CN112741787B

CN112741787B - Multi-component combined solution and preparation method and application thereof

Info

Publication number: CN112741787B
Application number: CN202011188880.1A
Authority: CN
Inventors: 章漳; 蒋丹丹; 蒋耀权; 王风楼; 黄�俊
Original assignee: Shanghai Natural Hall Group Co ltd
Current assignee: Shanghai Natural Hall Group Co ltd
Priority date: 2019-10-31
Filing date: 2020-10-30
Publication date: 2024-03-26
Anticipated expiration: 2040-10-30
Also published as: CN112741787A

Abstract

A multi-component combined solution comprising: the invention relates to a multi-component combined solution which is prepared from 1 to 10 weight percent of collagen, 1 to 10 weight percent of thiotaurine, 10 to 50 weight percent of hot spring water and 30 to 88 weight percent of deionized water, wherein the collagen and the thiotaurine are added by taking the deionized water and the hot spring water as solvents. The experimental result shows that the composition has obvious effects on the tissue structure of the epidermis model and the expression of the papilionaceous protein/epicutaneous protein, has positive regulation effect on dermal capillary endothelial cells, and is reflected to have correct response to capillary permeability control regulatory factors; has obvious effects of resisting lipid peroxidation and inflammation of skin keratinocyte.

Description

Multi-component combined solution and preparation method and application thereof

Technical Field

The invention relates to the field of daily chemical skin care, in particular to a multi-component combined solution and a preparation method thereof.

Background

In recent years, there is an increasing trend in the occurrence of sensitive skin in China, and one investigation in 2011 shows that about 46% of Chinese females and 30% of Chinese males have developed skin sensitivity to varying degrees (journal of clinical dermatology, 2011, 40 (4): 261). However, the reasons for the occurrence of sensitive skin are not unified at present, and may be related to factors such as skin barrier function damage, inflammatory reaction, nerve sensitivity enhancement, sebum change abnormality and the like. Because of the complexity of the cause and mechanism of sensitive skin, no specific treatment method exists at present. Studies have shown that cosmetic abuse may lead to sensitive skin, whereas the use of a soothing cosmetic helps to improve sensitive skin. Targeted daily skin care is therefore important, such as the use of compositions containing skin barrier repair, sebum antioxidant, anti-inflammatory soothing, to improve and alleviate discomfort in sensitive skin.

Collagen is the most abundant protein in animal bodies, is the main component of extracellular matrix, accounts for about 25-33% of total protein in the body, is equivalent to 6% of body weight, and has important effects on maintaining normal physiological functions and injury repair of cells, tissues and organs. Collagen has high strength and stability due to its special triple helix structure, and also has high biocompatibility and biodegradability, and is one of the hot spots for current bioengineering including tissue engineering material research. Collagen is an important biological resource and is widely used in the fields of cosmetic application such as wrinkle removal, moisture preservation, skin moisturizing, beauty treatment, hair care and the like: if the collagen molecule contains a large amount of hydrophilic groups, the collagen has good moisturizing effect, and the purpose of keeping skin moist can be achieved; the collagen can maintain the moisture of the horny layer and the integrity of a fiber structure, improve the living environment of skin cells and promote the metabolism of skin tissues, and has better anti-aging and anti-wrinkle effects; in addition, the collagen has good compatibility with surrounding tissue cells, and has the function of repairing tissues.

The thiotaurine can react with active oxygen preferentially in the presence of the active oxygen, so that the active oxygen is prevented from being oxidized with other components to generate the hypotaurine and taurine which are also biological molecules, the thiotaurine can display high-efficiency hydroxyl radical removal capability at very low concentration, can prevent primary stage oxidation of skin, inhibit skin damage caused by peroxidation of liposome of epidermis, and can supplement and strengthen the antioxidant capability of skin. Thiotaurine has the capability of removing active oxygen such as super oxidation, singlet oxygen and the like, namely removing sebum oxide, and also has a moisturizing effect.

The hot spring water is ground water which is gushed out of the ground due to the influence of the motion change of various substances in the earth, the action of ground temperature and the pressure of water vapor and the change of crust structure, and usually contains trace elements which have some remarkable physical therapy effect on human bodies. Modern science discovers that some hot spring water contains a large amount of mineral substances and various trace elements, and has the skin care effects of resisting oxidation, scavenging free radicals, resisting inflammation, relieving skin, improving skin aging and the like. With the continuous intensive research of the efficacy of hot spring water on skin, more and more cosmetic brands feature hot spring water as a product. The middle sea of the Mediterranean-Himalayan tropical region of the Tibet site has extremely developed internal structure, strong magma activity and deterioration effect, and is one of the regions with the strongest global geothermal activity. According to the record of the Tibet Hot spring, the position of the hot spring 677 in each Tibet city is the region with rich geothermal resources in China. There is a long history of local Tibetan use of hot spring water to treat skin disorders.

Although the raw materials are all single or nearly single in effect, the raw materials cannot have the effects of repairing skin barrier, resisting allergy, resisting oxidization and the like.

Disclosure of Invention

The present invention is directed to solving the above problems, and the present invention provides a multi-component combined solution comprising: collagen, thiotaurine, hot spring water and deionized water, wherein the weight percentage of the collagen is 1-10%, the thiotaurine is 1-10%, the hot spring water is 10-80%, and the deionized water is 30-88%.

The collagen protein is selected from bovine collagen, fish collagen, human recombinant type I collagen and human recombinant type III collagen.

The hot spring water is selected from hot spring water in Riduo Tibet areas, hot spring water in Layue Tibet areas and hot spring water in Palong Tibet areas.

A method for preparing a multi-component combined solution comprising the steps of:

A. removing impurities from hot spring water, heating the hot spring water to 80 ℃, keeping the temperature for 30min, and cooling;

B. heating deionized water to 80 ℃, keeping the temperature for 30min, and cooling;

C. adding hot spring water obtained in the step A into deionized water obtained in the step B according to the weight ratio (15:132-220:132) to form mixed solution;

D. adding thiotaurine into the mixed solution 1 prepared in the step C according to the weight ratio (4:392-49:392), and stirring, dispersing and dissolving to obtain a mixed solution 2;

E. and D, adding the collagen into the mixed solution 2 prepared in the step D according to the weight ratio of (1:99-11:99), and stirring, dispersing and dissolving to obtain a combined solution.

The impurity treatment in the step A sequentially comprises 5 mu mPP cotton filtration, active carbon filtration and 1 mu mPP cotton filter element filtration.

And E, stirring, dispersing and dissolving the glass rod, and dispersing the glass rod by a homogenizer, wherein the rotating speed of the homogenizer is 3000rpm.

A skin external preparation comprising the aforementioned multicomponent combined solution, the mass of the multicomponent combined solution accounting for 0.1% to 100% of the total mass of the skin external preparation; the skin external agent is prepared into spray for use when the ratio of the mass of the multicomponent combined solution to the total mass of the skin external agent is 100%; when the ratio of the mass of the multicomponent combined solution to the total mass of the skin external agent is less than 100%, the skin external agent further contains other types of water, which is preferably deionized water, distilled water, or ultrapure water.

The invention takes deionized water and hot spring water as solvents to add collagen and thiotaurine to form a multi-component combined solution. The experimental result shows that the composition has obvious effects on the tissue structure of the epidermis model and the expression of the papilionaceous protein/epicutaneous protein, has positive regulation effect on dermal capillary endothelial cells, and is reflected to have correct response to capillary permeability control regulatory factors; has obvious effects of resisting lipid peroxidation and inflammation of skin keratinocyte.

The multi-component combination solution of the present invention can therefore be effective against the disturbance of sensitive skin by repairing the skin barrier; has the functions of remarkably promoting the tight connection between capillary endothelial cells, maintaining the skin homeostasis, increasing the antioxidant and anti-inflammatory effects of the skin through the actions of resisting lipid peroxidation of keratinocytes and releasing inflammatory factors, and effectively achieving the effects of relieving and improving the sensitive skin state.

Drawings

FIGS. 1a-f show cell epidermal tissue structures, wherein FIG. 1a shows control cells, FIG. 1b shows UV-irradiated cells, FIG. 1c shows 0.05% collagen (additive amount) UV-irradiated cells, FIG. 1d shows 0.05% thiotaurine (additive amount) UV-irradiated cells, FIG. 1e shows 0.5% spring water (additive amount) UV-irradiated cells, and FIG. 1f shows 1% combination (combination 1) UV-irradiated cells.

Designating figure 1a as the abstract figure.

FIG. 2 is a graph showing the comparison of the regulation of basal layer-related gene expression by the untreated control group (NT) and the collagen-containing component combination experimental group having a final concentration of 1% in example III.

FIG. 3 is a graph showing comparison of regulation of expression of anti-aging-related genes in an untreated control group (NT) and a collagen-containing component-combined experimental group having a final concentration of 1% in example III.

FIG. 4 is a graph showing comparison of regulation of expression of anti-aging-related genes in an untreated control group (NT) and a collagen-containing component-combined experimental group having a final concentration of 1% in example III.

Detailed Description

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

The main components in the following experiments comprise collagen, thiotaurine, hot spring water and deionized water. Wherein the collagen can be bovine collagen, fish collagen, human recombinant type I collagen, type III collagen, etc. The hot spring water can be Tibet day hot spring water, layue hot spring water, pa Long Cangbu hot spring water, etc. According to the weight percentage, the collagen accounts for 1-10%, the thiotaurine accounts for 1-10%, the hot spring water accounts for 10-50%, and the deionized water accounts for 30-88%. Specifically, the components can be distributed according to Table 1.

TABLE 1

	Range	Combination 1	Combination 2	Combination 3	Combination 4	Combination 5
							Collagen protein	1-10%	1%	3%	5%	7%	10%
Thiotaurine	1-10%	1%	3%	5%	7%	10%
							Spring water	10-50%	10%	30%	50%	50%	50%
Deionized water	30-88%	88%	64%	40%	36%	30%

Embodiment one:

the proportions are given in Table 1 above, wherein the proportions of composition 3 are exemplified by 5% collagen, 5% thiotaurine, 50% hot spring water, and 40% deionized water. The preparation process comprises the following steps:

filtering untreated hot spring water to remove impurities in the water (passing through 5-micrometer PP cotton, granular activated carbon and a 1-micrometer PP cotton filter core in sequence), and detecting conductivity in the range of less than 500 mu S/cm.

Deionized water was heated to 80 degrees celsius, held for 30min, and cooled. 50g of hot spring water was then added to 40g of deionized water.

5g of thiotaurine was weighed out and added to the above aqueous solution, followed by stirring, dispersion and dissolution.

Weighing 5g of collagen, adding the collagen into the aqueous solution, stirring the mixture by using a glass rod for preliminary dispersion, dispersing and homogenizing the mixture by using a homogenizer, adjusting the revolution of the homogenizer to 3000rpm, and homogenizing the mixture for 5 minutes; the desired multicomponent combined solution is obtained.

Embodiment two:

the multi-component combined solution obtained in the above preparation manner was subjected to the following efficacy test. Wherein, according to the proportion, the single component samples are respectively collagen solution with the concentration of 0.05 percent, thiotaurine solution with the concentration of 0.05 percent and daily hot spring water solution with the concentration of 0.5 percent, which are respectively called as combination 1-combination 5:

1. efficacy testing based on epidermis model

The purpose of the test was to compare the effect of 3 components and their compositions on the epidermis model, and to evaluate their effect on skin barrier repair.

The testing method comprises the following steps:

after normal human keratinocytes in the logarithmic growth phase were digested and resuspended, 1.0Mcells/mL was inoculated into a suspension culture chamber Millicell of a matched 24-well plate, the chamber and receiving well were filled with culture medium, a control group and a sample treatment group were set respectively, cultured in an incubator at 37 ℃,5% co2, saturated humidity, and the culture was continued for 2 weeks with 1-fold change of medium every 24 hours, and then the epidermis was irradiated with UVB, followed by further culture for 24 hours. After the culture, the epidermis model is soaked in 4% formaldehyde for 24 hours, and then dehydrated and embedded in conventional paraffin sections. And (3) dewaxing and rehydrating paraffin sections, observing tissue structures of epidermis models of a control group and a sample treatment group through HE staining, photographing microscopic images, performing semi-quantitative analysis on the thickness of the epidermis by adopting imageJ software, simultaneously calculating the number of photodamaged cells with nuclear shrinkage phenomenon in each group of epidermis, and calculating the relative ratio of photodamaged cells of each sample group by taking the number of photodamaged cells in the epidermis of a control UV group as a reference. .

Test results:

the results obtained are presented in the following figures 1a-f and table 2. The composition has the advantages of remarkably thickened epidermis layer, complete tissue structure, clear grain layer differentiation, thicker stratum corneum, less photodamaged cells after UVB irradiation compared with other groups, and better activity and tolerance of epidermis cells.

Table 2:

conclusion:

the composition has a remarkable effect on the tissue structure of the skin model compared with the single component, and the tissue structure is closely related to the barrier function of the skin, so that the effect can play a role in repairing the skin barrier. Thus, the composition obtained according to the invention can be effective against the disturbance of sensitive skin by repairing the skin barrier.

It should be noted that deionized water itself does not have a skin barrier repair effect, a soothing effect on sensitive skin, and a homeostasis effect on skin, so the above efficacy test results are not tired.

Example III

Component combination gene detection containing collagen

Skin aging is a complex physiological phenomenon affected by a variety of factors, and aging mainly takes two forms, natural aging and photoaging. Natural aging is aging caused by genes or ages, and photoaging is aging caused by ultraviolet irradiation. Skin aging starts at age 25 years old, skin aging is obvious at age 40-50 years old, healthy skin has high water content, skin is smooth and fine, is full of elasticity, the water retention components of cells can be gradually reduced along with the aging and the influence of external environmental factors such as ultraviolet rays, pollution, chemical substances and the like, when the water content is lower than a certain level, the skin can be rough, serious conditions have external manifestations such as wrinkles and the like, and the internal manifestations are reduction of the expression of the regulation genes related to the moisture retention related factors, the aging of the fiber cells, the flow and deformation of elastin and the like, so that research on how to improve the expression of the genes related to the moisture retention and the anti-aging genes, and the effects of keeping the skin moist, the elasticity and restoring the physiological functions of the skin have important significance for delaying skin aging.

The invention utilizes the reconstructed human skin model to detect the regulation of the collagen-containing component combination on skin moisture retention, aging and other related genes, and verifies the influence of the collagen-containing component combination on aging.

The collagen-containing component comprises collagen, thiotaurine and hot spring water. The collagen may be bovine collagen, fish collagen, human recombinant type I collagen, type III collagen, etc. The hot spring water can be Tibet day hot spring water, layue hot spring water, pa Long Cangbu hot spring water, etc. The preferred combination of combination 3 is described. Recombinant human type III collagen is provided by Jiangsu Jiangshan poly-sourced biotechnology Co., ltd or Jiangsu Yuan Zhi biological medicine Co., ltd; fish hydrolyzed collagen is provided by Guangdong hundred vitamin technology Co., ltd; bovine soluble collagen is supplied by CLR-chemischen laboratory; thiotaurine is derived from japan mutual pharmaceutical corporation or Shanghai Hui Biotechnology Co., ltd; thermal spring aquatic land Tibet is provided by Shanghai Jia leaf Utility Co., ltd. Or Yunnan Kunming Qianzhen biotechnology Co., ltd.

	Combination 3
		Collagen protein	5%
Thiotaurine	5%
		Spring water	50%
Deionized water	40%

FIG. 2 is a graph showing the comparison of the regulation of basal layer-related gene expression by an untreated control group (NT) and a collagen-containing component combination experimental group having a final concentration of 1%.

FIG. 3 is a graph showing comparison of regulation of expression of anti-aging-related genes in untreated control (NT) and collagen-containing component-combined experimental groups at a final concentration of 1%.

FIG. 4 is a graph showing comparison of regulation of expression of anti-aging-related genes in untreated control (NT) and collagen-containing component-combined experimental groups at a final concentration of 1%.

1. Concentration of collagen-containing components

Preparation of the composition containing collagen at the experimental concentrations according to Table 1 below

TABLE 3 Table 3

Raw materials	Preparation method	Final concentration	Group category
				Collagen-containing component combinations	Adding culture medium for dilution	1%	Experimental group
/	/	0	Untreated control group (NT)

2. Gene regulation test of extracting RNA (ribonucleic acid) by using component combination containing collagen in 3D skin model

1) The preparation steps of the 3D skin model are as follows:

resuscitates human primary fibroblasts (NHDF): the freezing tube is taken out of the liquid nitrogen, rapidly put into a 37 ℃ water bath for melting, and centrifuged at 1000rpm/min for 10min. The culture was resuspended in DMEM medium, inoculated into T175 flasks and incubated in a carbon dioxide incubator at 37℃with a 2-day change.

After one week of incubation, the cells were digested with 0.05% trypsin, added to complete DMEM medium, transferred to a 50mL centrifuge tube, centrifuged at 1000rpm/min for 10min and the supernatant discarded.

The cells are resuspended by DMEM, mixed with collagen and placed in a carbon dioxide incubator at 37 ℃ for dermis stage culture for 3-4 days.

Resuscitates human primary keratinocyte (NHEK), inoculates on the skin model surface of the dermis stage, adds the culture medium and submerges the skin model to culture, change liquid every 2 days, set up the composition group experiment (1%, v/v) and untreated control group (NT) containing collagen of component combination of the collagen at last time of changing liquid, add the composition group containing collagen to culture medium of the experiment group according to the volume ratio.

After one week, the culture medium is changed into gas phase culture, liquid is changed every three days, and the composition containing collagen is added into the culture medium of the experimental group according to the volume ratio.

The skin was harvested after 8 days of gas phase culture for RNA extraction.

2) The extraction and preparation method of RNA comprises the following steps:

3D skin model tissue was cut approximately 2X 3 mm and placed in a 2mL EP tube.

1mL Trizol was added, 2 steel balls were added, homogenized with a tissue breaker, and sonicated for 5min at 50 Hz.

Centrifuge at 9000rpm, 4℃for 1min, transfer supernatant into a fresh 1.5mL EP tube.

Adding 200 mu L of chloroform, slightly reversing for mixing for several times, and standing at room temperature for 5 minutes; centrifugal at 9000rpm and 4℃for 15min.

The upper aqueous phase (about 400 μl) was transferred to a new 1.5mL EP tube, added with equal volume of isopropanol, mixed well and allowed to stand at room temperature for 10min.

Centrifugal for 10min at 9000rpm and 4 ℃.

The supernatant was discarded and the pellet was washed 1 time with pre-chilled 70% absolute ethanol and centrifuged at 8000rpm at 4℃for 5min.

The supernatant was discarded, precipitated in air for 5-10 minutes and dissolved in 30 μl DEPC (diethyl coked acid) water.

The RNA concentration was determined by spectrophotometry.

3) Analysis of Gene expression

And (3) carrying out RNA purification and gene quantitative analysis according to a QuantiGene R2.0 Plex Assay kit, analyzing gene expression by using a luminex platform, and detecting the expression level of related genes, wherein the results are shown in figures 1-3.

3. Regulation of expression of basal-layer-associated genes by combinations of collagen-containing components

COL7A1The VII type collagen encoded by the (collagen type VII alpha chain) gene is the main collagen expressed in the dermis junction, and the dermis junction is mainly composed of collagen, laminin, endo-albumin and diafiltration. Collagen is a closely related class of macromolecules with different genes, and more than 20 vertebrate collagens are currently identified. All collagen molecules are encoded by three polypeptide subunits, known as the alpha chain, and also by the human genome, in up to 25 different genes. The collagen with different genes has characteristic tissue distribution. For example, the most abundant collagens such as type I and type III are widely distributed, and fewer collagens are type VII collagens (COL VII), COL7A1 serving as an anchor at the dermis and epidermis interface to firmly connect between the eukaryotic cells, and when mutations occur in the absence of the corresponding collagens or related genes, a variety of diseases, such as bullous diseases, etc. may result.

COL4A1 shares a bi-directional promoter with paralogous genes on opposite strands. The protein consists of an amino-terminal 7S domain, a triple helix forming a collagen domain, and a carboxy-terminal non-collagen domain. It acts as part of a heterotrimer and interacts with other extracellular matrix components such as passion fruit, proteoglycans, and laminins. In addition, proteolytic cleavage of the non-collagenous carboxy-terminal domain results in a biologically active fragment called arresten, which has anti-angiogenic and tumor-inhibiting properties. Mutation of this gene leads to pulmonary capillary disease, cerebrovascular disease, kidney and muscle defects. Multiple transcript variants result in alternative splicing.

As can be seen from fig. 2:

1% collagen-containing component combination was added to the basal layer-associated gene as compared with untreated control (NT)COL7A1Up-regulated by 1.76 times.

Compared with untreated control group (NT), 5% highland barley glucan solution was added, basal layer related genesCOL7A1Up-regulated by 1.77 times.

4. Collagen-containing composition for regulating expression of anti-aging-related genes

HBEGF is a member of the EGF family that binds to and activates the EGF receptor growth factor. HBEGF plays a key role in pathophysiological processes, including cancer. After the partial thickness of human skin is damaged, the EGF can generate the EGF, promote the cell proliferation, is beneficial to the wound healing of the skin, and simultaneously EGF is the EGF, promotes the proliferation of the EGF cells and maintains the young state of the skin.

CSF2, known as granulocyte-macrophage colony stimulating factor, is a broad-spectrum cytokine that has a pro-proliferative effect on early hematopoietic progenitor cells. The cell line is mainly used for myeloid progenitor cells, and plays a biological effect through combination with high-affinity receptors, so that the myeloid progenitor cells rapidly enter a cell cycle to differentiate into granulocytes and macrophagous lines, finally become mature cells, and can prolong the service life of the mature cells and enhance the functions of the mature cells. Colony stimulating factor 2 is an important cell longevity factor and is closely related to skin aging.

The latent transforming growth factor beta binding protein 1 encoded by the LTBPI gene belongs to the latent TGF-beta binding protein (LTBP) family. Secretion and activation of TGF-beta is regulated by its association with latency-associated proteins and latency T G F-beta binding proteins. Transforming growth factor (transforminggrowth factor-beta, TGF-beta) is a multi-effect and complex-function cytokine involved in cell growth, tissue repair and wound healing, generation of blood vessels and scars, and generation and metastasis of tumors. Thus, latent transforming growth factor beta binding protein 1 is closely related to skin wound healing, tissue repair and anti-aging.

As can be seen from fig. 3:

(1) The anti-aging gene HBEGF was upregulated 1.67-fold by the addition of 1% collagen-containing component combination compared to the untreated control (NT).

(2) The anti-aging gene CSF2 was up-regulated 3.54-fold by the addition of 1% collagen-containing component combination compared to untreated control (NT).

(3) The anti-aging gene CSF2 was up-regulated 1.49-fold by the addition of 1% collagen-containing component combination compared to the untreated control (NT).

5. Expression modulation of moisture-retention related genes by combinations of collagen-containing components

CD44 is a cell surface transmembrane glycoprotein that, in addition to promoting cell differentiation and proliferation of epidermal cells, is also capable of binding to hyaluronic acid and is thus also known as a receptor for hyaluronic acid. Cells bind hyaluronic acid to extracellular spaces via CD44 to maintain high intercellular water content. In addition, CD44 plays an important role in the synthesis of hyaluronic acid, and the amount of CD44 expressed in skin cells determines the content of hyaluronic acid in the skin. However, as the age increases, CD44 in the body gradually decreases and the vitality also decreases, which is also the root cause of the dry and water-deficient skin. By activating the expression of CD44, the work efficiency of self-renewal and hyaluronic acid production can be improved, and the moisturizing ability of the skin can be improved.

As can be seen from fig. 4:

(1) The moisturizing modulator CD44 was up-regulated 1.71-fold by the addition of 1% collagen-containing component combination compared to the untreated control (NT).

According to the expression test results, 1% of the composition combination containing the collagen can up-regulate the expression of basal layer related genes, extracellular matrix protein related genes, anti-aging related genes and moisturizing related genes, so that the service life of mature cells is prolonged, the functions of the mature cells are enhanced, the moisturizing capability of skin is improved, tissue repair is accelerated, and the like, and finally the effect of delaying skin aging is achieved.

Example IV

When the multicomponent combined solution of the invention is applied to external skin preparations:

when the external preparation for skin is a toning lotion, the specific composition thereof is shown in the following table 4:

TABLE 4 Table 4

Referring to Table 4, the skin external preparation of the above ratio was prepared according to the preparation method of the conventional toning lotion in the art.

The skin external agent is essence, and the specific composition is shown in the following table 5:

TABLE 5

Referring to Table 5, the skin external preparation of the above ratio was prepared according to a preparation method of essence conventional in the art.

When the external preparation for skin is an emulsion or a cream, the specific compositions thereof are shown in Table 6 below.

TABLE 6

Referring to Table 6, the skin external preparation of the above ratio was prepared according to the preparation method of the emulsion and the cream conventional in the art.

In yet another embodiment, the multi-component composition solution according to the present invention is used in the form of a spray when the ratio of the mass of the multi-component composition solution to the total mass of the external skin preparation is 100%.

In the above embodiments, the recombinant human type III collagen is provided by a biological technology Co., ltd. From Jiangsu Jiangshan or Jiangsu Yuan Zhi biological medicine Co., ltd; fish hydrolyzed collagen is provided by Guangdong hundred vitamin technology Co., ltd; bovine soluble collagen is provided by CLR-Chemisches Laboratorium; thiotaurine is derived from japan mutual pharmaceutical corporation or Shanghai Hui Biotechnology Co., ltd; thermal spring aquatic land Tibet is provided by Shanghai Jia leaf Utility Co., ltd. Or Yunnan Kunming Qianzhen biotechnology Co., ltd.

The above description of the embodiments is only for aiding in the understanding of the method of the present invention and its core ideas. It should be noted that it will be apparent to those skilled in the art that various modifications and adaptations of the invention can be made without departing from the principles of the invention and these modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.

Claims

1. The application of the multi-component combined solution in preparing the external skin preparation is characterized in that,

the multicomponent composition solution is used as an active ingredient with skin care effect in skin external preparation, the mass of the multicomponent composition solution accounts for 1% of the total mass of the skin external preparation, the skin care effect is that the skin barrier is repaired after UV photodamage,

the multi-component combined solution consists of 5% of collagen, 5% of thiotaurine, 50% of hot spring water and 40% of deionized water, wherein the hot spring water is daily hot spring water in Tibet areas,

the preparation method of the multi-component combined solution comprises the following steps:

A. removing impurities from the hot spring water, heating the hot spring water to 80 ℃, keeping the temperature for 30min, and cooling;

B. heating the deionized water to 80 ℃, keeping the temperature for 30min, and cooling;

C. adding the hot spring water obtained in the step A into the deionized water obtained in the step B according to the weight ratio to form a mixed solution 1;

D. c, adding the thiotaurine into the mixed solution 1 prepared in the step C, and stirring, dispersing and dissolving to obtain a mixed solution 2;

E. and D, adding collagen into the mixed solution 2 prepared in the step D, and stirring, dispersing and dissolving to obtain a combined solution.

2. Use of the multicomponent combination solution according to claim 1 for the preparation of an external skin preparation, wherein the collagen is selected from the group consisting of bovine collagen, fish collagen, human recombinant type I collagen and human recombinant type III collagen.

3. Use of the multicomponent combined solution according to claim 1 for the preparation of an external skin preparation, wherein the impurity treatment of step a comprises, in order, 5 μmpp cotton filtration, activated carbon filtration and 1 μmpp cotton filter cartridge filtration.

4. Use of the multicomponent combined solution according to claim 3 for preparing a skin external preparation, wherein the stirring dispersion dissolution of step E comprises a glass rod stirring dispersion, a homogenizer dispersion in this order, wherein the homogenizer rotation speed is 3000rpm.