CN113637721A - Method and device for measuring moisturizing effect of cosmetics - Google Patents
Method and device for measuring moisturizing effect of cosmetics Download PDFInfo
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Abstract
The invention discloses a method and a device for measuring moisturizing efficacy of cosmetics, wherein the method comprises the following steps: the HaCaT cells are divided into three parts, and one part of the HaCaT cells is untreated; one part is subjected to dehydration treatment to obtain dehydrated damaged cells; adding one part of the extract into a cosmetic to be detected, culturing for a period of time, and then performing dehydration treatment to obtain repair cells; respectively measuring absorbance values of HaCaT cells, water loss damaged cells and repair cells at 570nm by adopting an MTT method to respectively obtain a, b and c; the moisturizing effect of the cosmetic to be tested is (c-b)/ax100%. The method has the advantages of high accuracy of the measured result, simple equipment and easy operation; the moisturizing effect of medicaments, cosmetics and cosmetic raw materials is effectively evaluated at a cell level by simulating biological skin cells.
Description
Technical Field
The invention relates to a method and a device for measuring efficacy of cosmetics, in particular to a method and a device for measuring moisturizing efficacy of cosmetics.
Background
The water in the horny layer of the skin has important significance for maintaining the softness and the elasticity of the skin, maintaining the normal metabolism of the skin and preventing the skin from aging and generating wrinkles. The water content of stratum corneum of healthy skin is 10-30%, and if the water content of skin is less than 10%, the skin will be dry, rough and possibly chapped. Under normal conditions, the stratum corneum has the functions of water absorption and barrier, and the lipid substances secreted by sweat glands and sebaceous glands cover the stratum corneum, so that the water in the stratum corneum of the skin is not easy to lose; however, when the skin is dry and cold in climate, low in environmental humidity and impaired in normal barrier function of the skin, the skin is prone to lack of water, and various problems such as dryness and roughness are caused.
Moisturizing toning lotion is a mainstream product of the toning lotion sold in the market, and meanwhile, moisturizing is also the most basic step in the skin care process. With the formal release of the regulations on supervision and management of cosmetics, the guide principles on the declaration and evaluation of cosmetic efficacy, and the like, it is clearly pointed out that the declaration of cosmetic efficacy needs to be supported by sufficient evidence. The existing method for measuring the moisturizing effect generally refers to the evaluation guidelines of moisturizing effect of cosmetics. Patent CN201310624204.8 discloses a moisture-retention degree detection system for cosmetics, wherein resistance increases with the decrease of water content according to the regular change between the resistance and the water content of the cosmetics, and the method is to evaluate the moisture-retention performance of skin care products by measuring the water content of the horny layer of human skin by a capacitance method; patent CN201310624195.2 discloses a method for testing moisture retention of cosmetics, which is to determine the moisture retention effect of a cosmetic sample to be tested by making a stable humidity difference on two sides of a filter paper coated with the cosmetic sample to be tested. The method is characterized in that the water loss amount of biological skin cells cannot be really simulated through a physical method, the obtained data cannot be verified on a human body, and compared with the actual moisturizing effect, the method has obvious errors.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a method for measuring the moisturizing efficacy of cosmetics with high accuracy; another object of the present invention is to provide a device for measuring the moisturizing efficacy of a cosmetic.
The technical scheme is as follows: the method for measuring the moisturizing effect of the cosmetics comprises the following steps:
(1) the HaCaT cells are divided into three parts, and one part of the HaCaT cells are not subjected to any treatment; one part is subjected to dehydration treatment to obtain dehydrated damaged cells; adding one part of the extract into a cosmetic to be detected, culturing for a period of time, and performing dehydration treatment to obtain repair cells;
(2) respectively measuring absorbance values of HaCaT cells, water loss damaged cells and repair cells at 570nm by adopting an MTT method to obtain a, b and c; the moisturizing effect of the cosmetic to be tested is as follows:
further, repeating the steps (1) and (2) n times, wherein the moisturizing efficacy of the cosmetic to be tested is the average value of the n times.
Further, in step (1), the HaCaT cells are HaCaT cells before F13 generation.
Further, in the step (1), the water loss treatment method is to culture HaCaT cells by using a mixed culture solution prepared by sterile NaCl and MEM culture solutions; the concentration of NaCl in the mixed culture solution is 1-4%, and the culture time is 0.5-1 h.
Further, in the step (3), b/a is 40-65%, b/a is a cell viability value of the dehydration damaged cells, and the accuracy of measuring the moisturizing effect value of the cosmetic is influenced by too large and too small cell viability values.
The device applying the method for measuring the moisturizing effect of the cosmetics comprises a cell acquisition module, a cell dehydration treatment module, a to-be-measured cosmetic testing module and a moisturizing effect evaluation module; the cell collection module collects HaCaT cells and divides the HaCaT cells into three groups, wherein one group is continuously remained in the cell collection module and is the HaCaT cells; the cell dehydration treatment module is used for performing dehydration treatment on a group of HaCaT cells to obtain dehydration damaged cells; the cosmetic testing module to be tested performs cosmetic protection on a group of HaCaT cells and then performs dehydration treatment to obtain repair cells; the moisturizing efficacy evaluation module comprises an enzyme-linked immunosorbent assay instrument and a data processing module, wherein the enzyme-linked immunosorbent assay instrument is used for respectively testing absorbance values of HaCaT cells, water loss damaged cells and repairing cells to obtain a, b and c, the data processing module is used for obtaining the moisturizing efficacy of the cosmetic to be tested through a moisturizing efficacy calculation formula (c-b)/a multiplied by 100%, and outputting a measurement result.
Furthermore, n cell culture cavities are arranged in the cell collection module, the cell dehydration treatment module and the cosmetic test module to be tested, and a temperature regulation device and a time reminding device are arranged in each cell culture cavity
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages: the accuracy of the measured result is high, the used equipment is simple, and the operation is easy; the moisturizing effect of medicaments, cosmetics and cosmetic raw materials is effectively evaluated at a cell level by simulating biological skin cells.
Drawings
FIG. 1 is a comparison of cell viability for each group of examples 1, 6-10;
fig. 2 is a schematic view of a moisturizing efficacy measuring apparatus according to the present invention;
FIG. 3 is a microscopic image of each group of cells in example 1.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
Example 1
Experimental materials: MEM basal medium, Fetal Bovine Serum (FBS), HaCaT cells, trypsin, NaCl; the main equipment is as follows: an enzyme-labeling instrument; a liquid transferring gun; a one-ten-thousandth balance; a multi-well cell culture plate; an ultra-clean bench; preparation of NaCl solution: 5.0000g of NaCl powder were weighed out precisely, dissolved in 50mL of MEM minimal medium, mixed well and filtered through a 0.22 μm filter for use. Preparing a sodium hyaluronate solution: 0.1000g of sodium hyaluronate powder is precisely weighed, dissolved in 50mL of PBS, mixed uniformly and filtered through a 0.22 mu m filter membrane for later use.
The method for measuring the moisturizing effect of the cosmetics comprises the following steps:
(1) HaCaT cells: recovering HaCaT cells before F13 generation in a clean bench, digesting when the cells are fused in a cell culture dish to more than 80%, uniformly mixing, counting, and adding 5.0 × 103Each well of 96-well plate was prepared by CO-treating 3X 3 wells, divided into 3 groups and labeled as group A, group B and group C, each three groups were cultured in 100. mu.L of 10% FBS-containing MEM at 37 ℃ and 5% CO per well2Culturing until about 70% of cells adhere to the wall, wherein the group A is a HaCaT cell group;
loss of water damages cells: placing group B in a clean bench, sucking out the culture medium in the previous step with vacuum pump or pipette, and replacing with fresh MEM culture solution prepared with 10% sterile NaCl at concentration of 5% CO at 37 deg.C and 3% concentration of 100 μ l culture solution per well2Culturing for 0.5h to obtain the dehydration damaged cells.
Repairing cells: using sodium hyaluronate as a cosmetic to be detected to protect HaCaT cells; placing group C in a clean bench, sucking out the culture medium in HaCaT cell plating plate by vacuum pump or pipette gun, changing to 90 μ l MEM culture solution +10 μ l sodium hyaluronate culture medium, 37 deg.C, and 5% CO2Culturing for 16h, sucking out culture medium with vacuum pump or pipette, adding 10% sterile NaCl solution prepared fresh MEM (100 μ l per well), controlling NaCl concentration at 3%, 37 deg.C, and 5% CO2Culturing for 0.5h to obtain the repair cells.
(2) Respectively testing absorbance (OD) values of HaCaT cells, water loss damaged cells and repaired cells under the wavelength of 570nm by an MTT method and using an enzyme-linked immunosorbent assay instrument to respectively obtain a, b and c, and respectively obtaining a moisturizing effect calculation formula:
wherein the cell viability value calculation formula of the water loss injured cells is as follows:
loss of water cell viability (%) (loss of water cell group OD)570value/HaCaT cell group OD570Value x 100%, i.e
The cell viability value of the repaired cells is calculated by the formula:
repair cell viability (%). repair cell group OD570value/HaCaT cell group OD570Value x 100%, i.e.
Moisturizing efficacy calculation formula: repair cell viability (%) -loss of water damage cell viability (%).
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 1 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, # indicates p < 0.001, i.e., the difference present was very significant, and a indicates p < 0.01, i.e., the difference present was significant. It can be seen that the mean value of cell viability of group B is 61.15%, which is very different from group a, and the mean value of cell viability of group C is 82.41%, which is different from group B; the moisturizing effect of the cosmetic sodium hyaluronate to be detected is 21.26%, and the moisturizing effect of the cosmetic sodium hyaluronate to be detected is better.
TABLE 1 OD of each group570And the value of cell viability
As shown in fig. 3, the morphological difference of each group of cells was observed by photographing with an inverted biomicroscope, group a is the cell morphology under normal condition, and compared with group a, the cells of group B are damaged by dehydration, the cell morphology is shrunk, and the membrane structure is damaged; the cells in group C partially collapsed morphologically compared to group A, but had better cell morphology and reduced damage compared to group B. The above results demonstrate the moisturizing effect of the cosmetic sodium hyaluronate to be tested.
Example 2
Fig. 1 is a schematic diagram of a device for measuring moisturizing efficacy of cosmetics, which includes a cell collection module, a cell dehydration treatment module, a to-be-measured cosmetic test module, and a moisturizing efficacy evaluation module; cell dehydration processing module and the cosmetics processing module that awaits measuring link to each other with cell collection module, and the efficiency of moisturizing evaluation module links to each other with cell collection module, cell dehydration processing module, the cosmetics test module that awaits measuring and the efficiency of moisturizing evaluation module, all is equipped with the cell culture chamber in cell collection module, cell dehydration processing module, the cosmetics test module that awaits measuring, is equipped with temperature regulation apparatus and time reminding device in the cell culture chamber, and the cell culture chamber is n.
The cell collecting module collects HaCaT cells and divides the HaCaT cells into three groups, each group of cells are respectively and automatically added into the cell culture cavity,the cell culture cavity contains cell culture solution; a group of the cell collection module is reserved for culture, and the remaining two groups of the cell collection module are respectively conveyed to the cell dehydration processing module and the cosmetic processing module to be detected through the conveying belt; a temperature regulating device and a time reminding device are arranged in the cell culture cavity to control the cell culture environment and time, wherein the temperature of the cell collection module is 37 ℃, and CO is2The concentration was 5%.
The cell dehydration treatment module receives the cell culture cavity, sucks out cell culture solution in the cell culture cavity through a vacuum pump arranged in the cell dehydration treatment module, and adds dehydration treatment solution which is mixed solution of NaCl and MEM culture solution; a temperature regulating device and a time reminding device are arranged in the cell culture cavity to control the cell culture environment and time, wherein the temperature of the cell dehydration treatment module is 37 ℃, and CO is2The concentration was 5%.
The to-be-tested cosmetic treatment module receives the cell culture cavity, the cell culture solution in the cell culture cavity is sucked out through a vacuum pump arranged in the to-be-tested cosmetic treatment module, the mixed solution of the to-be-tested cosmetic and the MEM culture solution is added, after the to-be-tested cosmetic treatment module is cultured for a period of time, the mixed solution in the cell culture cavity is sucked out through the vacuum pump, and the water loss treatment solution is added and is the mixed solution of NaCl and the MEM culture solution; a temperature regulating device and a time reminding device are arranged in the cell culture cavity to control the cell culture environment and time, wherein the temperature of the cosmetic treatment module to be detected is 37 ℃, and CO is2The concentration was 5%.
After the cell culture is finished, the cell collection module, the cell dehydration treatment module and the cosmetic treatment module to be tested respectively convey the cell culture cavity to the moisturizing effect evaluation module; an enzyme-linked immunosorbent assay instrument and a data processing module are arranged in the moisturizing effect evaluation module, the enzyme-linked immunosorbent assay instrument respectively measures absorbance values of each group of cell culture cavities under the wavelength of 570nm to obtain absorbance values of HaCaT cells, dehydration damaged cells and repair cells, the absorbance values are sent to the data processing module, the data processing module processes the absorbance values of each group to obtain the moisturizing effect of the cosmetic to be measured, and a measurement result is output.
The moisturizing effect calculation formula of the data processing module is as follows:
example 3: comparison of different modeling times of cell water loss model
Experimental materials: MEM basal medium, Fetal Bovine Serum (FBS), HaCaT cells, trypsin, NaCl; the main equipment is as follows: an enzyme-labeling instrument; a liquid transferring gun; a one-ten-thousandth balance; a multi-well cell culture plate; an ultra-clean bench; a cell culture dish; preparation of NaCl solution: 5.0000g of NaCl powder were weighed out precisely, dissolved in 50mL of MEM minimal medium, mixed well and filtered through a 0.22 μm filter for use.
HaCaT cells: recovering HaCaT cells before F13 generation in a clean bench, digesting when the cells are fused in a cell culture dish to more than 80%, uniformly mixing, counting, and adding 5.0 × 103Each well of 96-well plate was inoculated with 100. mu.L of 10% FBS MEM culture medium at 37 ℃ in 5% CO2Culturing until about 70% of cells adhere to the wall.
Loss of water damages cells: dividing the cultured HaCaT cells into 10 groups, wherein each group has 3 multiplied by 3 multiple holes, and the 10 groups are respectively treated by sterile NaCl solutions with different concentrations; in the clean bench, the culture medium solution in 10 groups was aspirated by a vacuum pump or pipette, and replaced with 100. mu.l of fresh MEM culture medium prepared with 10% sterile NaCl at concentrations of 0.00%, 1.00%, 1.50%, 1.80%, 1.90%, 2.00%, 2.10%, 2.20%, 2.50%, and 3.00%, respectively. Each group of 3X 3 multiple wells is divided into 3 groups according to the time for NaCl solution treatment, and the culture is carried out for 0.5h, 1h and 2h under the conditions of 37 ℃ and 5% CO2 respectively.
Cell viability assay: the cell viability assay was performed as in example 1, with the specific results shown in Table 2.
TABLE 2 cell viability values at different NaCl concentrations and treatment times
The cell viability value of the dehydration damaged cells is usually controlled to be 40-65%, the contrast effect can be influenced by too low or too high cell viability value, and the table 2 shows that when the treatment time is 0.5h, the NaCl concentration is 1.80-2.50%, and the cell viability value can reach 40-65%; when the treatment time is 1-2 h, the NaCl concentration is 1.50-1.80%, and the cell viability value can reach 40-65%; therefore, the treatment time is 0.5-1h, and the NaCl concentration is 1.50-2.50% to meet the requirement of the cell viability value. In addition, when the treatment time is 0.5h, the selection range of NaCl concentration is the largest, and thus the treatment time is the most preferable.
Example 4: comparison of different modeling concentrations for cellular water loss model
Experimental materials: MEM basal medium, Fetal Bovine Serum (FBS), HaCaT cells, trypsin, NaCl; the main equipment is as follows: an enzyme-labeling instrument; a liquid transferring gun; a one-ten-thousandth balance; a multi-well cell culture plate; an ultra-clean bench; a cell culture dish; preparation of NaCl solution: 5.0000g of NaCl powder were weighed out precisely, dissolved in 50mL of MEM minimal medium, mixed well and filtered through a 0.22 μm filter for use.
HaCaT cells: in a clean bench, HaCaT cells before F13 generation were thawed, digested when they reached 80% or more fusion in a cell culture dish, and then mixed and counted, and then cultured in a 96-well plate of 5.0X 103 cells/well in 100. mu.L 10% FBS MEM medium at 37 ℃ and 5% CO2 until cells adhered to the wall at about 70%.
Loss of water damages cells: dividing the cultured HaCaT cells into 10 groups, wherein each group has 4 multiple holes, and the 10 groups are respectively treated by sterile NaCl solutions with different concentrations; in the clean bench, the culture medium solution in 10 groups was aspirated by a vacuum pump or pipette, and replaced with 100. mu.l of fresh MEM culture medium prepared with 10% sterile NaCl at concentrations of 0.00%, 1.00%, 1.50%, 1.80%, 2.00%, 2.30%, 2.50%, 3.00%, 3.50%, and 4.00%, respectively. 37 ℃ and 5% CO2Cultured under the conditions for 0.5 hour.
Cell viability assay: the cell viability assay was performed as in example 1, with the specific results shown in Table 3.
TABLE 3 Absorbance values and cell viability comparison for different NaCl concentrations
The cell viability value of the dehydration damaged cells is controlled to be 40-65%, and as can be seen from Table 3, the cell viability value can be met when the treatment time is 0.5 hour and the NaCl concentration is 2.30-3.00%. Because the reproducibility of the cell experiment is poor, the results of the embodiment 3 and the embodiment 4 are combined to determine that the optimal NaCl concentration is 3.00 percent, the optimal processing time is 0.5 hour, and the error caused by the poor reproducibility of the cell experiment can be greatly reduced.
Example 5: verification of cellular water loss model
Experimental materials: MEM basal medium, Fetal Bovine Serum (FBS), HaCaT cells, trypsin, NaCl; the main equipment is as follows: an enzyme-labeling instrument; a liquid transferring gun; a one-ten-thousandth balance; a multi-well cell culture plate; an ultra-clean bench; a cell culture dish; preparation of NaCl solution: 5.0000g of NaCl powder were weighed out precisely, dissolved in 50mL of MEM minimal medium, mixed well and filtered through a 0.22 μm filter for use.
HaCaT cells: in a clean bench, HaCaT cells before F13 generation were thawed, digested when they reached 80% or more fusion in a cell culture dish, and then mixed and counted, and then cultured in a 96-well plate of 5.0X 103 cells/well in 100. mu.L 10% FBS MEM medium at 37 ℃ and 5% CO2 until cells adhered to the wall at about 70%.
Loss of water damages cells: dividing the cultured HaCaT cells into 2 groups, a blank group and a water-loss group, wherein each group comprises 3 multiple holes, sucking out the culture medium solution in the 2 groups by using a vacuum pump or a pipette gun in an ultra-clean workbench, and replacing the culture medium solution with 100 mu l of fresh MEM culture solution prepared by 10% sterile NaCl and NaC in each holeThe concentration of the system is controlled at 3.00 percent, 37 ℃ and 5 percent CO2Culturing for 0.5 h.
Cell viability and expression assay of AQP3 gene: cell viability assay method as in example 1, cells were tested for expression of AQP3 gene using Real-Time qPCR. AQP3 is aquaporin 3, and the detection of the expression of AQP3 gene is used for verifying the water loss condition of water loss damaged cells.
TABLE 4 OD of blank and dehydration-damaged cells570Cell viability and AQP3 expression level
As can be seen from the above table, the NaCl concentration is 3.00%, the culture time is 0.5h, the activity value of each parallel group is about 50%, and the activity value meets the requirement of 40-65%, compared with the blank group, the cell activity of the water loss group is reduced by 48.93%, the expression level of AQP3 is reduced by 21.76%, the fact that the cells in the cell water loss model have water loss injury is proved, and meanwhile, the positive correlation between the cell activity value and the cell water loss injury is verified, namely, the cell water loss model established by the invention is established.
Example 6
Substantially the same procedure as in example 1 except that in the repairing cell treatment, a culture solution was prepared using the scutellaria baicalensis extract 1 in place of the sodium hyaluronate solution, wherein the concentration of the scutellaria baicalensis extract 1 in the culture solution was 200 μ g/mL.
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 5 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, and # indicates p < 0.001, i.e., the difference was very significant, from which it can be seen that the cell viability of the B group was 51.55% in average and the difference from the a group was very significant, and the cell viability of the C group was 73.82% in average; the moisturizing effect of the scutellaria baicalensis extract 1 to be tested is 22.27%, and the moisturizing effect is good.
TABLE 5 OD570 and cell viability values of each group
Example 7
Substantially the same procedure as in example 1 except that in the repairing cell treatment, a culture solution was prepared using the scutellaria baicalensis extract 2 in place of the sodium hyaluronate solution, wherein the concentration of the scutellaria baicalensis extract 2 in the culture solution was 200 μ g/mL.
TABLE 6 OD of each group570And the value of cell viability
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 6 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, # indicates p < 0.001, i.e., the difference present was very significant, and a indicates p < 0.01, i.e., the difference present was significant. It can be seen that the mean cell viability of group B was 65.74%, which is very different from group a, and the mean cell viability of group C was 90.96%, which is different from group B; the cosmetic sodium hyaluronate to be tested has the moisturizing effect of 25.22 percent and good moisturizing effect.
Example 8
Substantially the same procedure as in example 1 except that in the repairing cell treatment, a culture solution was prepared using a licorice extract in place of the sodium hyaluronate solution, wherein the concentration of the licorice extract in the culture solution was 200. mu.g/mL.
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 7 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, and # indicates p < 0.001, i.e., the difference was very significant, from which it can be seen that the cell viability of the B group was 65.74% in average and the difference from the a group was very significant, and the cell viability of the C group was 72.94% in average; the liquorice extract of the cosmetic to be detected has the moisturizing effect of 7.20 percent and poor moisturizing effect.
TABLE 7 OD570 and cell viability values of each group
Example 9
Substantially the same procedure as in example 1, except that in the repairing cell treatment, a culture solution was prepared using a ganoderma lucidum extract in place of the sodium hyaluronate solution, wherein the concentration of the ganoderma lucidum extract in the culture solution was 200. mu.g/mL.
TABLE 8 OD570 and cell viability values of each group
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 8 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, # indicates p < 0.001, i.e., the difference present was very significant, and a indicates p < 0.01, i.e., the difference present was significant. It can be seen that the mean value of cell viability of the group B is 51.55%, which is very different from that of the group A, and the mean value of cell viability of the group C is 74.34%, which is different from that of the group B; the cosmetic sodium hyaluronate to be detected has the moisturizing effect of 22.79% and a good moisturizing effect.
Example 10
The procedure was substantially the same as in example 1 except that in the repairing cell treatment, a culture solution was prepared using the sikamizi decoction extract in place of the sodium hyaluronate solution, wherein the concentration of the sikamizi decoction extract in the culture solution was 200. mu.g/mL.
And (4) analyzing results: OD of group A, group B and group C570The analysis was compared to cell viability. As shown in table 9 and fig. 1, the GraphPad _ Prism t-test was statistically analyzed, # indicates p < 0.001, i.e., the difference present was very significant, and a indicates p < 0.01, i.e., the difference present was significant. It can be seen that the mean cell viability of group B was 51.55%, which is very different from group a, and the mean cell viability of group C was 82.90%, which is different from group B; the moisturizing effect of the cosmetic sodium hyaluronate to be detected is 31.35 percentPreferably.
TABLE 9 OD of each group570And the value of cell viability
The invention adopts HaCaT cells to simulate biological skin for measuring the moisturizing effect of cosmetics, reduces biological skin carriers, simplifies the procedures of human body tests, leads the measured result to be closer to the real result and improves the accuracy of the measured result. Through tests on various cosmetics and raw materials, the test result is found to be the same as the effect reported in the literature.
Claims (8)
1. A method for measuring the moisturizing efficacy of cosmetics is characterized by comprising the following steps:
(1) the HaCaT cells are divided into three parts, and one part of the HaCaT cells are not subjected to any treatment; one part is subjected to dehydration treatment to obtain dehydrated damaged cells; adding one part of the extract into a cosmetic to be detected, culturing for a period of time, and performing dehydration treatment to obtain repair cells;
(2) respectively measuring absorbance values of HaCaT cells, water loss damaged cells and repair cells at 570nm by adopting an MTT method to obtain a, b and c; the moisturizing effect of the cosmetic to be tested is as follows:
2. the method for measuring moisturizing efficacy of cosmetic according to claim 1, wherein steps (1) and (2) are repeated n times, and the moisturizing efficacy of the cosmetic to be measured is an average value of n times.
3. The method for measuring moisturizing efficacy of cosmetic compositions according to claim 1, wherein in step (1), the HaCaT cells are HaCaT cells before generation F13.
4. The method for measuring moisturizing efficacy of cosmetic compositions according to claim 1, wherein in step (1), the dehydration treatment is performed by culturing HaCaT cells in a mixed culture solution prepared from sterile NaCl and MEM culture solution.
5. The method for measuring moisturizing efficacy of cosmetics according to claim 4, wherein the concentration of NaCl in the mixed culture solution is 1-4%, and the culture time is 0.5-1 h.
6. The method for measuring moisturizing efficacy of cosmetic compositions according to claim 1, wherein in step (3), b/a is 40% to 65%.
7. The device for measuring the moisturizing effect of the cosmetics is characterized by comprising a cell collection module, a cell dehydration treatment module, a to-be-measured cosmetic testing module and a moisturizing effect evaluation module; the cell collection module collects HaCaT cells and divides the HaCaT cells into three groups, wherein one group is continuously remained in the cell collection module and is the HaCaT cells; the cell dehydration treatment module is used for performing dehydration treatment on a group of HaCaT cells to obtain dehydration damaged cells; the cosmetic testing module to be tested performs cosmetic protection on a group of HaCaT cells and then performs dehydration treatment to obtain repair cells; the moisturizing efficacy evaluation module comprises an enzyme-linked immunosorbent assay instrument and a data processing module, wherein the enzyme-linked immunosorbent assay instrument is used for respectively testing absorbance values of HaCaT cells, water loss damaged cells and repairing cells to obtain a, b and c, the data processing module is used for obtaining the moisturizing efficacy of the cosmetic to be tested through a moisturizing efficacy calculation formula (c-b)/a multiplied by 100%, and outputting a measurement result.
8. The device for measuring the moisturizing effect of the cosmetics according to claim 7, wherein n cell culture chambers are arranged in the cell collection module, the cell dehydration treatment module and the cosmetic testing module to be tested, and a temperature adjusting device and a time reminding device are arranged in each cell culture chamber.
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