CN115554187A

CN115554187A - Mussel mucin skin repair emulsion capable of achieving wet heat sterilization and preparation method thereof

Info

Publication number: CN115554187A
Application number: CN202211552905.0A
Authority: CN
Inventors: 徐林; 金玉; 徐念沁; 曹晶晶; 徐松泉; 居丽娜; 姜豪; 刘谋治; 黎文清
Original assignee: Nanjing Tzong Blotechnology Co ltd
Current assignee: Nanjing Tzong Blotechnology Co ltd
Priority date: 2022-12-06
Filing date: 2022-12-06
Publication date: 2023-01-03
Anticipated expiration: 2042-12-06
Also published as: CN115554187B

Abstract

The invention discloses a mussel mucin skin repairing emulsion capable of being sterilized by heat and moisture and a preparation method thereof, and the mussel mucin skin repairing emulsion comprises the following components in parts by weight: mussel mucin 0.1-0.2 weight portions, trehalose 3-15 weight portions, humectant 2-20 weight portions, emollient 1.5-2 weight portions, solubilizer 0.01-0.03 weight portions, thickener 0.2-1 weight portions, pH regulator 0.1-0.5 weight portions, and purified water 80-95 weight portions. The invention adopts the trehalose as the protective agent, so that the mussel mucin keeps good biological activity after moist heat sterilization, simultaneously, the emulsion system is stable, and the anti-inflammatory effect of the mussel mucin is enhanced by the synergistic effect of the trehalose and other components. The emulsion of the invention has good film forming property and anti-inflammatory effect, and has good industrial production value and economic value.

Description

Mussel mucin skin repair emulsion capable of achieving wet heat sterilization and preparation method thereof

Technical Field

The invention belongs to the technical field of biology, and relates to a mussel mucin preparation, in particular to a mussel mucin skin repairing emulsion capable of being sterilized by wet heat and a preparation method thereof.

Background

Mussel Adhesive Protein (MAP) is extracted from the foot silk gland of common mussel (blue mussel, mytilus edulis) and is a polyphenol single protein biomaterial. The mussel mucin molecule consists of 70-90 repetitive polypeptide fragments containing 20% lysine and 10% dopa groups (3, 4-dihydroxyphenylalanine). Lysine is positively charged, attracting negatively charged epidermal cells, fibroblasts, vascular endothelial cells, neural cell anchorage and creeping substitutions by electrostatic action; the auto-oxidation crosslinking of the dopa group is embodied in that the dopa group can be combined with oxygen in the air to form dopaquinone, and the dopaquinone and unoxidized dopa can be crosslinked to form a high polymer, so that the healing of a wound surface is promoted. Therefore, the mussel mucin is an excellent choice for wound repair dressing. For example, chinese patent document CN114569515A discloses a hydrogel for repairing damaged skin barrier after medical arts and a preparation method thereof, wherein mussel-like mucin is adopted to combine with tangerine extract, citronella essential oil, valerian essential oil and the like in a scientific ratio, and the prepared hydrogel has a good effect on repairing skin barrier after medical arts and crafts.

Mussel mucin is sensitive to temperature and humidity, the denaturation temperature is low, and high temperature can cause the structure to be dissociated and the bioactivity to be lost, so that the existing mussel mucin dressing type sterile products are sterilized by irradiation. For example, chinese patent document CN114917403A discloses a mussel-like mucin gel and its preparation method and application, wherein the gel is lyophilized into powder or the sponge and the auxiliary solvent are separated, the mussel-like mucin exists in the lyophilized powder or sponge, and the product can be preserved at room temperature by low-dose radiation sterilization.

Trehalose, also known as rhaponticum and mycose, is a non-reducing disaccharide composed of two glucose molecules. Trehalose is studied as the most stable class of natural disaccharides. Because of the non-reducibility, the product has very good stability to heat and acid and alkali. When the amino acid and the protein coexist, the Maillard reaction does not occur even when the mixture is heated. Thus, research into trehalose as a protein protectant has been of great interest, for example: chinese patent document CN101007840A discloses a method for keeping activity of active protein at normal temperature, which uses two chemical substances to form a composite component for improving stability of active protein, thereby achieving the purpose of storing and transporting active protein at normal temperature, wherein the two components are dodecyl polyglycol ether (Brij-35) and alpha-D-glucopyranosyl-alpha-D-glucopyranoside (trehalose ).

However, the research finds that the protein protective agent added with trehalose is mostly used for medicinesThe protection of the lyophilized powder of the product or preparation, even for the high temperature protection of the protein, does not exceed 42 ℃. For example: chinese patent document CN113694189A discloses a freeze-drying protective agent for protease K and its preparation method, which comprises adding Ca ²⁺ And one or more of sucrose, trehalose, maltose, glycine, L-serine and alpha-alanine are used as components of the freeze-drying protective agent, so that the freeze-drying survival rate of the proteinase K in the vacuum freeze-drying process can be effectively improved, and the storage stability of the proteinase K freeze-dried powder is improved. The Chinese patent document CN113717272A discloses a freeze-drying protective agent for improving the stability of recombinant human collagen, wherein the protective agent comprises, by weight, 500-2000 parts of trehalose, 100-500 parts of polyvinylpyrrolidone (PVP-K30) and 100-200 parts of mannitol, and takes 100 parts of recombinant human collagen as 100 parts; in the freeze-drying process of the recombinant human collagen, the freeze-drying protective agent is added, so that the stability of the recombinant human collagen freeze-dried powder at normal temperature can be obviously improved, the purity and activity of the recombinant human collagen freeze-dried powder are prevented from being reduced, and the prepared freeze-dried powder can be placed at normal temperature for a long time.

For medical and beauty products such as skin repair emulsion and the like, the components of the skin repair emulsion mostly contain high polymer materials, 60 Co-gamma rays commonly used for radiation sterilization have certain destructiveness on the high polymer materials, and meanwhile, the skin repair emulsion can also damage the packaging materials of the products to a certain degree. Therefore, how to realize the wet heat sterilization of medical and American products containing the mussel mucin and ensure the bioactivity of the mussel mucin in the products is an urgent problem to be solved.

Disclosure of Invention

Aiming at the problems of realizing the wet heat sterilization of medical and beauty products containing mussel mucin and simultaneously ensuring the bioactivity of the mussel mucin in the products, the invention provides the mussel mucin skin repair emulsion with the wet heat sterilization and the preparation method thereof. The specific technical scheme is as follows:

firstly, the invention provides a mussel mucoprotein skin care emulsion capable of being subjected to wet heat sterilization, which comprises the following components in parts by weight: mussel mucin 0.1-0.2 weight portions, trehalose 3-15 weight portions, humectant 2-20 weight portions, emollient 1.5-2 weight portions, solubilizer 0.01-0.03 weight portions, thickener 0.2-1 weight portions, pH regulator 0.1-0.5 weight portions, and purified water 80-95 weight portions.

Preferably, the mass ratio of the trehalose to the mussel mucin is 15-150.

Further preferably, the mass ratio of trehalose to mussel mucin is 50.

The mussel mucin skin care emulsion capable of being sterilized by heat and moisture can be prepared by mixing the mussel mucin skin care emulsion with a humectant, wherein the humectant is one or more of glycerin, propylene glycol, butanediol, pentanediol, sorbitol, polyethylene glycol, betaine, sodium hyaluronate, allantoin and urea; the emollient is one or more of cocoa butter, vaseline, water soluble silicon wax, liquid paraffin, lanolin, stearate, lecithin, squalane, polydimethylsiloxane, and cetostearyl alcohol; the solubilizer is one or more of hydrogenated castor oil, polyoxyethylene castor oil, fatty alcohol polyoxyethylene ether, polyoxyethylene sorbitan fatty acid ester and polyglycerol fatty acid ester; the thickening agent is one or more of carbomer, sodium polyacrylate, methylcellulose, ethyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose, soluble starch, xanthan gum and gelatin; the pH regulator is one of triethanolamine, aminomethyl propanol, sodium hydroxide and potassium hydroxide.

Preferably, the humectant is one or more of glycerin, propylene glycol, butanediol, betaine, sodium hyaluronate and allantoin; the emollient is one or more of cocoa butter, lecithin, squalane and polydimethylsiloxane; the solubilizer is one or more of hydrogenated castor oil, polyoxyethylene castor oil and fatty alcohol-polyoxyethylene ether; the thickening agent is one or more of carbomer, sodium polyacrylate, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose and carboxymethyl cellulose; the pH regulator is one of triethanolamine and aminomethyl propanol.

The mussel mucin skin repairing emulsion capable of being sterilized by heat and moisture is prepared from trehalose, glycerol, propylene glycol, butanediol, betaine and allantoin in a mass ratio of 1-30, preferably 2-10; the mass ratio of the trehalose to the lecithin, the squalane and the polydimethylsiloxane is 1-100, and preferably 3-20.

Secondly, the invention provides a preparation method of the mussel mucin skin repair emulsion capable of being sterilized by wet heat, which comprises the following steps:

s1: adding trehalose, a thickening agent and a humectant into purified water, stirring at the speed of 200-300 rpm, heating to 40-85 ℃, dissolving all solids, and continuously stirring for 30-60 minutes;

s2: adding mussel mucin into the material obtained in the step S1, and continuously stirring at the speed of 200-300 rpm for 30-60 minutes;

s3: heating the material obtained in the step S2 to 80-85 ℃, adding a solubilizer and an emollient, stirring at 200-300 rpm to fully mix the material, and homogenizing at 2000-4000 rpm for 5-10 minutes to obtain a homogeneous material;

s4: cooling the homogeneous material obtained in the step S3 to 40-45 ℃, adding a pH regulator to neutralize the homogeneous material to ensure that the pH value of the material is between 5.0 and 7.0, and then continuously stirring at the speed of 200-300 rpm until the homogeneous material is uniformly dispersed;

s5: and vacuumizing for many times until bubbles are completely eliminated, then cooling the material to be treated to below 38 ℃, filling into a packaging material, and performing damp-heat sterilization to obtain the mussel mucoprotein skin repairing emulsion.

In the preparation method of the mussel mucin skin-repairing emulsion capable of achieving wet heat sterilization, in the step S2, before mussel mucin is added, the temperature of the material is reduced to 40-50 ℃.

In the preparation method of the mussel mucin skin care emulsion capable of being sterilized by moist heat, in step S6, the conditions for moist heat sterilization are as follows: the sterilization temperature is 105-121 ℃, and the sterilization time is 15-60 minutes.

The invention has the beneficial effects that:

1) The invention takes the trehalose as the main body of the protective agent of the mussel mucin, combines with proper humectant and emollient to inhibit the denaturation of the mussel mucin at high temperature (above 80 ℃), so that the prepared mussel mucin skin repairing emulsion can be sterilized by moist heat, thereby avoiding the destructiveness of irradiation sterilization on high polymer materials and packing materials and the potential pollution to people and environment, and reducing the production cost; and the mussel mucin in the product still keeps good biological activity after moist heat sterilization, and has good anti-inflammatory effect.

2) The mussel mucin of the invention forms non-covalent interaction with other components, enhances the electrostatic interaction and promotes the autoxidation crosslinking of the self-dopa, thereby improving the anti-inflammatory efficacy of the self-dopa. Wherein, the combination of trehalose, betaine and allantoin promotes the formation of lysine positive charges and stabilizes the electrostatic interaction of mussel mucin; the trehalose, the glycerol, the propylene glycol, the butanediol and the pentanediol are combined to form a non-covalent interaction with a catechol structure of the dopa through a hydrogen bond, so that the oxidation of the dopa is catalyzed to dopaquinone, and the auto-oxidation crosslinking effect of the dopa is promoted; meanwhile, the oil phase components of the emollients such as lecithin, squalane, polydimethylsiloxane and the like can form non-covalent interaction with the hydrophobic groups of the mussel mucin through a hydrophobic effect, so that the hydrophobic interaction of the mussel mucin is improved; through these three effects between the components, the mussel mucin skin care emulsion of the invention exhibits an anti-inflammatory effect far exceeding that of the case where each component is added alone.

3) The preparation method of the invention comprises the steps of mixing and dissolving the added components at high temperature, adding the mussel mucin after the temperature is reduced, and fully mixing the mussel mucin and the components under stirring. In the process, the mussel mucin is combined with water molecules to perform a solvation exclusion effect on trehalose, so that trehalose molecules are distributed around mussel mucin molecules to play a protection role, and the denaturation of the mussel mucin is inhibited in the subsequent heating process.

4) According to the invention, the specific ratio of trehalose to mussel mucin and the addition amount of the mussel mucin are adjusted, so that the mussel mucin skin repair emulsion achieves the optimal film forming property and anti-inflammation property, further accelerates the healing of skin wound surfaces, and has good industrial production value and economic value.

Drawings

Fig. 1 is a film forming property investigation result of mussel mucin skin care emulsion with different formulas.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments.

Example 1

The embodiment is a mussel mucin skin repairing emulsion capable of being sterilized by heat and moisture and a preparation method thereof, and the mussel mucin skin repairing emulsion comprises the following components in parts by weight: mussel mucin 0.1-0.2 weight portions, trehalose 3-15 weight portions, humectant 2-20 weight portions, emollient 1.5-2 weight portions, solubilizer 0.01-0.03 weight portions, thickener 0.2-1 weight portions, pH regulator 0.1-0.5 weight portions, and purified water 80-95 weight portions. The mass ratio of the trehalose to the mussel mucin is 15.

The mussel mucoprotein skin-repairing emulsion capable of being sterilized by heat and moisture in the embodiment comprises one or more of glycerin, propylene glycol, butylene glycol, pentanediol, sorbitol, polyethylene glycol, betaine, sodium hyaluronate, allantoin and urea; preferably one or more of glycerol, propylene glycol, butylene glycol, betaine, sodium hyaluronate and allantoin. The emollient is one or more of cacao butter, vaseline, water soluble silicone wax, liquid paraffin, lanolin, stearate, lecithin, squalane, polydimethylsiloxane and cetostearyl alcohol; preferably one or more of cocoa butter, lecithin, squalane and polydimethylsiloxane. The solubilizer is one or more of hydrogenated castor oil, polyoxyethylene castor oil, fatty alcohol polyoxyethylene ether, polyoxyethylene sorbitan fatty acid ester and polyglycerol fatty acid ester; preferably one or more of hydrogenated castor oil, polyoxyethylene castor oil and fatty alcohol-polyoxyethylene ether. The thickening agent is one or more of carbomer, sodium polyacrylate, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose, soluble starch, xanthan gum and gelatin; preferably one or more of carbomer, sodium polyacrylate, methylcellulose, ethylcellulose, hydroxyethyl cellulose and carboxymethyl cellulose. The pH regulator is one of triethanolamine, aminomethyl propanol, sodium hydroxide and potassium hydroxide; preferably one of triethanolamine or aminomethyl propanol. Wherein the mass ratio of trehalose to glycerol, propylene glycol, butanediol, betaine and allantoin is 1-30, preferably 2-10; the mass ratio of the trehalose to the lecithin, the squalane and the polydimethylsiloxane is 1-100, and preferably 3-20.

The preparation method of the mussel mucin skin care emulsion capable of being sterilized by heat and moisture in the embodiment comprises the following steps:

s2: adding mussel mucin into the material obtained in the step S1, and continuously stirring at the speed of 200-300 rpm for 30-60 minutes; in order to better protect the mussel mucin by the trehalose, the material can be cooled to 40-50 ℃ before the mussel mucin is added.

S3: heating the material obtained in the step S2 to 80-85 ℃, adding a solubilizer and an emollient, stirring at the speed of 200-300 rpm to fully mix the material, and homogenizing at the stirring speed of 2000-4000 rpm for 5-10 minutes to obtain a homogeneous material;

s5: and vacuumizing for many times until bubbles are completely eliminated, then cooling the material to be treated to below 38 ℃, filling into a packaging material, and performing damp-heat sterilization to obtain the mussel mucoprotein skin repairing emulsion. The conditions of moist heat sterilization are as follows: the sterilization temperature is 105-121 ℃, and the sterilization time is 15-60 minutes.

Example 2

This example is to examine the stability of the mussel mucin skin care emulsion described in example 1 after moist heat sterilization and the bioactivity of mussel mucin. The formulation of the mussel mucin skin care emulsion for examination of this example is shown in table 1, prepared as described in example 1. The mussel mucoprotein skin-repairing emulsion prepared is sequentially placed at the temperature of-15 ℃,5 ℃,25 ℃,40 ℃ and 55 ℃ for 30 days, 60 days and 90 days, the phase separation condition of the emulsion after moist heat sterilization is observed, and the test results are shown in table 2.

Table 1:

table 2:

the test results in Table 2 show that the emulsions 1 to 3 prepared in this example were stored at 25 ℃,40 ℃, 55 ℃,5 ℃ and-15 ℃ for 20 to 90 days without phase separation; the emulsion 4 without trehalose is subjected to phase separation at each investigation temperature, which shows that the addition of trehalose plays a role in stabilizing the emulsion, and the prepared mussel mucoprotein skin repair emulsion can also keep excellent stability under the condition of moist heat sterilization.

The anti-inflammatory properties of the emulsions 1 to 3 prepared as described in this example were further examined. Tumor necrosis factor (TNF-alpha) and interleukin-6 (IL-6) are the most common and important inflammatory mediators in the inflammatory reaction process, and can cause inflammation symptoms such as red swelling and hot pain. By reducing the levels of these factors, the extent of the inflammatory response can be controlled. The test uses Lipopolysaccharide (LPS) intervention in macrophages to establish an inflammatory cell model and to evaluate the anti-inflammatory effects of mussel mucoprotein skin-rejuvenating emulsions by measuring the levels of TNF-alpha and IL-6 therein. The test groups were added with culture media of emulsions 1 to 3 and LPS (1. Mu.g/ml), respectively, the negative control group was added with a cell culture medium containing LPS (1. Mu.g/ml), and the contents of TNF-. Alpha.and IL-6 were measured by ELISA assay kit, and the results are shown in Table 3.

Table 3:

the test results in Table 3 show that emulsions 1-3 prepared in this example have significant anti-inflammatory effects, indicating that the anti-inflammatory efficacy of mussel mucin does not lose biological activity after moist heat sterilization.

Example 3

This example further examines the effect of moist heat sterilization on the anti-inflammatory effect of mussel mucin, and also examines the synergistic effect between the components. The formulations of emulsions 5 to 10 for the examination of the anti-inflammatory effect of this example are shown in Table 4, and the preparation process is likewise carried out as described in example 1, wherein emulsions 5, 7, 9 are sterilized without moist heat and emulsions 6, 8, 10 are sterilized with moist heat.

Method for examining anti-inflammatory Effect the anti-inflammatory effect of mussel mucoprotein skin care emulsions was assessed by measuring the levels of tumor necrosis factor (TNF-. Alpha.) and interleukin-6 (IL-6) as in example 2. The test results are shown in fig. 5.

Table 4:

table 5:

as can be seen from the anti-inflammatory test results in table 5, emulsion 5, which was not moist heat sterilized, had a significant anti-inflammatory effect, whereas emulsion 6, which was sterilized consistent with the formulation of emulsion 5, had little anti-inflammatory effect, indicating that the anti-inflammatory efficacy of mussel mucin was lost following moist heat sterilization.

Compared with emulsion 5, emulsion 7 added with trehalose has a certain increase in anti-inflammatory effect, which shows that trehalose has an obvious enhancement effect on the anti-inflammatory effect of mussel mucin; the formula of the emulsion 8 is consistent with that of the emulsion 7, and the anti-inflammatory effect of the emulsion is equivalent to that of the emulsion 7 after moist heat sterilization, which shows that the trehalose plays a role in protecting the mussel mucin during moist heat sterilization, so that the anti-inflammatory effect of the mussel mucin is maintained. Emulsion 9 and emulsion 10 both have significantly improved anti-inflammatory effects compared to emulsion 7 and emulsion 8, which demonstrates the synergistic interaction between the components of the mussel mucin skin care emulsions of the invention, resulting in further enhancement of the anti-inflammatory efficacy of the mussel mucin.

Example 4

This example is to further examine the protective effect of trehalose dosage on mussel mucin. The formulations of the emulsions 11 to 15 used in this example for the examination are shown in Table 6, and the preparation thereof is likewise carried out as described in example 1. The addition amount of mussel mucin of each emulsion is unchanged, the dosage of trehalose is different, and the optimal solution of trehalose addition is searched through the anti-inflammatory effect of each prepared emulsion. The anti-inflammatory effect of each emulsion was measured and the results are shown in table 7.

Table 6:

table 7:

as can be seen from the anti-inflammatory test results in table 7, emulsion 11 had no anti-inflammatory effect, further demonstrating the high temperature protective effect of trehalose on mussel mucins. From the anti-inflammatory effects of the emulsions 12-15, it is known that the anti-inflammatory effect of the mussel mucin skin care emulsion is correspondingly enhanced with the increase of the trehalose dosage. When the addition amount of trehalose was increased from 7.5% to 10.0%, the anti-inflammatory effect remained unchanged. Therefore, when the mass ratio of trehalose to mussel mucin is 50% or more, the protective effect of trehalose on mussel mucin is most strong, and the anti-inflammatory effect of the mussel mucin skin repair lotion is the best, but considering that the skin feel of the lotion is sticky due to excessive addition of trehalose, the mass ratio of trehalose to mussel mucin is limited to 50-150.

Example 5

This example is to investigate the effect of mussel mucin dosage on the film-forming properties of mussel mucin skin care emulsions and the effect of trehalose on the film-forming properties of mussel mucin.

The formulations of the emulsions 16 to 19 used in this example for the examination are shown in Table 8, and the preparation thereof is likewise carried out as described in example 1. The trehalose addition amount of each emulsion was unchanged, the mussel mucin amount was different, and emulsion 19 was used as a control, with no trehalose added. Respectively taking 16-19.0 g of the prepared emulsion, smearing the emulsion on dry plate glass, and uniformly coating. The coated plate glass was left to dry naturally at 37 ℃. + -. 1 ℃ and after 8 hours the film formed was observed. The results of the film formation of each emulsion are shown in FIG. 1.

Table 8:

as can be seen from the results of the film formation test in FIG. 1, the emulsion 17 has the best film formation integrity, moderate thickness and the best permeability. Emulsion 16 and emulsion 18, with either too little or too much mussel mucin addition, had less film forming properties than emulsion 17; emulsion 19 without trehalose addition had significantly poorer film formation than emulsion 17, indicating that without trehalose protection, the mussel mucin emulsion had poor film formation after sterilization.

Example 6

This example is to examine the order of addition during the preparation and the influence of the preparation process parameters on the anti-inflammatory effect. The important point is to examine the sequence of charging mussel mucin and trehalose, and the temperature control during charging mussel mucin, the influence of the anti-inflammatory effect. In this example, based on the formulation of emulsion 14 in example 4, the emulsion was prepared by different formulation experimental methods, and the specific experimental design is as follows:

experiment 1: an emulsion, emulsion 14, was prepared as described in example 1.

Experiment 2: preparation method the method described in example 1 differs in that: the temperature of the material before adding the mussel mucin is 80-85 ℃, and when the mussel mucin is added, the temperature is not reduced to 40-50 ℃, so that emulsion 20 is prepared; the other steps are identical to the emulsion 14.

Experiment 3: preparation method the method described in example 1 differs in that: the order of addition of mussel mucin and trehalose is changed. The preparation method comprises the following steps: adding a thickening agent and a humectant into purified water, stirring at 200-300 rpm, heating to 40-50 ℃, dissolving the solid completely, and continuing stirring for 30-60 minutes; adding mussel mucin, and continuously stirring for 30-60 minutes; adding trehalose, and stirring until all the trehalose is dissolved. The feed liquid is heated to 80-85 ℃, the solubilizer and the emollient are added, the materials are fully mixed by stirring, and then the materials are homogenized for 5-10 minutes at 2000-4000 rpm. Cooling the feed liquid to 40-45 ℃, adding a pH regulator to regulate the pH value to 5.0-7.0, continuously stirring at 200-300 rpm for 10 minutes, and then vacuumizing for many times until bubbles are completely eliminated; and (3) when the temperature is reduced to below 38 ℃, filling the feed liquid into a packaging material, and sterilizing at 115 ℃ for 30 minutes to obtain the emulsion 21.

The prepared emulsions 14, 20, and 21 were subjected to an anti-inflammatory test, and the results are shown in table 9.

Table 9:

from the anti-inflammatory test results in table 9, it is clear that the anti-inflammatory effect of the emulsion 20 with the preparation temperature raised is significantly inferior to that of the emulsion 14, and the main reason is that the protective effect of trehalose on mussel mucin is not fully exerted in the process of adding the mussel mucin at 80-85 ℃, and the mussel mucin begins to denature at high temperature, which finally results in the reduction of the anti-inflammatory effect of the mussel mucin. The anti-inflammatory effect of emulsion 21 with the trehalose addition order adjusted is also reduced compared with emulsion 14, the preferential exclusion effect of mussel mucin on trehalose may be affected due to the non-covalent interaction of other components and mussel mucin, which leads to the reduction of the protective effect of trehalose on mussel mucin, and finally the anti-inflammatory effect of mussel mucin after moist heat sterilization is reduced. Therefore, the feeding sequence of the mussel mucin and the trehalose and the temperature control when the mussel mucin is added in the preparation process have influence on the anti-inflammatory effect of the product.

Example 7

This example is to examine the irritation of the mussel mucin skin rejuvenating emulsion of the invention. The specific examination method is as follows:

healthy and early adult albino rabbits (3 each) and females with weight not less than 2kg are respectively adopted. The hair on both sides of the spinal column of the animal was removed (about 10 cm. Times.15 cm area) 4h to 24h before the test, 0.5ml of the test substance (test group were emulsion 13, emulsion 14, and emulsion 15 prepared in example 4, respectively, and control group was physiological saline) was applied to both sides of the back of the rabbit, the contact site was covered with a 2.5 cm. Times.2.5 cm piece of absorbent gauze, and then the patch was fixed with a bandage for 4h. After the contact period has ended, the patch is removed, the contact site is marked with permanent ink, and the remaining test material is removed by rinsing with warm water and wiped dry. According to GB/T16886.10-2005 biological evaluation of medical devices part 10: stimulation and delayed type hypersensitivity tests the scoring system given in the animal skin irritation test describes and scores the skin erythema and edema response at each contact site for each prescribed time. The skin irritation test results are shown in table 10.

Table 10:

as is clear from the results of the animal skin irritation test in Table 10, none of the emulsions 13 to 15 showed any irritation reaction in the skin irritation test of albino rabbits. The skin repair emulsion capable of being sterilized by wet heat has good safety.

Example 8

This example is a study of the moisturizing properties of mussel mucin skin care emulsions according to the invention.

The emulsion 14 prepared in example 4 was selected for skin moisturization testing. 5 volunteers in experimentAfter the forearm of the two hands is cleaned in front, 2 measurement areas marked 3cm x 3cm are marked, each at least 1cm apart. After measuring the initial value of each region, the test sample was measured at (5.0. + -. 0.1) mg/cm ² The amount of the composition is used for single coating, and the moisture content of the skin of a test area of a volunteer is measured at four time nodes of 0.5h, 1h, 2h and 3 h. The results are shown in Table 11, where one measurement area was coated with emulsion 1 and the other measurement area was not coated with any sample, as a blank control.

Table 11:

as can be seen from the results of the skin moisture test in Table 11, the skin moisture content of the area of the volunteer coated with the mussel mucin skin repair emulsion is significantly increased compared with the blank area, which indicates that the mussel mucin skin repair emulsion disclosed by the invention can increase the skin moisture content and has good moisture retention.

The mussel mucin skin repairing emulsion disclosed by the invention selects trehalose as a protective agent, so that the mussel mucin keeps good film forming and anti-inflammatory effects after moist heat sterilization. Meanwhile, the trehalose has the effect of stabilizing an emulsification system due to the unique structure and the biological activity of the trehalose. Therefore, the mussel mucoprotein skin-repairing emulsion disclosed by the invention has good stability and anti-inflammatory efficacy. And the trehalose and other components are cooperated to enhance the anti-inflammatory effect of the mussel mucin by strengthening the electrostatic interaction and the hydrophobic interaction of the mussel mucin and the auto-oxidation crosslinking of dopa, so that the mussel mucin skin repair emulsion has an anti-inflammatory effect far superior to that of the mussel mucin skin repair emulsion when each component is singly added.

The preparation method of the invention ensures that the trehalose plays a role in protecting the mussel mucin by the specific adding sequence and adding temperature of the mussel mucin, the trehalose and other components. Meanwhile, the mussel mucin and the components form non-covalent interaction to improve the anti-inflammatory effect. On the basis, the ratio of trehalose to mussel mucin and the addition amount of the mussel mucin are adjusted, so that the mussel mucin skin repair emulsion achieves the optimal film forming and anti-inflammatory effects, further accelerates the healing of skin wounds, and has good industrial production value and economic value.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive. Furthermore, it should be understood that although the present specification describes embodiments, this does not include only one embodiment, and such description is for clarity only, and those skilled in the art should be able to make the specification as a whole, and the embodiments may be appropriately combined to form other embodiments understood by those skilled in the art.

Claims

1. A mussel mucin skin repair emulsion capable of being sterilized by wet heat is characterized in that: the mussel mucin skin repair emulsion comprises the following components in parts by weight:

0.1 to 0.2 portion of mussel mucin,

3 to 15 parts of trehalose, and the like,

2 to 20 portions of humectant, 2 to 20 portions,

1.5 to 2 portions of emollient,

0.01 to 0.03 portion of solubilizer,

0.2 to 1 portion of thickening agent,

0.1 to 0.5 portion of pH regulator,

80-95 parts of purified water.

2. A heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 1, characterized by: the mass ratio of the trehalose to the mussel mucin is 15.

3. A heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 2, characterized by: the mass ratio of the trehalose to the mussel mucin is 50.

4. A heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 1, characterized by:

the humectant is one or more of glycerol, propylene glycol, butanediol, pentanediol, sorbitol, polyethylene glycol, betaine, sodium hyaluronate, allantoin and urea;

the emollient is one or more of cocoa butter, vaseline, water soluble silicon wax, liquid paraffin, lanolin, stearate, lecithin, squalane, polydimethylsiloxane, and cetostearyl alcohol;

the solubilizer is one or more of hydrogenated castor oil, polyoxyethylene castor oil, fatty alcohol polyoxyethylene ether, polyoxyethylene sorbitan fatty acid ester and polyglycerol fatty acid ester;

the thickening agent is one or more of carbomer, sodium polyacrylate, methylcellulose, ethyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose, soluble starch, xanthan gum and gelatin;

the pH regulator is one of triethanolamine, aminomethyl propanol, sodium hydroxide and potassium hydroxide.

5. A moist heat sterilised mussel mucoprotein skin rejuvenating emulsion according to claim 4, characterized by:

the humectant is one or more of glycerol, propylene glycol, butanediol, betaine, sodium hyaluronate and allantoin;

the emollient is one or more of cocoa butter, lecithin, squalane and polydimethylsiloxane;

the solubilizer is one or more of hydrogenated castor oil, polyoxyethylene castor oil and fatty alcohol-polyoxyethylene ether;

the thickening agent is one or more of carbomer, sodium polyacrylate, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose and carboxymethyl cellulose;

the pH regulator is one of triethanolamine and aminomethyl propanol.

6. A heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 5, characterized by:

the mass ratio of trehalose to glycerol, propylene glycol, butanediol, betaine and allantoin is 1-30;

the mass ratio of the trehalose to the lecithin to the squalane to the polydimethylsiloxane is 1-100.

7. A moist heat sterilised mussel mucoprotein skin rejuvenating emulsion according to claim 6, characterized by:

the mass ratio of trehalose to glycerin, propylene glycol, butanediol, betaine and allantoin is 2-10;

the mass ratio of trehalose to lecithin, squalane and polydimethylsiloxane is 3-20.

8. A method of preparing a heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to any one of claims 1 to 7, characterized by: the method comprises the following steps:

s2: adding mussel mucin into the material obtained in the step S1, and continuously stirring for 30-60 minutes at the speed of 200-300 rpm;

9. A method of making a heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 8, characterized in that: in step S2, before adding the mussel mucin, the temperature of the material is reduced to 40-50 ℃.

10. A method of making a heat and moisture sterilizable mussel mucin skin rejuvenating emulsion according to claim 8, characterized in that: in step S6, the conditions of the moist heat sterilization are: the sterilization temperature is 105-121 ℃, and the sterilization time is 15-60 minutes.