CN115317432A

CN115317432A - Facial mask containing bifidobacterium lactis fermentation product

Info

Publication number: CN115317432A
Application number: CN202210458633.1A
Authority: CN
Inventors: 张义淦
Original assignee: Shenzhen Duoweisheng Health Food Co ltd
Current assignee: Shenzhen Duoweisheng Health Food Co ltd
Priority date: 2022-04-24
Filing date: 2022-04-24
Publication date: 2022-11-11
Anticipated expiration: 2042-04-24
Also published as: CN115317432B

Abstract

The invention provides a mask which comprises a fermentation product of bifidobacterium lactis, wherein the preservation number of the bifidobacterium lactis is CGMCC NO.24307, and the bifidobacterium lactis is preserved in China general microbiological culture collection management center at 1 month and 14 days in 2022. The facial mask also comprises Tremella extract, hydrolyzed collagen, semen Tritici Aestivi culture extract, etc. The facial mask provided by the invention has the effects of resisting aging, locking water, repairing and relieving.

Description

Facial mask containing bifidobacterium lactis fermentation product

Technical Field

The invention belongs to the field of cosmetics, and particularly relates to a mask containing a bifidobacterium lactis fermentation product.

Background

The probiotics can be colonized in a human body, the flora of a certain part of a host is changed, the nutrition absorption is promoted to keep the intestinal tract healthy by regulating the immune function of the mucous membrane and the system of the host or regulating the balance of the flora in the intestinal tract, and the probiotics are closely related to human beings. In fact, probiotics not only play a role in maintaining the balance of intestinal flora, but also are widely applied to the field of cosmetics at present due to various active ingredients rich in fermentation products.

The mask as an indispensable step for daily skin care occupies a large market in cosmetics, and the product types are rich and diversified, and various effects such as: the mask products for moisturizing, whitening, removing freckles and resisting aging have no overlapping layers. The facial mask can be generally divided into two types, one is a chemical facial mask and the other is a natural facial mask, according to the components of the facial mask. The chemical facial mask is mainly compounded by chemical raw materials to achieve the skin care effect of the facial mask, and the natural facial mask is mainly compounded by natural extracts, wherein the application range of the plant extracts is particularly wide. However, there are some problems with both of the above two types of masks: chemical raw materials adopted in the chemical facial mask easily contain irritant components such as alcohol, preservatives and the like, and extracts in the natural facial mask are too complex and are prone to sensitization, so that the preparation of the facial mask by using active probiotics can be one of the development directions for improving the conditions.

Chinese patent 201910715219.2 discloses a biological fiber mask of probiotic extracts and a preparation method thereof, wherein the mask liquid specifically comprises the following components: aloe, pearl powder, skim milk powder, glucose, soluble starch, ethyl maltol, sodium alginate, food-grade sodium benzoate, protease, lipase, bacterial powder, egg white and honey. According to the invention, the aloe extract and the mask matrix solution are added into the lactobacillus culture dish, and the liquorice powder and the potato extract powder are added in the preparation process, so that the biological fiber mask prepared by the method has the advantages of strong skin-friendly ability, high mildness, high antibiosis and anti-inflammation, and high essence absorption rate in use. However, the ingredients are complex and are likely to cause a burden on sensitive skin.

Chinese patent 201711297075.0 discloses a whitening mask liquid containing probiotic fermentation liquid and a preparation method thereof. The clam is treated by enzymolysis and biological fermentation to prepare antioxidant peptide with high antioxidant activity, and then 2-aminobutanol is used as a stabilizer and cosmetic auxiliary materials to prepare the whitening mask liquid with high antioxidant activity. The whitening mask liquid prepared by the method disclosed by the invention is stable in physical and chemical properties, high in free radical clearance rate and green and safe in product. But the preparation method is complex and the controllability of the fermentation process is not strong.

The method is a problem to be solved in the field, and is characterized in that proper strains are excavated and a fermentation method of the strains is optimized, so that fermentation products of the strains can be better applied to the field of facial masks.

Disclosure of Invention

In order to solve the above problems, the present invention provides a mask comprising a bifidobacterium lactis fermentation product.

In the present invention, the fermentation product of bifidobacterium lactis is also referred to as a yeast bifidus fermentation product in the cosmetic field.

In one aspect, the present invention provides a mask.

The mask comprises a fermentation product of bifidobacterium lactis, the preservation number of the bifidobacterium lactis is CGMCC NO.24307, and the bifidobacterium lactis is preserved in China general microbiological culture collection management center in 2022 at 1 month and 14 days.

The preparation method of the bifidobacterium lactis fermentation product comprises the following steps: fermenting the bifidobacterium lactis, removing solid impurities such as thalli, suspended particles and the like by using a disk centrifuge, and then carrying out microfiltration to remove soluble impurities which can cause denaturation of fermentation liquor to obtain fermentation supernatant.

The fermentation condition of the bifidobacterium lactis is 37 +/-1 ℃, and the bifidobacterium lactis is anaerobically cultured for 72 +/-3 hours.

The facial mask comprises the following components:

water (W)	85-95％
		Tremella extract	1.00-2.00％
Glycerol	0.50-1.50％
		Hydrolyzed collagen	1.00-2.00％
Bifidobacterium lactis fermentation product	2.00-3.00％
		Bacillus/glutamic acid fermentation product filtrate	0.50-1.50％

Preferably, the mask comprises the following components:

water (W)	90-95％
		Tremella extract	1.50-2.00％
Glycerol	1.00-1.50％
		Hydrolyzed collagen	1.50-2.00％
Bifidobacterium lactis fermentation product	2.50-3.00％
		Bacillus/glutamic acid fermentation product filtrate	1.00-1.50％

Further preferably, the facial mask comprises the following components:

water (I)	92.50％
		Tremella extract	1.50％
Glycerol	1.00％
		Hydrolyzed collagen	1.50％
Bifidobacterium lactis fermentation product	2.50％
		Bacillus/glutamic acid fermentation product filtrate	1.00％

The facial mask is in the form of but not limited to: applying facial mask and face sticking film.

When the mask is in a form of a smearing mask, the mask may further comprise other components such as an excipient.

In another aspect, the invention provides a method for preparing a mask.

The preparation method is used for preparing the mask

The preparation method comprises the preparation of a bifidobacterium lactis fermentation product, wherein the preservation number of the bifidobacterium lactis is CGMCC NO.24307.

The preparation method also comprises compounding according to the formula.

The mask disclosed by the invention is rich in trehalose which belongs to a micromolecular humectant, has good affinity with membrane protein, can increase the absorption of skin on nutrient components, increases the hydration function of cells, and has good moisturizing performance;

in the present invention, hydrolyzed collagen has at least the following effects:

1. having multiple repeats of RGD (consisting of arginine, glycine, aspartic acid) can bind to at least 8 different integrins (integrins). Through combination with the recognition point of the integrin, the fibroblast cell takes part in signal conduction, greatly promotes division and proliferation of fibroblast cells, improves the differentiation speed and the differentiation quantity of the cells, and finally realizes the replacement process of replacing old cells with new cells;

2. the collagen protein has a large number of amino acid fragment sequences, can particularly recognize tripeptide sequences in the molecular chaperone Hsp47 (heat shock protein) of the collagen protein, effectively, firmly and stably has a triple-helix structure through targeted combination, and the collagen protein can support the skin only under the triple-helix structure, so that the skin is durably tender and glossy;

3. the GER series segment has stronger adhesion to the fibroblast and can effectively repair the damage of the fibroblast, thereby improving the activity of the fibroblast.

In the invention, the bifidobacterium lactis fermentation product has at least the following effects:

1. the product contains beneficial skin care components such as amino acids, small peptides, polysaccharides, B vitamins, etc., and has effects of enhancing stratum corneum metabolism, removing cell free radicals, inhibiting lipid peroxidation, activating organism repairing performance, inhibiting cell tyrosinase activity, repairing, whitening, and resisting oxidation.

2. The fermented product of Bifidobacterium lactis can balance the ratio of various media, is beneficial to regulating immune system, and can relieve scytitis, and has antibacterial and antiinflammatory effects.

3. The product is rich in lactic acid and sodium lactate, is a main water-soluble acid in Natural Moisturizing Factors (NMF) of a human body, has strong hygroscopicity, effectively enhances the hydration of the horny layer, and prevents the water loss of the skin; lactic acid and sodium lactate constitute buffer solution, which has good affinity to skin and can regulate pH value of skin.

The invention has the beneficial effects that:

the facial mask provided by the invention has the effects of resisting aging, locking water, repairing and relieving. Through detection, the facial mask provided by the invention has the advantages that the bioactive substances with obvious immunological activity are extracted efficiently by adopting a biological fermentation technology, the core function active substances are scientifically compounded with various adjuvants, the skin tolerance can be comprehensively enhanced, the water can be supplemented and locked, and the facial mask has the advantages of high transdermal absorption speed, high bioavailability and the like.

Drawings

FIG. 1 is a graph showing Western Blotting results of active ingredient analysis of the mask pack of example 2.

Detailed Description

The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.

Example 1A method for preparing a fermented product of Bifidobacterium lactis

Bifidobacterium lactis CGMCC NO.24307 is fermented, mupirocin lithium salt and cysteine hydrochloride modified MRS agar medium (Qingdao Haibo organism) is selected as a fermentation medium, the fermentation condition is 37 ℃, the culture is carried out for 72h under anaerobic condition, and the fermentation product is detected.

The results show that: the product contains components beneficial to skin care, such as amino acid (14.26 mg/g), small peptide (3.29 mg/g), polysaccharide (13.24 mg/g), B vitamins (0.15 mg/g) and the like, can strengthen the metabolism of stratum corneum, remove cell free radicals, inhibit the peroxidation of lipid, activate the repair performance of organisms, inhibit the activity of cell tyrosinase, and have the functions of repair, whitening and oxidation resistance.

In the cosmetic field, the fermentation products of bifidobacterium lactis can also be classified as lactobacillus fermentation products.

EXAMPLE 2A method for preparing an active ingredient for skin care products

The fermentation product in the example 1 is processed to prepare the active ingredients of the bifidobacterium lactis fermentation product, and the specific steps are as follows: after fermentation is finished, solid impurities such as thalli, suspended particles and the like are removed by adopting a disk centrifuge, and then, soluble impurities and the like which can cause fermentation liquor denaturation are removed by microfiltration, so that fermentation supernatant is obtained.

The detection results of the products are as follows: amino acid (13.83 mg/g), small peptide (3.15 mg/g), polysaccharide (12.38 mg/g), B vitamins (0.15 mg/g), lactic acid (11.84 mg/g) and sodium lactate (9.23 mg/g).

The detection and analysis method of various active ingredients comprises the following steps:

(1) Amino acids

And (3) drawing an amino acid standard curve: weighing 0.5g of amino acid, dissolving with water, diluting to 100mL to prepare 5g/L of amino acid standard solution, measuring peak areas corresponding to the amino acid standard solutions with different concentrations by HPLC, and drawing a standard curve. And (3) measuring the content of amino acid in the fermentation supernatant: absorbing 2 mu L of fermentation supernatant, diluting and fixing the volume by using a specific derivative buffer solution, and performing qualitative analysis by adopting an HPLC (high performance liquid chromatography) measuring method according to the retention time of amino acid and an ultraviolet absorption spectrum; and (4) carrying out quantitative determination on the amino acid by an external standard peak area method, and calculating the content of the amino acid.

(2) Small peptides

Quantitatively sampling, centrifugally separating high molecular protein by adopting a trichloroacetic acid TCA precipitation method to obtain mixed liquid of the small peptide and the amino acid, determining the total nitrogen content by using a Kjeldahl method to obtain the nitrogen content in the mixed liquid of the small peptide and the amino acid, converting the nitrogen content into absolute quantity, dividing the absolute quantity by the amount of a sampling sample, multiplying the absolute quantity by 6.25 to obtain the total content of the small peptide and the amino acid in fermentation supernatant, and deducting the content of the amino acid to obtain the content of the small peptide.

(3) Polysaccharides

Anthrone-sulfuric acid colorimetry, drawing of a glucose standard curve: quantitatively weighing a glucose standard substance, preparing a reference substance solution with the concentration of 0.5mg/mL, diluting the reference substance solution into different concentrations according to the weight ratio of glucose: mixing 0.1% anthrone-sulfuric acid solution at a volume ratio of 1:4, boiling for 10min, cooling rapidly, standing in dark for 10min, developing, measuring absorbance under specific wave by ultraviolet spectrophotometry, and drawing standard curve. Quantitatively measuring supernatant of the fermentation supernatant, and performing post-treatment such as concentration and drying to obtain crude polysaccharide. Quantitatively weighing the crude polysaccharide to prepare a solution with a specific concentration, measuring the absorbance of the solution by adopting an ultraviolet spectrophotometry after color development according to a method under a standard curve preparation item, and calculating the content.

(4) B vitamins

And (3) drawing a B vitamin standard curve: and quantitatively weighing, dissolving and fixing the volume, diluting to different concentrations, measuring peak areas corresponding to the vitamin standard solutions with different concentrations by adopting HPLC, and drawing a standard curve. And (3) measuring the content of B vitamins in the fermentation supernatant: quantitatively sucking fermentation supernatant, dissolving and fixing volume, and performing qualitative analysis by adopting an HPLC (high performance liquid chromatography) measuring method according to retention time and ultraviolet absorption spectrum of different vitamins; and (5) carrying out quantitative determination on the vitamins by an external standard peak area method, and calculating the content of the B vitamins.

(5) Lactic acid

Drawing a standard curve of L-lactic acid: quantitatively measuring an L-lactic acid standard substance, preparing a reference substance mother solution with a certain concentration, diluting the reference substance mother solution into different concentrations, measuring the peak area under 210nm by adopting HPLC, and drawing a standard curve. And (3) measuring the content of lactic acid in the fermentation supernatant: quantitatively sucking fermentation supernatant, diluting and fixing volume, and performing qualitative analysis by adopting an HPLC (high performance liquid chromatography) measuring method according to the retention time of lactic acid and an ultraviolet absorption spectrum; and (4) carrying out quantitative determination by an external standard peak area method, and calculating the content of the lactic acid.

(6) Sodium lactate

Drawing a standard curve of sodium lactate: quantitatively weighing 100mg of sodium lactate standard, diluting to 100mL, shaking up, preparing reference mother liquor, diluting to different concentrations, measuring peak area by HPLC, and drawing a standard curve. Measuring the content of sodium lactate in the fermentation supernatant: quantitatively sucking fermentation supernatant, diluting and fixing volume, and performing qualitative analysis by HPLC determination method according to retention time and ultraviolet absorption spectrum of sodium lactate; and (4) carrying out quantitative determination by an external standard peak area method, and calculating the content of the sodium lactate.

Example 3 Bifidobacterium lactis fermentation product antioxidant ability test

DPPH 4mg was weighed out and dissolved in 100mL of absolute ethanol.

Taking 1mL of Bifidobacterium lactis fermentation products with different dilution times (diluted by distilled water), adding 3mL of DPPH solution, reacting at room temperature in a dark place, and measuring OD after 6h ₅₁₇ (three replicates were averaged and reported as A). Control group 1:1mL sample +3mL absolute ethanol (average value calculated in three groups of replicates, OD measured) ₅₁₇ As the value of B); control group 2:1mL of water +3mL of DPPH solution (three replicates for calculating the mean OD measurement) ₅₁₇ As C value); the DPPH radical clearance calculation formula is as follows:

C-(A-B)/C×100％；

the DPPH free radical clearance of the fermentation product of the bifidobacterium lactis of the application is finally measured to be 42.7%.

Example 4A mask containing a fermentation product of Bifidobacterium lactis

Comprises the following components:

water (W)	92.50％
		Tremella fuciformis berkExtract of plant	1.50％
Glycerol	1.00％
		Hydrolyzed collagen	1.50％
Bifidobacterium lactis fermentation product	2.50％
		Bacillus/glutamic acid fermentation product filtrate	1.00％

A bifidobacterium lactis fermentation product was prepared according to the method of example 2.

Hydrolyzed collagen was purchased from warrior bairison biomedical ltd;

the split yeast fermentation lysate was purchased from Shanghai Biley chemical.

The bacillus/glutamic acid fermentation product filtrate is prepared by fermenting glutamic acid serving as a raw material under the conventional conditions of bacillus and then filtering to obtain fermentation product filtrate, wherein the bacillus selected in the embodiment is bacillus subtilis CGMCC 1.3358.

Example 5A mask containing a Bifidobacterium lactis fermentation product

Comprises the following components:

Hydrolyzed collagen was purchased from warrior biomedical ltd;

the secondary split yeast fermentation lysate was purchased from Shanghai Bilai chemical.

Example 6 mask bioactive substance assay comprising bifidobacterium lactis fermentation product

The mask provided in example 4 was submitted to a third party for active ingredient testing, the testing procedure was as follows:

company mandated samples were collected, extracted, concentrated, BCA quantified, and prepared for monitoring analysis.

Anti-EGF antibody [ EPR19899] (ab 206423) is selected as a main antibody, and Western Blotting is used for detecting active ingredients in a sample, so that the results show that: the sample contains a biologically active substance with significant immunological activity. As shown in the following figure, lane 6 shows the control in lane 1, and other products on the market in

lanes

2 and 3.

In conclusion, the mask is rich in active ingredients.

Example 7 transdermal Capacity test of active ingredients of facial mask containing fermented product of Bifidobacterium lactis

The mask provided in example 4 is sent to the institute of medicine of department of medicine of Shenzhen university for active substance transdermal capacity detection, and detection and analysis are carried out on the transdermal capacity of the active substance in the sample by using a Franz diffusion cell test scheme with reference to a chemical skin absorption in vitro test method (GB/T27818-2011). The results show that: the sample has a penetration level of 25% in 0.5h, 39% in 1h and 47% in 2h, which is close to the peak level, and the penetration levels of 4 and 12h are stable by 48%.

And (4) conclusion: the active ingredients of the facial mask containing the bifidobacterium lactis fermentation product have remarkable transdermal capacity.

Example 8 facial mask population experiment

80 volunteers (female, age 25-35) were recruited and divided into 2 groups of 40 persons each, using a facial mask not comprising bifidobacterium lactis fermentation product as a control group and the others as experimental groups, in winter in the experimental season, the initial detection time was 8 a.m.: 00, detecting the face of the part, respectively carrying out skin moisture content tests for 0h, 6h, 12h and 24h before and after the facial essence is used, and carrying out average calculation on test results, wherein the results are as follows:

group of	Before use (%)	0h(％)	6h(％)	12h(％)	24h(％)
						Control group	44.6	78.1	52.4	43.6	44.7
Experimental group	42.8	83.4	66.7	64.1	47.3

Claims

1. The mask is characterized by comprising a fermentation product of bifidobacterium lactis, wherein the preservation number of the bifidobacterium lactis is CGMCC NO.24307.

2. The mask pack according to claim 1, wherein the bifidobacterium lactis fermentation product is prepared by: fermenting the bifidobacterium lactis, removing solid impurities such as thalli, suspended particles and the like by using a disk centrifuge, and then carrying out microfiltration to remove soluble impurities which may cause the denaturation of fermentation liquor to obtain fermentation supernatant.

3. The mask according to claim 2, wherein the fermentation conditions of the bifidobacterium lactis are 37 ± 1 ℃ and anaerobic culture is carried out for 72 ± 3h.

4. The mask pack of claim 1, wherein said mask pack comprises the following components:

。

5. The mask pack according to claim 4, wherein the mask pack comprises the following components:

。

6. The mask pack according to claim 5, wherein the mask pack comprises the following components:

7. the preparation method of the mask is characterized by comprising the preparation of a bifidobacterium lactis fermentation product, wherein the preservation number of the bifidobacterium lactis is CGMCC NO.24307.

8. The method according to claim 7, wherein the culture conditions of Bifidobacterium lactis are as follows: anaerobic culturing at 37 + -1 deg.C for 72 + -3 h.

9. The method according to claim 8, wherein the culture medium of Bifidobacterium lactis is mupirocin lithium salt and cysteine hydrochloride modified MRS agar medium.

10. The method of claim 9, comprising formulating the facial mask formulation of any one of claims 4-6.