Antioxidant composition and preparation method and application thereof

Abstract

The invention discloses an antioxidant composition and a preparation method and application thereof, belonging to the technical field of skin care products. The composition comprises the following components by mass: 30-40 parts of prickle oil, 10-15 parts of walnut oil, 4-6 parts of rice bran oil, 1-3 parts of ampelopsis serrulata extract fraction, 1-3 parts of rosemary extract fraction, 5-10 parts of ampelopsis serrulata extract freeze-dried powder, 5-10 parts of rosemary extract freeze-dried powder, and 1-2 parts of tea polyphenols. The obtained composition has good antioxidant properties and good biocompatibility.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and more specifically, to an antioxidant composition and a preparation method and application thereof. Background Art

[0002] With the change of modern lifestyle, external factors such as environmental pollution and ultraviolet radiation are increasingly damaging the skin, which has led to higher demands for the antioxidant properties of cosmetics. As an important tool for daily skin care and beauty, the antioxidant properties of cosmetics are directly related to the health and beauty of the skin. Therefore, it is imperative to improve the antioxidant properties of cosmetics.

[0003] Although antioxidant cosmetics on the market have met consumers' needs for skin protection and beauty to a certain extent, they still have some obvious disadvantages that need to be paid attention to and improved. Some antioxidant cosmetics have insufficient antioxidant content or unreasonable ratios, resulting in limited antioxidant effects. They cannot effectively resist the damage of free radicals and are not effective in delaying skin aging and improving skin quality. In addition, some antioxidant cosmetics may cause allergic reactions or irritate the skin after use. This may be because the antioxidants or other ingredients used in the products are irritating or unsuitable for some people.

[0004] Therefore, it is necessary to provide a composition with good adaptability and significant antioxidant properties for addition to cosmetics. Summary of the invention

[0005] The invention provides an antioxidant composition, comprising 30-40 parts of prickle oil, 10-15 parts of walnut oil, 4-6 parts of rice bran oil, 1-3 parts of ampelopsis serrulata extract fraction, 1-3 parts of rosemary extract fraction, 5-10 parts of ampelopsis serrulata extract freeze-dried powder, 5-10 parts of rosemary extract freeze-dried powder and 1-2 parts of tea polyphenols.

[0006] Preferably, the composition comprises the following components in parts by weight: 32-38 parts of prickle oil, 11-14 parts of walnut oil; 4-6 parts of rice bran oil; 1-2 parts of Ampelopsis pilosa extract fraction, 1-2 parts of rosemary extract fraction, 5-8 parts of Ampelopsis pilosa extract freeze-dried powder, 5-8 parts of rosemary extract freeze-dried powder, and 1-2 parts of tea polyphenols.

[0007] Preferably, the rosemary extract fraction / lyophilized powder is extracted from rosemary seeds; and the Ampelopsis grossedentata extract fraction / lyophilized powder is extracted from Ampelopsis grossedentata fruit.

[0008] The present invention also provides a method for preparing the above-mentioned rosemary / Ampelopsis glauca extract fraction, comprising the following steps:

[0009] S1. Crush the rosemary / Ampelopsis serrulata fruits by ultrafine powder jet mill;

[0010] S2, passing steam into S1 powder at 100-200 kg / h, collecting fractions, and condensing;

[0011] S3. When the temperature of S2 is ≤40°C, seal the fraction with nitrogen and store it at 5°C for later use.

[0012] The present invention also provides a method for preparing the above-mentioned lyophilized powder of rosemary / Ampelopsis glauca extract, comprising the following steps:

[0013] S1, drying the extracted fraction residue, and then adding water to adjust the pH to 8.0±0.5;

[0014] S2. Add Bacillus Velez enzyme and papain into S1 and react for 3-5 hours.

[0015] S3. After the reaction in S2 is completed, filter, centrifuge, collect the supernatant, freeze-dry, package, and store at ≤-20℃ for later use.

[0016] Preferably, the mass ratio of the residue, water, Bacillus Velez subtilis and papain is 1:12:0.011:0.04.

[0017] The present invention also provides a method for preparing the Bacillus Velez enzyme, comprising the following steps:

[0018] S1, taking Bacillus Velezii, thawing it, activating it, and culture it to obtain a culture solution;

[0019] S2, adding the expanded culture solution into LB broth medium, and culturing for 24-48 hours to obtain a fermentation solution;

[0020] S3. Centrifuge the fermentation broth, take the supernatant, and concentrate it to 1 / 5-6 of the original volume to obtain the Bacillus Velez enzyme.

[0021] Preferably, the mass percentage of the expanded culture solution in the LB medium is 1.5-3%.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows: firstly, the present invention uses prickle fruit oil, walnut oil, rice bran oil, and scutellaria serrata extract and rosemary extract as main components and tea polyphenols, wherein prickle fruit oil contains relatively rich fatty acids and triterpenoid compounds, walnut oil contains rich vitamin E, folic acid, unsaturated fatty acids and other substances, and rice bran oil contains a large amount of vitamin E, vitamin B and trace elements, and the above ingredients have good antioxidant, skin improvement, moisturizing, blood circulation and other effects. On this basis, the fractions extracted from scutellaria serrata seeds and rosemary seeds, i.e., volatile polyphenols, aldehydes and short-chain esters, etc., are added to make its antioxidant effect even better. Secondly, the present invention uses Bacillus velez enzyme and papain to ferment and enzymolyze the residue, and the obtained enzymolysis solution has greatly increased the polypeptide and phenolic substances, and the residue is used as a raw material for secondary processing, which enhances the resource utilization rate and greatly reduces the cost.

[0023] It is found through experiments that the composition prepared by using the raw materials and process defined in the present invention can have significant antioxidant properties. DETAILED DESCRIPTION

[0024] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention is further described by the following examples. It should be understood that the examples of the present invention are only used to illustrate the technical effects of the present invention, but not to limit the scope of protection of the present invention. In the examples, the methods used are conventional methods unless otherwise specified.

[0025] Some of the raw materials in the present invention come from the following sources:

[0026] Prinsepia Utilis Oil: purchased from Jiangxi Global Natural Flavors Co., Ltd.

[0027] Walnut Oil: purchased from Jiangxi Global Natural Flavors Co., Ltd.

[0028] Rice Bran Oil: purchased from Jiangxi Global Natural Flavors Co., Ltd.

[0029] Ampelopsis Grossedentata Seed: purchased from Xi'an Noster Biotechnology Co., Ltd.

[0030] Rosemary Seed: purchased from Xi'an Noster Biotechnology Co., Ltd.

[0031] Ampelopsis grossedentata leaf extract: purchased from Xi'an Noster Biotechnology Co., Ltd.

[0032] Rosemary leaf extract: purchased from Xi'an Noster Biotechnology Co., Ltd.

[0033] Tea polyphenols: purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0034] Papain was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0035] Bacillus velezensis: purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0036] The preparation processes of the Ampelopsis grossedentata fraction, freeze-dried powder and the Rosemary fraction, freeze-dried powder are as follows:

[0037] First, the fruits of Agave sphenanthera or rosemary seeds are dried with hot air, crushed with a micro-airflow mill, sieved, steam is introduced at a rate of 100-200 kg / h, and then condensed, and fractions are collected, the fraction temperature is ≤40°C, and the fractions are nitrogen-packaged and stored at 5°C for later use; then, after the fractions are extracted, the residues from which the corresponding fractions have been extracted are dried with hot air, dried and sieved, water, Bacillus velezensis enzyme, and papain are added to react at 60-70°C for 4 hours, and then vacuum filtered, centrifuged at 5000 r / min, the supernatant is collected, and freeze-dried in a freeze dryer, and the powder is collected and placed in an ampoule, packaged, and stored at -20°C for later use.

[0038] The compositions of Examples 1-3 and Comparative Examples 1-8 and their formula weight parts are shown in Table 1.

[0039] Table 1

[0040]

[0041]

[0042] The raw material ratios for the preparation of Bacillus velezinase are shown in Table 2.

[0043] Table 2

[0044]

[0045] 1.DPPH detection

[0046] The antioxidant capacity of Examples 1-3 and Comparative Examples 1-4 was verified using DPPH free radicals.

[0047] Prepare 0.1mmol / L DPPH solution Take examples 1-3 and comparative examples 1-4 to prepare 10g / L sample solution, 1mg / mL positive control Vc solution and blank control solution. Take 0.1ml sample solution and 4.9ml DPPH solution respectively and place them in a dark room for reaction for 20 minutes, then measure the absorbance at 517nm wavelength and calculate the clearance rate DVC%.

[0048]

[0049] Table 3

[0050] Experimental Group Clearance Example 1 84％ Example 2 82％ Example 3 83％ Comparative Example 1 38％ Comparative Example 2 80％ Comparative Example 3 72％ Comparative Example 4 78％ Comparative Example 5 21％ Comparative Example 6 82％ Comparative Example 7 14％ Comparative Example 8 83％ VC 100％

[0051] According to the results of Examples 1-3 in Table 3, the compositions provided by the embodiments of the present invention have good antioxidant properties, especially Example 1 has a better effect.

[0052] By comparing the results of Example 1 with Comparative Examples 1 and 4, it can be seen that the combination of different components can produce a synergistic effect on the antioxidant effect, and the antioxidant effect achieved by the present invention cannot be achieved if any one of the components is omitted.

[0053] Comparing the results of Example 1 with Comparative Examples 2-3, it can be seen that the composition of different components can produce a synergistic effect on the antioxidant effect, and changing any one of them cannot achieve the antioxidant effect achieved by the present invention.

[0054] Comparison of Example 2 with Comparative Examples 5 and 7 shows that the Bacillus Velez subtilis enzyme has a more obvious effect on anti-oxidation, and the omission or deletion of the preparation process of the Bacillus Velez subtilis enzyme provided by the present invention cannot achieve the effect provided by the present invention.

[0055] Comparison of Example 2 with Comparative Examples 6 and 8 shows that increasing the proportion of the culture medium and the mass fraction of Bacillus Velezii can not significantly improve the antioxidant capacity compared with increasing the concentration, and its antioxidant capacity does not show a linear improvement with increasing concentration. Based on cost-effectiveness considerations, the use of the process, mass fraction and ratio provided by the present invention can achieve the best antioxidant performance shown in the present invention.

[0056] 2. Determination of antioxidant activity (AOA) in PUFA peroxidation system

[0057] Fe 2+ The peroxidation model of very low density lipoprotein (VLDL) and low density lipoprotein (DL) superunsaturated fatty acids (PUFA) at the C2 position of egg yolk phospholipids was induced, and the anti-lipid peroxidation effect of the samples was determined by the thiobarbituric acid method.

[0058] Yolk suspension: remove the white of fresh eggs, and prepare the egg yolk into a 1:1 suspension with an equal volume of pH 7.45, 0.1 mol / L sodium phosphate buffer (PBS). Stir magnetically for 10 min, and then dilute with PBS buffer to a suspension with a volume ratio of 1:25 (refrigerate for later use).

[0059] Determination method: Add 0.4mL of egg yolk suspension, 0.1mL of 10g / L sample solution, 0.4mL of FeSO4 (25mmol / L), and 3.1mL of PBS (0.1mo / L; pH 7.45) to a stoppered test tube, mix well and oscillate at 37℃ for 15min. After taking out, add 1mL of trichloroacetic acid (mass fraction 20%), centrifuge at 3500r / min for 10min, aspirate 2mL of supernatant, add 1mL of TBA (mass fraction 0.8%), stopper and place in a boiling water bath for 15min. After cooling, adjust to zero with a blank tube (replaced with 3mL of distilled water), and determine the absorbance colorimetrically at a wavelength of 532nm. The absorbance of the tube without sample (replaced with 0.1mL of 1mg / mLVc) is A0, and the absorbance of the sample tube is A.

[0060] The AOA of the samples was expressed as the inhibition rate of lipid peroxidation (LPO) of egg yolk lipoprotein.

[0061]

[0062] Note: AOA is the peroxidation inhibition rate of the sample solution in the yolk lipoprotein lipid peroxidation system; A is the average absorbance after adding the example or comparative example; A0 is the average absorbance of the blank group.

[0063] Table 4

[0064] Experimental Group Inhibition rate Example 1 82％ Example 2 85％ Example 3 83％ Comparative Example 1 45％ Comparative Example 2 79％ Comparative Example 3 64％ Comparative Example 4 76％ Comparative Example 5 21％ Comparative Example 6 83％ Comparative Example 7 7％ Comparative Example 8 85％ VC 100％

[0065] According to the results of Examples 1-3 in Table 4, the compositions provided by the embodiments of the present invention have good antioxidant properties, especially Example 2 has a better effect.

[0066] By comparing the results of Example 2 with Comparative Examples 1 and 4, it can be seen that the combination of different components can produce a synergistic effect on the antioxidant effect, and the antioxidant effect achieved by the present invention cannot be achieved if any one of the components is omitted.

[0067] Comparing the results of Example 2 with Comparative Examples 2-3, it can be seen that the composition of different components can produce a synergistic effect on the antioxidant effect, and changing any one of them cannot achieve the antioxidant effect achieved by the present invention.

[0068] Comparison of Example 2 with Comparative Examples 5 and 7 shows that the Bacillus Velez subtilis enzyme has a more obvious effect on anti-oxidation, and the omission or deletion of the preparation process of the Bacillus Velez subtilis enzyme provided by the present invention cannot achieve the effect provided by the present invention.

[0069] Comparison of Example 2 with Comparative Examples 6 and 8 shows that increasing the proportion of the culture medium and the mass fraction of Bacillus Velezii can not significantly improve the antioxidant capacity compared with increasing the concentration, and its antioxidant capacity does not show a linear improvement with increasing concentration. Based on cost-effectiveness considerations, the use of the process, mass fraction and ratio provided by the present invention can achieve the best antioxidant performance shown in the present invention.

[0070] 3. Safety Testing

[0071] CCK-8 experiments were performed to verify the cytotoxicity of Examples 1-3.

[0072] Human immortalized epidermal cells (HaCaT) were cultured at 3x10 3 The cells were inoculated into a 96-well cell culture plate at a density of 100 μg / cm2. After 24 hours, the culture medium was replaced with a culture medium containing 10% of the composition of Example 1-3, and an equal amount of water was added to the blank control group. After further incubation for 24 hours, the culture medium was replaced with a fresh culture medium without additives, and 10 μl of CCK-8 was added to treat the cells. The cells were incubated at 37°C for 1 hour in a light-shielded state. After that, the culture medium was removed and the absorbance of the sample at 450 nm was measured.

[0073] Formula (1)

[0074] Table 5

[0075] Experimental Group Cell viability Example 1 95.3％ Example 2 96.7％ Example 3 97.4％

[0076] According to the experimental results in Table 5, the cell survival rate was maintained above 95%. The combination of the present experiment had little effect on the viability of HaCat cells.

[0077] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present application.

Claims

1. An antioxidant composition comprising the following mass components: 30-40 parts of prickle oil; 10-15 parts of walnut oil; 4-6 parts of rice bran oil; 1-3 parts of Ampelopsis grossedentata extract fraction; 1-3 parts of rosemary extract fraction; 5-10 parts of freeze-dried powder of Ampelopsis grossedentata extract; 5-10 parts of rosemary extract freeze-dried powder; 1-2 parts of tea polyphenols; The extract fraction of Ampelopsis grossedentata and the freeze-dried powder of Ampelopsis grossedentata extract are extracted from the fruit of Ampelopsis grossedentata. The rosemary extract fraction and the rosemary extract freeze-dried powder are extracted from rosemary seeds; The preparation method of the Ampelopsis grossedentata extract fraction is as follows: The fruits of Ampelopsis grossedentata are dried, crushed, sieved, steamed at 100-200 kg / h, and then condensed, and fractions are collected, the fraction temperature is ≤40°C, and the fractions are nitrogen-packed and stored at 5°C for later use; The preparation method of the rosemary extract fraction is as follows: The rosemary seeds are dried, crushed, sieved, steamed at 100-200 kg / h, and then condensed to collect fractions, the fraction temperature is ≤40°C, the fractions are nitrogen packaged and stored at 5°C for later use; The preparation methods of the freeze-dried powder of Ampelopsis grossedentata extract and the freeze-dried powder of Rosemary extract are as follows: The material residue from which the corresponding fractions have been extracted is dried, and water, Bacillus Velez enzyme, and papain are added, reacted at 60-70°C for 3-5h, filtered, centrifuged, the supernatant is collected, freeze-dried, sealed, and stored at ≤-20°C for later use; The mass ratio of the slag, water, Bacillus Velez enzyme and papain is 1:12:0.011:0.04; When adding water after the slag is dried, the pH value needs to be adjusted to 8.0±0.5; The preparation method of the Bacillus Velez enzyme is as follows: S1: activating and expanding Bacillus velezii to obtain an expanded culture solution; S2: adding the culture solution to LB broth medium and culturing for 24-48 hours to obtain fermentation solution; S3 fermentation broth was centrifuged, and the supernatant was filtered and concentrated to obtain the Bacillus Velezii enzyme; in, The mass percentage of the expanded culture solution in S2 in the LB medium is 1.5-3%.

2. Use of the composition as claimed in claim 1 in preparing cosmetics.

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