CN115531272A - Antioxidant composition derived from alpine plants and preparation method and application thereof - Google Patents

Antioxidant composition derived from alpine plants and preparation method and application thereof Download PDF

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CN115531272A
CN115531272A CN202211357150.9A CN202211357150A CN115531272A CN 115531272 A CN115531272 A CN 115531272A CN 202211357150 A CN202211357150 A CN 202211357150A CN 115531272 A CN115531272 A CN 115531272A
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extract
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antioxidant
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CN115531272B (en
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张兴琪
李雪珍
苏潇雨
刘瑞学
冷群英
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Guangdong Bawei Biotechnology Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9755Gymnosperms [Coniferophyta]
    • A61K8/9761Cupressaceae [Cypress family], e.g. juniper or cypress
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/02Preparations for care of the skin for chemically bleaching or whitening the skin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
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Abstract

The invention discloses an antioxidant composition derived from alpine plants, which mainly comprises the following components in parts by weight: 1-30 parts of saussurea involucrate extract, 1-30 parts of alpine edelweiss extract, 1-30 parts of oriental cherry extract and 1-30 parts of gentian extract; the composition has good antioxidant effect, can exert good effect of removing free radicals when being applied to cosmetics, can promote the generation of collagen, effectively relieves the symptoms of rough skin, obvious wrinkles, color spots and the like, has the advantages of pure nature, no stimulation, safety and no toxicity, can meet the requirement of safe skin care, and has good popularization prospect and wide application market.

Description

Antioxidant composition derived from alpine plants and preparation method and application thereof
Technical Field
The invention discloses a composition, in particular to an antioxidant composition derived from alpine plants, a preparation method and application thereof, belonging to the technical field of cosmetics.
Background
The aging is a necessary rule of the human life process and a long-term proposition that the skin care world is not changed, the endogenous aging cannot be avoided, but the exogenous aging can be improved by a good living mode through regulating the living system. In recent years, with the enrichment of dietary culture and the increase of living pressure, a great number of people begin to have different levels of exogenous accelerated aging, which is specifically represented by the problems of increased wrinkles, color spots and the like.
Although antioxidants such as vitamin C are added to conventional cosmetic formulations, they tend to aggregate on the surface area of the human body and are not highly stable. CN108888565B discloses an antioxidant toner added with ginseng liposome and a preparation method thereof, wherein the formula comprises Aureobasidium pullulans polysaccharide, betaine, sodium hyaluronate, ellagic acid, edelweiss, ginseng total saponin freeze-dried powder, ginseng liposome suspension, VC ethyl ether, ginseng essential oil and the like.
CN110934209A discloses a composition with antioxidant function, health food and preparation method thereof, wherein the main preparation raw materials comprise: blueberry powder, green tea extract, saussurea involucrate extract culture, cherry powder, rosemary extract and the like. Although the granules disclosed by the patent have better antioxidant capacity, the granules need to be eaten and cannot be well applied to the field of cosmetics.
In northwest areas of China, most areas belong to plateau areas, the characteristic of more plants in local snow is created under severe conditions of long-term high cold, high radiation, oxygen deficiency, drought and the like, but part of high mountain antioxidant plants grow durably, and the local plants are rich in more active ingredients such as phenols, amino acids, flavones and the like due to long-term resistance to severe environments. However, no research on the antioxidant effect of mountain plants in cosmetics has been reported.
Therefore, there is a need to develop a composition derived from alpine plants with good antioxidant effect.
Disclosure of Invention
In view of the above disadvantages, the present invention aims to provide a composition derived from alpine plants and having antioxidant effect, which has good antioxidant effect, can exert good effect of scavenging free radicals, promote collagen generation, and effectively alleviate the symptoms of rough skin, obvious wrinkles, color spots, etc. when applied to cosmetics.
Therefore, the first technical scheme provided by the invention is as follows:
an antioxidant composition derived from alpine plants mainly comprises the following components in parts by weight: 1-30 parts of saussurea involucrate extract, 1-30 parts of alpine edelweiss extract, 1-30 parts of oriental cherry extract and 1-30 parts of gentian extract.
Furthermore, the composition with the antioxidant effect also comprises 1-30 parts of selaginella tamariscina extract.
Preferably, the composition with the antioxidant effect consists of the following components in parts by weight: 1-30 parts of saussurea involucrata extract, 1-30 parts of alpine edelweiss extract, 1-20 parts of oriental cherry extract, 1-20 parts of felwort extract and 1-30 parts of selaginella tamariscina extract.
The plant extracts in the invention have good effects, and the specific effects are as follows:
the saussurea involucrate extract is an ideal natural antioxidant, contains saussurea involucrate extract lactone, amino acid and flavonoid, can effectively remove free radicals, and has the effects of tissue repair, inflammation resistance, sun protection and the like.
The alpine edelweiss extract contains rich phenolic acids and mineral substances, has the effects of clearing heat and cooling blood, removing blood stasis and relieving pain, and has the effects of resisting wrinkles, relieving, whitening and nourishing skin.
The cherry extract is rich in flavone, sakura glycoside, sakura element, vitamins and the like, has good effects of shrinking pores and balancing grease, and has excellent oxidation resistance.
The selaginella extract is activated when meeting water, and has better antibacterial and moisturizing effects.
The gentian extract has the effects of relieving, calming and moistening the skin, and has the effects of whitening and moisturizing on the premise of regulating other plant extracts.
The plant extract composition of the invention. The saussurea involucrate extract and the alpine edelweiss extract are the first effective components, can play the effects of removing free radicals, resisting inflammation, relieving and the like, and can prevent long-term damage of the free radicals induced by physiological or non-physiological factors through tissue repair and abundant mineral substances contained in the tissue repair. The cherry blossom extract is a second effective component, can assist the first effective component to exert the effect, contains active substances such as cherry blossom extract glycoside, cherry blossom extract essence, abundant vitamins and the like, and has good effects of shrinking pores and balancing grease. Herba Selaginellae extract as third effective component has effects of promoting blood circulation, keeping moisture, and promoting skin absorption, and can be combined with the first and second effective components to enhance therapeutic effect. The gentian extract is a fourth active ingredient, can relieve, calm and moisten skin, and also has the effect of harmonizing other plant extracts.
The second technical scheme provided by the invention is as follows:
a method for preparing antioxidant composition derived from alpine plant comprises the following steps:
s1, raw material pretreatment: weighing herba Saussureae Involueratae, herba Leontopodii Hirsonii, oriental cherry blossom, herba Selaginellae, and radix Gentianae, pulverizing into powder with pulverizer, and mixing.
S2, extraction and separation: putting the powder obtained in the step S1 into an extraction tank, adding an extraction solvent for the first time, performing ultrasonic extraction for 1 time at 40-90 ℃ and 400W for 1 time, centrifuging for 5min at the speed of 4000r/min after extracting for 1-3h, and separating the first supernatant; adding the extraction solvent for the second time, adding mixed enzyme, performing enzymolysis at 40-80 deg.C for 1-3 hr, heating to 85 deg.C, maintaining for 15 min, separating the second supernatant, and mixing the first supernatant and the second supernatant to obtain extractive solution.
S3, concentrating and refining: and (3) concentrating the extracting solution obtained in the step (S2) under reduced pressure to 1/3 to 1/4 of the original mass, then adding the concentrated solution into macroporous adsorption resin for purification, eluting with an ethanol solution with the volume ratio concentration of 10-95%, collecting ethanol eluent, performing rotary evaporation concentration, adding butanediol with the total mass (the total mass of the concentrated solution and the butanediol) accounting for 30% of the total mass of the concentrated solution and the butanediol into the concentrated solution for dissolution, and performing membrane filtration to obtain the final composition.
Preferably, the amount of the mixed enzyme in S2 is 4000u/g, and the mixed enzyme is cellulase, pectinase and protease, wherein the ratio of the mixed enzyme to the protease is 1:1:1 in proportion.
Preferably, the extraction solvent in S2 is any one of water, ethanol, or an aqueous ethanol solution.
Preferably, the macroporous adsorption resin in S3 is AB-8 type or D101 type resin.
Preferably, the membrane used for membrane filtration in S3 is a microfiltration membrane.
The third technical scheme provided by the invention is as follows: the composition with the antioxidant effect is applied to cosmetics.
Preferably, the antioxidant cosmetic is a skin external preparation prepared by taking the composition with the antioxidant effect as an active component and adding conventional pharmaceutical or cosmetic auxiliary materials or auxiliary components.
Specifically, the antioxidant cosmetic is one or more of astringent, essence, cream, facial mask and gel.
Compared with the prior art, the invention has the following beneficial effects:
1. the antioxidant composition derived from the alpine plants disclosed by the invention refers to the habits and effects of various alpine plants, and the extracts of various plants are reasonable in selection and proportion, so that the antioxidant composition has a good antioxidant effect, can promote collagen synthesis, effectively relieves the symptoms of rough skin, obvious wrinkles, color spots and the like, has a certain whitening and moisturizing effect, and really meets the requirement of effective skin care.
2. The antioxidant composition derived from alpine plants disclosed by the invention has the advantages of pure nature, no stimulation, safety and no toxicity, can meet the requirements of safety skin care, and has good popularization prospect and wide application market.
Drawings
FIG. 1 is a graph showing the results of mean wrinkle score determination in experiment 5;
fig. 2 is a graph showing the results of measurement of skin firmness F4 in experiment 5.
Detailed Description
The following claims are hereby incorporated into the detailed description of the invention, with the understanding that the invention is not to be limited in any way, as any number of modifications may be made by one within the scope of the claims and the invention is still within the scope of the claims.
Example 1
An antioxidant composition derived from alpine plants, comprising the following components: 20g of saussurea involucrate extract, 20g of alpine edelweiss extract, 10g of cherry blossom extract, 10g of selaginella extract and 10g of gentian extract.
Example 2
An antioxidant composition derived from alpine plants, comprising the following components: 17.5g of saussurea involucrate extract, 17.5g of alpine edelweiss extract, 17.5g of cherry blossom extract, 8.75g of selaginella tamariscina extract and 8.75g of gentian extract.
Example 3
An antioxidant composition derived from alpine plants, comprising the following components: 14g of saussurea involucrate extract, 14g of alpine edelweiss extract, 14g of cherry blossom extract, 14g of selaginella extract and 14g of gentian extract.
Example 4
An antioxidant composition derived from alpine plants, comprising the following components: 30g of saussurea involucrate extract, 10g of alpine edelweiss extract, 10g of cherry blossom extract, 10g of selaginella extract and 10g of gentian extract.
Example 5
An antioxidant composition derived from alpine plants, comprising the following components: 5g of saussurea involucrate extract, 30g of alpine edelweiss extract, 10g of cherry blossom extract, 15g of selaginella extract and 10g of gentian extract.
Example 6
An antioxidant composition derived from alpine plants, comprising the following components: 5g of saussurea involucrate extract, 15g of alpine edelweiss extract, 30g of cherry blossom extract, 5g of selaginella extract and 15g of gentian extract.
Example 7
An antioxidant composition derived from alpine plants, comprising the following components: 15g of saussurea involucrate extract, 10g of alpine edelweiss extract, 10g of oriental cherry extract, 30g of selaginella tamariscina extract and 5g of gentian extract.
Example 8
An antioxidant composition derived from alpine plants, comprising the following components: 5g of saussurea involucrate extract, 10g of alpine edelweiss extract, 5g of oriental cherry extract, 20g of selaginella tamariscina extract and 30g of gentian extract.
Examples preparation examples:
s1, raw material pretreatment: weighing herba Saussureae Involueratae, herba Leontopodii Hirsonii, oriental cherry blossom, herba Selaginellae, and radix Gentianae, pulverizing into powder with a pulverizer, and mixing.
S2, extraction and separation: putting the powder obtained in the step S1 into an extraction tank, adding an extraction solvent for the first time, performing ultrasonic extraction for 1 time at 65 ℃ and with the power of 400w for 1.2 hours, and separating the supernatant for the first time, wherein the addition amount of the extraction solvent is 15 times of the total mass of the raw materials; adding an extraction solvent for the second time, wherein the addition amount is 15 times of the total mass of the raw materials, and adding cellulase: and (3) pectinase: the protease is 1:1:1 (based on the total mass of the raw materials), cooling to 55 ℃ for enzymolysis for 2h, then heating to 85 ℃, preserving the temperature for 15 min, separating the second supernatant, and combining the first supernatant and the second supernatant to obtain an extracting solution.
S3, concentrating and refining: and (3) concentrating the extracting solution obtained in the step (S2) under reduced pressure to 1/3 to 1/4 of the original mass, then adding the concentrated solution into macroporous adsorption resin for purification, eluting by using an ethanol solution with the volume ratio concentration of 70%, collecting an ethanol eluent, performing rotary evaporation and concentration, adding 30% of butanediol in the total mass (the total mass of the concentrated solution and the butanediol) into the concentrated solution for dissolution, and performing membrane filtration to obtain the final composition.
To demonstrate the effect of the present invention, comparative experimental examples of the present application are given below:
comparative examples 1-5 the preparation of any of the compositions was the same as in the examples.
Comparative example 1
An antioxidant composition derived from alpine plants, comprising the following components: 20g of edelweiss extract, 20g of oriental cherry extract, 20g of selaginella tamariscina extract and 10g of gentian extract.
Comparative example 2
An antioxidant composition derived from alpine plants, comprising the following components: 20g of saussurea involucrate extract, 20g of oriental cherry extract, 20g of selaginella tamariscina extract and 10g of gentian extract.
Comparative example 3
An antioxidant composition derived from alpine plants, comprising the following components: 20g of saussurea involucrate extract, 10g of alpine edelweiss extract, 20g of selaginella tamariscina extract and 20g of gentian extract.
Comparative example 4
An antioxidant composition derived from alpine plants, comprising the following components: 20g of saussurea involucrate extract, 20g of alpine edelweiss extract, 10g of oriental cherry extract and 20g of gentian extract.
Comparative example 5
An antioxidant composition derived from alpine plants, comprising the following components: 20g of saussurea involucrate extract, 20g of alpine edelweiss extract, 10g of cherry blossom extract and 20g of selaginella tamariscina extract.
The composition with antioxidant effect can be used for preparing antioxidant cosmetics. The antioxidant cosmetic is a skin external preparation prepared by taking the composition with the antioxidant effect as an active component and adding conventional pharmaceutical or cosmetic auxiliary materials or auxiliary components. When the composition is used specifically, the composition prepared by the invention is directly added into a cosmetic matrix, and the addition amount can be adjusted according to actual requirements, and is preferably 0.5-10 wt%.
Sample examples
Preparing the antioxidant moisturizing cream: the antioxidant compositions prepared in the examples were added to the formula to prepare a cream, and the formula of the antioxidant cream containing the antioxidant composition is given in table 1, and the amounts of the respective substances are expressed by mass fraction.
TABLE 1 antioxidant cream formulations
Figure BDA0003920443740000061
The preparation method of the antioxidant cream comprises the following steps: mixing, stirring and heating the components of the phase A to 80 ℃, and fully and uniformly dispersing the components for later use; mixing, stirring and heating the phase B to 85 ℃, and fully and uniformly dispersing; adding the treated phase A into the phase B, fully homogenizing, and after uniformly dispersing, starting stirring and cooling; cooling to below 70 ℃, adding the phase C, fully homogenizing, uniformly stirring, cooling to below 60 ℃, adding the phase D, and uniformly stirring; and (5) cooling to below 50 ℃, adding the phase E, uniformly stirring, and discharging after the detection is qualified.
To better illustrate the advantages of the present invention, the efficacy test of the composition with antioxidant efficacy provided by the present invention is given below:
experiment 1, antioxidant efficacy test-DPPH free radical scavenging experiment
Accurately weighing 41.67mg of DPPH reagent, fixing the volume to a 100mL brown volumetric flask, and fixing the volume to a 50mL brown volumetric flask by using anhydrous ethanol for 5mL to obtain DPPH test solution (0.1 mmol/L). The test solutions of 1mL of PPH were transferred and 0.5mL of the extracts obtained in examples 1 to 8 and comparative examples 1 to 5 were added to each of the solutions as shown in Table 4. Standing in dark for 30min after mixing, measuring absorbance at 517nm with anhydrous ethanol as blank control, setting vitamin C as control, calculating DPPH clearance rate of each extractive solution according to the following formula, and calculating half inhibitory concentration IC50 value: DPPH clearance (%) = [1- (As-Ar)/A0 ]. Times.100%
In the formula: as is the absorbance of the reaction system of the extract and DPPH; ar-absorbance of extract-absolute ethanol; a0-absorbance of blank control in absolute ethanol.
TABLE 1 reaction solution composition
Absorbance of the solution Addition amount of extract (mL) DPPH test solution (mL) Anhydrous ethanol (mL)
A0 0 1 0.5
Ar 0.5 0 1
As 0.5 1 0
The DPPH free radical scavenging rate reflects the antioxidant capacity to a certain extent, and the higher the DPPH free radical scavenging rate is, the better the antioxidant capacity is indicated, and the better the antioxidant effect is, so that the antioxidant effect can be judged by researching the DPPH free radical scavenging capacity of the composition. Vitamin C is a high-efficiency antioxidant accepted by the cosmetic industry, and the antioxidant capacity of the experiment is examined by taking vitamin C as a control.
As can be seen from Table 5, the control vitamin C has a DPPH half inhibitory concentration (dry weight) of 38.39. Mu.g/mL, whereas Experimental example 2 has a half inhibitory concentration of 58.46. Mu.g/mL, which is slightly greater than the IC50 value of vitamin C, indicating that the composition has good DPPH scavenging ability within the same scale. Meanwhile, as can be seen from the following table, the IC50 values of examples 1 to 8 are smaller than those of comparative examples 1 to 5, which indicates that the antioxidant capacity of the composition containing five plant extracts is superior to that of the composition lacking any plant extract, and the formula compatibility is reasonable, so that the synergistic effect can be achieved, and the good antioxidant effect can be exerted together.
TABLE 5 DPPH radical scavenging test results for antioxidant compositions
Sample name DPPH half clearance IC50 (μ g/mL)
Vitamin C 38.39
Example 1 67.25
Example 2 58.46
Example 3 69.54
Example 4 78.84
Example 5 74.26
Example 6 69.72
Example 7 88.95
Example 8 84.63
Comparative example 1 107.32
Comparative example 2 135.65
Comparative example 3 142.63
Comparative example 4 98..56
Comparative example 5 97.29
Experiment 2, antioxidation efficacy test-hydroxyl radical scavenging experiment
The test was carried out using the plant compositions prepared in Experimental examples 1-8 and comparative examples 1-5. The test was as follows:
hydroxyl radicals are generated by Fenton reaction. 1.0mLH is firstly added into a reaction system 2 O 2 Solution (9.0 mmol/L), 1mL salicylic acid-ethanol solution (9.0 mmol/L), 1mL antioxidant composition with the same concentration, and 1mL LH added 2 O 2 (8.8 mmol/L), reacting in 37 ℃ water bath for 30min, taking absolute ethyl alcohol as blank control, measuring the absorbance at 510nm, setting vitamin C as control, and calculating the clearance rate of each extracting solution to hydroxyl according to the following formula:
hydroxyl group clearance (%) = [1- (Ax-Ax 0)/A0 ]. Times.100%
In the formula: ax is the absorbance of an extracting solution reaction system; ax 0-without H 2 O 2 Absorbance of the sample (absolute ethanol instead of H) 2 O 2 ) (ii) a A0-absorbance of blank control with absolute ethanol.
TABLE 6 experimental results of the composition on scavenging ability of hydroxyl radicals
Sample name Clearance of hydroxyl group (%)
Vitamin C 85.3
Example 1 83.5
Example 2 82.6
Example 3 79.6
Example 4 78.3
Example 5 77.3
Example 6 79.5
Example 7 66.5
Example 8 65.3
Comparative example 1 60.2
Comparative example 2 58.6
Comparative example 3 53.9
Comparative example 4 68.5
Comparative example 5 70.3
Vitamin C was used as a positive control to examine its hydroxyl radical scavenging ability. As shown in table 6 above, when the same mass concentration of the composition was added, the example had a clearance rate similar to that of vitamin C, especially the clearance rate of example 1 was as high as 83.5%, which is close to that of vitamin C, indicating that the composition of the formula had a good hydroxyl radical scavenging ability. The clearance rates of the examples 1 to 8 are higher than those of the comparative examples 1 to 5, which shows that the oxidation resistance of the composition containing five plant extracts is better than that of the composition lacking any plant extract, the five plant extracts have a synergistic effect, the formula compatibility is reasonable, and the composition has good oxidation resistance.
Experiment 3, cell level-anti-wrinkle efficacy test
The composition prepared in example 1 was selected for testing as follows:
cytotoxicity assay by adjusting the concentration of HFF-1 cell suspension, adding to a 96-well plate, and incubating at 37 ℃ for 24h, 5% in CO2. A series of samples were prepared with concentration gradients, 100 ul/well in 96-well plates, and cultured at 37 ℃ for 24h in 5% CO2. The culture medium was discarded, washed 2 times with PBS, added with 50 ul/well MTT solution, and cultured at 37 ℃ for 2-6h with 5% CO2. Discarding the culture medium containing MTT, adding DMSO at 150 ul/well, shaking for 5-10min, mixing, and measuring OD at 570 nm. The concentration without obvious cytotoxicity is selected for carrying out protein content determination experiments.
Type i collagen content determination: adjusting the concentration of HFF-1 cell suspension, adding to a 96-well plate, and culturing at 37 ℃ under 5% CO2 for 24h. Discarding the old medium, inducing molding with UVB, adding 100ul of DMEM medium containing the sample into each well after irradiation is finished, setting the blank medium control group and the medium containing TGF-beta as positive control groups, and culturing at 37 deg.C and 5% CO2 for 24h + -2 h. After the culture is finished, the content of human type I collagen is measured according to the specification of an ELISA kit. And (3) taking the concentration of the standard substance as an abscissa and the OD450 value as an ordinate to prepare a standard curve, and substituting the standard curve according to the OD450 value of the sample to obtain the content of the collagen in the experimental group. Finally, the average value of each group of 3 multiple wells is selected as the result of the final collagen. Statistical and significance analysis was performed using GraphPad Prism or other statistical software, calculating a P value, with P < 0.05 indicating significant difference, otherwise no significant difference.
Up-regulation rate (%) = (T/C-1) × 100%
Wherein: mean value of collagen in T-Experimental group
C-blank/solvent group collagen mean
TABLE 7 results of collagen content experiment with compositions
Figure BDA0003920443740000091
The results of the composition on collagen content are shown in Table 7 above, where NC is the experimental blank control and PC is the positive control. As can be seen from the table, when the sample of example 1 in the safe concentration range is added, the mean value of collagen in the cells is 0.34ng/ml, compared with the NC group, the collagen content is up-regulated by 88.34%, which is slightly lower than that of the positive group, and the composition has good effect of promoting cell collagen synthesis and can exert good anti-wrinkle effect.
Experiment 4, patch experiment
The irritation of the antioxidant composition is evaluated through a human skin patch experiment. Cream bases containing 5.0% and 10.0% of the plant antioxidant compositions of examples 1-3 were prepared for closed patch application and the possibility of adverse reaction of the samples to the human body was examined. In the test, sample 1 is a cream to which 5.0% of the plant antioxidant composition is added, and sample 2 is a cream to which 10.0% of the plant antioxidant composition is added. Adverse skin reactions were graded as shown in table 7 below:
TABLE 7 adverse skin reaction grading
Figure BDA0003920443740000101
The cream prepared in the above examples 1 to 3 was subjected to the spot test of the subject, and 30 persons (between 20 and 50 years of age) were examined for 24 hours of spot application time, and skin reactions of the subject were observed when the subject removed the spot tester for 30min, 24 hours and 48 hours, and the results are shown in the following table.
Figure BDA0003920443740000102
As shown in the table, the cream added with the plant antioxidant composition has no adverse irritant reaction to human body and good safety.
Experiment 5, evaluation experiment in vitro
1. Experimental method
The antioxidant moisturizing cream prepared in application example 1 was used for 20 volunteers selected at random. The test population is as follows: 20 healthy volunteers suffering from wrinkles such as dry lines, fine lines, crow's feet, expression lines and the like are selected, and the external preparation is not applied, has no allergy to cosmetics, has no acute inflammation on the face, does not use hormone medicines or immunosuppressants, does not participate in other clinical tests. .
The test method comprises the following steps: after cleaning the skin of each group of volunteers with warm water, the product was uniformly applied to the face 1 time each day in the morning and evening, and after 2 and 4 weeks of continuous use, the skin was tested using a Visia facial image analyzer, a skin tightness test probe, and the like, respectively. Each time facial images were taken and wrinkle parameters, skin tightness were analyzed for 10 cycles per site, averaged. The results are shown in FIGS. 1-2 (note: D0, D14, D28 represent test results on days 0, 14, 28, respectively).
As shown in fig. 1-2, after 20 volunteers used the cream base added with 5% of the antioxidant composition, the average value of the wrinkle values was respectively reduced by 18.87% and 24.48% on days D14 and D28, which indicates that the composition can effectively improve the obvious and deepened wrinkles and the like, so as to achieve the effect of resisting wrinkles; the skin tightness F4 average values are respectively reduced by 6.84% and 23.62% at D14 and D28, which shows that the skin tightness F4 average values can effectively improve skin relaxation and promote expression of related elastin in skin, and has good tightening effect. ( Note: the higher the wrinkle score, the more visible the skin wrinkles; the smaller the skin firmness F4, the more firm the skin is )
In conclusion, the anti-oxidation composition is prepared by scientifically mixing the snow lotus, the alpine edelweiss, the oriental cherry, the gentian and the like and extracting and purifying technologies such as ultrasound, biological enzymolysis, column chromatography and the like, has good effects of eliminating free radicals, promoting collagen generation, tightening and resisting wrinkles and the like, can effectively relieve rough skin, obvious wrinkles, color spots and the like, and improves the skin state deeply.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.

Claims (10)

1. An antioxidant composition derived from alpine plants is characterized by mainly comprising the following components in parts by weight: 1-30 parts of saussurea involucrate extract, 1-30 parts of alpine edelweiss extract, 1-30 parts of cherry blossom extract and 1-30 parts of gentian extract.
2. The antioxidant composition derived from alpine plant according to claim 1, further comprising tamariskoid spikemoss herb extract 1-30 parts.
3. The alpine plant-derived antioxidant composition according to claim 2, comprising the following components in parts by weight: 1-30 parts of saussurea involucrata extract, 1-30 parts of alpine edelweiss extract, 1-20 parts of oriental cherry extract, 1-20 parts of felwort extract and 1-30 parts of selaginella tamariscina extract.
4. A method for preparing the composition having antioxidant effect as set forth in any one of claims 1 to 3, comprising the steps of:
s1, raw material pretreatment: weighing herba Saussureae Involueratae, herba Leontopodii Kaempferi, flos Rosae Laevigatae, herba Selaginellae, and radix Gentianae, pulverizing into powder with a pulverizer, and mixing.
S2, extraction and separation: putting the powder obtained in the step S1 into an extraction tank, adding an extraction solvent for the first time, carrying out ultrasonic extraction for 1 time at 40-90 ℃ and with the power of 400w, centrifuging for 5 minutes at the speed of 4000r/min after extracting for 1-3 hours, and separating supernatant for the first time; adding the extraction solvent for the second time, adding mixed enzyme, performing enzymolysis at 40-80 deg.C for 1-3 hr, heating to 85 deg.C, maintaining for 15 min, separating the second supernatant, and mixing the first supernatant and the second supernatant to obtain extractive solution.
S3, concentrating and refining: and (3) concentrating the extracting solution obtained in the step (S2) under reduced pressure to 1/3 to 1/4 of the original mass, then adding the concentrated solution into macroporous adsorption resin for purification, eluting with an ethanol solution with the volume ratio concentration of 10-95%, collecting ethanol eluent, performing rotary evaporation concentration, adding butanediol with the total mass (the total mass of the concentrated solution and the butanediol) accounting for 30% of the total mass of the concentrated solution and the butanediol into the concentrated solution for dissolution, and performing membrane filtration to obtain the final composition.
5. The method of preparing an antioxidant composition derived from alpine plants according to claim 4, wherein the mixed enzyme of cellulase, pectinase and protease is added to the S2 in an amount of 4000u/g, and the ratio of the mixed enzyme to the protease is 1:1:1, and mixing the components in a ratio of 1.
6. The method of preparing an antioxidant composition derived from alpine plants according to claim 4, wherein the extraction solvent in S2 is any one of water, ethanol, or an aqueous ethanol solution.
7. The method for preparing the antioxidant composition derived from alpine plants according to claim 4, wherein the macroporous adsorption resin in S3 is AB-8 type or D101 type resin.
8. Use of the composition having antioxidant effect according to any one of claims 1 to 3, wherein the antioxidant composition is used in cosmetics.
9. The use of claim 8, wherein the antioxidant cosmetic is a skin external preparation prepared by using the composition with antioxidant effect as an active component and adding pharmaceutically or cosmetically conventional adjuvants or auxiliary components.
10. The use of claim 9, wherein the antioxidant cosmetic is one or more of a lotion, a serum, a cream, a mask, and a gel.
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