) is ethylene glycol; The reaction temperature in step (4) is 80-140°C.

2. The preparation method according to claim 1, characterized in that: The total molar concentration of one or more of the organic acid, amino acid and amine reagents in step (1) in solution A is 0.2-0.6 mol / L.

3. The preparation method according to claim 1, characterized in that: The copper salt in step (2) is copper sulfate or copper nitrate.

4. The preparation method according to claim 1, characterized in that: The molar concentration of the copper salt in the solution B in step (2) is 0.03-0.1 mol / L.

5. The preparation method according to claim 1, characterized in that: The reagent for adjusting pH in step (3) is sodium hydroxide or anhydrous sodium acetate, and the adjusted pH is 5-7.

6. The preparation method according to claim 1, characterized in that: In step (4), the surfactant is selected from one or more of sodium dodecyl sulfate, polyvinyl pyrrolidone and hexadecyl trimethyl ammonium bromide, and the molar ratio of the surfactant to the copper salt is 1:2.5-15.

7. The preparation method according to any one of claims 1 to 6, characterized in that: The reaction pressure in step (4) is 2-3 MPa, and the reaction time is 24-48 h.

8. A complex with antioxidant and whitening effects prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the complex prepared by the preparation method according to any one of claims 1 to 7 in the preparation of cosmetics.

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