CN113230166A

CN113230166A - Mask liquid, preparation method and application thereof

Info

Publication number: CN113230166A
Application number: CN202110413450.3A
Authority: CN
Inventors: 吴迪; 刘继涛; 安全; 霍彤; 刘平平; 王昌涛
Original assignee: Beiqingjiahua Huangshan Technology Co ltd; Yunnan Baiyao Group Co Ltd
Current assignee: Yunnan Baiyao Group Health Products Co ltd; Yunnan Baiyao Group Shanghai Technology Co ltd
Priority date: 2021-04-16
Filing date: 2021-04-16
Publication date: 2021-08-10
Also published as: WO2022218173A1

Abstract

The invention provides a mask liquid, which does not contain any chemical additive component and is prepared from pure plant raw materials and water, wherein the pure plant raw materials comprise the following components: pineapple, oat, highland barley, coix seed and vegetable oil; also provides a preparation method and application thereof. The facial mask liquid disclosed by the invention is natural in formula components, and is not added with potential irritant components such as essence and preservative. Experiments prove that the mask liquid has a promoting effect on proliferation of fibroblasts, has a certain transdermal absorption effect, can obviously improve the content of collagen, and achieves the anti-aging effect.

Description

Mask liquid, preparation method and application thereof

Technical Field

The invention belongs to the field of cosmetics, and particularly relates to a mask liquid, and a preparation method and application thereof.

Background

Natural cosmetics are nowadays more and more favored by people. The natural cosmetics are natural in formula components, and are not added with potential irritant components such as essence, preservative and the like.

Enzymolysis is a technology widely applied to the fields of food, medicine and cosmetics, and can modify macromolecular components into small molecules and generate other active substances such as polysaccharide, polypeptide and the like. The enzymes contained in plants are various and comprise protease, cellulase, amylase, esterase and the like, macromolecular protein can be enzymolyzed into oligopeptide by utilizing the enzymes contained in the plants, starch and cellulase are hydrolyzed into oligosaccharide, and fat is enzymolyzed into glycerol and fatty acid. The products are applied to cosmetics, so that the functional components can be absorbed by the skin more conveniently, and a certain effect is finally achieved.

Disclosure of Invention

Therefore, the invention aims to overcome the defects in the prior art and provide the facial mask liquid, and the preparation method and the application thereof.

In order to achieve the above purpose, the first aspect of the present invention provides a facial mask solution, which does not contain any chemical additive components and is prepared from pure plant raw materials and water, wherein the pure plant raw materials comprise the following components: pineapple, oat, highland barley, coix seed and vegetable oil;

preferably, the vegetable oil is selected from one or more of the following: olive oil, prinsepia utilis royle oil, peony seed oil, macadamia nut oil, babassu seed oil and Chinese toon flower oil; preferably olive oil.

The mask liquid according to the first aspect of the invention comprises the following raw material components in parts by weight:

1-10 parts of pineapple, 0.1-10 parts of oat, 0.5-5 parts of highland barley and 0.5-5 parts of coix seed;

preferably, 1-5 parts of pineapple, 0.2-6 parts of oat, 0.5-3 parts of highland barley and 0.5-3 parts of coix seed;

more preferably, 1-2 parts of pineapple, 0.5-5 parts of oat, 0.5-2 parts of highland barley and 0.5-2 parts of coix seed.

The second aspect of the present invention provides a method for preparing the facial mask solution of the first aspect, which may comprise the following steps:

(1) cleaning, peeling and slicing pineapples, mixing with water, pulping, and standing to obtain an extracting solution, namely enzyme solution;

(2) physically crushing and sieving oat, highland barley and coix seed, adding the crushed oat, highland barley and coix seed into the extracting solution obtained in the step (1), heating and stirring, cooling to room temperature, centrifuging and taking supernatant to obtain enzymolysis treatment solution;

(3) and (3) adding vegetable oil into the supernatant obtained in the step (2) and homogenizing to obtain the facial mask liquid.

The preparation method according to the second aspect of the invention, wherein in the step (1), the proportion of the pineapple to the water is 1: 1-500 g/mL, preferably 1: 5-500 g/mL, more preferably 1: 10-400 g/mL, and further preferably 1: 50-200 g/mL; and/or

The standing time is 1-5 hours, preferably 2-4 hours, and most preferably 3 hours.

According to the preparation method of the second aspect of the invention, in the step (2), the oat, the highland barley and the coix seed are ground and then sieved by a 80-mesh sieve.

The preparation method according to the second aspect of the invention, wherein in the step (2), the mass ratio of the oat to the extracting solution obtained in the step (1) is 1-50: 100, preferably 1-20: 100, more preferably 1-10: 100, and further preferably 1-5: 100;

the mass ratio of the highland barley to the extracting solution obtained in the step (1) is 1-10: 100, preferably 1-5: 100, more preferably 1-3: 100, and still more preferably 1.25-2: 100; and/or

The mass ratio of the coix seed to the extracting solution obtained in the step (1) is 1-10: 100, preferably 1-5: 100, more preferably 1-3: 100, and further preferably 1.25-2: 100.

The preparation method according to the second aspect of the present invention, wherein, in the step (2), the heating temperature is 20 to 150 ℃, preferably 25 to 100 ℃, more preferably 30 to 100 ℃, and most preferably 40 ℃;

the heating time is 1-10 hours, preferably 1-8 hours, more preferably 2-6 hours, and most preferably 4 hours;

the stirring speed is 10-1000 r/min, preferably 10-800 r/min, more preferably 10-600 r/min, and most preferably 200 r/min; and/or

The centrifugal rotation speed is 10-10000 r/min, preferably 100-8000 r/min, more preferably 200-6000 r/min, and most preferably 1200 r/min.

The preparation method according to the second aspect of the present invention, wherein in the step (3), the vegetable oil is added in an amount of 1 to 50%, preferably 1 to 40%; more preferably 10 to 30%, and still more preferably 10 to 20%;

the homogenizing speed is 3000-8000 r/min, preferably 3000-7000 r/min, and more preferably 4000-6000 r/min; the further excellent is 4000-5000 r/min; and/or

The homogenization time is 1 to 60 minutes, preferably 1 to 50 minutes, more preferably 10 to 40 minutes, and further preferably 20 to 30 minutes.

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

(4) sterilizing the facial mask liquid obtained in the step (3);

preferably, the sterilization temperature is 80-100 ℃, and preferably 80 ℃; and/or the sterilization time is 1-5 hours, preferably 2 hours.

In a third aspect of the invention, there is provided a facial mask comprising the facial mask fluid of the first aspect or the facial mask fluid prepared according to the method of the second aspect.

According to a specific embodiment of the present invention, the preparation method of the facial mask solution of the present invention comprises the following steps:

(1) mixing pineapple with water and pulping. The mixture was allowed to stand at room temperature for 3 hours.

(2) Physically pulverizing herba Avenae Fatuae, semen Avenae Nudae and Coicis semen, adding into the extractive solution (1), heating, stirring, centrifuging, and collecting supernatant.

(3) And (2) adding vegetable oil and homogenizing to obtain the mask liquid.

(4) Sterilizing at 100 deg.C for 2 hr, and packaging under aseptic condition.

In the step (1), the proportion of the pineapple to the water is 1: 1-500 g/mL, preferably 1: 5-500 g/mL, more preferably 1: 10-400 g/mL, and most preferably 1:100 g/mL;

in the step (2), the ratio of the oat to the extracting solution (1) is 1: 1-300 g/mL, preferably 1: 20-200 g/mL, more preferably 1: 20-150 g/mL, and most preferably 1:100 g/mL;

the proportion of the highland barley to the extracting solution (1) is 1: 1-200 g/mL, preferably 1: 10-150 g/mL, more preferably 1: 30-100 g/mL, and most preferably 1:80 g/mL;

the proportion of the coix seed to the extracting solution (1) is 1: 1-200 g/mL, preferably 1: 10-150 g/mL, more preferably 1: 50-100 g/mL, and most preferably 1:80 g/mL;

the heating temperature is 1-150 ℃, preferably 20-100 ℃, more preferably 30-100 ℃, and most preferably 40 ℃;

the heating time is 1 to 10 hours, preferably 1 to 8 hours, more preferably 2 to 6 hours, and most preferably 4 hours

The stirring speed is 10-1000 r/min, preferably 10-800 r/min, more preferably 10-600 r/min, and most preferably 200 r/min.

In the step (3), the addition amount of the vegetable oil is 1-50%, preferably 1-40%; more preferably 10 to 30%, most preferably 20%; (ii) a

The homogenizing speed is 3000-8000 r/min, preferably 3000-7000 r/min, and more preferably 4000-6000 r/min; optimally 5000 r/min;

the homogenization time is 1 to 60 minutes, preferably 1 to 50 minutes, more preferably 10 to 40 minutes, and most preferably 30 minutes.

The facial mask liquid of the invention can have the following beneficial effects:

the facial mask liquid disclosed by the invention is natural in formula components, and is not added with potential irritant components such as essence and preservative. Experiments prove that the mask liquid has a promoting effect on proliferation of fibroblasts, has a certain transdermal absorption effect, can obviously improve the content of collagen, and achieves the anti-aging effect.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 shows the HPLC profile of 220nm sample 1 in example 1.

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

FIG. 3 shows an HPLC chromatogram of the 220nm blank set in example 1.

FIG. 4 shows the results of toxicity of different concentrations of the facial mask fluid of the present invention on fibroblasts in Experimental example 1.

Fig. 5 shows the results of transdermal absorption of the facial mask fluid of the present invention in test example 2.

FIG. 6 shows the results of the test of the effect of the facial mask fluid of the present invention on the collagen content in fibroblasts in Experimental example 3.

Detailed Description

The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

This section generally describes the materials used in the testing of the present invention, as well as the testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible. It will be apparent to those skilled in the art that the materials and methods of operation used in the present invention are well within the skill of the art, provided that they are not specifically illustrated.

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

materials:

pineapple is purchased from red star farm in Zhanjiang province of Guangdong province;

oat (fried) was purchased from Zhangjiakou Jianjun oat food Co., Ltd;

highland barley from Beijing Tongrentang

Coix seed from Beijing Tongrentang

Olive oil was purchased from yunan yuancheng long-curried trades ltd;

fibroblasts, melanocytes, purchased from the stem cell bank of the department of Chinese;

red blood cells, purchased from Beijing Xinglong animal breeders;

SDS, purchased from alatin;

sodium chloride, purchased from national medicine.

The instrument comprises the following steps:

homogenizer, purchased from IKA, model: t25. .

Enzyme-linked immunoassay instrument, purchased from Thermo Fisher Scientific OY, model 1510-00662C.

A medical centrifuge, purchased from ruijiang analytical instruments ltd, tin-free.

Example 1

This example illustrates the selection of bromelain solution according to the invention.

The experiment researches the influence of enzyme solutions obtained from different varieties of pineapples on the molecular weight of the product polypeptide when the pineapples are used as the enzyme solution and the oats, the highland barley and the coix seeds are used as substrates.

The experimental method comprises the following steps:

(1) cleaning, peeling and slicing different varieties of pineapples, mixing the pineapples with water according to the mass ratio of 1:1, and pulping to obtain the pineapple enzyme liquid 1 and 2. Wherein sample 1 is a basil pineapple and sample 2 is a golden diamond pineapple.

(2) Carrying out physical crushing on oat, and mixing the oat with water according to the mass ratio of 1:25 to obtain a pretreatment solution (2).

(3) The pretreatment solution (2) was added with (1) at a mass ratio of enzyme solution to substrate of 1: 50. The obtained mixture was numbered as sample 1 and sample 2, respectively, and the blank group was the pretreatment solution (2) without adding the bromelain solution.

(4) And (4) preserving the temperature of the mixed solution in the step (3) for 1 hour at 35 ℃, and then putting the mixed solution in a water bath with the temperature of 95 ℃ for 0.5 hour to inactivate the enzyme.

(5) And (4) centrifuging the mixed solution in the step (4) for 20 minutes at 5000r/min to obtain a supernatant.

(6) And measuring the molecular weight of the polypeptide of the supernatant by using high performance liquid chromatography. The specific determination method is as follows:

(1) chromatographic conditions

A chromatographic column: tsk gel 2000SWXL 300 mm. times.7.8 mm

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

Detection wavelength: UV 220nm

Flow rate: 1ml/min

Column temperature: 25 deg.C

Sample preparation: a sample with a concentration of 5mg/ml is prepared by using a mobile phase as a solvent, and is filtered by a microporous membrane (0.45 mu m) and then is injected.

Standard samples: bovine serum albumin (Mr 67000), B12(Mr 1335) and oxidized glutathione (Mr 614) were mixed to obtain a mixed standard, and the content of each substance was 5 mg/ml.

(2) Determination of the Standard Curve

Preparing three standard samples according to 5mg/ml, preparing a standard curve according to the chromatographic conditions, and solving a linear regression equation by adopting a least square method, wherein the linear regression equation is as follows: y is-0.3913 x +7.4563, and the regression coefficient R2 is 0.9982

The regression coefficient R2 of the equation is 0.9982, so that the standard curve linear relation of the mixed standard sample is good, and the accuracy of calculation can be improved. In the formula, X represents elution time and Y represents logarithm of molecular weight. Thus, when the facial mask sample is subjected to HPLC analysis, the corresponding molecular weight can be calculated through the appearance time of the elution peak of each substance in the elution map.

(3) Molecular weight measurement results

Fig. 1 to 3 show HPLC profiles of 220nm sample 1, sample 2 and blank set in example 1, respectively. The time to peak, area and molecular weight for sample 1, sample 2 and the blank are shown in table 1 below.

TABLE 1 Peak out time, area and molecular weight for sample 1, sample 2 and blank

Experimental results and discussion:

in the blank group, the content of the polypeptide with the molecular weight of 262946.30Da is 6.14%, the content of the polypeptide with the molecular weight of 69342.58Da is 71.11%, the content of the polypeptide with the molecular weight of 2266.84Da is 6.95%, the content of the polypeptide with the molecular weight of 1661.12Da is 81.29%, and the content of the polypeptide with the molecular weight of 142.01Da is 11.76%; in sample 1, the content of the polypeptide with the molecular weight of 1394.14Da is 43.35%, the content of the polypeptide with the molecular weight of 955.95Da is 41.72%, and the content of the polypeptide with the molecular weight of 146.21Da is 14.93%; in sample 2, the content of the polypeptide with molecular weight of 1413.49Da was 87.19%, and the content of the polypeptide with molecular weight of 146.83Da was 12.81%. The results showed that the supernatant obtained by adding bromelain solution had an increased proportion of polypeptides with a molecular weight of 1600Da or less compared to the blank, and in sample 1, the content of polypeptides with a molecular weight of 1000 or less was 56.65% under the action of the protease in the pineapple. Therefore, the bromelain can degrade high molecular weight polypeptide in oat into lower molecular weight polypeptide, and the molecular weight distribution results of the polypeptide of the sample 1 and the polypeptide of the sample 2 are compared, so that the pineapple preparation enzyme solution used for preparing the sample 1 in the subsequent experiment is finally selected.

Example 2

This example illustrates the preparation of a facial mask solution according to the present invention.

1. Raw material treatment

Pineapple: cleaning, peeling and cutting into blocks.

Oat, highland barley and coix seed: pulverizing, and sieving with 80 mesh sieve.

2. Preparation of facial mask liquid

(1) The mass ratio of the pineapple to water is 1:50, mixing, pulping, standing at room temperature for 3 hours to obtain an extracting solution (1).

(2) The addition ratio of oat, highland barley and coix seed to the extract (1) is shown in table 1, and a mixed solution is formed.

TABLE 1 addition ratio of pretreatment solution

Raw materials	Parts by mass
		Extract
1	100
		Oat	5
Highland barley	2
		Coix seed	2

(3) Heating the mixed solution obtained in the step (2) at 70 ℃ for 4 hours, and stirring at 100 r/min; cooling to room temperature, centrifuging at 500r/min, and collecting supernatant.

(4) And (4) adding olive oil with the mass of 10% of the supernatant into the supernatant obtained in the step (3), and homogenizing for 30 minutes at the rotation speed of a homogenizer of 4000 r/min.

3. Sterilizing facial mask liquid

Filling the facial mask liquid into facial mask bag (containing facial mask cloth).

Sterilizing at 80 deg.C for 2 hr.

Example 3

1. Raw material treatment

Pineapple: cleaning, peeling and cutting into blocks.

2. Preparation of facial mask liquid

(1) Mixing oat flour and water according to the mass ratio of 1:200, and standing for 3 hours at room temperature to obtain an extracting solution (1).

(2) The addition ratio of oat, highland barley, and coix seed to the extract (1) is shown in table 2, to form a mixture.

TABLE 2 addition ratio of pretreatment solution

Raw materials	Parts by mass
		Extracting solution (1)	100
Oat	1
		Highland barley	1.25
Coix seed	1.25

(3) Heating the mixed solution obtained in the step (2) at 40 ℃ for 4 hours, and stirring at 500 r/min; cooling to room temperature, centrifuging at 2000r/min, and collecting supernatant.

(4) And (4) adding olive oil accounting for 20% of the mass of the supernatant into the supernatant obtained in the step (3), and homogenizing for 20 minutes at the rotation speed of the homogenizer of 5000 r/min.

3. Sterilizing facial mask liquid

Sterilizing at 80 deg.C for 2 hr.

Test example 1

This test example is intended to illustrate the effect of the facial mask solution prepared in example 2 on the proliferation of fibroblasts.

Fibroblast cytotoxicity test (MTT)

Logarithmic phase cells were collected, cell suspension concentration was adjusted, 100. mu.L (96 well plates) was added to each well, and cells to be tested were plated to 5000 cells/well, (marginal wells filled with sterile PBS), 5% CO₂And incubating at 37 ℃ until the cells grow to a certain density, changing the liquid, adding samples to be detected with different concentration gradients, and taking the culture solution without the medicine to be detected as a control. 5% CO₂After 24 hours incubation at 37 deg.C, 20. mu.L of MTT solution (5mg/mL, i.e., 0.5% MTT tetrazolium salt) was added to each well and incubation was continued for 4 hours. The drug was allowed to react well with MTT, and the culture was discarded after centrifugation, and after washing 2-3 times with PBS carefully, MTT-containing culture (FM-Fibroblast Medium from ScienCell) was added. The culture was terminated and the culture medium in the wells was carefully aspirated. Add 150. mu.L of dimethyl sulfoxide into each well, and shake for 10min at low speed on a shaking bed to dissolve the crystals sufficiently. The absorbance of each well was measured at 490nm OD in an ELISA.

Cell viability ═ 100% (assay well OD value-blank OD value)/(cell control OD value-blank OD value).

As a result: and comparing the test results of different concentration gradients of the sample. The toxicity of different concentrations of the facial mask solution on fibroblasts was examined. The results are shown in FIG. 4. The experimental result shows that the facial mask liquid has no obvious cytotoxicity detected, and the cell survival rate is obviously increased within the range of 5-40 percent of the sample concentration and is about 140-180 percent. The facial mask liquid has the function of promoting the proliferation of fibroblasts.

Test example 2

This test example is provided to illustrate the results of transdermal absorption of the facial mask solution prepared in example 2

(1) Preparation of rat skin

The nude mice are killed by taking off the cervical vertebrae, the back hairs are quickly shaved off by a shaver, the back skin is peeled off, subcutaneous fat and blood vessels are removed, the nude mice are repeatedly washed to be clean by distilled water, washed by physiological saline for a plurality of times and stored in a refrigerator at the temperature of 80 ℃ below zero for standby (used up within 5 days).

(2) In vitro transdermal absorption experiment of Franz diffusion cell

The experimental process is that a proper amount of water is added into a thermostatic bath of the in vitro infiltration diffusion device. Starting a power supply and magnetically stirring the mixture in the constant temperature tank, and setting the water temperature in the constant temperature tank to be 37 +/-0.1 ℃. Fixing the prepared rat skin between two diffusion cells by using an iron clamp, adding 5mL of receiving liquid into a receiving cell of a vertical diffusion cell, putting the receiving cell into a thermostatic bath of an in vitro permeation diffusion device for preheating, and setting the stirring speed of the receiving cell to be 400 r/min. Feeding liquid is respectively added into the feeding tanks, and the upper openings are sealed by preservative films. At the beginning, when the samples permeate for 0, 2, 4, 6, 8 and 24 hours (specific time intervals are determined according to actual samples), 500 μ l of samples are respectively taken and placed in a centrifuge tube with a plug, and the same amount of receiving liquid is supplemented into a receiving pool and air bubbles in the pool are removed at the same time of each sampling. The sample sodium hyaluronate content will be determined. The cumulative permeation amount Q (mg/cm2) was calculated according to the following equation. Wherein the diameter of the bottom of the diffusion cell is 1.50cm, and the contact area of the sample is 1.77cm²。

Note that Q is the cumulative permeation, S is the transdermal diffusion area, V is the volume of the receiving chamber of the modified Franz diffusion cell, Cn is the concentration of the receiving solution at the nth sampling, Ci is the concentration of the receiving solution at the ith sampling, and 0.5 is the sampling volume. After calculation by the above formula, a plot of accumulated amount versus time is made. The receiving solution is 0.9% NaCl solution (normal saline); the supply liquids are respectively samples to be detected.

As a result: the polysaccharide is used as an index for investigation, and the test results are shown in FIG. 5. Under the condition of ensuring the consistency of the total sugar content of the liquid to be detected, the polysaccharide transmittance is continuously increased along with the prolonging of the action time within 0-24h of the action time of the mask liquid, and the transdermal process of the polysaccharide tends to be stable at 8 h. The mask liquid has a certain transdermal absorption effect.

Test example 3

This test example is intended to illustrate the effect of the facial mask solution prepared in example 3 on the collagen content of fibroblasts.

In this test example, the collagen content in fibroblasts was measured using a type I collagen (Col I) test kit, HYPERLINK "http:// www.njjcbio.com/products, ash ═ 701" \\ t "/Users/lpp/Documents \ \ x/_ blank".

(1) Preparation of the experiment

a. And (3) diluting the standard: the lyophilized standard powder was made up to 150. mu.l (160ng/ml) with standard diluent and mixed for 30 s. Five clean EP tubes were loaded with 150. mu.l of standard dilutions, labeled 80ng/ml, 40ng/ml, 20ng/ml, 10ng/ml, and 5ng/ml, respectively. Adding 150 μ l of the stock solution of the standard substance into a tube marked as 80ng/ml, uniformly mixing, taking out 150 μ l of the stock solution, adding the stock solution into the next tube, and repeating the steps until the last tube is obtained. (zero-hole direct addition of standard diluent)

b. Dilution of biotin antigen: centrifuging the concentrated biotin antigen at 6000-10000rpm for 30s, then taking 1ml of biotin antigen diluent to the concentrated biotin antigen, mixing for 15s until the biotin antigen is completely dissolved, then pouring all the liquid into a diluent bottle, and mixing uniformly to obtain the biotin antigen working solution.

c. Dilution of avidin-HRP: and (3) centrifuging the concentrated biotin antigen at 6000-.

Sample treatment:

d. diluting a washing solution: pouring 25 times of concentrated solution in the kit into a 500ml volumetric flask, and using distilled water to fix the volume to 500ml to obtain the working solution.

The plate washing method comprises the following steps: firstly absorbing the liquid in the enzyme labeling hole, patting the enzyme labeling hole dry on absorbent paper, adding 300 mul of washing liquid into each hole, slightly shaking for 30s, then throwing off the enzyme labeling hole dry, patting the enzyme labeling hole dry on the absorbent paper, and repeating the steps for 4-5 times.

(2) Detailed experimental procedures

a. The kit was allowed to equilibrate at room temperature for half an hour prior to use

b. Blank wells were not loaded, only color reagent A, B and stop solution were added for zeroing.

c. Standard sample wells: 50. mu.l of diluted standard is added to each well, 50. mu.l of standard/sample diluent is added to zero wells, and then 50. mu.l of biotin antigen working solution is added.

d. Sample well: the sample was added in 50. mu.l followed by the biotin antigen working solution in 50. mu.l.

Gently shake, cover with a sealing membrane, incubate at 37 ℃ for 30 min.

e. Diluting 25 times of the concentrated washing solution with distilled water for later use.

f. First washing: carefully peeling off the sealing plate membrane, discarding the liquid, spin-drying, adding 200 μ l of washing solution into each hole, standing for 30s, discarding, repeating the steps for 5 times, and draining.

g. Add 50. mu.l avidin-HRP to the standard and sample wells, shake gently, cover the cover plate membrane, incubate for 30min at 37 ℃ in the incubator.

h. And (3) second washing: carefully uncovering the sealing plate membrane, discarding liquid, spin-drying, filling washing liquid into each hole, standing for 30s, discarding, repeating the steps for 5 times, and patting dry.

i. Color development: 50 mul of color developing agent A is added into each hole, 50 mul of color developing agent B is added into each hole, the mixture is evenly mixed by gentle shaking, and color development is carried out for 10 minutes at 37 ℃ in a dark place.

j. And (4) terminating: stop the reaction by adding 50. mu.l of stop solution to each well (at this time, blue color immediately turns yellow)

k. And (3) determination: the absorbance (OD value) of each well was measured in turn at a wavelength of 450nm while the blank well was set to zero. The measurement should be performed within 10min after the stop solution.

Calculating: and (3) calculating a standard curve and a regression equation according to the concentration and the OD value, calculating by using special software, and selecting a logistic curve (four parameters) as a fitting model. The schematic drawing of the standard is shown in the following table 3:

TABLE 3 schematic drawing of the standard

Concentration of

Blank

S5

S4

S3

S2

S1

S0

OD value

VB

V5

V4

V3

V1

V0

Calibration value

VB-VB

V5-VB

V4-VB

V3-VB

V2-VB

V1-VB

V0-VB

As a result: the results of the test of the effect of the facial mask solution on the collagen content in fibroblasts are shown in fig. 6. The model group is a cell group which is irradiated by only UVA and is not added with a sample, the blank group is a cell group which is not irradiated and is not added with a sample, and the positive control group is a cell group which is added with a positive control sample VC with the mass concentration of 0.1g/L after UVA irradiation.

Experiments show that UVA irradiation can seriously damage collagen of cells, and a sample group is compared with a model group, a blank group and a positive control group.

Although the present invention has been described to a certain extent, it is apparent that appropriate changes in the respective conditions may be made without departing from the spirit and scope of the present invention. It is to be understood that the invention is not limited to the described embodiments, but is to be accorded the scope consistent with the claims, including equivalents of each element described.

Claims

1. The mask liquid is characterized by not containing any chemical additive component and being prepared from pure plant raw materials and water, wherein the pure plant raw materials comprise the following components: pineapple, oat, highland barley, coix seed and vegetable oil;

2. The mask liquid according to claim 1, wherein the mask liquid comprises the following raw material components in parts by weight:

3. The preparation method of the mask liquid according to claim 1 or 2, wherein the method comprises the following steps:

(1) cleaning, peeling and slicing pineapples, mixing with water, pulping, and standing to obtain an extracting solution;

(2) physically crushing and sieving oat, highland barley and coix seed, adding the crushed oat, highland barley and coix seed into the extracting solution obtained in the step (1), heating and stirring, cooling to room temperature, and centrifuging to obtain a supernatant;

4. The method according to claim 3, wherein in the step (1), the proportion of the pineapple to the water is 1: 1-500 g/mL, preferably 1: 5-500 g/mL, more preferably 1: 10-400 g/mL, and further preferably 1: 50-200 g/mL; and/or

5. The method according to claim 3 or 4, wherein in the step (2), the oat, the highland barley and the coix seed are ground and then sieved by a 80-mesh sieve.

6. The method according to any one of claims 3 to 5, wherein in the step (2), the mass ratio of the oat to the extracting solution obtained in the step (1) is 1-50: 100, preferably 1-20: 100, more preferably 1 to 10:100, more preferably 1-5: 100;

the mass ratio of the highland barley to the extracting solution obtained in the step (1) is 1-10: 100, preferably 1-5: 100, more preferably 1 to 3:100, more preferably 1.25-2: 100; and/or

The mass ratio of the coix seeds to the extracting solution obtained in the step (1) is 1-10: 100, preferably 1-5: 100, more preferably 1 to 3:100, and more preferably 1.25 to 2: 100.

7. The method according to any one of claims 3 to 6, wherein in step (2), the heating temperature is 20 to 150 ℃, preferably 25 to 100 ℃, more preferably 30 to 100 ℃, and most preferably 40 ℃;

8. The method according to any one of claims 3 to 7, wherein in step (3), the vegetable oil is added in an amount of 1 to 50%, preferably 1 to 40%; more preferably 10 to 30%, and still more preferably 10 to 20%;

9. The method according to any one of claims 3 to 8, characterized in that the method further comprises the steps of:

(4) sterilizing the facial mask liquid obtained in the step (3);

10. A facial mask, characterized in that it comprises the facial mask fluid of claim 1 or 2 or a facial mask fluid prepared according to the method of any one of claims 4 to 9.