WO2022218172A1 - Preparation method for mask liquid and product thereof - Google Patents

Abstract

The present invention provides a preparation method for a mask liquid, and further provides a mask product thereof. In the mask liquid prepared in the method of the present invention, the mask liquid has no cytotoxicity on B16 cells, has a certain transdermal absorption effect, has a relatively strong free radical scavenging effect, and has the capability of significantly enhancing superoxide dismutase activity in fibroblasts, as well as having a certain anti-oxidation effect.

Description

A kind of preparation method of facial mask liquid and product thereof

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of Chinese Invention Patent Application No. CN202110412295.3 filed on April 16, 2021, which is incorporated herein by reference in its entirety.

technical field

The invention belongs to the field of cosmetics, and in particular relates to a preparation method of a facial mask liquid and a product thereof.

Background technique

Natural cosmetics are more and more popular nowadays. Natural cosmetics, in addition to natural ingredients, do not add flavors, preservatives and other potentially irritating ingredients.

As a technology widely used in the fields of food, medicine and cosmetics, enzymolysis can transform macromolecular components into small molecules, which are accompanied by the production of other active substances such as polysaccharides and polypeptides. Plants contain a variety of enzymes, including protease, cellulase, amylase, esterase, etc. The enzymes contained in plants can be used to hydrolyze macromolecular proteins into oligopeptides, and starch and cellulose into oligosaccharides. , lipolysis into glycerol, fatty acid. The application of these products in cosmetics is more conducive to the skin's absorption of functional ingredients, and finally achieves a certain effect.

SUMMARY OF THE INVENTION

Therefore, the object of the present invention is to overcome the defects in the prior art, and provide a preparation method of facial mask liquid and products thereof.

Before describing the content of the present invention, the terms used herein are defined as follows:

The term "SOD" refers to: superoxide dismutase.

The term "PBS" refers to: Phosphate Buffered Saline.

The term "t-test" refers to: T-test.

The term "VE" means: Vitamin E.

The term "B16 cells" refers to: B16 mouse melanoma cells.

The term "DPPH" refers to: 1,1-diphenyl-2-trinitrophenylhydrazine.

In order to achieve the above object, the first aspect of the present invention provides a preparation method of facial mask liquid, the preparation method comprises the following steps:

(1) prepare a pretreatment solution after cleaning, peeling and dicing pineapple, papaya, and Luo Han Guo;

(2) after the oat is pulverized, mixed with water, heated and stirred to obtain the oat extract;

(3) mixing the pretreatment solution obtained in step (1) with the oat extract obtained in step (2), heating and stirring, and filtering to obtain a supernatant;

(4) adding pueraria root powder and olive oil to the supernatant liquid obtained in step (3), and homogenizing to obtain the facial mask liquid.

According to the method of the first aspect of the present invention, wherein, in step (1), the proportion by weight of each raw material component in the pretreatment solution is:

30-80 parts of pineapple, 1-50 parts of papaya, 1-50 parts of Luo Han Guo, and 50 parts of water;

Preferably, 40-60 parts of pineapple, 1-30 parts of papaya, 1-30 parts of Luo Han Guo, and 50 parts of water;

More preferably, 50 parts of pineapple, 5-20 parts of papaya, 5-20 parts of Luo Han Guo, and 50 parts of water.

According to the method of the first aspect of the present invention, wherein, in step (1), the preparation method of the pretreatment solution comprises the following steps:

After cleaning, peeling and dicing pineapple, papaya, and Luo Han Guo, they are mixed with water and beaten, and left to stand to obtain a pretreatment solution;

Preferably, the standing time is 0.5 to 8 hours, preferably 1 to 4 hours, and most preferably 2 hours.

The method according to the first aspect of the present invention, wherein, in step (2), the ratio of the oat flour to water is 1:10-200, preferably 1:10-150, more preferably 1:10-100;

The heating temperature is 50-150°C, preferably 60-120°C, more preferably 70-110°C, further preferably 70-102°C;

The heating time is 0.5-8 hours, preferably 0.5-5 hours, more preferably 0.5-4 hours, further preferably 1-3 hours; and/or

The stirring rate is 100-800 r/min, preferably 100-500 r/min, more preferably 100-300 r/min, further preferably 120-200 r/min.

According to the method of the first aspect of the present invention, wherein, in step (3), the mixing ratio of the pretreatment solution obtained in step (1) and the oat extract obtained in step (2) is 1:1～500g/g, preferably 1:10 to 300 g/g, more preferably 1:20 to 200 g/g.

The method according to the first aspect of the present invention, wherein, in step (3), the heating temperature is 20-150°C, preferably 25-100°C, more preferably 30-90°C, further preferably 40-65°C;

The heating time is 0.1 to 5 hours, preferably 0.5 to 3 hours, more preferably 0.5 to 2 hours, further preferably 1 to 1.5 hours; and/or

The stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 20-300 r/min, further preferably 100-250 r/min.

The method according to the first aspect of the present invention, wherein, in step (4), the mass ratio of the pueraria root powder to the supernatant is 1:10～200, preferably 1:20～150, more preferably 1:50～ 100; and/or

The mass of the olive oil is 1-10% of the mass of the supernatant, preferably 1-5%, more preferably 2-4%.

According to the method of the first aspect of the present invention, wherein, in step (4), the homogenization time is 1-60 minutes, preferably 1-30 minutes, more preferably 1-20 minutes, still more preferably 10-20 minutes .

The method according to the first aspect of the present invention, wherein the method further comprises the steps of:

(5) sterilizing the mask liquid obtained in step (4);

Preferably, the sterilization temperature is 80-100°C, more preferably 75-80°C; and/or the sterilization time is 1-5 hours, more preferably 2 hours.

The second aspect of the present invention provides the facial mask liquid prepared according to the method of the first aspect.

A third aspect of the present invention provides a facial mask or a face-sticking product, which includes the facial mask liquid described in the second aspect.

The fourth aspect of the present invention provides the application of the facial mask liquid described in the second aspect in the preparation of an antioxidant product.

According to the application of the fourth aspect of the present invention, the antioxidant has the ability to scavenge free radicals.

According to the application of the fourth aspect of the present invention, the antioxidant has the ability to significantly enhance the activity of superoxide dismutase in fibroblasts.

A fifth aspect of the present invention provides a method for improving skin antioxidant capacity, the method comprising: administering to a subject in need:

The facial mask described in the second aspect; or

The facial mask or face-sticking product described in the third aspect.

The method of the present invention comprises the following steps:

(1) Mix and beat pineapple, papaya, and Luo Han Guo with water to prepare a pretreatment solution.

(2) Mix oats with water and heat to prepare oat pretreatment solution.

(3) Add (1) to (2), heat and stir, and filter.

(4) Add pueraria root powder to (3) extract. Then, vegetable oil is added and homogenized to obtain the facial mask liquid.

(5) Sterilize at 100℃ for 2h and fill under aseptic conditions.

In step (1), the ratio of the pineapple to water is 1:1～1000g/mL, preferably 1:10～800g/mL, more preferably 1:10～500g/mL, most preferably 1:100g/mL mL;

The ratio of papaya to water is 1:1～2000g/mL, preferably 1:10～500g/mL, more preferably 1:20～300g/mL, most preferably 1:200g/mL;

The ratio of Luo Han Guo to water is 1:1～2000g/mL, preferably 1:10～1000g/mL, more preferably 1:50～500g/mL, most preferably 1:200g/mL;

In step (2), the ratio of the oats to water is 1:1～500g/mL, preferably 1:5～200g/mL, more preferably 1:20～150g/mL, most preferably 1:100g/mL mL;

The heating temperature is 1-200°C, preferably 10-150°C, more preferably 20-150°C, most preferably 102°C;

The heating time is 1-10 hours, preferably 1-8 hours, more preferably 1-5 hours, most preferably 3 hours

The stirring rate is 10-500 r/min, preferably 10-300 r/min, more preferably 10-200 r/min, and most preferably 120 r/min.

In step (3), the heating temperature is 1-100°C, preferably 1-90°C, more preferably 1-80°C, most preferably 40°C;

The heating time is 0.5-3 hours, preferably 0.5-2 hours, more preferably 0.5-1.5 hours, most preferably 1 hour

The stirring rate is 10-500 r/min, preferably 10-400 r/min, more preferably 10-300 r/min, most preferably 250 r/min.

In step (4), the ratio of the pueraria root powder to the extract is 1:1～200g/mL, preferably 1:5～150g/mL, more preferably 1:10～100g/mL, most preferably 1:1 50g/mL;

The amount of vegetable oil added is 1-30%, preferably 1-20%; more preferably 1-10%, most preferably 4%;

The homogenization speed is 1000-10000r/min, preferably 1000-8000r/min, more preferably 2000-5000r/min; the most preferably 3000r/min;

The homogenization time is 1 to 60 minutes, preferably 1 to 50 minutes, more preferably 10 to 40 minutes, and most preferably 20 minutes.

Sterilize at 100°C for 2h and fill under aseptic conditions.

The preparation method of the present invention can have but is not limited to the following beneficial effects:

The invention provides a method for preparing a facial mask solution. The facial mask solution prepared by the method has no cytotoxicity to B16 cells and has a certain transdermal absorption effect, and the facial mask solution has a strong free radical scavenging effect and has Certain antioxidant effect.

Brief Description of Drawings

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, wherein:

Figure 1 shows the HPLC profile of sample 1 at 220 nm in Example 1.

FIG. 2 shows the HPLC profile of sample 2 at 220 nm in Example 1. FIG.

FIG. 3 shows the HPLC profile of sample 3 at 220 nm in Example 1. FIG.

FIG. 4 shows the HPLC profile of the blank group at 220 nm in Example 1. FIG.

FIG. 5 shows the toxicity results of the facial mask liquid of the present invention with different concentrations on fibroblasts in Test Example 1. FIG.

FIG. 6 shows the transdermal absorption results of the mask liquid of the present invention in Test Example 2. FIG.

FIG. 7 shows the test results of the effect of the facial mask liquid of the present invention on the content of collagen in fibroblasts in Test Example 3. FIG.

Best way to implement your invention

The present invention is further described below through specific examples, however, it should be understood that these examples are only used for more detailed description, and should not be construed to limit the present invention in any form. If no specific technique or condition is indicated in the examples, the technique or condition described in the literature in the field or the product specification is used. The reagents or instruments used without the manufacturer's indication are conventional products that can be purchased through regular channels.

This section provides a general description of the materials and test methods used in the tests of the present invention. While many of the materials and methods of operation used for the purposes of the present invention are known in the art, the present invention is described in as much detail as possible. It will be clear to those skilled in the art that, in the context, if not specifically stated, the materials and methods of operation used in the present invention are well known in the art.

The reagents and instruments used in the following examples are as follows:

Reagents:

Pineapples were purchased from Hongxing Farm, Zhanjiang City, Guangdong Province;

Papaya was purchased from Dole Red Heart Papaya Co., Ltd.;

Luohanguo was purchased from Fudonghai Luohanguo Co., Ltd.;

Oats (fried and cooked) were purchased from Zhangjiakou Jianjun Oat Food Co., Ltd.;

Pueraria lobata (dried powder) was purchased from Beijing Tongrentang (Group) Co., Ltd., Beijing, China;

VE, purchased from Sigma;

Fibroblasts and B16 cells were purchased from Saibaikang (Shanghai) Biotechnology Co., Ltd.

Reagents:

Anhydrous ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.

instrument:

Homogenizer, purchased from IKA Co., Ltd., model: T25.

The medical centrifuge was purchased from Wuxi Ruijiang Analytical Instrument Co., Ltd., model RJ-TGL-10B.

Example 1

This example is used to illustrate that the present invention can reduce the molecular weight of the polypeptide in the pineapple enzyme solution.

This experiment explored the effect of the ratio of enzyme solution and substrate on the molecular weight of oat polypeptide when pineapple was used as enzyme solution and oat was used as substrate. The experimental method is as follows:

(1) After cleaning, peeling and cutting the pineapple, it is mixed and beaten with water in a mass ratio of 1:1 to obtain a pineapple enzyme solution.

(2) Mix oat and water in a mass ratio of 1:50.

(3) Adding (1) to the pretreatment liquid (2) according to different mass ratios. The ratios of (1) and (2) are 1:200, 1:100, and 1:50, respectively, and the numbers of the obtained mixed solutions are sample 1, sample 2, and sample 3, respectively. The blank group is the pre-prepared sample without pineapple enzyme solution. Treatment fluid (2).

(4) The mixed solution in (3) was incubated at 35°C for 1 hour, and then placed in a 95°C water bath for 0.5 hour to inactivate the enzyme.

(5) Centrifuge the mixed solution in (4) at 5000 rpm for 20 minutes to obtain a supernatant.

(6) Determination of the polypeptide molecular weight of the supernatant by high performance liquid chromatography.

(7) The specific measurement method is as follows:

(a) Chromatographic conditions

Chromatographic column: TsK gel 2000 SWXL 300mm×7.8mm

Mobile phase: 0.05mol/L phosphate buffer (pH 7)+0.3mol/LNaCl

Detection wavelength: UV 220nm

Flow rate: 1ml/min

Column temperature: 25℃

Sample preparation: use the mobile phase as the solvent to prepare a sample with a concentration of 5 mg/ml, and then filter it with a microporous membrane (0.45 μm) for injection.

Standard sample: bovine serum albumin (Mr 67000), B12 (Mr 1335), and oxidized glutathione (Mr 614) were prepared into mixed standards, and the content of each substance was 5mg/ml.

(b) Determination of standard curve

The three standard samples were prepared at 5 mg/ml, and the standard curve was prepared according to the above chromatographic conditions. The least square method was used to obtain the regression equation of the straight line: y=-0.3913x+7.4563, and its regression coefficient R ² =0.9982.

It can be seen from the regression coefficient R ² =0.9982 of the equation that the standard curve of the mixed standard sample has a better linear relationship, which can improve the accuracy of the calculation. In the relationship, X represents the elution time, and Y represents the logarithm of the molecular weight. In this way, when the mask sample is analyzed by HPLC, its corresponding molecular weight can be calculated by the appearance time of the elution peak of each substance in the elution pattern.

(c) Molecular weight measurement results

Figures 1 to 3 show the HPLC chromatograms of sample 1, sample 2 and blank group at 220 nm in Example 1, respectively. The peak time, area and molecular weight of sample 1, sample 2 and blank group are shown in Table 1 below.

Table 1 Peak time, area and molecular weight of sample 1, sample 2 and blank group

Experimental results: In the blank sample, the content of the polypeptide with a molecular weight of 191804.31Da was 38.99%, and the content of the polypeptide with a molecular weight of 2028.36Da was 59.51%. In sample 1, the content of polypeptides with molecular weight of 2029.56Da is 100%; in sample 2, the content of polypeptides with molecular weight of 2041.56 is 100%; in sample 3, the content of polypeptides with molecular weights of 2056.85Da is 100%. The results showed that compared with the blank group, the proportion of polypeptides with a molecular weight of about 2000 Da in the supernatant obtained by adding the pineapple enzyme solution was greatly increased. The protease in pineapple can degrade 38.99% of the 191804.31Da polypeptide molecules in the blank group into about 2000Da polypeptide molecules. This shows that bromelain can degrade high molecular weight polypeptides in oat into lower molecular weight polypeptides.

Example 2

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel and cut into pieces.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 2. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo into pieces, mix and beat with water according to the ratio in Table 2, and let stand for 2 hours, which is the pretreatment solution.

Table 2 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	50
pawpaw	20
Monk fruit	20

(2) Mixing oat flour and water according to a ratio of 1:50 g/mL, heating at 70° C. for 3 hours, stirring at 200 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) according to a ratio of 1:200 g/g, heat at 40° C. for 1 hour, stir at 250 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) Add pueraria root powder, the ratio of pueraria root powder to supernatant liquid is 1:50g/mL, add olive oil with 2% mass of supernatant liquid, and homogenize for 10 minutes in a homogenizer.

3. Sterilization treatment of mask liquid

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 75°C for 2 hours.

Example 3

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel and cut into pieces.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 3. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo into pieces, mix and beat with water according to the ratio in Table 3, and let stand for 2 hours, which is the pretreatment solution.

Table 3 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	50
pawpaw	5
Monk fruit	5

(2) Mixing oat flour and water at a ratio of 1:50 g/mL, heating at 102° C. for 1 hour, stirring at 120 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) in a proportion of 1:200 g/g, heat at 65°C for 1.5 hours, stir at 100 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) adding pueraria root powder, the ratio of pueraria root powder and supernatant liquid is 1:100g/mL, adding olive oil with 4% mass of supernatant liquid, and homogenizing for 20 minutes in a homogenizer.

3. Sterilization treatment of mask liquid

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 80°C for 2 hours.

Example 4

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel, and dice.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 4. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo into pieces, mix and beat with water according to the ratio in Table 4, and let stand for 2 hours, which is the pretreatment solution.

Table 4 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	50
pawpaw	15
Monk fruit	15

(2) Mixing oat flour and water at a ratio of 1:10 g/mL, heating at 80° C. for 2 hours, stirring at 150 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) according to a ratio of 1:20 g/g, heat at 50° C. for 1.5 hours, stir at 150 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) Add pueraria root powder, the ratio of pueraria root powder to supernatant is 1:70g/mL, add olive oil with 3% mass of supernatant, and homogenize for 20 minutes in a homogenizer.

3. Mask liquid sterilization treatment

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 80°C for 2 hours.

Example 5

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel, and dice.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 5. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo into pieces, mix and beat with water according to the ratio in Table 5, and let stand for 2 hours, which is the pretreatment solution.

Table 5 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	50
pawpaw	10
Monk fruit	10

(2) Mix oat flour and water at a ratio of 1:100 g/mL, heat at 95°C for 1.5 hours, stir at 150 rad/min, and cool to room temperature to obtain an oat extract

(3) Mix the pretreatment solution in (1) and the oat extract in (2) in a proportion of 1:100 g/g, heat at 60°C for 1 hour, stir at 200 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) adding pueraria root powder, the ratio of pueraria root powder and supernatant liquid is 1:90g/mL, adding olive oil with 3% mass of supernatant liquid, and homogenizing for 15 minutes in a homogenizer.

3. Mask liquid sterilization treatment

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 80°C for 2 hours.

Example 6

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel, and diced.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 6. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo into pieces, mix and beat with water according to the ratio in Table 6, and let stand for 3 hours, which is the pretreatment solution.

Table 6 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	60
pawpaw	30
Monk fruit	30

(2) Mixing oat flour and water at a ratio of 1:150 g/mL, heating at 110° C. for 4 hours, stirring at 100 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) in a proportion of 1:10 g/g, heat at 90° C. for 2 hours, stir at 300 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) adding pueraria root powder, the ratio of pueraria root powder and supernatant liquid is 1:20g/mL, adding olive oil with 5% mass of supernatant liquid, and homogenizing for 5 minutes in a homogenizer.

3. Mask liquid sterilization treatment

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 100°C for 1 hour.

Example 7

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel, and dice.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 7. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo, mix and beat with water according to the ratio in Table 7, and let stand for 4 hours, which is the pretreatment solution.

Table 7 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	55
pawpaw	25
Monk fruit	25

(2) Mixing oat flour and water at a ratio of 1:120 g/mL, heating at 110° C. for 0.5 hour, stirring at 500 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) in a proportion of 1:300 g/g, heat at 100° C. for 0.5 hour, stir at 500 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) Add pueraria root powder, the ratio of pueraria root powder to supernatant is 1:150g/mL, add olive oil with 5% mass of supernatant, and homogenize for 30 minutes in a homogenizer.

3. Mask liquid sterilization treatment

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 80°C for 2 hours.

Example 8

This example is used to illustrate the preparation method of the facial mask liquid of the present invention.

1. Raw material processing (food grade)

Pineapple: Wash, peel, and dice.

Papaya: Wash, peel, and dice.

Luo Han Guo: Wash, peel and cut into pieces.

Oatmeal: After crushing, pass through 80 mesh for use.

2. Preparation of mask liquid

(1) The addition ratio of pineapple, papaya, and Luo Han Guo pretreatment solution is shown in Table 8. After cleaning, peeling, and cutting pineapple, papaya, and Luo Han Guo, mix and beat with water according to the ratio in Table 8, and let stand for 8 hours, which is the pretreatment solution.

Table 8 Addition ratio of pretreatment solution

raw material	parts by mass
water	50
Pineapple	80
pawpaw	50
Monk fruit	50

(2) Mixing oat flour and water in a ratio of 1:200 g/mL, heating at 50° C. for 8 hours, stirring at 100 rad/min, and cooling to room temperature to obtain an oat extract.

(3) Mix the pretreatment solution in (1) and the oat extract in (2) in a proportion of 1:500 g/g, heat at 100° C. for 0.5 hour, stir at 800 rad/min, filter impurities, and leave the supernatant (opaque ).

(4) adding pueraria root powder, the ratio of pueraria root powder and supernatant liquid is 1:200g/mL, adding olive oil with 10% mass of supernatant liquid, and homogenizing for 60 minutes in a homogenizer.

3. Mask liquid sterilization treatment

The mask liquid is filled into the mask bag (including the mask cloth).

Sterilize at 100°C for 1 hour.

Test Example 1

This test example is used to illustrate the cytotoxicity of the facial mask liquid prepared in Example 2 on fibroblasts.

The fibroblasts in logarithmic growth phase were seeded in a 96-well culture plate and cultured overnight at 37°C in a 5% CO ₂ environment. The culture medium was aspirated and discarded, samples of different concentrations were added, and five parallels were made for each sample. The cell control group was left untreated and cultured for 24 hours. Add 100uL MTT (1mg/ml) solution, put it into 37°C, 5% CO ₂ environment to culture, stop the culture after 4 hours; add 150uL DMSO after discarding the solution, shake slowly for 10min, measure the absorbance value of each well at 490nm wavelength. Cell viability=(OD value of assay well-OD value of blank control)/(OD value of cell control group-OD value of blank control)*100%.

Results: The toxicity test results of different concentrations of facial mask liquid on fibroblasts are shown in Figure 5. It can be seen from Figure 5 that no obvious cytotoxicity was detected in the mask liquid, and the cell survival rate was between 105.09% and 110.53% in the range of the sample concentration of 0.078% to 10%. It shows that the mask liquid has obvious promoting effect on the proliferation of fibroblasts.

Test Example 2

This test example is used to illustrate the transdermal absorption results of the facial mask liquid prepared in Example 3

(1) Preparation of mouse skin

Nude mice were sacrificed by decapsulation of their cervical vertebrae, their back hair was quickly shaved with a razor, the back skin was peeled off, and subcutaneous fat and blood vessels were removed, rinsed repeatedly with distilled water until clean, and then rinsed with normal saline several times, and placed in a -80°C refrigerator. Refrigerate for later use (use within 5 days).

(2) Franz diffusion cell for in vitro transdermal absorption experiments

Experimental process: add appropriate amount of water to the constant temperature tank of the in vitro osmosis diffusion device. Turn on the power and magnetic stirring in the thermostat, and set the water temperature in the thermostat to 37±0.1°C. The prepared rat skin was fixed between the two-chamber diffusion cells with iron clips, and 5 mL of the accepting solution was added to the accepting cell of the vertical diffusion cell, and then placed in the thermostatic bath of the in vitro osmotic diffusion device to preheat, and the stirring speed of the accepting cell was set to 400 r/ min. Add the supply liquid to the supply pool respectively, and seal the upper opening with plastic wrap. Start to record the time. When the sample penetrates for 0, 2, 4, 6, 8, and 24 hours (the specific time interval is determined according to the actual sample), take a sample of 500 μl and place it in a stoppered centrifuge tube. An equal amount of receiver solution was removed and air bubbles were removed from the cell. The samples will be assayed for sodium hyaluronate content. Then, the cumulative transmittance Q (mg/cm ² ) was calculated according to the following formula. The diameter of the bottom of the diffusion cell is 1.50 cm, and the contact area of the sample is 1.77 cm ² .

Results: Taking polysaccharide as the inspection index, the test results are shown in Figure 6. Under the condition that the total sugar content of the liquid to be tested is consistent, the permeation rate of polysaccharide increases continuously with the prolongation of the action time within the action time of the mask solution for 0-24h, and the process of polysaccharide permeation tends to be stable at 8h. It shows that the mask liquid has a certain transdermal absorption effect.

Test Example 3

This test example is used to illustrate the free radical scavenging ability of the facial mask liquid of Example 3.

DPPH free radical scavenging ability test method

Tube A1: Take 3mL of sample and mix it with 3mL of DPPH solution (2×10 ^-4 mol/L); Tube A2: Take 3mL of water and mix it with the same volume of DPPH solution; Tube A3: Take 3mL of absolute ethanol Mix with an equal volume of sample; measure the absorbance of A1, A2 and A3 tubes at a wavelength of 517 nm for 30 min of reaction.

DPPH free radical scavenging rate=[(A2+A3)-A1]/A2×100%.

Results: This test example is used to illustrate the ability of the mask liquid of the present invention to scavenge DPPH free radicals. The test results are shown in Figure 7 and Table 9. It can be seen from the chart that the free radical scavenging ability of this mask liquid reaches 55.02% when the concentration is 25%, indicating that the mask has a strong free radical scavenging effect and a certain antioxidant effect.

Table 9 Free radical scavenging ability of facial mask liquid with different concentrations

concentration	clearance rate	standard deviation
3.13%	14.18%	0.005939697
6.25%	24.29%	0.017819091
12.50%	43.80%	0.022485996
25%	55.02%	0.03026417
50%	62.39%	0.028991378
100%	77.95%	0.02298097

Test Example 4

This test example is used to illustrate the effect of the facial mask liquid prepared in Examples 2 to 8 on the activity of superoxide dismutase (SOD) in fibroblasts.

The test scheme is shown in Table 10

Table 10 Test plan

The specific operation steps are as follows:

(1) Seeding: Seeding fibroblasts in a 6-well plate at a seeding density of 2.2×10 ⁵ cells/well, and incubating in an incubator (37° C., 5% CO ₂ ) overnight.

(2) Administration: According to the test protocol in Table 6, when the cell plating rate in the 6-well plate reaches 40% to 60%, group administration is performed, and 2 mL of sample is added to each well, and each group has 3 duplicate wells. After the administration was completed, the 6-well plate was placed in an incubator (37°C, 5% CO ₂ ) for 24 hours.

(3) UVA irradiation: According to the test grouping, 30J/cm ² of UVA irradiation was performed on the groups with UVA irradiation.

(4) After irradiation, put it into an incubator (37°C, 5% CO ₂ ) for 24 hours, and wash the cells with PBS for 3 times;

(5) detection: collect cells, and detect according to the operating instructions of the SOD kit;

(6) Result analysis: The SOD activity of each group was summarized, and statistical analysis was carried out.

Results: The effects of the mask liquids prepared in Examples 2 to 8 on the activity of superoxide dismutase (SOD) in fibroblasts are shown in Table 11. It was found that UVA irradiation would significantly reduce the activity of SOD. The facial mask liquid prepared by the invention can significantly enhance the SOD activity in fibroblasts. Superoxide dismutase (SOD) is an antioxidant metalloenzyme that exists in living organisms. It can catalyze the dismutation of superoxide anion free radicals to generate oxygen and hydrogen peroxide, and plays a crucial role in the balance of oxidation and antioxidants in the body. It is inseparable from the occurrence and development of many diseases. From the experimental results, it can be concluded that the product prepared by the present invention has anti-oxidative and anti-aging effects.

Table 11 Changes in intracellular SOD activity

Remarks: When the t-test method is used for statistical analysis, the NC group is compared with the BC group, the significance is represented by #, P-value<0.05 is represented as #, P-value<0.01 is represented as ##; PC group, sample group Compared with the NC group, significance is indicated by *, P-value<0.05 is indicated by *, and P-value<0.01 is indicated by **.

Although this invention has been described to a certain extent, it will be apparent that suitable changes in various conditions may be made without departing from the spirit and scope of the invention. It is to be understood that the invention is not limited to the embodiments described, but is to be included within the scope of the claims, which include equivalents for each of the elements described.

Claims (15)

1. A preparation method of facial mask liquid, characterized in that, the preparation method comprises the following steps:

   (1) prepare a pretreatment solution after cleaning, peeling and dicing pineapple, papaya, and Luo Han Guo;

   (2) after the oat is pulverized, mixed with water, heated and stirred to obtain the oat extract;

   (3) mixing the pretreatment solution obtained in step (1) with the oat extract obtained in step (2), heating and stirring, and filtering to obtain a supernatant;

   (4) adding pueraria root powder and olive oil to the supernatant liquid obtained in step (3), and homogenizing to obtain the facial mask liquid.
2. The method according to claim 1, wherein in step (1), the proportion by weight of each raw material component in the pretreatment solution is:

   30-80 parts of pineapple, 1-50 parts of papaya, 1-50 parts of Luo Han Guo, and 50 parts of water;

   Preferably, 40-60 parts of pineapple, 1-30 parts of papaya, 1-30 parts of Luo Han Guo, and 50 parts of water;

   More preferably, 50 parts of pineapple, 5-20 parts of papaya, 5-20 parts of Luo Han Guo, and 50 parts of water.
3. The method according to claim 1 or 2, wherein in step (1), the preparation method of the pretreatment solution comprises the following steps:

   After cleaning, peeling and dicing pineapple, papaya, and Luo Han Guo, they are mixed with water and beaten, and left to stand to obtain a pretreatment solution;

   Preferably, the standing time is 0.5 to 8 hours, preferably 1 to 4 hours, and most preferably 2 hours.
4. The method according to any one of claims 1 to 3, wherein in step (2), the ratio of the oat flour to water is 1:10-200, preferably 1:10-150, more preferably 1:10～100;

   The heating temperature is 50-150°C, preferably 60-120°C, more preferably 70-110°C, further preferably 70-102°C;

   The heating time is 0.5-8 hours, preferably 0.5-5 hours, more preferably 0.5-4 hours, further preferably 1-3 hours; and/or

   The stirring rate is 100-800 r/min, preferably 100-500 r/min, more preferably 100-300 r/min, further preferably 120-200 r/min.
5. The method according to any one of claims 1 to 4, wherein in step (3), the mixing ratio of the pretreatment solution obtained in step (1) and the oat extract obtained in step (2) is 1: 1 to 500 g/g, preferably 1:10 to 300 g/g, more preferably 1:20 to 200 g/g.
6. The method according to any one of claims 1 to 5, wherein in step (3), the heating temperature is 20-150°C, preferably 25-100°C, more preferably 30-90°C, More preferably it is 40～65 ℃;

   The heating time is 0.1 to 5 hours, preferably 0.5 to 3 hours, more preferably 0.5 to 2 hours, further preferably 1 to 1.5 hours; and/or

   The stirring rate is 10-800 r/min, preferably 10-500 r/min, more preferably 20-300 r/min, further preferably 100-250 r/min.
7. The method according to any one of claims 1 to 6, wherein in step (4), the mass ratio of the pueraria root powder to the supernatant is 1:10～200, preferably 1:20～150 , more preferably 1:50 to 100; and/or

   The mass of the olive oil is 1-10% of the mass of the supernatant, preferably 1-5%, more preferably 2-4%.
8. The method according to any one of claims 1 to 7, wherein in step (4), the homogenization time is 1 to 60 minutes, preferably 1 to 30 minutes, more preferably 1 to 20 minutes , more preferably 10 to 20 minutes.
9. The method according to any one of claims 1 to 8, wherein the method further comprises the steps of:

   (5) sterilizing the mask liquid obtained in step (4);

   Preferably, the sterilization temperature is 80-100°C, more preferably 75-80°C; and/or the sterilization time is 1-5 hours, more preferably 2 hours.
10. The facial mask liquid prepared by the method according to any one of claims 1 to 9.
11. A facial mask or face sticking product, characterized in that, the facial mask or face sticking product comprises the facial mask liquid according to claim 10 .
12. The application of the facial mask liquid according to claim 10 in the preparation of anti-oxidant products.
13. The use according to claim 12, wherein the antioxidant has the ability to scavenge free radicals.
14. The use according to claim 12, wherein the antioxidant has the ability to significantly enhance the activity of superoxide dismutase in fibroblasts.
15. A method for improving skin antioxidant capacity, characterized in that the method comprises: administering to a subject in need:

    The facial mask solution according to claim 10; or

    The facial mask or face stick product according to claim 11.

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