阅读说明：本技术 一种促进微生态平衡益生菌复合物及其制备方法 (Probiotic compound for promoting microecological balance and preparation method thereof ) 是由 黄楚明 黄楚佳 黄楚杰 于 2021-07-29 设计创作，主要内容包括：本发明公开了一种促进微生态平衡益生菌复合物及其制备方法,属于化妆品技术领域。且该益生菌复合物包括益生菌、益生元、保湿剂和水；所述保湿剂包括神经酰胺、透明质酸和磷酸酯衍生物；所述磷酸酯衍生物为磷酸酯、吡咯烷酮和聚二甲基硅油的反应物。且本发明引入了益生元,该益生元为复合益生元,为益生菌提供充足的“食物”,促进益生菌的长期有效稳定储存,促进益生菌对皮肤的修复作用的发挥；同时,引入了神经酰胺、透明质酸和磷酸酯衍生物的保湿剂,具有优异的吸水、保湿和成膜作用,促进角质保持湿润,驱除皮肤内的寄生虫,与益生菌发挥协同作用,形成薄膜维护屏障,促进皮脂修复肌底屏障功能。(The invention discloses a probiotic compound for promoting microecological balance and a preparation method thereof, belonging to the technical field of cosmetics. And the probiotic compound comprises probiotics, prebiotics, humectant and water; the humectant comprises ceramide, hyaluronic acid and phosphate ester derivatives; the phosphate derivative is a reactant of phosphate, pyrrolidone and polydimethylsiloxane oil. In addition, the prebiotics are introduced and are composite prebiotics, so that sufficient food is provided for the probiotics, the long-term effective and stable storage of the probiotics is promoted, and the restoration effect of the probiotics on the skin is promoted; meanwhile, the moisturizing agent introduced with the ceramide, the hyaluronic acid and the phosphate derivative has excellent water absorption, moisturizing and film forming effects, promotes cutin to keep moist, expels parasites in skin, and has a synergistic effect with probiotics to form a film maintenance barrier and promote sebum to repair the muscular basal barrier.)

1. A probiotic compound for promoting microecological balance, characterized by: the method comprises the following steps: probiotics, prebiotics, humectants, and water;

the humectant comprises ceramide, hyaluronic acid and phosphate ester derivatives;

the phosphate derivative is prepared by the following steps:

x1, mixing dihydroxy-terminated polydimethylsiloxane oil and N, N-dimethylformamide, adjusting the pH value of a reaction solution to 8-9 under the protection of light and nitrogen at 67 ℃, dropwise adding N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone, performing reflux reaction for 6 hours, continuously keeping the pH value of the reaction solution to 8-9, dropwise adding epoxy chloropropane, performing reflux reaction for 4 hours, performing reduced pressure rotary evaporation, washing until a washing solution is neutral, and performing vacuum drying to obtain an intermediate 2;

and X2, mixing the intermediate 2 with N, N-dimethylformamide, controlling the reaction temperature to 83 ℃, dropwise adding a sodium dihydrogen phosphate aqueous solution, keeping the pH of the reaction solution at 4-5, continuously reacting for 5h after complete dropwise addition, and performing rotary evaporation, recrystallization and vacuum drying to obtain the phosphate derivative.

2. A probiotic compound for promoting microecological balance according to claim 1, characterized in that: the compound comprises the following components in percentage by weight of the total compound: 25-65% of probiotics, 2-7% of prebiotics, 1.5-6.5% of humectant and the balance of water.

3. A probiotic compound for promoting microecological balance according to claim 1, characterized in that: the probiotics are selected from one or more of the following bacteria in any ratio: lactobacillus, propionibacterium, bacillus, saccharomycete, bifidobacterium and streptococcus thermophilus.

4. A probiotic compound for promoting microecological balance according to claim 2, characterized in that: the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1-3:1-3: 1-3.

5. A probiotic compound for promoting microecological balance according to claim 1, characterized in that: the N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone is prepared by the following steps:

mixing epoxy chloropropane, alpha-pyrrolidone and glacial acetic acid, carrying out reflux reaction at 88 ℃ for 12h under the protection of light and nitrogen, and then carrying out rotary evaporation under reduced pressure to obtain the N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone.

6. A probiotic compound for promoting microecological balance according to claim 1, characterized in that: the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 15-25% of ceramide, 20-40% of hyaluronic acid and the balance of phosphate derivatives.

7. The method for preparing probiotic compound for promoting microecological balance according to claim 1, wherein the probiotic compound comprises the following components: the method comprises the following steps:

A. activating probiotics, inoculating the activated probiotics into a fermentation culture medium, culturing at 37 ℃ to obtain strain fermentation liquor, centrifuging, removing supernatant, and collecting bacterial sludge;

B. and (3) stirring and mixing the bacterial sludge, the prebiotics, the humectant and the water at the temperature of 5-15 ℃ until the mixture is uniform, thus obtaining the probiotic compound for promoting the microecological balance.

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to a probiotic compound for promoting micro-ecological balance and a preparation method thereof.

Background

The skin is the largest organ of the human body, and is colonized by billions of microorganisms (bacteria, yeast, fungi, viruses, etc.) collectively called "microbiota" or "microflora". The term "skin microbiome" refers to all these microorganisms, their genomes and their interactions with the environment. Among these microbial flora, there can be divided into a resident flora and a transient flora. The resident flora consists of commensal bacteria, i.e. survive on the host without causing any damage. The transient flora is mainly composed of harmless fungi, viruses and bacteria, and is called saprophytic bacteria. The flora is not permanent and will vary throughout the day depending on the activity being performed and the changes in the surrounding conditions and the degree of exposure of the individual to these conditions. These microorganisms are essential to life. In addition to their role in generating body odour, they are also closely associated with maintaining healthy skin. Under normal conditions, i.e. when the hygiene conditions are good and when the resident flora, immune response and barrier function of the skin are intact, the resident microorganisms and transient microorganisms do not cause diseases or dysfunctions. The skin microbiota is thus able to act as a barrier and protect its host. When the skin microbiota is unbalanced, the function of the muscle bottom barrier can be damaged and lost, so that harmful bacteria grow, and the skin is easy to suffer from diseases such as oil, acne and the like.

However, the use of skin care and cosmetic products, which are currently popular, disrupts the balance of skin microflora. Application of, for example, foundations interferes with the balance of skin microbiota, particularly by interfering with bacterial diversity. The invisible cosmetic film on skin from skin care bottle can temporarily change the water-fat balance of skin and cause desquamation, which can destroy the balance of skin microflora.

Therefore, there is a need to develop a probiotic compound that promotes microbial homeostasis, promotes the rapid restoration of the balance of skin microbiota, and promotes the rapid restoration of the fundal barrier function.

Disclosure of Invention

The invention aims to provide a probiotic compound for promoting microecological balance and a preparation method thereof, so as to provide the probiotic compound which has a strong moisturizing effect, promotes the water-fat balance of skin, restores the balance of skin microbial flora, promotes the rapid recovery of the function of the muscular basal barrier, avoids skin diseases, quickly relieves the skin problems and protects the skin.

The technical problems to be solved by the invention are as follows: solves the problem of disrupting the balance of skin microbiota due to the use of current skin care and cosmetic products.

The purpose of the invention can be realized by the following technical scheme:

a probiotic compound for promoting microecological balance, comprising: probiotics, prebiotics, a humectant and water.

Further, the probiotic compound comprises the following components in percentage by weight of the total compound: 25-65% of probiotics, 2-7% of prebiotics, 1.5-6.5% of humectant and the balance of water.

Further, the complex is in a liquid state, solution or emulsion, and can be sprayed for use.

Further, the probiotic is a live or inactivated microorganism, which is used to increase the diversity of bacteria in the skin microbiota and promote the ecological balance of the skin microbiota.

Further, the probiotics are selected from one or more of the following bacteria in any ratio: lactobacillus, propionibacterium, bacillus, saccharomycete, bifidobacterium and streptococcus thermophilus.

Furthermore, the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1-3:1-3:1-3: 1-3. The prebiotics can provide food for probiotics, can regulate probiotics, and simultaneously can generate a large amount of physiological active substances, regulate the pH value of the skin surface, kill pathogenic bacteria, and inhibit the generation and absorption of endogenous harmful substances.

Further, the moisturizer includes ceramide, hyaluronic acid, and phosphate ester derivatives.

Further, the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 15-25% of ceramide, 20-40% of hyaluronic acid and the balance of phosphate derivatives.

Furthermore, the phosphate derivative is a reactant of phosphate, pyrrolidone and polydimethylsiloxane, has the water absorption and water retention characteristics of the pyrrolidone and phospholipid, and therefore has excellent moisture retention property, and meanwhile, the molecular structure of the phosphate derivative has a polydimethylsiloxane structure, so that the phosphate derivative has a good insect expelling effect, is compounded with ceramide and hyaluronic acid to form a humectant, promotes cutin to keep moist, expels parasites in skin, and has a synergistic effect with probiotics to form a film maintenance barrier and promote sebum repair of the muscle base barrier function.

Further, the molecular structure of the phosphate ester derivative is as follows:

further, the phosphate derivative is prepared by the following steps:

s1, adding epoxy chloropropane, alpha-pyrrolidone and glacial acetic acid into a three-neck flask in sequence, stirring uniformly, carrying out light-shielding treatment by using tinfoil, heating a reaction system to 88 ℃ by using an oil bath kettle under the nitrogen protection state, carrying out reflux reaction for 12h, then carrying out reduced pressure rotary evaporation to remove a solvent to obtain N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone, and utilizing the ring-opening reaction of secondary amine and an epoxy group, wherein the dosage ratio of epoxy chloropropane, alpha-pyrrolidone and glacial acetic acid is 0.1mol:0.21-0.23mol:80-160 mL;

the reaction formula is shown as follows:

s2, adding dihydroxy-terminated polydimethylsiloxane oil and N, N-dimethylformamide into a three-neck flask, uniformly stirring, keeping out of the sun with tinfoil, heating a reaction system to 67 ℃ with an oil bath kettle under the nitrogen protection state, adjusting the pH value of the reaction liquid to 8-9 with a solution of 35% by mass of sodium hydroxide under the stirring state, dropwise adding N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone with a constant-flow dropping funnel, performing reflux reaction for 6 hours, keeping the pH value of the reaction liquid to 8-9, dropwise adding epichlorohydrin with the constant-flow dropping funnel, performing reflux reaction for 4 hours, performing reduced pressure rotary evaporation, washing with deionized water until the washing liquid is neutral, performing vacuum drying to constant weight to obtain an intermediate 2, and performing Williamson synthesis reaction by using dihydroxy-terminated polydimethylsiloxane oil and a chlorohydrocarbon derivative, wherein the dosage ratio of the dihydroxy-terminated polydimethylsiloxane, the N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone, the epichlorohydrin and the N, N-dimethylformamide is 0.01mol:0.011-0.14mol:0.01mol:50-150 mL;

the reaction formula is shown as follows:

s3, adding the intermediate 2 and N, N-dimethylformamide into a three-neck flask, controlling the reaction temperature to be 83 ℃, dropwise adding 30-50% of sodium dihydrogen phosphate aqueous solution by using a constant-pressure dropping funnel at the dropping speed of 1-3 drops/second, adjusting the pH of the reaction solution to be 4-5 by using 10% of hydrochloric acid solution, continuing to react for 5 hours after the dropwise adding is completed, removing the solvent by rotary evaporation, recrystallizing for 2-3 times by using ethyl acetate/ethanol, drying in vacuum to constant weight to obtain a phosphate derivative, and performing ring-opening reaction on cyclohexyl and hydroxyl in sodium dihydrogen phosphate under an acidic condition, wherein the using ratio of the intermediate 2, N-dimethylformamide to sodium dihydrogen phosphate is 0.1mol:50-150mL:0.12-0.13 mol.

The reaction formula is shown as follows:

a preparation method of a probiotic compound for promoting microecological balance comprises the following steps:

A. activating probiotics via slant culture medium, inoculating to fermentation culture medium at 37 deg.C, and culturing to make thallus concentration reach logarithmic growth phase of 0.5-2 × 10⁹CFU/mL to obtain strain fermentation liquor, centrifuging, removing supernatant, and collecting strain mud;

B. the bacterial sludge, the prebiotics, the humectant and the water are stirred and mixed evenly at the speed of 150-200r/min at the temperature of 5-15 ℃, so as to obtain the probiotic compound for promoting the microecological balance.

The invention has the beneficial effects that:

the invention utilizes live or inactivated microorganisms as probiotics as main components of the probiotic compound, and the probiotics are used for increasing the diversity of bacteria in skin microbiota and promoting the ecological balance of the skin microbiota; the mixture of fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin type fructification is introduced as a prebiotic, the prebiotic is a composite prebiotic, provides sufficient food for the probiotic, can adjust probiotic bacteria, promotes long-term effective and stable storage of the probiotic bacteria, and simultaneously has the functions of generating a large amount of physiological active substances, adjusting the pH value of the surface of skin, killing pathogenic bacteria, inhibiting the generation and absorption of endogenous harmful substances and promoting the restoration of the skin by the probiotic bacteria; meanwhile, the moisturizing agent is introduced, the moisturizing agent is ceramide, hyaluronic acid and phosphate ester derivatives, has excellent water absorption and moisturizing effects, and the hyaluronic acid and the phosphate ester derivatives have excellent film forming effects, promote cutin to keep moist, expel parasites in skin, and play a synergistic effect with probiotics to form a film maintenance barrier and promote sebum to repair the muscle bottom barrier function. The derivative is a combination of phosphate, pyrrolidone and polydimethylsiloxane, and has the water absorption and water retention characteristics of the pyrrolidone and phosphate, so that the derivative has excellent moisture retention property, and meanwhile, the molecular structure of the derivative has a polydimethylsiloxane structure, so that the derivative has a good insect repelling effect.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

The phosphate derivative is prepared by the following steps:

s1, sequentially adding 0.1mol of epoxy chloropropane, 0.21mol of alpha-pyrrolidone and 80mL of glacial acetic acid into a three-neck flask, uniformly stirring, carrying out light-shielding treatment by using tinfoil, heating a reaction system to 88 ℃ by using an oil bath kettle under the nitrogen protection state, carrying out reflux reaction for 12 hours, and then carrying out reduced pressure rotary evaporation to remove a solvent to obtain N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone;

s2, adding 0.01mol of dihydroxy-terminated polydimethylsiloxane oil and 50mLN, N-dimethylformamide into a three-neck flask, uniformly stirring, carrying out light-shielding treatment by using tinfoil, heating a reaction system to 67 ℃ by using an oil bath kettle in a nitrogen protection state, adjusting the pH value of a reaction solution to 8 by using a solution of 35% by mass of sodium hydroxide in a stirring state, dropwise adding 0.011mol of N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone by using a constant-flow dropping funnel, carrying out reflux reaction for 6 hours, keeping the pH value of the reaction solution to 8, dropwise adding 0.01mol of epichlorohydrin by using the constant-flow dropping funnel, carrying out reflux reaction for 4 hours, carrying out reduced pressure rotary evaporation, washing with deionized water until a washing solution is neutral, and carrying out vacuum drying until the weight is constant, thereby obtaining an intermediate 2;

s3, adding 0.1mol of intermediate 2 and 50mLN, N-dimethylformamide into a three-neck flask, controlling the reaction temperature to 83 ℃, dropwise adding 30% by mass of sodium dihydrogen phosphate aqueous solution by using a constant-pressure dropping funnel at a dropping speed of 1 drop/second, adjusting the pH of the reaction solution to 4 by using 10% by mass of hydrochloric acid solution, continuing to react for 5 hours after complete dropwise addition, removing the solvent by rotary evaporation, recrystallizing for 2 times by using ethyl acetate/ethanol, and drying in vacuum to constant weight to obtain the phosphate derivative, wherein the mole number of the sodium dihydrogen phosphate is 0.12 mol.

Example 2:

the phosphate derivative is prepared by the following steps:

s1, sequentially adding 0.1mol of epoxy chloropropane, 0.23mol of alpha-pyrrolidone and 160mL of glacial acetic acid into a three-neck flask, uniformly stirring, carrying out light-shielding treatment by using tinfoil, heating a reaction system to 88 ℃ by using an oil bath kettle under the nitrogen protection state, carrying out reflux reaction for 12 hours, and then carrying out reduced pressure rotary evaporation to remove a solvent to obtain N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone;

s2, adding 0.01mol of dihydroxy-terminated polydimethylsiloxane oil and 150mLN, N-dimethylformamide into a three-neck flask, uniformly stirring, carrying out light-shielding treatment by using tinfoil, heating a reaction system to 67 ℃ by using an oil bath kettle in a nitrogen protection state, adjusting the pH value of a reaction solution to 8.3 by using a solution of 35% by mass of sodium hydroxide in a stirring state, dropwise adding 0.14mol of N- (3-chloro-2-hydroxypropyl) -2-pyrrolidone by using a constant-current dropping funnel, carrying out reflux reaction for 6h, keeping the pH value of the reaction solution to 8.3, dropwise adding 0.01mol of epichlorohydrin by using the constant-current dropping funnel, carrying out reflux reaction for 4h, carrying out reduced pressure rotary evaporation, washing by using deionized water until a washing solution is neutral, and carrying out vacuum drying until the weight is constant to obtain an intermediate 2;

s3, adding 0.1mol of intermediate 2 and 150mLN, N-dimethylformamide into a three-neck flask, controlling the reaction temperature to 83 ℃, dropwise adding 40% by mass of sodium dihydrogen phosphate aqueous solution by using a constant-pressure dropping funnel at a dropping speed of 1 drop/second, adjusting the pH of the reaction solution to 4.3 by using 10% by mass of hydrochloric acid solution, continuing to react for 5 hours after complete dropwise addition, removing the solvent by rotary evaporation, recrystallizing for 2 times by using ethyl acetate/ethanol, and drying in vacuum to constant weight to obtain the phosphate derivative, wherein the mole number of the sodium dihydrogen phosphate is 0.13 mol.

Example 3:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 25% of probiotics, 2% of prebiotics, 1.5% of humectant and the balance of water; wherein the probiotic is lactobacillus; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1:1:1: 1; the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 15% of ceramide, 20% of hyaluronic acid and the balance of the phosphate derivative prepared in example 1;

the probiotic compound for promoting microecological balance is prepared by the following steps:

A. activating probiotics via slant culture medium, inoculating to fermentation culture medium at 37 deg.C, and culturing to make thallus concentration reach logarithmic phase of 0.5 × 10⁹CFU/mL to obtain strain fermentation liquor, centrifuging, removing supernatant, and collecting strain mud;

B. the bacterial sludge, the prebiotics, the humectant and the water are stirred and mixed evenly at the speed of 150r/min at the temperature of 5 ℃, and the probiotic compound for promoting the micro-ecological balance is obtained.

Example 4:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 30% of probiotics, 4% of prebiotics, 4% of humectant and the balance of water; wherein the probiotics are composite bacteria formed by mixing lactobacillus, propionibacterium, bacillus, saccharomycetes, bifidobacterium and streptococcus thermophilus according to the mass ratio of 1:2:1.5:3:1: 2; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1:1.6:2: 1; the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 20% of ceramide, 30% of hyaluronic acid and the balance of the phosphate derivative prepared in example 2;

the probiotic compound for promoting microecological balance is prepared by the following steps:

A. activating probiotics via slant culture medium, inoculating to fermentation culture medium at 37 deg.C, and culturing to make thallus concentration reach logarithmic growth phase of 1 × 10⁹CFU/mL to obtain strain fermentation liquor, centrifuging, removing supernatant, and collecting strain mud;

B. the bacterial sludge, the prebiotics, the humectant and the water are stirred and mixed evenly at the speed of 200r/min at the temperature of 10 ℃, and the probiotic compound for promoting the micro-ecological balance is obtained.

Example 5:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 65% of probiotics, 7% of prebiotics, 6.5% of humectant and the balance of water; wherein the probiotics are selected from bifidobacteria and streptococcus thermophilus which are mixed according to the mass ratio of 1: 1; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 3:1:3: 2; the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 25% of ceramide, 40% of hyaluronic acid and the balance of the phosphate derivative prepared in example 1;

the probiotic compound for promoting microecological balance is prepared by the following steps:

A. activating probiotics via slant culture medium, inoculating to fermentation culture medium at 37 deg.C, and culturing to make thallus concentration reach logarithmic growth phase 2 × 10⁹CFU/mL to obtain strain fermentation liquor, centrifuging, removing supernatant, and collecting strain mud;

B. the bacterial sludge, the prebiotics, the humectant and the water are stirred and mixed evenly at the speed of 200r/min at the temperature of 15 ℃, and the probiotic compound for promoting the micro-ecological balance is obtained.

Comparative example 1:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 25% of probiotics, 2% of prebiotics, 1.5% of humectant and the balance of water; wherein the probiotic is lactobacillus; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1:1:1: 1; the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 15% of ceramide, 20% of hyaluronic acid and the balance of simethicone;

the probiotic compound for promoting microecological balance is prepared by the following steps: refer to the procedure for preparation in example 1.

Comparative example 2:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 30% of probiotics, 4% of humectant and the balance of water; wherein the probiotics are composite bacteria formed by mixing lactobacillus, propionibacterium, bacillus, saccharomycetes, bifidobacterium and streptococcus thermophilus according to the mass ratio of 1:2:1.5:3:1: 2; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 1:1.6:2: 1; the humectant comprises the following components in percentage by weight based on the total weight of the humectant: 20% of ceramide, 30% of hyaluronic acid and the balance of the phosphate derivative prepared in example 2;

the probiotic compound for promoting microecological balance is prepared by the following steps: refer to the procedure for preparation in example 4.

Comparative example 3:

a probiotic compound for promoting microecological balance comprises the following components in parts by weight: 65% of probiotics, 7% of prebiotics and the balance of water; wherein the probiotics are selected from bifidobacteria and streptococcus thermophilus which are mixed according to the mass ratio of 1: 1; the prebiotics are formed by mixing fructo-oligosaccharide, isomaltooligosaccharide, xanthan gum and inulin-type fructan according to the mass ratio of 3:1:3: 2;

the probiotic compound for promoting microecological balance is prepared by the following steps: refer to the procedure for preparation in example 5.

Example 6:

the probiotic complexes obtained in examples 3 to 5 and comparative examples 1 to 3 were tested according to the following test methods:

first, experiment for recovering function of muscle-bottom barrier

(1) The probiotic compound obtained in examples 3-5 and comparative examples 1-3 was applied, diluted 10 times with water, and sprayed on the face of the subject;

(2) the subject: 40 men, 100 women, 20-55 years of age, and a mean age (30 ± 14) of age, wherein 14 people are over-cleansed resulting in an impaired skin barrier, 28 people are under-exfoliated resulting in an impaired skin barrier, 32 people are under-exposed to laser wrinkle removal surgery resulting in an impaired skin barrier, 32 people are under-exposed to laser speckle removal surgery resulting in an impaired skin barrier, and 34 people are under-exposed to laser acne removal surgery resulting in an impaired skin barrier. All subjects were randomized into 7 groups, one control and 6 experimental.

(3) The experimental method comprises the following steps: the experimental groups used the samples prepared in examples 1-3 and comparative examples 1-3; control 1 was not smeared with any sample. After six groups of skin lesions were cleaned, the test article was used. The composition is administered once daily for 20 days.

(4) And (3) safety evaluation: in the clinical use process, adverse reactions do not appear through careful observation, the adverse reactions are carefully recorded and tracked, and if the patient is found to be uncomfortable, the experiment should be immediately interrupted and measures are actively taken.

(5) Skin barrier function test: the stratum corneum water content and the transepidermal water loss were measured using a skin water tester (VapoMeter) and a transepidermal water loss meter (vapometerttewl) of delf and finland corporation. The skin is cleaned before detection, and the skin enters a test environment 30 minutes in advance to be quietly tested, and non-skin damage parts of cheeks are respectively detected.

(6) The statistical method comprises the following steps: statistical analysis statistical processing was performed using the t-test using SPSS3.0 software.

The statistical results are shown in table 1 below.

TABLE 1

As shown in table 1, the skin had a higher transdermal water loss and a lower stratum corneum water content after the skin barrier had been compromised prior to treatment. After the treatment, the effect of improving the transdermal water loss using the complexes of examples 3 to 5 was superior to that using comparative examples 1 to 3, the increase in the moisture content of the stratum corneum using the complexes of examples 3 to 5 was significantly superior to that using comparative examples 1 to 3, and it was demonstrated that the skin barrier function improving effect of the subjects was superior to that of examples 3 to 5 compared to comparative examples 1 to 3.

Secondly, detecting the skin moisturizing function:

the probiotic compound obtained in examples 3-5 and comparative examples 1-3 was diluted 15 times.

(1) 70 dry skin volunteers (skin moisture content less than 10%) aged 30-50 were selected and randomized into 7 groups of 10 persons each, one of which was blank.

(2) Cleaning the inner sides of the forearms of both hands of the subject uniformly, sitting still in a laboratory, drinking water and beverage without intentional confusion, exposing the forearms, keeping the forearms relaxed, naturally drying for 0.5 hour, and selecting 3cm on the inner sides of the forearms of both hands respectively²The area of (A) serves as a test area and a control areaThe front inner sides of the two arms of the group and the experimental group 1-5 are respectively smeared with 0.2mL of the corresponding test article, the empire state is measured by a skin epidermal layer water content tester at each time point of 1, 2, 4 and 6h after the smearing, and the water content of the tested area of the front inner sides of the two arms of each test subject is recorded.

(3) And taking the average value of the moisture content of the tested areas on the inner sides of the two forearm of the testee as the moisture content test result of the skin of the testee, and taking the average value of the moisture content test results of the skins of the testees of all groups as the moisture content test result of the skin of the testee of the group. The long-term moisturizing effect of the invention is judged by comparing the difference of the skin moisture content of each group before and after being smeared with the test article and the difference of the skin moisture content of the blank control group. And the results of the moisture content test of the skin are shown in table 2.

TABLE 2

	1h	2h	4h	6h
					Blank group	28.3	28.5	28.9	28.7
Example 3	55.3	49.4	45.7	42.5
					Example 4	54.6	48.9	44.4	40.8
Example 5	56.1	50.3	46.0	41.3
					Comparative example 1	45.9	37.2	29.8	28.3
Comparative example 2	46.2	37.4	33.0	28.1
					Comparative example 3	39.2	30.3	28.5	28.3

As shown in table 2, the moisture content of the skin of the subjects in the blank group hardly changed within 6h, after 6h, the moisture content of the skin of the subjects using examples 3 to 5 was significantly higher than that of the skin of the subjects using comparative examples 1 to 3, and within 6h, the decrease in the moisture content of the skin of the subjects using comparative example 3 was significant, and the moisture content retention rate of the skin of the subjects using examples 3 to 5 was the best, indicating that the probiotic compound of the present invention has a significant long-term moisturizing function on the skin.

In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

The foregoing is illustrative and explanatory only and is not intended to be exhaustive or to limit the invention to the precise embodiments described, and various modifications, additions, and substitutions may be made by those skilled in the art without departing from the scope of the invention or exceeding the scope of the claims.