Anti-free radical composition as well as preparation method and application thereof

Abstract

The invention relates to the technical field of skin care, in particular to an anti-free radical composition as well as a preparation method and application thereof. The invention relates to an anti-free radical composition, which is prepared from 0.1 to 0.5 part of Chinese herbaceous peony root extract composition, 0.001 to 0.01 part of essence, 0.01 to 0.1 part of peach resin extract composition, 0.01 to 0.1 part of rye seed extract composition, 1 to 3 parts of leukocyte extract composition, 0.5 to 1.5 parts of eryngium maritimum callus culture filtrate composition and 0.5 to 1.5 parts of crithmum maritimum callus culture filtrate composition. The crithmum maritimum callus culture filtrate is compounded with the paeonia lactiflora root extract, the peach resin extract, the rye seed extract, the leukocyte extract and the coastal eryngii callus culture filtrate, so that the effects of resisting free radicals, resisting ageing and the like on skin are achieved.

Description

Technical Field

[0001] The invention relates to the technical field of skin care, in particular to an anti-free radical composition and a preparation method and application thereof. Background Art

[0002] The impact of free radicals on human health is mainly reflected in the fact that their excessive generation can cause oxidative stress, damage cells, and then cause a variety of diseases. For example, in biological systems, excessive generation of free radicals can cause cell membrane damage, protein denaturation, DNA damage, etc., thus causing aging, cancer, cardiovascular disease and other problems.

[0003] In recent years, studies have found that peach resin, coastal eryngium and sea fennel have a certain scavenging effect on human free radicals, especially the callus extract of sea fennel can also have a repairing effect on blue light damage. For example, a Chinese patent discloses a composition and application for repairing blue light damage containing sea fennel callus culture filtrate, patent number 202010430066.X, authorization publication number 111700827B, which records: specifically relates to a composition and application for repairing blue light damage containing sea fennel callus culture filtrate. The composition of the present invention includes sea fennel callus culture filtrate, lactobacillus / North American holy grass fermentation product extract, fern extract, aster extract, epigallocatechin gallate glucoside, and several components synergistically enhance the synergy, which can minimize the free radicals generated under blue light irradiation and repair the damage caused by blue light; at the same time, it can also solve the problem of skin aging caused by blue light radiation.

[0004] The disadvantage of the prior art is that the amount of the filtrate of the callus culture of sea fennel is relatively large. According to the above technical solution, the amount must reach 11wt% to achieve the best effect. As is known, sea fennel can only be planted on reefs on the coast of temperate zones, and it is currently impossible to plant it on a large scale in my country. Therefore, there is an urgent need for a composition that can reduce the amount of the filtrate of the callus culture of sea fennel and still resist free radicals and repair blue light damage. Summary of the invention

[0005] In view of the deficiencies in the prior art, one of the objects of the present invention is to provide an anti-free radical composition, a second object of the present invention is to provide a method for preparing a skin care product containing the anti-free radical composition, and a third object of the present invention is to provide an application of the anti-free radical composition in skin anti-aging.

[0006] One of the purposes of the present invention is achieved through the following technical solutions:

[0007] An anti-free radical composition comprises the following raw materials in parts by weight:

[0008] 0.1-0.5 parts of a peony (PAEONIA ALBIFLORA) root extract composition, 0.001-0.01 parts of essence (daily use), 0.01-0.1 parts of a peach (PRUNUS PERSICA) resin extract composition, 0.01-0.1 parts of a rye (SECALE CEREALE) seed extract composition, 1-3 parts of a leukocyte extract composition, 0.5-1.5 parts of an Eryngium maritimum callus culture filtrate composition, and 0.5-1.5 parts of a sea fennel callus culture filtrate composition; wherein:

[0009] The peony root extract composition is a mixture of water, butylene glycol, peony root extract and 1,2-hexanediol in a weight ratio of 80:1:18:1;

[0010] The peach resin extract composition is a mixture of water, peach resin extract, 1,2-hexanediol and ethylhexylglycerin in a weight ratio of 59:18:13:10;

[0011] The rye seed extract composition is a mixture of water, 1,2-pentanediol and rye seed extract in a weight ratio of 70:18:12;

[0012] The leukocyte extract composition is a mixture of leukocyte extract, butylene glycol, phenoxyethanol and water in a weight ratio of 18:1:11:70;

[0013] The Eryngium maritima callus culture filtrate composition is prepared by mixing Eryngium maritima callus culture filtrate, butanediol and water in a weight ratio of 90:2:8;

[0014] The cristata callus culture filtrate composition is prepared by mixing cristata callus culture filtrate, butanediol and water in a weight ratio of 90:2:8.

[0015] The above extracts and culture filtrate have skin conditioning effects, while butylene glycol, 1,2-hexanediol, ethylhexylglycerin, and 1,2-pentanediol are humectants and phenoxyethanol is an antibacterial agent.

[0016] Preferably, the method for preparing the filtrate of the Crimson Fennel callus culture comprises the following steps:

[0017] Step 1, subjecting adult sea fennel to high temperature stress treatment;

[0018] Step 2, returning the adult sea fennel to normal temperature for daily maintenance for 21 days, and selecting the individuals that show no signs of wilting (having live buds and branches that are not wilted) as tissue culture mother plants;

[0019] Step 3, cutting stem segments from the budding part of the mother plant and cutting them into thin slices as explants for callus induction;

[0020] Step 4, performing suspension culture on the callus tissue;

[0021] Step 5, filtering the suspended matter to obtain the Ceratonia suffruticosa callus culture filtrate.

[0022] Preferably, the high temperature stress treatment method is: increase the temperature from 25°C to 35°C within 0.5h, stay for 2 hours, then increase the temperature from 35°C to 45°C, and maintain 45°C for 1h. This method is a gradient temperature increase high temperature stress. The present application reduces the intermediate residence time and the final high temperature maintenance time according to the characteristics of sea fennel that it is not adapted to high temperature environment and has a small volume, so as to prevent the survival rate from being too low to obtain effective explants.

[0023] Preferably, in step 3, the length of the stem segment cut is 3 cm, and the distance between the two ends of the stem segment and the budding site is at least 1 cm. As is known, the cell activity near the budding site is better, so this site is selected as the explant.

[0024] Preferably, step 3 further includes a step of disinfecting the cut stem segments, the method of which is: first soaking the stem segments in 70wt% alcohol for 15s, then soaking the stem segments in 2wt% sodium hypochlorite for 15min, and finally rinsing with clean water for 5 times.

[0025] Preferably, in step 3, the method for inducing callus tissue is: spreading the slices on the surface of a culture medium, the culture medium is MS culture medium mixed with 1.9 mg / L 6-BA and 0.5 mg / L NAA, and culturing for 6 days. Experiments have shown that the callus induction rate exceeds 80%.

[0026] Preferably, in step 4, the suspension culture method is: MS culture medium and water are mixed in a weight ratio of 5:995 and heated to dissolve, and after returning to room temperature, 3% sucrose, 0.005% 2,4-D and 0.001% KT are added according to the weight of the solution, and the culture is placed on a shaker at a speed of 100 r / min for 30 days.

[0027] The second object of the present invention is achieved by the following technical solutions:

[0028] A method for preparing a skin care product containing an anti-free radical composition comprises the following steps:

[0029] S1, weighing phase A, phase B, phase C, phase D and anti-free radical composition;

[0030] S2, add phase A into the emulsifying pot, stir at a speed of 15-20r / min, heat to 82-85℃, homogenize at a speed of 2500-3000r / min for 3-5min, keep warm for 20min, then defoam and cool down;

[0031] S3, after the emulsification pot is cooled to 75°C, add phase B and stir at a speed of 15-20r / min for 3-5min;

[0032] S4, add phase C into the emulsifier and stir at 15-20r / min for 3-5min;

[0033] S5, heat phase D in a pre-melting pot to 60-65°C and stir until there are no particles;

[0034] S6, after the emulsification pot is cooled to 65°C, add phase D and stir at a speed of 15-20 r / min for 5-10 min;

[0035] S7, after the emulsifying pot is cooled to 45°C, the anti-free radical composition is added and stirred at a speed of 15-20 r / min for 5-10 minutes.

[0036] Preferably, the phase A comprises, by weight, 14.49-44.969 parts of water, 0.01-0.3 parts of allantoin, 0.3-0.6 parts of carbomer, 0.1-0.3 parts of niacinamide and 1-3 parts of butylene glycol; the phase B comprises, by weight, 0.1-0.3 parts of polysilicone-11, 49.3-66.1 parts of water, 0.3-2 parts of hydrogenated lecithin, 0.1-1 parts of 1,2-hexanediol and 0.2-0.6 parts of p-hydroxyacetophenone; the phase C comprises, by weight, 0.05-0.1 parts of aminomethyl propanol and 0.05-0.3 parts of water; the phase D comprises, by weight, 0.2-0.6 parts of p-hydroxyacetophenone, 1-3 parts of butylene glycol and 0.2-0.6 parts of 1,2-hexanediol.

[0037] The third object of the present invention is achieved by the following technical solutions:

[0038] The invention discloses an application of an anti-free radical composition in skin anti-aging. The anti-free radical composition is made into a skin care product and applied on the skin surface.

[0039] The beneficial effects of the present invention are as follows: the filtrate of callus culture of sea fennel is compounded with peony root extract, peach resin extract, rye seed extract, leukocyte extract and eryngium littoralis callus culture filtrate to have the effects of anti-free radical and anti-aging on the skin. DETAILED DESCRIPTION

[0040] The following is further described in conjunction with specific implementation methods:

[0041] Unless otherwise specified, the raw materials in the following technical solutions are purchased through normal channels and comply with the relevant provisions of the "Technical Specifications for Safety of Cosmetics".

[0042] Example 1

[0043] Preparation of Crimson Fennel Callus Culture Filtrate:

[0044] Step 1, subjecting adult sea fennel to high temperature stress treatment; the high temperature stress treatment method is: raising the temperature from 25°C to 35°C within 0.5h, staying for 2 hours, then raising the temperature from 35°C to 45°C, and maintaining 45°C for 1h.

[0045] Step 2, returning the adult sea fennel to normal temperature for daily maintenance for 21 days, and selecting the individuals that show no signs of wilting as tissue culture mother plants;

[0046] Step 3, cut the stem segment from the budding part of the mother plant, the length of the cut stem segment is 3 cm, and the distance between the two ends of the stem segment and the budding part is at least 1 cm. First, soak the stem segment in 70wt% alcohol for 15s, then soak the stem segment in 2wt% sodium hypochlorite for 15min, and finally rinse with water 5 times. Cut the stem segment into slices as explants for callus induction, and the method of callus induction is as follows: spread the slices on the surface of the culture medium, the culture medium is MS culture medium mixed with 1.9mg / L 6-BA and 0.5mg / L NAA, and culture for 6d.

[0047] Step 4, the callus tissue is subjected to suspension culture. The suspension culture method is: MS medium and water are mixed at a weight ratio of 5:995, heated to dissolve, 3% sucrose, 0.005% 2,4-D and 0.001% KT are added to the solution after returning to room temperature, and the solution is placed on a shaker and shaken at a speed of 100 r / min for 30 days.

[0048] Step 5, filtering the suspended matter to obtain the Ceratonia suffruticosa callus culture filtrate.

[0049] Preparation of anti-free radical composition:

[0050] Weigh the following raw materials in parts by weight:

[0051] 0.1 part of peony root extract composition, 0.001 part of essence (daily use), 0.01 part of peach resin extract composition, 0.01 part of rye seed extract composition, 1 part of leukocyte extract composition, 0.5 part of Eryngium maritima callus culture filtrate composition and 0.5 part of sea fennel callus culture filtrate composition; wherein:

[0052] The peony root extract composition is a mixture of water, butylene glycol, peony root extract and 1,2-hexanediol in a weight ratio of 80:1:18:1;

[0053] The peach resin extract composition is a mixture of water, peach resin extract, 1,2-hexanediol and ethylhexylglycerin in a weight ratio of 59:18:13:10;

[0054] The rye seed extract composition is a mixture of water, 1,2-pentanediol and rye seed extract in a weight ratio of 70:18:12;

[0055] The leukocyte extract composition is a mixture of leukocyte extract, butylene glycol, phenoxyethanol and water in a weight ratio of 18:1:11:70;

[0056] The Eryngium maritima callus culture filtrate composition is prepared by mixing Eryngium maritima callus culture filtrate, butanediol and water in a weight ratio of 90:2:8;

[0057] The cristata callus culture filtrate composition is prepared by mixing cristata callus culture filtrate, butanediol and water in a weight ratio of 90:2:8.

[0058] Preparation of skin care products:

[0059] S1, weigh: the phase A comprises, by weight, 44 parts of water, 0.01 parts of allantoin, 0.3 parts of carbomer, 0.1 parts of niacinamide and 1 part of butylene glycol; the phase B comprises, by weight, 0.1 parts of polysilicone-11, 50 parts of water, 0.3 parts of hydrogenated lecithin, 0.1 parts of 1,2-hexanediol and 0.2 parts of p-hydroxyacetophenone; the phase C comprises, by weight, 0.05 parts of aminomethyl propanol and 0.05 parts of water; the phase D comprises, by weight, 0.2 parts of p-hydroxyacetophenone, 1 part of butylene glycol and 0.2 parts of 1,2-hexanediol; and the anti-free radical composition prepared above.

[0060] S2, add phase A into the emulsifying pot, stir at a speed of 15r / min, heat to 82℃, homogenize at a speed of 2500r / min for 3min, keep warm for 20min, then defoam and cool down;

[0061] S3, after the emulsification pot is cooled to 75°C, phase B is added and stirred at 15 r / min for 3 min;

[0062] S4, add phase C into the emulsifier and stir at 15 r / min for 3 min;

[0063] S5, heat phase D in a pre-melting pot to 60°C and stir until there are no particles;

[0064] S6, after the emulsification pot is cooled to 65°C, phase D is added and stirred at 15 r / min for 5 min;

[0065] S7, after the temperature of the emulsifying pot is lowered to 45° C., the anti-free radical composition prepared above is added, and the mixture is stirred at a speed of 15 r / min for 5 min.

[0066] Example 2

[0067] Preparation of Crimson Fennel Callus Culture Filtrate:

[0068] Step 1, subjecting adult sea fennel to high temperature stress treatment; the high temperature stress treatment method is: raising the temperature from 25°C to 35°C within 0.5h, staying for 2 hours, then raising the temperature from 35°C to 45°C, and maintaining 45°C for 1h.

[0069] Step 2, returning the adult sea fennel to normal temperature for daily maintenance for 21 days, and selecting the individuals that show no signs of wilting as tissue culture mother plants;

[0070] Step 3, cut the stem segment from the budding part of the mother plant, the length of the cut stem segment is 3 cm, and the distance between the two ends of the stem segment and the budding part is at least 1 cm. First, soak the stem segment in 70wt% alcohol for 15s, then soak the stem segment in 2wt% sodium hypochlorite for 15min, and finally rinse with water 5 times. Cut the stem segment into slices as explants for callus induction, and the method of callus induction is as follows: spread the slices on the surface of the culture medium, the culture medium is MS culture medium mixed with 1.9mg / L 6-BA and 0.5mg / L NAA, and culture for 6d.

[0071] Step 4, the callus tissue is subjected to suspension culture. The suspension culture method is: MS medium and water are mixed at a weight ratio of 5:995, heated to dissolve, 3% sucrose, 0.005% 2,4-D and 0.001% KT are added to the solution after returning to room temperature, and the solution is placed on a shaker and shaken at a speed of 100 r / min for 30 days.

[0072] Step 5, filtering the suspended matter to obtain the Ceratonia suffruticosa callus culture filtrate.

[0073] Preparation of anti-free radical composition:

[0074] Weigh the following raw materials in parts by weight:

[0075] 0.5 parts of peony root extract composition, 0.01 parts of essence (daily use), 0.1 parts of peach resin extract composition, 0.1 parts of rye seed extract composition, 3 parts of leukocyte extract composition, 1.5 parts of Eryngium maritima callus culture filtrate composition and 1.5 parts of sea fennel callus culture filtrate composition; wherein:

[0076] The peony root extract composition is a mixture of water, butylene glycol, peony root extract and 1,2-hexanediol in a weight ratio of 80:1:18:1;

[0077] The peach resin extract composition is a mixture of water, peach resin extract, 1,2-hexanediol and ethylhexylglycerin in a weight ratio of 59:18:13:10;

[0078] The rye seed extract composition is a mixture of water, 1,2-pentanediol and rye seed extract in a weight ratio of 70:18:12;

[0079] The leukocyte extract composition is a mixture of leukocyte extract, butylene glycol, phenoxyethanol and water in a weight ratio of 18:1:11:70;

[0080] The Eryngium maritima callus culture filtrate composition is prepared by mixing Eryngium maritima callus culture filtrate, butanediol and water in a weight ratio of 90:2:8;

[0081] The cristata callus culture filtrate composition is prepared by mixing cristata callus culture filtrate, butanediol and water in a weight ratio of 90:2:8.

[0082] Preparation of skin care products:

[0083] S1, weigh: the phase A comprises, by weight, 15 parts of water, 0.3 parts of allantoin, 0.6 parts of carbomer, 0.3 parts of niacinamide and 3 parts of butylene glycol; the phase B comprises, by weight, 0.3 parts of polysilicone-11, 66 parts of water, 2 parts of hydrogenated lecithin, 1 part of 1,2-hexanediol and 0.6 parts of p-hydroxyacetophenone; the phase C comprises, by weight, 0.1 parts of aminomethyl propanol and 0.3 parts of water; the phase D comprises, by weight, 0.6 parts of p-hydroxyacetophenone, 3 parts of butylene glycol and 0.6 parts of 1,2-hexanediol; and the anti-free radical composition prepared above.

[0084] S2, add phase A into the emulsifying pot, stir at a speed of 20r / min, heat to 85°C, homogenize at a speed of 3000r / min for 5min, keep warm for 20min, then defoam and cool down;

[0085] S3, after the emulsification pot is cooled to 75°C, phase B is added and stirred at 20 r / min for 5 min;

[0086] S4, add phase C into the emulsifier and stir at 20 r / min for 5 min;

[0087] S5, heat phase D in a pre-melting pot to 65°C and stir until there are no particles;

[0088] S6, after the emulsification pot is cooled to 65°C, phase D is added and stirred at 20 r / min for 10 min;

[0089] S7, after the temperature of the emulsifying pot is lowered to 45° C., the anti-free radical composition prepared above is added, and the mixture is stirred at a speed of 20 r / min for 10 min.

[0090] Example 3

[0091] Preparation of Crimson Fennel Callus Culture Filtrate:

[0092] Step 1, subjecting adult sea fennel to high temperature stress treatment; the high temperature stress treatment method is: raising the temperature from 25°C to 35°C within 0.5h, staying for 2 hours, then raising the temperature from 35°C to 45°C, and maintaining 45°C for 1h.

[0093] Step 2, returning the adult sea fennel to normal temperature for daily maintenance for 21 days, and selecting the individuals that show no signs of wilting as tissue culture mother plants;

[0094] Step 3, cut the stem segment from the budding part of the mother plant, the length of the cut stem segment is 3 cm, and the distance between the two ends of the stem segment and the budding part is at least 1 cm. First, soak the stem segment in 70wt% alcohol for 15s, then soak the stem segment in 2wt% sodium hypochlorite for 15min, and finally rinse with water 5 times. Cut the stem segment into slices as explants for callus induction, and the method of callus induction is as follows: spread the slices on the surface of the culture medium, the culture medium is MS culture medium mixed with 1.9mg / L 6-BA and 0.5mg / L NAA, and culture for 6d.

[0095] Step 4, the callus tissue is subjected to suspension culture. The suspension culture method is: MS medium and water are mixed at a weight ratio of 5:995, heated to dissolve, 3% sucrose, 0.005% 2,4-D and 0.001% KT are added to the solution after returning to room temperature, and the solution is placed on a shaker and shaken at a speed of 100 r / min for 30 days.

[0096] Step 5, filtering the suspended matter to obtain the Ceratonia suffruticosa callus culture filtrate.

[0097] Preparation of anti-free radical composition:

[0098] Weigh the following raw materials in parts by weight:

[0099] 0.3 parts of peony root extract composition, 0.005 parts of essence (daily use), 0.05 parts of peach resin extract composition, 0.05 parts of rye seed extract composition, 2 parts of leukocyte extract composition, 1 part of Eryngium maritima callus culture filtrate composition and 1 part of Crimson Fennel callus culture filtrate composition; wherein:

[0100] The peony root extract composition is a mixture of water, butylene glycol, peony root extract and 1,2-hexanediol in a weight ratio of 80:1:18:1;

[0101] The peach resin extract composition is a mixture of water, peach resin extract, 1,2-hexanediol and ethylhexylglycerin in a weight ratio of 59:18:13:10;

[0102] The rye seed extract composition is a mixture of water, 1,2-pentanediol and rye seed extract in a weight ratio of 70:18:12;

[0103] The leukocyte extract composition is a mixture of leukocyte extract, butylene glycol, phenoxyethanol and water in a weight ratio of 18:1:11:70;

[0104] The Eryngium maritima callus culture filtrate composition is prepared by mixing Eryngium maritima callus culture filtrate, butanediol and water in a weight ratio of 90:2:8;

[0105] The cristata callus culture filtrate composition is prepared by mixing cristata callus culture filtrate, butanediol and water in a weight ratio of 90:2:8.

[0106] Preparation of skin care products:

[0107] S1, weigh: the phase A comprises, by weight, 30 parts of water, 0.1 parts of allantoin, 0.4 parts of carbomer, 0.2 parts of niacinamide and 2 parts of butylene glycol; the phase B comprises, by weight, 0.2 parts of polysilicone-11, 55 parts of water, 1.5 parts of hydrogenated lecithin, 0.5 parts of 1,2-hexanediol and 0.5 parts of p-hydroxyacetophenone; the phase C comprises, by weight, 0.07 parts of aminomethyl propanol and 0.2 parts of water; the phase D comprises, by weight, 0.4 parts of p-hydroxyacetophenone, 2 parts of butylene glycol and 0.4 parts of 1,2-hexanediol; and the anti-free radical composition prepared above.

[0108] S2, add phase A into the emulsifying pot, stir at 18r / min, heat to 83°C, homogenize at 2800r / min for 4min, keep warm for 20min, then defoam and cool down;

[0109] S3, after the emulsification pot is cooled to 75°C, phase B is added and stirred at 18 r / min for 4 min;

[0110] S4, add phase C into the emulsifier and stir at 18 r / min for 4 min;

[0111] S5, heat phase D in a pre-melting pot to 63°C and stir until there are no particles;

[0112] S6, after the emulsification pot is cooled to 65°C, phase D is added and stirred at 18 r / min for 8 min;

[0113] S7, after the temperature of the emulsifying pot is lowered to 45° C., the anti-free radical composition prepared above is added, and the mixture is stirred at a speed of 18 r / min for 8 min.

[0114] Experimental Example 1

[0115] The sea fennel callus culture filtrate in Example 4 described in the specification of a composition for repairing blue light damage containing sea fennel callus culture filtrate and its application (patent number 202010430066.X, authorization publication number 111700827B) is replaced with the sea fennel callus culture filtrate prepared in Example 3. The amount of sea fennel callus culture filtrate in this technical solution is 5g / 100g.

[0116] Experimental Example 2

[0117] The sea fennel callus culture filtrate in Example 3 is replaced with the sea fennel callus culture filtrate of the prior art (that is, a composition for repairing blue light damage containing sea fennel callus culture filtrate and the sea fennel callus culture filtrate in Example 4 recorded in the application instructions).

[0118] Blue light damage test

[0119] The purpose of the experiment is to verify that the filtrate of the sea fennel callus culture obtained by the present invention still has the effect of resisting blue light damage at a lower concentration.

[0120] Experimental method: In a carbon dioxide incubator, 37°C, 5% CO2, fibroblasts (HFF-1) were cultured in a 10 cm diameter culture dish, the culture medium contained 10% fetal bovine serum, and when the cell confluence reached 80-90%, the fibroblasts were seeded in a 96-well cell culture plate at a cell density of 1×10 4 The test can be carried out. Experimental Example 1 was configured to a sample solution with a mass concentration of 1.5 mg / mL. The experiment was divided into a control group and an experimental group. The culture medium in the control group was replaced with a fresh culture medium, and the culture medium in the experimental group was replaced with a culture medium containing the solution prepared in Experimental Example 1, and the culture was continued for 12 hours. The control group was partially shielded from light, and the experimental group was placed under a blue light lamp for continuous irradiation for 6 hours. Then 200 μL of MTT (1 mg / mL) solution was added to each well, and the cells were continued to be incubated at 37°C for 3 hours; 200 μL of DMSO was added to each well, shaken to dissolve it, and then the OD value was measured at 490 nm on a microplate reader. Cell activity = OD experimental group / OD control group.

[0121] The experimental results are shown in Table 1.

[0122] Table 1 Cell activity calculation table

[0123] sample Cell viability Control group 100％ Experimental Example 1 88.02％

[0124] This experiment proves that the filtrate of the callus culture of C. cristata prepared by the present invention can produce almost the same effect as the similar product at a high concentration (11 g / 100 g) at a lower concentration (5 g / 100 g).

[0125] Anti-free radical test

[0126] The purpose of the experiment is to verify that the skin care product prepared by the present invention has an anti-free radical effect on the skin.

[0127] Experimental method: Take 2mL of ethanol and 2mL of 0.3mM (3×10-5mol / L) DPPH solution and place them in test tube A0, let them stand for 30min, and use a UV spectrophotometer to measure the absorbance at a wavelength of 517nm. Prepare sample solutions with a mass concentration of 1.2mg / mL in Examples 1-3 and Experimental Example 2, take 2mL of the solutions prepared in Examples 1-3 and Experimental Example 2 and place them in reaction test tube Ai, add 2mL of DPPH ethanol solution; take 2.0mL of ethanol and 2.0mL of the solutions prepared in Examples 1-3 and Experimental Example 2 and place them in test tube Aj. Let each test tube stand for 30min, and use a UV spectrophotometer to measure the absorbance of each test tube at a wavelength of 517nm. Scavenging rate = [A0-(Ai-Aj)] / A0, A0 is the absorbance value of DPPH ethanol solution; Ai is the absorbance value of the sample solution after scavenging DPPH free radicals; Aj is the absorbance value of the sample solution.

[0128] The experimental results are shown in Table 2.

[0129] Table 2 DPPH removal rate statistics of each sample

[0130] sample Example 1 Example 2 Example 3 Experimental Example 2 Clearance rate (%) 87.11 88.45 88.91 79.31

[0131] Obviously, the skin care product prepared by the present invention has a better anti-free radical effect than similar products.

[0132] Anti-aging test

[0133] The purpose of the experiment is to verify that the skin care product prepared by the present invention has anti-aging effect.

[0134] Experimental method: HaCaT cells were inoculated in a 96-well plate and cultured to a density of 80%. A blank group, a model group, and an experimental group were set up, and 6 replicate wells were set up in each group to take the average. The blank group was not irradiated with UVB, and 10 μL of culture medium was added for 2 hours, and then cultured at 37°C for 24 hours before being used in the experiment. The HaCaT cells in the model group were treated with 10 μL of culture medium for 2 hours, and then irradiated with 60 mJ / cm2 of UVB, and cultured at 37°C for 24 hours before being used in the experiment. The HaCaT cells in the experimental group were treated with 10 μL of culture medium (skin care product content 0.5 mg / mL) for 2 hours, and then irradiated with 60 mJ / cm2 of UVB, and cultured at 37°C for 24 hours before being used in the experiment. Centrifuge for 10 minutes at 4°C and 12000 r / min, and collect the supernatant. The protein in the supernatant was quantified using the BCA protein quantification kit to determine the final concentration of 10 μg / mL. The expression level of secretory protein MMP-1 was determined according to the instructions of the enzyme-linked immunosorbent assay (ELISA) kit, and the inhibition rate of MMP-1 (matrix metalloproteinase) was calculated.

[0135] The experimental results are shown in Table 3.

[0136] Table 3 Statistical table of the inhibition rate of each sample on MMP-1

[0137] sample Example 1 Example 2 Example 3 Experimental Example 2 Inhibition rate of MMP-1 (%) 5.57 5.62 5.59 5.05

[0138] By comparison, it is found that the skin care product prepared by the present invention has better anti-aging effect than similar products.

[0139] The above embodiments and descriptions are only for illustrating the principles and best embodiments of the present invention. Without departing from the spirit and scope of the present invention, the present invention may be subject to various changes and improvements, all of which fall within the scope of the present invention to be protected.

Claims

1. An anti-free radical composition, characterized in that: The invention comprises the following raw materials in parts by weight: 0.1-0.5 parts of peony root extract composition, 0.001-0.01 parts of essence, 0.01-0.1 parts of peach resin extract composition, 0.01-0.1 parts of rye seed extract composition, 1-3 parts of leukocyte extract composition, 0.5-1.5 parts of Eryngium maritima callus culture filtrate composition and 0.5-1.5 parts of sea fennel callus culture filtrate composition; wherein: The peony root extract composition is a mixture of water, butylene glycol, peony root extract and 1,2-hexanediol in a weight ratio of 80:1:18:1; The peach resin extract composition is a mixture of water, peach resin extract, 1,2-hexanediol and ethylhexylglycerin in a weight ratio of 59:18:13:10; The rye seed extract composition is a mixture of water, 1,2-pentanediol and rye seed extract in a weight ratio of 70:18:12; The leukocyte extract composition is a mixture of leukocyte extract, butylene glycol, phenoxyethanol and water in a weight ratio of 18:1:11:70; The Eryngium maritima callus culture filtrate composition is prepared by mixing Eryngium maritima callus culture filtrate, butanediol and water in a weight ratio of 90:2:8; The cristata callus culture filtrate composition is prepared by mixing cristata callus culture filtrate, butanediol and water in a weight ratio of 90:2:

8.

2. The anti-free radical composition according to claim 1, characterized in that: The preparation method of the filtrate of the sea fennel callus culture comprises the following steps: Step 1, subjecting adult sea fennel to high temperature stress treatment; Step 2, returning the adult sea fennel to normal temperature for daily maintenance for 21 days, and selecting the individuals that show no signs of wilting as tissue culture mother plants; Step 3, cutting stem segments from the budding part of the mother plant and cutting them into thin slices as explants for callus induction; Step 4, performing suspension culture on the callus tissue; Step 5, filtering the suspended matter to obtain the Ceratonia suffruticosa callus culture filtrate.

3. The anti-free radical composition according to claim 2, characterized in that: The high temperature stress treatment method is: raising the temperature from 25°C to 35°C within 0.5h, staying for 2 hours, then raising the temperature from 35°C to 45°C, and maintaining 45°C for 1h.

4. The anti-free radical composition according to claim 2, characterized in that: In step 3, the stem segment is cut to a length of 3 cm, and both ends of the stem segment are at least 1 cm away from the budding site.

5. The anti-free radical composition according to claim 2, characterized in that: The step 3 also includes a step of disinfecting the cut stem segments, wherein the stem segments are first soaked in 70wt% alcohol for 15s, and then soaked in 2wt% sodium hypochlorite for 15min, and finally rinsed with clean water for 5 times.

6. The anti-free radical composition according to claim 2, characterized in that: In step 3, the method for inducing callus tissue is: spreading the slices on the surface of the culture medium, the culture medium is MS culture medium mixed with 1.9 mg / L 6-BA and 0.5 mg / L NAA, and culturing for 6 days.

7. The anti-free radical composition according to claim 2, characterized in that: In step 4, the suspension culture method is: MS culture medium and water are mixed in a weight ratio of 5:995, heated to dissolve, and after returning to room temperature, 3% sucrose, 0.005% 2,4-D and 0.001% KT are added according to the weight of the solution, and the culture is placed on a shaker at a speed of 100 r / min for 30 days.

8. A method for preparing a skin care product, comprising the following steps: S1, weighing phase A, phase B, phase C, phase D and the anti-free radical composition according to claim 1; S2, add phase A into the emulsifying pot, stir at a speed of 15-20r / min, heat to 82-85℃, homogenize at a speed of 2500-3000r / min for 3-5min, keep warm for 20min, then defoam and cool down; S3, after the emulsification pot is cooled to 75°C, add phase B and stir at a speed of 15-20r / min for 3-5min; S4, add phase C into the emulsifier and stir at 15-20r / min for 3-5min; S5, heat phase D in a pre-melting pot to 60-65°C and stir until there are no particles; S6, after the emulsification pot is cooled to 65°C, add phase D and stir at a speed of 15-20 r / min for 5-10 min; S7, after the temperature of the emulsifying pot is lowered to 45°C, the anti-free radical composition is added and stirred at a speed of 15-20 r / min for 5-10 minutes.

9. The method for preparing the skin care product according to claim 8, characterized in that: The phase A comprises, by weight, 14.49-44.969 parts of water, 0.01-0.3 parts of allantoin, 0.3-0.6 parts of carbomer, 0.1-0.3 parts of niacinamide and 1-3 parts of butylene glycol; the phase B comprises, by weight, 0.1-0.3 parts of polysiloxane-11, 49.3-66.1 parts of water, 0.3-2 parts of hydrogenated lecithin, 0.1-1 parts of 1,2-hexanediol and 0.2-0.6 parts of p-hydroxyacetophenone; the phase C comprises, by weight, 0.05-0.1 parts of aminomethyl propanol and 0.05-0.3 parts of water; the phase D comprises, by weight, 0.2-0.6 parts of p-hydroxyacetophenone, 1-3 parts of butylene glycol and 0.2-0.6 parts of 1,2-hexanediol.

10. An application of an anti-free radical composition in skin anti-aging, characterized in that: The anti-free radical composition according to claim 1 is made into a skin care product and applied to the skin surface.