Human albumin antioxidant essence and preparation method thereof

Abstract

The invention relates to the technical field of protein cosmetics, in particular to human albumin antioxidant essence and a preparation method thereof. The human albumin antioxidant essence is prepared from the following components in parts by mass: 5 to 35 parts of human albumin; 10 to 25 parts of a ginseng extract; 0.2 to 0.8 part of a humectant; 0.01 to 0.1 part of a solubilizer; 0.2-1 part of fatty acid; 0.2 to 2 parts of amino acid; and the balance of deionized water. The human albumin anti-oxidation essence provided by the invention adopts the human albumin and the ginseng extract as active ingredients, has good biocompatibility and effectiveness, and has important significance on skin oxidation resistance and skin improvement.

Description

Technical Field

[0001] The present invention relates to the technical field of protein cosmetics, and in particular to a human albumin antioxidant essence and a preparation method thereof. Background Art

[0002] With the continuous development of social economy and the improvement of people's living standards, the problem of skin aging has received more and more attention. Among many factors, oxidative damage caused by excessive free radicals is considered to be the main factor of skin problems such as skin aging. Free radicals are the products of human metabolism. Under normal circumstances, the antioxidant enzymes in the body will prevent free radicals from damaging cells. However, endogenous factors such as work and rest disorders and mental stress and exogenous factors such as ultraviolet rays and pollution break the balance of free radicals and destroy the skin's own antioxidant system, causing the activity level of antioxidant enzymes to decrease and causing lipid peroxide accumulation, non-enzymatic free radical scavengers to be exhausted, leading to skin damage, and damaged skin antioxidant defense system will lead to the generation of more free radicals. Therefore, solving skin problems requires improving antioxidant and free radical resistance.

[0003] Existing cosmetics use chemical reagents or plant extracts as main ingredients. Chemical reagents have poor biocompatibility and are prone to cause inflammation, redness, swelling, allergies, pain and other problems. Plant extracts are subject to the extraction process, resulting in low and fluctuating active ingredient content, which leads to a high amount of plant extracts and increased costs. Therefore, it is difficult for existing cosmetics to meet both biocompatibility and antioxidant effects at the same time, and it is difficult to meet the skin's antioxidant needs. Summary of the invention

[0004] In order to solve the deficiencies of the prior art, the present invention provides a human albumin antioxidant essence and a preparation method thereof. The human albumin antioxidant essence provided by the present invention uses human albumin and ginseng extract as active ingredients, has good biocompatibility and effectiveness, and is of great significance for skin anti-oxidation and skin improvement.

[0005] The present invention is achieved through the following technical solutions: In a first aspect, the present invention provides a human albumin antioxidant essence, which comprises the following components by weight and does not contain a preservative: Human albumin 5-35 parts; 10-25 parts of ginseng extract; Moisturizer 0.2~0.8 parts; Solubilizer 0.01~0.1 part; 0.2~1 part fatty acid; Amino acids 0.2~2 parts; The balance was deionized water.

[0006] Human albumin (HA), also known as human serum albumin (HSA), is a monomeric protein produced by the liver. Mature human albumin contains 585 amino acid residues and a molecular weight of 66 KD. Human albumin is characterized by containing 17 disulfides and only one unpaired cysteine ​​(Cys34). Cysteine ​​can be free in reduced albumin or as a mixture of disulfide and cysteine. Oxidation reactions are closely related to human health. Many diseases are caused by human oxidation reactions. At the same time, free radicals in human oxidation reactions are an important type of oxidative component. Excessive free radicals in the body will cause lipid peroxidation, which in turn leads to aging and the occurrence of diseases. Human albumin has antioxidant properties and can scavenge free radicals (such as hypochlorous acid (HOCl) and peroxynitrite (ONOOH)) and provide sulfhydryl groups (-SH).

[0007] The Chinese Pharmacopoeia (2020 edition) records that ginseng (Ginseng Radix et Rhizoma Rubra, GRRR) is the dried root and rhizome of Panax ginseng CA Mey. of the Araliaceae family. Ginseng is warm in nature, sweet and slightly bitter in taste, and has the effects of greatly replenishing vital energy, restoring pulse and strengthening deficiency, and replenishing qi and blood. Because ginseng contains ginsenosides, ginsenodiol, ginsenotriol, protopanaxadiol, protopanaxadiol and other ingredients, it has the effects of anti-cholesterol, improving skin nutrition, and can play a role in delaying skin aging.

[0008] Preferably, the human albumin comprises recombinant human albumin.

[0009] Preferably, the mass fraction of human albumin is 14 to 18 parts.

[0010] Preferably, the preparation method of the ginseng extract comprises the following steps: After the ginseng is crushed, it is soaked in ethanol with a volume concentration of 60% to 95% at a solid-liquid ratio of 1:(5 to 10) for 8 to 14 hours, reflux extraction is performed 2 to 3 times, and the filtrate is combined. After the ethanol is removed from the filtrate, it is extracted 1 to 3 times with pre-saturated ethyl acetate at a ratio of 1:1, and the ethyl acetate layer is distilled to obtain an extract. The extract is vacuum dried at 40°C to 60°C and -0.1 to 0.1 Mpa, and then crushed by a grinder. The powder obtained by passing through a 20-mesh sieve is the ginseng extract.

[0011] Preferably, the ginseng extract comprises tetracyclic triterpenoid compounds R g1 , R b1 , R e , R b2 One or more of.

[0012] Preferably, the moisturizing agent includes one or more of PEG-400, PEG-1000, and PEG-2000. Further preferably, the moisturizing agent is PEG-1000.

[0013] Preferably, the mass fraction of the moisturizing agent is 0.4 to 0.6 parts.

[0014] Preferably, the solubilizer includes one or more of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. Further preferably, the solubilizer is polysorbate 80.

[0015] Preferably, the mass fraction of the solubilizer is 0.04 to 0.06 parts.

[0016] Preferably, the fatty acid comprises one or more of linoleic acid, linolenic acid, palmitic acid, and myristic acid. Further preferably, the fatty acid is linolenic acid.

[0017] Preferably, the mass fraction of the fatty acid is 0.6-0.8 parts.

[0018] Preferably, the amino acid includes one or more of arginine, histidine, and lysine. Further preferably, the amino acid is arginine.

[0019] Preferably, the mass fraction of the amino acid is 1.0-1.3 parts.

[0020] Preferably, the pH of the human albumin antioxidant essence is 6.6-7.0.

[0021] In a second aspect, the present invention provides a method for preparing the human albumin antioxidant essence as described in the first aspect, comprising the following steps: Ginseng extract, moisturizer, solubilizer, fatty acid and amino acid are dissolved in deionized water to obtain a mixed solution, the mixed solution is added to human albumin, the balance is supplemented with deionized water, and after stirring, sterilization and filtration, human albumin antioxidant essence is obtained.

[0022] Preferably, the mixture is stirred at 40-60 rpm for 10-20 min and sterilized by filtration through filters with pore sizes of 0.22 µm and 0.1 µm, respectively.

[0023] The beneficial effects achieved by one or more technical solutions of the present invention are as follows: The human albumin antioxidant essence of the present invention uses human albumin and ginseng extract as main components. Compared with commercially available cosmetics, the effective ingredients are simplified, while the antioxidant activity and biocompatibility are improved. The storage is simple, and the pH value meets the requirements of human skin.

[0024] The human albumin antioxidant essence of the present invention has a simple preparation process, and is excellent in indicators such as moisturizing property, spreadability, and moisturizing property, and has application prospects.

[0025] The present invention further optimizes the preparation process of ginseng extract and increases the content of tetracyclic triterpenoid compounds (R g1 , R b1 , R e ) total content, which is beneficial to enhance the antioxidant effect of human albumin antioxidant essence. DETAILED DESCRIPTION

[0026] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in combination with specific embodiments and comparative examples.

[0027] Definition: Recombinant human albumin refers to human albumin prepared by recombinant DNA technology for treatment and prevention.

[0028] Source of materials: Recombinant human albumin: Tonghua Anrui Special Biopharmaceutical Co., Ltd., catalog number: ART101S; Ginseng was purchased from Kangmei Xinkaihe (Jilin) ​​Pharmaceutical Co., Ltd., item number: 20231120; equipment: High-speed pulverizer (FW100, Tianjin Test Instrument Co., Ltd.); DZF desktop vacuum oven (DZF-6050, Shanghai Yiheng Scientific Instrument Co., Ltd.); Water bath (HH.S21-8, Shanghai Boxun Medical Biological Instrument Co., Ltd.); Multifunctional fluorescence microplate reader (SPMax 3500FL, Shanghai Shanpu Biotechnology Co., Ltd.); Analytical balance (BSA224S, Sartorius Scientific Instruments Beijing Co., Ltd.); High performance liquid chromatography (Vanquish F, Thermo Fisher Scientific China Ltd.); pH meter (PB-10, Sartorius Scientific Instruments Beijing Co., Ltd.).

[0029] Unless otherwise specified, other reagents and instruments used were commercially available.

[0030] Example 1 Preparation of ginseng extract Accurately weigh 1.0 g of ginseng, crush it and pass it through a 20-mesh sieve to obtain a powder. Use 95% ethanol to soak the dried ginseng powder at a solid-liquid ratio of 1:5 for 12 h, reflux extraction at water bath temperature for 20 min, extract twice, and combine the filtrates; distill to remove ethanol until there is no alcohol taste, add deionized water to suspend to 100 mL, extract with pre-saturated ethyl acetate at a ratio of 1:1, extract twice, take the ethyl acetate layer for distillation operation, remove ethyl acetate, vacuum dry (temperature: 50°C, vacuum degree: 0 Mpa), and then grind it with a grinder, pass it through a 20-mesh sieve, and the obtained powder is recorded as ginseng extract 1.

[0031] Accurately weigh 1.0 g of ginseng, crush it and pass it through a 20-mesh sieve to obtain a powder. Use 95% ethanol to soak the dried ginseng powder for 12 h at a solid-liquid ratio of 1:10, reflux extraction at water bath temperature for 20 min, extract twice, and combine the filtrates; distill to remove ethanol until there is no alcohol taste, add deionized water to suspend to 100 mL, extract with pre-saturated ethyl acetate at a ratio of 1:1, extract twice, take the ethyl acetate layer for distillation operation, remove ethyl acetate, vacuum dry (temperature: 50°C, vacuum degree: 0 Mpa), and then grind it with a grinder, pass it through a 20-mesh sieve, and the obtained powder is recorded as ginseng extract 2.

[0032] Accurately weigh 1.0 g of ginseng, crush it and pass it through a 20-mesh sieve to obtain a powder. Use 60% ethanol to soak the dried ginseng powder at a solid-liquid ratio of 1:10 for 12 h, reflux extraction at water bath temperature for 20 min, extract three times, and combine the filtrates; distill to remove ethanol until there is no alcohol taste, add deionized water to suspend to 100 mL, extract with pre-saturated ethyl acetate at a ratio of 1:1, extract three times, take the ethyl acetate layer for distillation operation, remove ethyl acetate, vacuum dry (temperature: 50 ℃, vacuum degree: 0 MPa), and then grind it with a grinder, pass it through a 20-mesh sieve, and the obtained powder is recorded as ginseng extract 3.

[0033] Accurately weigh 1.0 g of ginseng, crush it and pass it through a 20-mesh sieve to obtain a powder. Use 95% ethanol to soak the dried ginseng powder at a solid-liquid ratio of 1:10 for 12 h, reflux extraction at water bath temperature for 20 min, extract twice, and combine the filtrates; distill to remove ethanol until there is no alcohol taste, add deionized water to suspend to 100 mL, and extract with pre-saturated ethyl acetate at a ratio of 1:1 for 3 times. Take the ethyl acetate layer for distillation to remove ethyl acetate, vacuum dry (temperature: 50 ℃, vacuum degree: 0 MPa), and then grind it with a grinder, pass it through a 20-mesh sieve, and the obtained powder is recorded as ginseng extract 4.

[0034] Accurately weigh 1.0 g of ginseng, crush it and pass it through a 20-mesh sieve to obtain a powder. Use 95% ethanol to soak the dried ginseng powder at a solid-liquid ratio of 1:10 for 12 h, reflux extraction at water bath temperature for 20 min, extract twice, and combine the filtrates; distill to remove ethanol until there is no alcohol taste, add deionized water to suspend to 100 mL, extract with pre-saturated ethyl acetate at a ratio of 1:1, extract once, take the ethyl acetate layer for distillation operation, remove ethyl acetate, vacuum dry (temperature: 50 ℃, vacuum degree: 0 MPa), and then grind it with a grinder, pass it through a 20-mesh sieve, and the obtained powder is recorded as ginseng extract 5.

[0035] Tetracyclic triterpenoids (R g1 , R b1 , R e ) The total content is the evaluation index of ginseng extract. The detection method refers to the content test under ginseng in Part I of the 2020 edition of the "Chinese Pharmacopoeia". The data results are shown in Table 1.

[0036] Table 1 R of different ginseng extracts g1 , R b1 , R e Ingredients

[0037] As shown in Table 1, the total content of tetracyclic triterpenoid compounds in ginseng extract 3 obtained by using 60% ethanol as the extraction solvent is as low as 2.85%, and the total content of tetracyclic triterpenoid compounds in ginseng extracts extracted by 95% ethanol is above 3%, so 95% ethanol is a better extraction solvent. The total content of tetracyclic triterpenoid compounds in ginseng extract 4 is 3.26%, but since ginseng extract 4 is only 0.01% higher than ginseng extract 2 in the case of one more extraction process, considering the time and process cost, the extraction process of ginseng extract 2 is preferably the best extraction process. In the subsequent examples, ginseng extract 2 is used for human albumin antioxidant essence.

[0038] Example 2 Preparation of human albumin antioxidant essence Ginseng extract (ginseng extract 2 in Example 1), moisturizer, solubilizer, fatty acid and amino acid were dissolved in 50 mL of deionized water, and the dissolved mixed solution was slowly added to recombinant human albumin, and the balance was supplemented with deionized water. The mixture was stirred at 50 rpm for 15 min to dissolve, and sterilized and filtered through filters with pore sizes of 0.22 µm and 0.1 µm, respectively. The filtrate obtained was the human albumin antioxidant essence.

[0039] The specific ingredients (by mass, with the remainder made up with deionized water) and pH of different groups of human albumin antioxidant essence are shown below.

[0040] Group 1: recombinant human albumin: 5 parts, ginseng extract: 10 parts, polysorbate 80: 0.01 parts, PEG-1000: 0.2 parts, linolenic acid: 0.2 parts, arginine: 0.2 parts, pH 6.68.

[0041] Group 2: 10 parts of recombinant human albumin, 15 parts of ginseng extract, 0.03 parts of polysorbate 80, 0.4 parts of PEG-1000, 0.5 parts of linolenic acid, 0.8 parts of arginine, pH 6.80.

[0042] Group 3: 15 parts of recombinant human albumin, 20 parts of ginseng extract, 0.06 parts of polysorbate 80, 0.55 parts of PEG-1000, 0.8 parts of linolenic acid, 1.2 parts of arginine, pH 6.82.

[0043] Group 4: 20 parts of recombinant human albumin, 10 parts of ginseng extract, 0.10 parts of polysorbate 80, 0.8 parts of PEG-1000, 1.0 parts of linolenic acid, 2 parts of arginine, pH 6.95.

[0044] Group 5: 15 parts of recombinant human albumin, 15 parts of ginseng extract, 0.01 parts of polysorbate 80, 0.55 parts of PEG-1000, 0.5 parts of linolenic acid, pH 6.48.

[0045] Group 6: 30 parts of recombinant human albumin, 25 parts of ginseng extract, 0.06 parts of polysorbate 80, 0.2 parts of PEG-1000, 0.8 parts of linolenic acid, 0.6 parts of arginine, pH 6.82.

[0046] Group 7: 25 parts of recombinant human albumin, 20 parts of ginseng extract, 0.05 parts of polysorbate 80, 0.4 parts of PEG-2000, 1.0 parts of linolenic acid, pH 6.42.

[0047] Group 8: 15 parts of recombinant human albumin, 20 parts of ginseng extract, 0.06 parts of polysorbate 20, 0.55 parts of PEG-400, 0.8 parts of linoleic acid, 0.8 parts of lysine, pH 6.91.

[0048] Group 9: 10 parts of recombinant human albumin, 15 parts of ginseng extract, 0.03 parts of polysorbate 40, 0.6 parts of PEG-2000, 0.5 parts of linolenic acid, 1.0 parts of histidine, pH 6.91.

[0049] Group 10: 15 parts of recombinant human albumin, 10 parts of ginseng extract, 0.05 parts of polysorbate 60, 0.5 parts of PEG-2000, 0.5 parts of palmitic acid, 0.8 parts of arginine, pH 6.83.

[0050] Group 11: 20 parts of recombinant human albumin, 10 parts of ginseng extract, 0.01 parts of polysorbate 80, 0.6 parts of PEG-400, 0.8 parts of myristic acid, 1.0 parts of histidine, pH 6.94.

[0051] Comparative Example 1 20 parts of ginseng extract (ginseng extract 2 in Example 1), 0.06 parts of polysorbate 80, 0.55 parts of PEG-10000 and 0.8 parts of linolenic acid were added to 50 mL of deionized water to make up the balance and dissolved, stirred at 50 rpm for 15 min to dissolve, and sterilized and filtered through filters with pore sizes of 0.22 µm and 0.1 µm, respectively. The filtrate obtained was the antioxidant essence, recorded as group 12, and the measured pH was 6.76.

[0052] Test Case The human albumin antioxidant essence groups 1 to 11 obtained in Example 2 and the antioxidant essence group 12 obtained in Comparative Example 1 were tested, and the commercially available A liquid (Ziyingsheng Ginseng Intensive Repair Essence, item number: G622708AB18288) was used as group 13, and the commercially available B liquid (Dabao SOD Honey, item number: 202310) was used as group 14 for comparison.

[0053] In vitro antioxidant assay 1) DPPH scavenging ability determination: Preparation of DPPH solution: Accurately weigh 20 mg of DPPH, dissolve it in anhydrous ethanol and dilute it to a 500 mL brown volumetric flask to obtain a DPPH solution with a mass fraction of 0.004%. Store it away from light for later use.

[0054] Vitamin C was selected as the positive control. The experimental group (group 1-14) and the positive control solution were prepared with a starting concentration of 1 mg / mL, and the concentration range was 0.002-1 mg / mL. The 96-well plate was divided into a sample group and a control group, with 3 replicates in each group. A blank control was set up for both the sample group and the control group. The blank group was anhydrous ethanol, which was added to the DPPH solution in an equal volume of 1:1. The 96-well plate was reacted in the dark at room temperature for 30 min, shaken and evenly mixed, and the absorbance was measured at a wavelength of 517 nm. The half inhibitory concentration (IC 50 ) value (μg / mL). IC 50 The value is an important indicator to measure the strength of antioxidant activity. It indicates the concentration required for the antioxidant capacity to achieve half of the scavenging effect. The calculation formula for free radical scavenging rate is shown in formula (1): Free radical scavenging rate (%) = ( )×100%(1) Wherein: A1 is the measured OD value of the sample group, A2 is the measured OD value of the sample control group, and A0 is the measured OD value of the blank group.

[0055] 2) ATBS + Determination of scavenging capacity: ABTS + For the scavenging capacity determination, vitamin C was selected as the positive control. The experimental groups (groups 1 to 14) and the positive control solutions were prepared with a starting concentration of 1 mg / mL, and the concentration range was 0.002 to 1 mg / mL. + The scavenging capacity assay kit was operated according to the instructions. After the 96-well plate was reacted in the dark for 6 min, the absorbance was measured at a wavelength of 405 nm. The test was repeated three times. The calculation formula of free radical scavenging rate was the same as formula (1).

[0056] 3) Determination of free radical OH scavenging ability In the free radical OH scavenging ability assay, vitamin C was selected as the positive control. The experimental group (group 1-14) and the positive control solution were prepared with a starting concentration of 4 mg / mL, and the concentration range was 0.010-1 mg / mL. The operation was carried out according to the instructions of the free radical OH scavenging ability assay kit. The color developer was used to form a red substance, and its color was proportional to the amount of free radical OH. The absorbance was measured at a wavelength of 550 nm, and the test was repeated three times. The calculation formula for the free radical scavenging rate is the same as formula (1).

[0057] The results of in vitro antioxidant tests are shown in Table 2. 50 The smaller the value, the stronger its antioxidant capacity.

[0058] Table 2 Antioxidant test results

[0059] Note: Compared with the vitamin C group, P<0.05, * has significant difference.

[0060] As shown in Table 2, the DPPH scavenging activity, ABTS+ scavenging activity and free radical·OH scavenging activity of groups 1 to 12 were 11.5±0.26~14.2±0.25, 15.4±0.28~23.5±0.29 and 76.99±0.28~85.04±0.22, respectively, which were better than vitamin C as the positive control (15.4±0.31, 35.5±0.26, 160.1±0.39).

[0061] DPPH scavenging activity IC of groups 1 to 4 50The values ​​(11.5±0.26~12.9±0.22) were similar to those of groups 13~14 (12.3±0.32~12.6±0.41), among which the IC 50 The value (11.5±0.28) was significantly lower than that of groups 13~14. + Method, IC of Group 1~4 50 The values ​​(15.4±0.28~20.5±0.31) were similar to those of groups 13~14 (18.4±0.32~19.0±0.41), and the best IC 50 The value (15.4±0.28) was significantly lower than that of groups 13~14. The free radical OH method IC of groups 1~4 50 The values ​​(76.99±0.28~81.25±0.37) were similar to the free radical OH method IC values ​​of groups 13~14. 50 Compared with the values ​​(80.55±0.37~82.41±0.49), it can be seen that the two have similar free radical scavenging abilities, and the IC 50 The value (76.99±0.28) was significantly lower than that of groups 13~14.

[0062] Compared with groups 1 to 11, group 12 without recombinant human albumin had better DPPH scavenging ability and ABTS + And the free radical ·OH scavenging is weak, which further indicates that recombinant human albumin has good antioxidant capacity, and its antioxidant capacity comes from its rich free thiol groups. In summary, the human albumin essence prepared by the present invention has significant antioxidant activity.

[0063] Based on the test results of antioxidant activity, human albumin antioxidant essence groups 1 to 4 were selected, with spreadability, moisturizing, stickiness, moisturizing and antioxidant properties as indicators (as shown in Table 3). According to the requirements of the "Technical Specifications for Safety of Cosmetics" (2015 edition), 50 female employees of Tonghua Anrui Biopharmaceutical Co., Ltd. were selected to conduct a product questionnaire survey. The survey results are shown in Table 4.

[0064] Table 3 Index evaluation table of human albumin antioxidant essence

[0065] Table 4 Index evaluation results of human albumin antioxidant essence

[0066] The results of the questionnaire showed that more than 95% of the subjects believed that human albumin antioxidant essence had better moisturizing effect, more than 90% of the subjects believed that the skin had better spreadability after using human albumin antioxidant essence, and 85% had better moisturizing effect.

[0067] Antioxidant capacity test in mice Establishment of mouse photooxidation model: 40 SPF male mice, weighing 30 g ± 2 g, aged 6-8 weeks, were raised in a general-grade animal breeding room, 5 mice per cage. After the experiment, the mice were randomly divided into 4 groups: 10 mice in each group: Group A: blank control group (no irradiation, no essence group), Group B: UVA + UVB irradiation, Group C: UVA + UVB irradiation + essence group (using essence component 3), Group D: UVA + UVB irradiation + commercial B liquid group (using commercial B liquid).

[0068] Use the UVA / UVB irradiance monitor of Shanghai Sigma High-Tech Co., Ltd. to calibrate the irradiance of SS03AB ultraviolet light therapy device. UVA / UVB dose = UVA / UVB irradiance × time (seconds) Referring to the animal model method established by Li Zhangjun et al. (Journal of Xi'an Jiaotong University (Medical Edition), 2016 (37), 01, 144-147), ultraviolet light box was used for irradiation. Six UVA lamps were installed, each with a wavelength range of 320-400 nm and an irradiation height of 30 cm. There were also three UVB lamps, 40 W, with a wavelength range of 290-320 nm. The nine lamps were made into a light box with an irradiation height of 30 cm. The hair on the back of the mouse was shaved with a baby hair clipper, and then the hair was removed twice with Veet depilatory cream. The essence was applied externally 1 hour before irradiation. Each group was irradiated with UVA+UVB once every other day, and the irradiation intensity of UVB was 0.18 mW / cm 2 , UVA is 1.25 mW / cm 2 The irradiation time was 30 minutes each time from the first to the seventh week, and then extended by 10 minutes each week. At the 12th week, the irradiation time was 80 minutes each time, and it was terminated at the 14th week. UV irradiation (UVA irradiation during the modeling period reached a cumulative amount of 148.5 J / cm 2 , UVB radiation measurement cumulatively reached 21.38 J / cm 2 ).

[0069] After the experiment, the mice were killed and part of the skin from the irradiated area on the back of the mice was taken for determination of superoxide dismutase (SOD) index.

[0070] Superoxide dismutase (SOD) detection was performed using a superoxide dismutase (SOD) assay kit. Sample pretreatment: Accurately weigh the skin tissue, add 9 times the volume of physiological saline at a ratio of weight (g): volume (mL) = 1:9, use dissecting scissors to appropriately chop the skin tissue, and homogenize at low temperature. After tissue homogenization, centrifuge at 3000 rpm for 10 min, and aspirate the supernatant (i.e., 10% homogenate supernatant) for testing. Refer to the instructions for use of the kit, incubate at 37 °C for 20 min, and measure with an enzyme reader at 560 nm.

[0071] Calculation formula: SOD activity (U / mg mass) = [inhibition percentage ÷ (1-inhibition percentage) × V total] ÷ (W × V sample ÷ V total) × F = 11.11 × inhibition percentage ÷ (1-inhibition percentage) ÷ W × F Inhibition percentage = (ΔA blank - ΔA assay) ÷ ΔA blank × 100% Vreaction: total reaction volume, 0.2 mL; Vsample: volume of sample added to the reaction system, 0.018 mL; Vsample: volume of extract added, 1 mL; W: sample mass, mg; 500: F: sample dilution factor Table 5 SOD enzyme content in the skin of mice after ultraviolet irradiation

[0072] Note: Compared with the blank group, P<0.05, * is significantly different.

[0073] As shown in Table 5, the SOD enzyme content measured in the essence group (Group C) was significantly higher than that in the commercially available group (Group D), which was 65.2 U / mg. The SOD enzyme content was significantly increased compared with the blank group, indicating that human albumin essence has significant antioxidant activity in vivo.

[0074] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A human albumin antioxidant essence, characterized in that: Calculated by mass, it includes the following components and does not contain preservatives: Human albumin 5-35 parts; 10-25 parts of ginseng extract; Moisturizer 0.2~0.8 parts; Solubilizer 0.01~0.1 part; 0.2~1 part fatty acid; Amino acids 0.2~2 parts; The balance was deionized water.

2. The human albumin antioxidant essence according to claim 1, characterized in that: The human albumin includes recombinant human albumin.

3. The human albumin antioxidant essence according to claim 1, characterized in that: The preparation method of the ginseng extract comprises the following steps: After the ginseng is crushed, it is soaked in 95% ethanol at a solid-liquid ratio of 1:(5-10) for 8-14 hours, reflux extraction is performed 2-3 times, and the filtrate is combined. After the ethanol is removed from the filtrate, it is extracted 1-3 times with pre-saturated ethyl acetate at a ratio of 1:1, and the ethyl acetate layer is distilled to obtain an extract, which is vacuum dried at 40°C-60°C and -0.1-0.1 Mpa, and then crushed by a grinder. The powder obtained by passing through a 20-mesh sieve is the ginseng extract.

4. The human albumin antioxidant essence according to claim 1, characterized in that: The ginseng extract includes tetracyclic triterpenoid compounds R g1 , R b1 , R e , R b2 One or more of.

5. The human albumin antioxidant essence according to claim 1, characterized in that: The moisturizing agent includes one or more of PEG-400, PEG-1000, and PEG-2000.

6. The human albumin antioxidant essence according to claim 1, characterized in that: The solubilizer includes one or more of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

7. The human albumin antioxidant essence according to claim 1, characterized in that: The fatty acids include one or more of linoleic acid, linolenic acid, palmitic acid, and myristic acid.

8. The human albumin antioxidant essence according to claim 1, characterized in that: The amino acids include one or more of arginine, histidine and lysine.

9. A method for preparing the human albumin antioxidant essence according to any one of claims 1 to 8, characterized in that: The following steps are involved: Ginseng extract, moisturizer, solubilizer, fatty acid and amino acid are dissolved in deionized water to obtain a mixed solution, the mixed solution is added to human albumin, the balance is supplemented with deionized water, and after stirring, sterilization and filtration, human albumin antioxidant essence is obtained.

10. The preparation method according to claim 9, characterized in that: Stir at 40-60 rpm for 10-20 min and sterilize by filtration through filters with pore sizes of 0.22 µm and 0.1 µm, respectively.

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