Anti-allergy repairing barrier composition and preparation method and application thereof
阅读说明:本技术 一种抗敏修护屏障组合物及其制备方法和应用 (Anti-allergy repairing barrier composition and preparation method and application thereof ) 是由 陈嘉玲 林艺青 于 2020-11-12 设计创作,主要内容包括:本发明公开了一种抗敏修护屏障组合物及其制备方法和应用。按重量百分比计,该抗敏修护屏障组合物包含:寡肽-1 0.1~20%;积雪草提取物0.1~50%;凤凰单丛茶茶叶挥发油0.1~30%;反式-4-叔丁基环己醇0.1~20%;神经酰胺2脂质体0.1~20%;低分子量透明质酸钠0.1~10%;高分子量透明质酸钠0.1~10%;尿素0.1~10%;去离子水余量。本发明在上百种原料中选取合适的原料,经过多种原料的科学配比,上百次功效验证实验后证明,寡肽-1、积雪草提取物、凤凰单丛茶茶叶挥发油、反式-4-叔丁基己醇、神经酰胺2、透明质酸钠、尿素等组合后能达到舒缓皮肤敏感状态,修护皮肤屏障,抗炎等功效。(The invention discloses an anti-allergy repairing barrier composition, and a preparation method and application thereof. The anti-allergy repair barrier composition comprises the following components in percentage by weight: 10.1-20% of oligopeptide; 0.1-50% of centella asiatica extract; 0.1-30% of the phoenix Dancong tea volatile oil; trans-4-tert-butylcyclohexanol 0.1-20%; 0.1-20% of ceramide 2 liposome; 0.1-10% of low molecular weight sodium hyaluronate; 0.1-10% of high molecular weight sodium hyaluronate; 0.1-10% of urea; the balance of deionized water. According to the invention, appropriate raw materials are selected from the above hundred raw materials, and scientific proportioning of various raw materials is carried out, and hundreds of efficacy verification experiments prove that the oligopeptide-1, the centella extract, the phoenix Dancong tea leaf volatile oil, the trans-4-tert-butyl hexanol, the ceramide 2, the sodium hyaluronate, the urea and the like can achieve the effects of relieving the skin sensitivity state, repairing the skin barrier, resisting inflammation and the like after being combined.)
1. An anti-sensitivity repair barrier composition characterized by: the composite material is prepared from the following raw materials in percentage by weight:
2. the anti-sensitivity repair barrier composition of claim 1, wherein: the composite material is prepared from the following raw materials in percentage by weight:
3. the anti-sensitivity repair barrier composition of claim 1, wherein: the centella asiatica extract is prepared by the following process:
1) crushing and sieving: crushing the dried centella asiatica by using a wall breaking machine, and sieving the crushed centella asiatica by using a sieve of 50-200 meshes;
2) extraction: taking 50g of centella asiatica, adding 10-30 times of water, extracting for 1-2 hours at the temperature of 60-80 ℃, repeatedly extracting for 3 times, and combining the extracting solutions;
3) concentration and filtration: filtering the extract to remove filter residue, and concentrating;
4) adsorption and elution: adsorbing the concentrated solution by using D101B macroporous resin, eluting for 1-3 times by using 50% ethanol, combining the eluate, and carrying out reduced pressure concentration to separate the ethanol to obtain the centella extract.
4. The anti-sensitivity repair barrier composition of claim 1, wherein: the Fenghuang Dancong tea leaf volatile oil is prepared by the following process:
1) steam distillation: putting the phoenix Dancong tea leaves into a steam distillation generator, and distilling and extracting volatile oil of the tea leaves;
2) separation: separating the mixture of the prepared tea volatile oil and water to obtain the Fenghuang Dancong tea volatile oil.
5. The method of making the anti-sensitivity repair barrier composition of any one of claims 1-4, wherein: is prepared by the following processes:
mixing oligopeptide-1, herba Centellae extract, FENGHUANGSINGUOCHA volatile oil, trans-4-tert-butylcyclohexanol, ceramide 2 liposome, low molecular weight sodium hyaluronate, high molecular weight sodium hyaluronate, urea, and deionized water at a certain proportion, and stirring.
6. Use of an anti-sensitivity repair barrier composition in a skin care product, characterized in that: according to the weight percentage, the anti-allergy repairing barrier composition accounts for 0.1-10% of the total weight of the skin care product; and 90-99.9% of conventional auxiliary materials.
7. Use of an anti-sensitivity repair barrier composition according to claim 6 in a skin care product, characterized in that: the anti-allergy repairing barrier composition accounts for 0.5-5% of the total amount of the skin care product in percentage by weight.
8. Use of an anti-sensitivity repair barrier composition according to claim 6 in a skin care product, characterized in that: the conventional auxiliary materials comprise a surfactant, a thickening agent, a conditioning agent, a humectant, a preservative, a chelating agent, essence and deionized water.
9. Use of an anti-sensitivity repair barrier composition according to claim 6 in a skin care product, characterized in that: the skin care product can be made into various dosage forms of toner, smoothing toner, firming toner, nourishing lotion, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing lotion, facial mask, pressed powder, body lotion, body cream, body oil, body lotion and body essence.
Technical Field
The invention relates to an anti-allergy repairing composition, belongs to the technical field of skin care products, and particularly relates to an anti-allergy repairing barrier composition as well as a preparation method and application thereof.
Background
Sensitive Skin (Sensitive Skin SS) is a highly reactive, poorly tolerated, and relatively allergic Skin condition, i.e., a condition of subjective sensory symptoms induced by susceptibility to a variety of factors. The cause of the sensitive skin is relatively complex, and the currently accepted causes in the industry are three, namely, the barrier of the skin is damaged; second, the nerve response of the skin is enhanced; and thirdly, inflammatory reaction. Sensitive skin is a condition of the skin, which is mainly characterized by being drier, easily tense and red; the stimulation can produce the feelings of desquamation, pruritus, burning, stabbing pain and the like.
With the aggravation of environmental pollution, the pressure of people on life and work is increased, and the proportion of sensitive skin is increased due to the random use of the eight-flower cosmetics without thinking. Research data show that the incidence rate of sensitive skin is 31% in young people of 18-25 years old, and the proportion of women is higher.
At present, cosmetics which can effectively achieve an anti-allergy effect and repair a skin barrier are not available in the market, and people suffering from sensitive skin symptoms urgently need products which can solve the problem.
Disclosure of Invention
The invention aims to provide an anti-allergy repairing barrier composition and a preparation method thereof, and the anti-allergy repairing barrier composition can achieve skin care effects of relieving skin sensitivity, repairing skin barriers, resisting inflammation and the like after being combined.
The invention also aims to provide application of the anti-allergy repairing barrier composition, which can be applied to various skin care formulations and has the skin care effects of relieving the skin sensitivity state, repairing the skin barrier, resisting inflammation and the like.
The purpose of the invention and the technical problem to be solved are realized by adopting the following technical scheme. The invention provides an anti-allergy repairing barrier composition which is prepared from the following raw materials in percentage by weight:
further, the anti-allergy repairing barrier composition is prepared from the following raw materials in percentage by weight:
further, the centella asiatica extract is prepared by the following processes:
1) crushing and sieving: crushing the dried centella asiatica by using a wall breaking machine, and sieving the crushed centella asiatica by using a sieve of 50-200 meshes;
2) extraction: taking 50g of centella asiatica, adding 10-30 times of water, extracting for 1-2 hours at the temperature of 60-80 ℃, repeatedly extracting for 3 times, and combining the extracting solutions;
3) concentration and filtration: filtering the extract to remove filter residue, and concentrating;
4) adsorption and elution: adsorbing the concentrated solution by using D101B macroporous resin, eluting for 1-3 times by using 50% ethanol, combining the eluate, and carrying out reduced pressure concentration to separate the ethanol to obtain the centella extract.
Further, the Fenghuang Dancong tea leaf volatile oil is prepared by the following process:
1) steam distillation: putting the phoenix Dancong tea leaves into a steam distillation generator, and distilling and extracting volatile oil of the tea leaves;
2) separation: separating the mixture of the prepared tea volatile oil and water to obtain the Fenghuang Dancong tea volatile oil.
Preferably, the preparation method of the anti-sensitivity repairing barrier composition is prepared by the following processes:
mixing oligopeptide-1, herba Centellae extract, FENGHUANGSINGUOCHA volatile oil, trans-4-tert-butylcyclohexanol, ceramide 2 liposome, low molecular weight sodium hyaluronate, high molecular weight sodium hyaluronate, urea, and deionized water at a certain proportion, and stirring.
In addition, the invention also discloses an application of the anti-allergy repairing barrier composition in a skin care product, wherein the anti-allergy repairing barrier composition accounts for 0.1-10% of the total amount of the skin care product in percentage by weight; and 90-99.9% of conventional auxiliary materials.
Further, the anti-allergy repairing barrier composition is applied to a skin care product, and the anti-allergy repairing barrier composition accounts for 0.5-5% of the total amount of the skin care product in percentage by weight.
Further, the conventional auxiliary materials comprise a surfactant, a thickening agent, a conditioning agent, a humectant, a preservative, a chelating agent, essence and deionized water.
Further, the anti-allergy repairing barrier composition is applied to a skin care product, and the skin care product can be prepared into various dosage forms of toner, smoothing toner, firming toner, nourishing lotion, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, facial mask, powder cake, body lotion, body cream, body oil, body lotion and body essence.
By means of the technical scheme, the invention has the following advantages and beneficial effects:
1) according to the invention, appropriate raw materials are selected from the above hundred raw materials, and scientific proportioning of various raw materials is carried out, and hundreds of experiments for efficacy verification prove that the oligopeptide-1, the centella extract, the phoenix Dancong tea leaf volatile oil, the trans-4-tert-butylcyclohexanol, the ceramide 2, the sodium hyaluronate (low molecular weight and high molecular weight), the urea and the like can be combined to achieve the effects of relieving the sensitive state of the skin, repairing the skin barrier, resisting inflammation and the like;
2) the oligopeptide-1 and the centella asiatica extract can promote the proper proliferation of the stratum corneum and repair damaged cells, so that the effect of repairing skin barriers is achieved;
3) the combination of ceramide 2, sodium hyaluronate (low molecular weight and high molecular weight), urea and allantoin can achieve the optimal moisturizing effect, and the water content of the skin is improved, so that the skin barrier is firmed;
4) the active ingredients in the volatile oil of the Fenghuang Dancong tea can achieve the effects of sterilization and bacteriostasis and inhibit the expression of inflammatory factors of organisms, thereby achieving the anti-inflammatory effect;
5) the trans-4-tert-butyl cyclohexanol can reduce nerve sensitivity and has tranquilizing and soothing effects.
Drawings
Fig. 1 is a graph comparing the moisturizing effect of barrier repair-containing compositions of the present examples with blank and commercial products.
FIG. 2 is a graph showing the effect of the anti-allergy test of example 2.
Detailed Description
The present invention will be described in more detail below with reference to specific preferred embodiments and examples of effect tests, but the present invention is not limited to the following examples.
The anti-sensitivity repairing barrier composition comprises the following components in percentage by weight: 10.1-20% of oligopeptide; 0.1-50% of centella asiatica extract; 0.1-30% of the phoenix Dancong tea volatile oil; trans-4-tert-butylcyclohexanol 0.1-20%; 0.1-20% of ceramide 2 liposome; 0.1-10% of low molecular weight sodium hyaluronate; 0.1-10% of high molecular weight sodium hyaluronate; 0.1-10% of urea; the balance of deionized water.
The raw material components are respectively introduced as follows:
1, oligopeptide-1:
the polypeptide polymer consisting of glycine, histidine and lysine can promote cell proliferation and differentiation, make the skin smooth and tender, strengthen the skin barrier, accelerate the skin repair process, increase the intercellular substance, keep the skin moisture and reduce the skin moisture loss;
② centella extract:
extract of whole plant of centella asiatica of Umbelliferae genus Leptoradix contains asiaticoside, ginseng nixin, isoginseng nixin, madecassoside, etc., and flavonoid glycoside of carotene, chlorophyll, quercetin and glucose. Can promote collagen synthesis and cardiovascular formation in vivo, and stimulate granulation growth, so as to promote wound healing and metabolism.
③ the volatile oil of the phoenix Dancong tea:
the tea is prepared from Fenghuang Dancong tea from Fenghuang mountain in Guangdong Chaozhou, and its fresh tea is extracted with plant volatile oil, and has anti-inflammatory activity.
Trans-4-tert-butylcyclohexanol:
artificially synthesized, can inhibit the over-expression of vanillic acid subtype Transient receptor potential 1 channel protein (TRPV-1), reduce the neural sensitivity and achieve the effect of relieving and calming.
Ceramide 2 liposome:
ceramides, which are a major part of the intercellular matrix and play an important role in maintaining the moisture balance of the stratum corneum, have a strong ability to associate with water molecules, and maintain skin moisture by forming a network structure in the stratum corneum.
Sixthly, sodium hyaluronate (high molecular weight and small molecular weight):
natural moisturizing factors; the sodium hyaluronate with large molecular weight (150-180 ten thousand daltons) has better moisturizing effect; the sodium hyaluronate with small molecular weight (1000-2000 daltons) can permeate into the epidermis of the skin to promote the supply of skin nutrition and the excretion of waste, thereby preventing skin aging and playing the roles of maintaining beauty and keeping young;
and seventh, urea: a humectant.
The preparation process of the centella asiatica extract can be expressed as follows:
1) crushing and sieving: crushing the dried centella asiatica by using a wall breaking machine, and sieving the crushed centella asiatica by using a sieve of 50-200 meshes;
2) extraction: taking 50g of centella asiatica, adding 10-30 times of water, extracting for 1-2H at the temperature of 60-80 ℃, repeatedly extracting for 3 times, and combining the extracting solutions;
3) concentration and filtration: filtering the extract to remove filter residue, and concentrating;
4) adsorption and elution: adsorbing the concentrated solution by using D101B macroporous resin, eluting for 1-3 times by using 50% ethanol, combining the eluate, and carrying out reduced pressure concentration to separate the ethanol to obtain the centella extract.
The preparation process of the phoenix Dancong tea volatile oil can be expressed as follows:
1) steam distillation: putting the phoenix Dancong tea leaves into a steam distillation generator, and distilling and extracting volatile oil of the tea leaves;
2) separation: separating the mixture of the prepared tea volatile oil and water to obtain the Fenghuang Dancong tea volatile oil.
Examples 1 to 3
An anti-allergy repairing barrier composition comprises the following raw material components shown in the following table 1.
TABLE 1 formulation components (% by weight) of anti-allergy repair barrier composition
Raw material components
Example 1
Example 2
Example 3
Oligopeptide-1
10%
0.1%
20%
Centella asiatica extract
20%
0.1%
50%
Phoenix Dancong tea leaf volatile oil
20%
0.1%
30%
Trans-4-tert-butylcyclohexanol
4%
20%
0.1%
Ceramide 2 liposome
4%
20%
0.1%
Low molecular weight sodium hyaluronate
5%
0.1%
10%
High molecular weight sodium hyaluronate
5%
10%
0.1%
Urea
5%
10%
0.1%
Deionized water
Balance of
Balance of
Balance of
The anti-sensitivity repair barrier compositions of examples 1-3 above were prepared as follows:
mixing oligopeptide-1, herba Centellae extract, FENGHUANGSINGUOCHA volatile oil, trans-4-tert-butylcyclohexanol, ceramide 2 liposome, low molecular weight sodium hyaluronate, high molecular weight sodium hyaluronate, urea, and deionized water at a certain proportion, and stirring.
Examples 4-6 below are skin care preparations for external use containing the anti-sensitivity repair barrier composition of example 1
Example 4 skin gel with anti-allergy repair Barrier composition
A skin care gel containing the anti-sensitivity repair barrier composition of example 1 was formulated as shown in table 2 below.
Table 2 formulation of skin gel containing the anti-sensitivity repair barrier composition of example 1 (% by weight)
The skin care gel containing the anti-sensitivity repair barrier composition of example 4 above was prepared as follows:
1) firstly, dissolving a thickening agent in deionized water;
2) dissolving a skin conditioner, a preservative and deionized water;
3) mixing the two solutions and stirring; degassing;
4) adding pH regulator, active component, essence, solubilizer, etc., stirring, degassing, and discharging.
Example 5 (containing anti-allergy)Skin lotion of repairing barrier composition
A skin lotion containing the anti-sensitivity repairing barrier composition of example 1 was formulated as shown in table 3 below.
Table 3 skin lotion formulation containing the anti-sensitivity repair barrier composition of example 1 (% by weight)
Components
Raw materials
Example 5
Active ingredient
Example 1 anti-allergy repair Barrier composition
7%
Preservative
Phenoxyethanol
0.1%
Aromatic agent
Essence for daily use
0.05%
Others
Deionized water
Balance of
The skin lotion containing the anti-sensitivity repairing barrier composition of example 5 above was prepared by the following process:
adding all the components into deionized water, stirring and mixing, and discharging.
Example 6 (skin cream with anti-allergy repairing Barrier composition)
A skin cream containing the anti-sensitivity repair barrier composition of example 1 was formulated as shown in table 4 below.
Table 4 formulation of skin cream containing the anti-allergy repair barrier composition of example 1 (% by weight)
The cream containing the anti-allergy repair barrier composition of example 6 above was prepared by the following process:
1) firstly heating to 60 ℃ and dissolving the skin conditioner and the solubilizer in deionized water;
2) cooling to below 40 deg.C, adding active ingredient, antiseptic, pH regulator, thickener, and antiseptic, mixing, stirring, and homogenizing for 10 min;
3) and (6) cooling and discharging.
The following are contents of the effect test examples.
Examples of Effect test
Comparative example 1:
compared with the example 1, the preparation method comprises 20% of the volatile oil of the tea leaves of the phoenix Dancong tea, 5% of trans-4-tert-butylcyclohexanol, 5% of ceramide 2 liposome, 6% of low molecular weight sodium hyaluronate, 6% of high molecular weight sodium hyaluronate and 6% of urea except that the oligopeptide-1 and centella extract are not contained, and the specific preparation process is the same as that of the example 1;
comparative example 2:
compared with the example 1, the preparation method comprises 20% of centella asiatica extract, 25% of phoenix single fir tea leaf volatile oil, 5% of trans-4-tert-butylcyclohexanol, 5% of ceramide 2 liposome, 6% of low molecular weight sodium hyaluronate, 6% of high molecular weight sodium hyaluronate and 6% of urea, and the specific preparation process is the same as that of the example 1 except that the oligopeptide-1 is not contained;
comparative example 3:
compared with the example 1, except that the centella asiatica extract is not contained, the preparation method comprises the following specific steps of oligopeptide-110%, the volatile oil of the tea leaves of the phoenix Dancong tea 10%, the trans-4-tert-butylcyclohexanol 3%, the ceramide 2 liposome 3%, the low molecular weight sodium hyaluronate 7%, the high molecular weight sodium hyaluronate 3% and the urea 5%, and the specific preparation process is the same as that of the example 1;
comparative example 4:
contains oligopeptide-110%;
comparative example 5:
contains herba Centellae extract 20%;
comparative example 6:
compared with the example 1, except that the volatile oil of the tea leaves of the phoenix single fir, the preparation method comprises the following steps of-110% of oligopeptide, 10% of centella asiatica extract, 5% of trans-4-tert-butylcyclohexanol, 5% of ceramide 2 liposome, 6% of low molecular weight sodium hyaluronate, 6% of high molecular weight sodium hyaluronate and 6% of urea, and the specific preparation process is the same as that of the example 1;
comparative example 7:
compared with the example 1, except for not containing trans-4-tert-butylcyclohexanol, the preparation method comprises oligopeptide-115%, snow grass extract 15%, phoenix single-fir tea volatile oil 10%, ceramide 2 liposome 5%, low molecular weight sodium hyaluronate 6%, high molecular weight sodium hyaluronate 6% and urea 6%, and the specific preparation process is the same as that of the example 1;
comparative example 8:
compared with the embodiment 1, the preparation method comprises the following steps of except for not containing ceramide 2 liposome, low molecular weight sodium hyaluronate, high molecular weight sodium hyaluronate and urea, and comprising-110% of oligopeptide, 10% of centella asiatica extract and 6% of trans-4-tert-butylcyclohexanol, and the specific preparation process is the same as that of the embodiment 1;
comparative example 9:
compared with example 1, except for not containing ceramide 2 liposome, the preparation method comprises the following steps of-110% of oligopeptide, 10% of centella asiatica extract, 6% of trans-4-tert-butylcyclohexanol, 10% of low molecular weight sodium hyaluronate, 10% of high molecular weight sodium hyaluronate and 10% of urea, and the specific preparation process is the same as that of example 1;
comparative example 10:
compared with the embodiment 1, the preparation method comprises the following steps of except that the low molecular weight sodium hyaluronate is not contained, oligopeptide-110%, centella asiatica extract 10%, trans-4-tert-butylcyclohexanol 6%, ceramide 2 liposome 10%, high molecular weight sodium hyaluronate 10% and urea 10%, and the specific preparation process is the same as that of the embodiment 1;
comparative example 11:
compared with the embodiment 1, the preparation method comprises the following specific steps of except that the high molecular weight sodium hyaluronate is not contained, oligopeptide-110%, centella asiatica extract 10%, trans-4-tert-butylcyclohexanol 6%, ceramide 2 liposome 10%, low molecular weight sodium hyaluronate 10% and urea 10%, and the specific preparation process is the same as that of the embodiment 1;
comparative example 12:
compared with example 1, except that no urea is contained, the preparation method comprises the following specific steps of oligopeptide-110%, centella asiatica extract 10%, trans-4-tert-butylcyclohexanol 6%, ceramide 2 liposome 10%, low molecular weight sodium hyaluronate 10% and high molecular weight sodium hyaluronate 10%, and the specific preparation process is the same as that of example 1.
Effect evaluation 1: HaCat-MTT inhibition of cytotoxicity assay
MTT assay principle:
succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline formazan, whereas dead cells do not. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and the light absorption value is measured by an enzyme-labeling instrument at 490nm wavelength, and the MTT crystal formation amount is in direct proportion to the number of cells in a certain cell number range. The number of living cells was judged from the measured absorbance value (OD value), and the larger the OD value, the stronger the cell activity.
The concentration of HaCat (immortalized human keratinocyte) cell suspension used in the experiment is 3 x 105cell/ml, 100. mu.L/well in 96-well plates at 37 ℃ with 5% CO2The culture is carried out. After adding 100. mu.L/well sample solution PBS solution (negative control) and culturing for 24H, adding 60. mu.g/mLSDS solution and culturing for 24H (the cell viability is 60% in the presence of 60. mu.g/mLSDS), then adding 20. mu.L/well 5mg/ml MTT solution, placing in an incubator and reacting for 4H, removing supernatant, adding 150. mu.L/well DMSO, shaking in a shaking table for 10min, and measuring the absorbance at 490nm by using an enzyme linked immunosorbent assay instrument.
In this experiment, the greater the absorbance value, the greater the ability of the sample to protect cells from SDS, indicating greater sample repair ability. As shown in table 5 below.
TABLE 5 concentration and OD values of different samples
From table 5, the following conclusions can be drawn:
1) comparison of comparative examples 1, 4-5 with the negative control shows that: the OD value of comparative example 1, which contained no oligopeptide-1, centella asiatica extract, was close to that of the negative control; the OD values of comparative examples 4-5 containing only oligopeptide-1 and only centella extract are obviously higher than that of negative control, which shows that oligopeptide-1 and the centella extract have the capability of preventing cells from being damaged by SDS, and have the effects of repairing damaged cells and repairing skin barriers;
2) examples 1-3 compared with examples 2-3, it can be seen that after the oligopeptide-1 and the centella asiatica extract are compounded and combined, the OD value is obviously increased, which shows that the damaged capability of the composition for repairing cells is enhanced; and when the oligopeptide content is 110 percent and the centella asiatica extract content is 20 percent, the repairing capacity is strongest, and the proportion is proved to be the optimal proportion.
Effect evaluation 2: anti-inflammatory assayAnd test for cell barrier repair ability
Inhibiting the expression of inflammatory factor (interleukin IL-6) and promoting the expression of Vascular Endothelial Growth Factor (VEGF).
The inflammatory factor interleukin IL-6 may cause intracellular inflammatory production.
The vascular endothelial growth factor VEGF can promote the proliferation of HaCat and repair the skin barrier under proper concentration.
Selecting HaCat cell suspension with concentration of 3 x 105cell/ml, 100. mu.L/well in 6-well plates at 37 ℃ with 5% CO2Culturing until the wall adheres. Adding a mixed solution of 3 mL/hole sample solution and SDS solution, placing the SDS solution (positive control) and the PBS solution (negative control) in an incubator for further 48H, taking out, centrifuging, and removing precipitates.
The method is characterized in that a multi-liquid phase protein quantification technology is adopted, microspheres with specific fluorescent signals are linked with specific capture antibodies, the capture antibodies are used for capturing different to-be-detected objects in a solution, and the to-be-detected objects are quantitatively detected according to the fluorescent signal intensity. Table 6 shows the quantitative determination of the inhibition of inflammatory factors (interleukin IL-6).
TABLE 6 quantitative determination of the inhibition of inflammatory factor (interleukin IL-6)
From table 6, the following conclusions can be drawn:
1) examples 1-3 and comparative examples 1-6 show that the content of IL-6 in a test sample in the volatile oil of the Fenghuang Dancong tea leaves is lower than that of a positive control, and the test sample proves that the volatile oil of the Fenghuang Dancong tea leaves has the capability of inhibiting the generation of inflammatory factors and has a good anti-inflammatory effect;
2) examples 1-3 and comparative examples 1-3 show that the IL-6 content of a sample lacking one of oligopeptide-1 and centella asiatica extract is higher than that of a negative control, but lower than that of a positive control, and the compounding of oligopeptide-1, centella asiatica extract and Fenghuangdong tea volatile oil can exert a better anti-inflammatory effect. And the best effect is achieved when the oligopeptide accounts for 110 percent, the centella extract accounts for 20 percent, and the Fenghuang Dancong tea volatile oil accounts for 20 percent.
TABLE 7 quantitative determination of Vascular Endothelial Growth Factor (VEGF)
Sample (I)
Concentration of
VEGF(pg/mL)
Example 1
0.1%
153.52
Example 2
0.1%
97.53
Example 3
0.1%
84.45
Comparative example 1
0.1%
20.74
Comparative example 2
0.1%
30.12
Comparative example 3
0.1%
29.47
Comparative example 6
0.1%
21.25
Comparative example 7
0.1%
23.46
Comparative example 8
0.1%
30.67
Comparative example 9
0.1%
19.45
Comparative example 10
0.1%
24.00
Comparative example 11
0.1%
25.14
Comparative example 12
0.1%
34.45
SDS (Positive control)
60μg/ml
24.23
PBS (negative control)
10%
51.58
From table 7, the following conclusions can be drawn:
1) examples 1-3 and comparative examples 1-11 show that the content of VEGF in a sample lacking any component is significantly lower than that of a negative control, and the content of VEGF in examples 1-3 is significantly higher than that of the negative control, which proves that the anti-allergic repair barrier composition has the ability of promoting the expression of Vascular Endothelial Growth Factor (VEGF) and has good cell barrier repair ability;
2) the VEGF content of the composition in the example 1 is obviously higher than that of the compositions in the examples 2-3, and the compatibility rationality of the barrier repairing composition is proved.
Effect test example 3 (human body moisture test)
Selecting 30 subjects with the ages of 18-60 years according with requirements; the basal value of the forearm test area is between 15 and 45(Corneometer Unit, c.u.) after pre-examination by a capacitance skin moisture tester.
In the test process, a subject needs to wipe the marked tested part on the inner side of the arm with a clean paper towel in a constant temperature and humidity (temperature: 20-22 ℃, humidity: 40-60%) environment, sit still for 30min, and test a blank value according to (2.0 +/-0.1 mg)/cm2The amount of (c) is applied to the sample.
And waiting for 1h, 2h, 4h and 8h, and then carrying out skin moisture value test. As shown in figure 1 and table 8 below.
TABLE 8 results of moisture value (C.U) of human moisture test
Sample (I)
0H
1H
2H
4H
8H
Example 4
32.5
60.1
59.2
57.2
45.7
Example 4 moisture Rate of Change
/
84.9%
82.2%
76.0%
40.61%
Market contest
33.1
52.1
45.1
40.3
35.1
Rate of change of moisture for a commercial offering
/
57.4%
36.3%
21.8%
6.0%
Blank control
30.1
31.2
33.4
29.8
32.3
Rate of change of blank moisture
/
3.7%
11.0%
-1.0%
7.3%
From the results of fig. 1 and table 8, it can be seen that:
after the test subject uses the barrier repairing composition-containing example 4, the moisture value is obviously higher than that of the market competitive products and blank, and the significant difference exists between the blank and the market competitive products, so that the test subject proves that the barrier repairing composition-containing example 4 has the moisturizing effect, the moisturizing effect is better than that of the market competitive products, and the moisturizing effect is better.
Effect test example 4 (human body anti-allergy test)
The samples were: example 2 and a commercially available well-marketed moisturizing gel.
Table 9 comparison of example 2 and commercially available tombstock moisturizing gels before and after use
From the results of fig. 2 and table 9, it can be seen that:
after 1 month of sample application, the red zone of the subject was reduced by 46%, and the facial skin sensitivity was decreased.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, so that any simple modification, equivalent change and modification made to the above embodiment according to the technical spirit of the present invention are within the scope of the technical solution of the present invention.
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