阅读说明：本技术 一种具有抗蓝光作用的组合物 (Composition with blue light resisting effect ) 是由 熊丽丹 肖青 唐洁 李利 于 2021-10-27 设计创作，主要内容包括：本发明公开了一种具有抗蓝光作用的组合物,其组成包括叶黄素和叶绿素盐,所述叶黄素与叶绿素盐的质量百分比为1～4：1。本发明组合物,通过叶黄素/叶绿素铜钠盐复配,获得优异的抗蓝光(400-500nm)的作用,显著优于单独使用叶黄素和叶绿素铜钠盐,具有协同增效、使得所抗波段蓝移而作用于危害程度更大的短波蓝光(400-450nm)。同时,利用该抗蓝光复配,制备成的眼贴膜、面膜等化妆品,可保护眼周、面部皮肤的蓝光损伤,减少相应部位的氧化应激、色素沉着,减少黑眼圈、细纹,且粘附性强,不易脱落,在活动时间也能方便使用。(The invention discloses a composition with a blue light resisting effect, which comprises lutein and chlorophyll salt, wherein the mass percentage of the lutein to the chlorophyll salt is (1-4): 1. the composition obtains excellent blue light resistance (400 + 500nm) effect through lutein/sodium copper chlorophyllin complex formulation, is obviously superior to the effect of singly using lutein and sodium copper chlorophyllin, has synergistic interaction, and enables the blue shift of the resistant waveband to act on the short-wave blue light (400 + 450nm) with larger harm degree. Meanwhile, cosmetics such as eye masks and facial masks prepared by the blue light resistant compound can protect blue light damage of skin around eyes and faces, reduce oxidative stress and pigmentation of corresponding parts, reduce black eye circles and fine lines, have strong adhesion, are not easy to fall off, and can be conveniently used during activity time.)

1. Use of lutein in combination with chlorophyll salts for the preparation of a combined composition having an anti-blue effect.

2. Use according to claim 1, characterized in that: the mass ratio of the lutein to the chlorophyll salt is 1-4: 1, preferably 2: 1.

3. use according to claim 1 or 2, characterized in that: the chlorophyll salt is sodium copper chlorophyllin, sodium magnesium chlorophyllin, potassium copper chlorophyllin and/or sodium zinc chlorophyllin, preferably sodium copper chlorophyllin.

4. A combination composition having an anti-blue light effect, characterized by: the drug contains lutein and chlorophyll salt for simultaneous or separate administration, wherein the mass ratio of the lutein to the chlorophyll salt is (1-4): 1.

5. the combination composition according to claim 1, characterized in that: the mass ratio of the lutein to the chlorophyll salt is 2: 1.

6. the combination pharmaceutical composition according to claim 4 or 5, characterized in that: the chlorophyll salt is sodium copper chlorophyllin, sodium magnesium chlorophyllin, potassium copper chlorophyllin and/or sodium zinc chlorophyllin, preferably sodium copper chlorophyllin.

7. The composition with the blue light resisting effect is characterized by comprising lutein and chlorophyll salts, wherein the mass ratio of the lutein to the chlorophyll salts is (1-4): 1.

8. the composition of claim 7, wherein: the mass ratio of the lutein to the chlorophyll salt is 2: 1.

9. the composition of claim 7, wherein: the chlorophyll salt is sodium copper chlorophyllin, sodium magnesium chlorophyllin, potassium copper chlorophyllin and/or sodium zinc chlorophyllin, preferably sodium copper chlorophyllin.

10. The composition according to any one of claims 7 to 9, characterized in that: the external preparation is prepared by taking lutein and chlorophyll salt as active ingredients and adding pharmaceutically acceptable auxiliary materials.

11. The composition of claim 10, wherein: the external preparation is granules, powder, pills, capsules or solutions, and microcapsules are preferred.

12. The composition of claim 11, wherein: the microcapsule is prepared from the following raw materials in parts by weight:

1-5 parts of lutein/sodium copper chlorophyllin, 1-5 parts of gelatin, 1-5 parts of carrageenan and 1-3 parts of glycerol.

13. The composition of claim 12, wherein: the microcapsule is prepared from the following raw materials in parts by weight:

1 part of lutein/sodium copper chlorophyllin, 1 part of gelatin, 1 part of carrageenan and 1-3 parts of glycerol.

14. A process for preparing a composition according to any one of claims 7 to 13, characterized in that: it comprises the following steps:

(1) weighing the raw materials according to the proportion;

(2) adding lutein/sodium copper chlorophyllin into water solution containing gelatin and carrageenan, stirring at 450r/min at 40 deg.C for 30min, adjusting pH to 4, stirring for 15min, adding glycerol, adjusting pH to 7, stirring at 300r/min at 4 deg.C for 30min, and freeze drying.

15. Use of the composition of any one of claims 7 to 13 for preparing a medicament or skin care product with blue light resisting effect.

16. A skin care product with blue light resisting effect is characterized in that: the composition is prepared from the following raw materials in parts by weight:

the composition of claim 7, wherein the composition comprises 1-2% of carrageenan, 0.5-1% of hydroxymethyl cellulose, 0.5-1% of chitosan and the balance of water.

17. The skin care product of claim 7, wherein: the composition is prepared from the following raw materials in parts by weight:

the composition of claim 7, wherein the composition comprises 1.5% of carrageenan, 0.5% of hydroxymethyl cellulose, 0.5% of chitosan, and the balance of water.

18. A method of making the skin care product of claim 16, comprising: it comprises the following steps:

a. weighing the raw materials according to the proportion;

b. mixing carrageenan, hydroxymethyl cellulose, chitosan and water, stirring at 450r/min at 40 deg.C for 3 hr, adding the composition of claim 7, stirring for 1 hr, and filtering.

Technical Field

The invention particularly relates to a composition with an anti-blue-light effect.

Background

Visible light accounts for approximately 50% of solar radiation, one third of which is natural blue light. Blue light penetrates deeper into the skin than UVA radiation, which represents only 5% of sunlight. Blue light, especially short-wave blue light (400-450nm), triggers oxidative stress, which accelerates skin aging and increases pigmentation, especially in the periocular region. Short-wave blue light can also cause DNA damage. The skin around human eyes is very thin, is 1/4 the thickness of cheek skin, sebaceous glands and sweat glands are rarely distributed, and simultaneously, a thick muscle layer is lacked, so that the human eyes are more easily damaged by blue light, and the problems of fine lines, black eyes and the like are easily caused.

Lutein is an antioxidant, has strong antioxidant function, and can protect eyes and skin from free radical damage. Besides, the lutein can effectively filter the short-wave blue light which most seriously damages the retina, dispel the short-wave blue light after absorbing ultraviolet rays, inhibit the recession phenomenon caused by photooxidation and protect macular tissues in the retina from being damaged. Research shows that lutein can reduce the risk of senile macular degeneration, and the lutein is more recommended to be used in preventing cataract and protecting heart and skin. According to the invention, the full scanning is carried out at 380-500 nm wavelength in the early stage, the lutein absorption peak range is 440-500 nm, the maximum absorption peak exists at the wavelength of 460nm, the wavelength of the short-wave blue light is 400-450nm, and the narrow wavelength range for effectively filtering the short-wave blue light by independently using lutein is seen, so that the exertion of the blue light resistant effect of lutein is limited, therefore, a method for expanding the blue light resistant wavelength range of lutein is still to be developed to improve the blue light resistant effect of lutein, but no research report about the effect of enhancing the blue light resistant effect of lutein on short wave is provided at present, and no lutein product with better short-wave blue light prevention effect is disclosed.

Disclosure of Invention

In order to solve the above problems, the present invention provides the use of lutein in combination with chlorophyllin salt for the preparation of a combination composition with anti-blue light effect.

Further, the mass ratio of the lutein to the chlorophyll salt is 1-4: 1, preferably 2: 1.

further, the chlorophyll salt is sodium copper chlorophyll, sodium magnesium chlorophyll, potassium copper chlorophyll and/or sodium zinc chlorophyll, preferably sodium copper chlorophyll.

The invention also provides a combined pharmaceutical composition with blue light resisting effect, which contains lutein and chlorophyll salt for simultaneous or separate administration, wherein the mass ratio of the lutein to the chlorophyll salt is (1-4): 1.

further, the mass ratio of the lutein to the chlorophyll salt is 2: 1.

further, the chlorophyll salt is sodium copper chlorophyll, sodium magnesium chlorophyll, potassium copper chlorophyll and/or sodium zinc chlorophyll, preferably sodium copper chlorophyll.

The invention also provides a composition with a blue light resisting effect, which comprises lutein and chlorophyll salt, wherein the mass ratio of the lutein to the chlorophyll salt is (1-4): 1.

further, the mass ratio of the lutein to the chlorophyll salt is 2: 1.

further, the chlorophyll salt is sodium copper chlorophyll, sodium magnesium chlorophyll, potassium copper chlorophyll and/or sodium zinc chlorophyll, preferably sodium copper chlorophyll.

Furthermore, the external preparation is prepared by taking lutein and chlorophyll salt as active ingredients and adding pharmaceutically acceptable auxiliary materials.

Further, the external preparation is a granule, powder, pill, capsule or solution, preferably a microcapsule.

Furthermore, the microcapsule is prepared from the following raw materials in parts by weight:

1-5 parts of lutein/sodium copper chlorophyllin, 1-5 parts of gelatin, 1-5 parts of carrageenan and 1-3 parts of glycerol.

Furthermore, the microcapsule is prepared from the following raw materials in parts by weight:

1 part of lutein/sodium copper chlorophyllin, 1 part of gelatin, 1 part of carrageenan and 1-3 parts of glycerol.

The present invention also provides a process for preparing the aforementioned composition, comprising the steps of:

(1) weighing the raw materials according to the proportion;

(2) adding lutein/sodium copper chlorophyllin into water solution containing gelatin and carrageenan, stirring at 450r/min at 40 deg.C for 30min, adjusting pH to 4, stirring for 15min, adding glycerol, adjusting pH to 7, stirring at 300r/min at 4 deg.C for 30min, and freeze drying.

The invention also provides an application of the composition in preparing medicines or skin care products with blue light resisting effect.

The invention also provides a skin care product with blue light resisting effect, which is prepared from the following raw materials in parts by weight:

1-2% of the composition, 0.5-1% of carrageenan, 0.5-1% of hydroxymethyl cellulose, 0.5-1% of chitosan and the balance of water.

Further, the feed additive is prepared from the following raw materials in parts by weight:

1.5 percent of the composition, 0.5 percent of carrageenan, 0.5 percent of hydroxymethyl cellulose, 0.5 percent of chitosan and the balance of water.

The invention finally provides a process for preparing the aforementioned skin care product, which comprises the steps of:

a. weighing the raw materials according to the proportion;

b. mixing carrageenan, hydroxymethyl cellulose, chitosan and water, stirring at 450r/min at 40 deg.C for 3 hr, adding the composition of claim 1, stirring for 1 hr, and filtering to obtain the final product

The composition obtains excellent blue light resistance (400 + 500nm) effect through lutein/sodium copper chlorophyllin complex formulation, is obviously superior to the effect of singly using lutein and sodium copper chlorophyllin, has synergistic interaction, and enables the blue shift of the resistant waveband to act on the short-wave blue light (400 + 450nm) with larger harm degree. Meanwhile, cosmetics such as eye masks and facial masks prepared by the blue light resistant compound can protect blue light damage of skin around eyes and faces, reduce oxidative stress and pigmentation of corresponding parts, reduce black eye circles and fine lines, have strong adhesion, are not easy to fall off, can be conveniently used in activity time, and have practical popularization and application values.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

1380-500 nm wavelength full scan

FIG. 2 micrographs and particle size distributions of microcapsules

FIG. 3 microcapsule stability under different conditions

FIG. 4 xanthophyll/sodium copper chlorophyllin microcapsules cytotoxicity toxicology

FIG. 5 is a physical image and a scanning electron microscope image of lutein/sodium copper chlorophyllin microcapsule gel

FIG. 6 influence of lutein/sodium copper chlorophyllin loaded microcapsule gel on cell survival rate

FIG. 7 is the transdermal condition of lutein/sodium copper chlorophyllin microcapsule gel

FIG. 8 Effect of lutein/sodium copper chlorophyllin loaded microcapsules on blue light induced ROS

FIG. 9 Effect of lutein/sodium copper chlorophyllin microcapsule gel on pigmentation caused by blue light

FIG. 10 effect of lutein/sodium copper chlorophyllin microcapsule gel on blue light-induced fibroblast aging

FIG. 11 typical effect diagram of lutein/sodium copper chlorophyllin loaded microcapsule gel

Detailed Description

Example 1 preparation of lutein/sodium copper chlorophyllin microcapsules of the invention

The formula is as follows: 0.25g of lutein/sodium copper chlorophyllin (mass ratio of 2: 1), 0.25g of gelatin, 0.25g of carrageenan and 0.25g of glycerol.

The preparation method comprises the following steps: dissolving gelatin and carrageenan in water to obtain 50mL solution, adding lutein and sodium copper chlorophyllin, stirring at 450r/min at 40 deg.C for 30 min. The pH was adjusted to 4 and stirring was continued for 15 min. Adding glycerol, adjusting pH to 7, 300r/min, and stirring at 4 deg.C for 30 min. And (5) freeze-drying to obtain the product.

Example 2 preparation of lutein/sodium copper chlorophyllin microcapsules of the invention

The formula is as follows: 0.25g of lutein/sodium copper chlorophyllin (mass ratio of 2: 1), 0.25g of gelatin, 0.25g of carrageenan and 0.75g of glycerol.

The preparation method comprises the following steps: same as example 1

Example 3 preparation of lutein/sodium copper chlorophyllin microcapsules of the invention

The formula is as follows: 0.5g of lutein/sodium copper chlorophyllin (mass ratio of 2: 1), 0.25g of gelatin, 0.25g of carrageenan and 0.3g of glycerol.

The preparation method comprises the following steps: same as example 1

Example 4 preparation of lutein/sodium copper chlorophyllin microcapsules of the invention

The formula is as follows: 0.5g of lutein/sodium copper chlorophyllin (mass ratio of 2: 1), 0.5g of gelatin, 0.25g of carrageenan and 0.1g of glycerol.

The preparation method comprises the following steps: same as example 1

Example 5 preparation of an eye gel patch of the invention

The formula is as follows: 1.5% of lutein/sodium copper chlorophyllin microcapsule prepared in example 1, 0.5% of carrageenan, 0.5% of hydroxymethyl cellulose, 0.5% of chitosan and the balance of deionized water;

the preparation method comprises the following steps:

a. weighing the raw materials according to the proportion;

b. mixing carrageenan, hydroxymethyl cellulose, chitosan and water, stirring at 450r/min at 40 deg.C for 3 hr, adding lutein/sodium copper chlorophyllin microcapsule, stirring for 1 hr, and filtering with semipermeable membrane to obtain gel patch.

Example 6 preparation of an eye gel patch of the invention

The formula is as follows: 1% of lutein/sodium copper chlorophyllin microcapsule prepared in example 1, 0.5% of carrageenan, 0.5% of hydroxymethyl cellulose, 0.5% of chitosan and the balance of deionized water;

the preparation method comprises the following steps: same as example 5

Example 7 preparation of an eye gel patch of the invention

The formula is as follows: 1% of lutein/sodium copper chlorophyllin microcapsule prepared in example 1, 1% of carrageenan, 1% of hydroxymethyl cellulose, 1% of chitosan and the balance of deionized water;

the preparation method comprises the following steps: same as example 5

Example 8 preparation of an eye gel patch of the invention

The formula is as follows: 2% of lutein/sodium copper chlorophyllin microcapsule prepared in example 1, 1% of carrageenan, 1% of hydroxymethyl cellulose, 1% of chitosan and the balance of deionized water;

the preparation method comprises the following steps: same as example 5

Example 9 preparation of an eye gel patch of the invention

The formula is as follows: 2% of lutein/sodium copper chlorophyllin microcapsule prepared in example 1, 0.5% of carrageenan, 0.5% of hydroxymethyl cellulose, 0.5% of chitosan and the balance of deionized water;

the preparation method comprises the following steps: same as example 5

The advantageous effects of the present invention are described below by way of test examples.

Test example 1 blue light resistance study of the present invention

1. Lutein/sodium copper chlorophyllin complex

Respectively dissolving lutein, sodium copper chlorophyllin and a lutein/sodium copper chlorophyllin compound by using DMSO, and performing full scanning at a wavelength of 380-500 nm to obtain a lutein absorption peak range of 440-500 nm, wherein the maximum absorption peak exists at a wavelength of 460 nm; the maximum absorption peak range of the sodium copper chlorophyllin is 405-420 nm, and the maximum absorption peak exists at the wavelength of 420 nm; when the ratio of lutein to sodium copper chlorophyllin is 1: 1 to 4: 1 proportion, particularly the proportion is 2: 1, the absorption peak is subjected to blue shift and moves to 380-500 nm, and under the same concentration (25 mu g/mL), the absorption peak is obviously increased, and the absorption intensity is improved by 3-10 times. See figure 1 for details.

2. Preparation of lutein/sodium copper chlorophyllin microcapsules

The optimal condition and process for preparing the lutein/sodium copper chlorophyllin microcapsule are determined by a screening experiment and comprise the following steps:

dissolving 0.25g gelatin and 0.25g carrageenan in water to prepare 50mL solution, adding 0.25g lutein and sodium copper chlorophyllin, stirring at 450r/min at 40 deg.C for 30 min. The pH was adjusted to 4 and stirring was continued for 15 min. Adding 0.25g of glycerol, adjusting the pH to 7, stirring at 300r/min and stirring at 4 ℃ for 30 min. Freeze drying to obtain;

the morphological characteristics of the obtained lutein/sodium copper chlorophyllin microcapsules are shown in fig. 2, and can be seen from fig. 2: the microcapsule has regular shape, uniform distribution and particle size of 5 μm. The encapsulation efficiency was 48.21%.

3. Microcapsule stability:

relative humidity: mixing the following components in a mass ratio of 2: the lutein/sodium copper chlorophyllin mixture of 1 and the lutein/sodium copper chlorophyllin microcapsule prepared above were stored under different relative humidity (30%, 80%) in dark at constant temperature, and the lutein/sodium copper chlorophyllin content was tested every ten days. The relative stability at 30% humidity, with increasing humidity, the lutein/sodium copper chlorophyllin content of untreated microcapsules decreased rapidly, while the content decreased slowly after treatment.

Illumination: mixing the following components in a mass ratio of 2: the lutein/sodium copper chlorophyllin mixture of 1 and the lutein/sodium copper chlorophyllin microcapsule prepared above are respectively placed in a transparent bottle and a brown bottle for constant temperature preservation, and the lutein/sodium copper chlorophyllin content is tested every ten days. The lutein/sodium copper chlorophyllin microcapsule is relatively stable under the dark condition, the content of the lutein/sodium copper chlorophyllin which is not treated into the microcapsule under the illumination condition is rapidly reduced, and the content is slowly reduced after the lutein/sodium copper chlorophyllin microcapsule is treated.

③ temperature: mixing the following components in a mass ratio of 2: the lutein/sodium copper chlorophyllin mixture of 1 and the lutein/sodium copper chlorophyllin microcapsule prepared above were stored in the dark at different relative temperatures (5 ℃, 25 ℃, 45 ℃), and the lutein/sodium copper chlorophyllin content was tested every ten days. The treatment temperature is relatively stable under the condition of 5 ℃, the content of an untreated group is reduced faster than that of a treated group under the condition of 25 ℃, and the content of the untreated group is further reduced rapidly than that of the treated group under the condition of 45 ℃. See in particular fig. 3.

4. Xanthophyll/sodium copper chlorophyllin microcapsule toxicology

In 96-well plates according to 4X 10³Keratinocyte and melanocyte are inoculated at the density of each hole, after the cells are completely attached to the wall, solutions (0, 12.5, 25, 50 and 10uM) of lutein/sodium copper chlorophyllin (the mass ratio is 2: 1) with different concentrations are added for intervention, each group has 5 multiple holes, and a blank control group (without cells and only with culture medium) and a negative control group are arranged at the same time. After intervention for 24h, adding CCK-8 solution with the volume of 10 ul/hole by using a discharging gun, incubating for 1-2 hours at 37 ℃ in a dark place, and detecting the OD value at 450nm by using an enzyme-linked immunosorbent assay. Data were derived, and the cell proliferation rate = (% (OD450 experimental group-OD 450 blank group)/(OD 450 negative control group-OD 450 blank group) × 100% the lutein/sodium copper chlorophyllin microcapsules prepared above were not cytotoxic to keratinocytes and melanocytes to 100 μ g/mL, and the cell survival rate was greater than 90%. See figure 4 for details.

5. Preparation of adhesive gel film

0.5-1% of carrageenan, 0.5-1% of hydroxymethyl cellulose, 0.5-1% of chitosan, 450r/min, stirring for 3h at 40 ℃, adding 1-2% of the prepared lutein/sodium copper chlorophyllin microcapsule, continuously stirring for 1h, and filtering by a semipermeable membrane to prepare the gel with adhesive force. The prepared gel film has good viscosity and can not fall off under the ordinary moving state. The scanning electron micrograph shows that the surface is smooth and the morphology is good. See figure 5 for details.

6. Blue light resistance of gel film

(ii) cell survival rate

Collecting keratinocyte and melanocyte at a ratio of 3 × 10 per well⁵The individual cells were seeded in 6-well plates at 37 ℃ with 5% CO₂The culture was carried out overnight in an incubator. When the cell density reaches 70% of the dish bottom, covering the cell with a piece of burl paper, irradiating with 100J/cm2 blue light, covering with a blank gel without lutein/sodium copper chlorophyllin microcapsule, covering the treated cell with the prepared lutein/sodium copper chlorophyllin microcapsule gel, removing the waste liquid, washing with PBS once, adding CCK-8 solution with the volume of 10 ul/hole, incubating at 37 ℃ in the dark for 1-2 hours, and detecting the OD value at 450nm by an enzyme-labeling instrument. And (6) deriving data and calculating the cell survival rate.

At the cell level, 100J/cm²The blue light causes the death of keratinocytes and melanocytes by about 50 percent, and the lutein/sodium copper chlorophyllin microcapsule gel can resist the blue light to about 90 percent of the cell survival rate, and has obvious effect. See figure 6 for details.

(ii) Release Rate

The negative lutein/sodium copper chlorophyllin microcapsule gel can be slowly released in the skin, the microcapsule is observed by a fluorescent section, the medicine penetrates through the horny layer after 10min, enters the epidermal layer after 30min, and penetrates through the epidermal basal layer and enters the dermal layer after 60 min. See figure 7 for details.

③ free radical caused by resisting blue light

Collecting keratinocyte and melanocyte at a ratio of 3 × 10 per well⁴The individual cells were seeded in 24-well plates at 37 ℃ with 5% CO₂The culture was carried out overnight in an incubator. When the cell density reaches 70% of the bottom of the dish, covering the cell with a piece of burry paper, irradiating with 100J/cm2 blue light, covering with a blank gel without lutein/sodium copper chlorophyllin microcapsule, and covering the treated cell with the prepared lutein/sodium copper chlorophyllin microcapsule gel, removing the waste liquid, washing with PBS once, adding DCFH-DA dye solution, incubating at 37 ℃ in the dark for 30min, washing with PBS once, and observing and taking a picture under a fluorescence microscope.

100J/cm²The blue light causes the increase of ROS in keratinocytes, and the lutein/sodium copper chlorophyllin microcapsule can resistROS caused by blue light, and the effect is obvious. See figure 8 for details.

Anti-pigmentation caused by blue light

Melanocytes at 3X 10 per well⁵The individual cells were seeded in 6-well plates at 37 ℃ with 5% CO₂The culture was carried out overnight in an incubator. When the cell density reaches 70% of the bottom of the dish, covering the cell with a piece of burley paper, irradiating with 100J/cm2 blue light, covering with a blank gel without lutein/sodium copper chlorophyllin microcapsules, covering the cell with the prepared lutein/sodium copper chlorophyllin microcapsule gel, removing waste liquid, washing with PBS once, digesting the cell with 0.25% pancreatin at 37 ℃, adding a FBS-containing culture medium to terminate digestion, slightly blowing down all adherent cells, and collecting in a 1.5mL centrifuge tube. Cells were collected by centrifugation at 1000rpm for 3min and the supernatant was removed. After adding 1mL of PBS and mixing, washing once, and observing and photographing. After PBS is removed, 350uL of 1MNaOH solution is added into each tube, water bath is carried out for 30min at 80 ℃ until melanin is completely dissolved, an enzyme-linked immunosorbent assay (OD) value at 405nm is detected, and the melanin content is calculated. 100J/cm²The blue light causes the content of melanin in melanocytes to be increased, and the lutein/sodium copper chlorophyllin microcapsule can resist the content of the melanin caused by the blue light, and the effect is obvious. See figure 9 for details.

Resisting cell aging caused by blue light

Fibroblast cells were 3X 10 per well⁵The individual cells were seeded in 6-well plates at 37 ℃ with 5% CO₂The culture was carried out overnight in an incubator. When the cell density reaches 70% of the bottom of the dish, covering the cells with the tinfoil paper at 100J/cm²Blue light irradiation, blank gel covering without lutein/sodium copper chlorophyllin microcapsule, covering the cells treated by the prepared lutein/sodium copper chlorophyllin microcapsule gel, removing waste liquid, washing with PBS once, adding 1mL of beta-galactosidase staining fixing solution, and fixing at room temperature for 15 min. The cell fixative was aspirated, cells were washed 3 times with PBS, and 1mL of staining solution was added to each well. Incubation was overnight at 37 ℃ and the 6-well plate was sealed with parafilm to prevent evaporation. And (4) observing under a common optical microscope.

100J/cm²The blue light causes the beta-galactosidase content of the fibroblasts to be increased, representing aging, so as to load lutein/chlorophyllThe copper sodium salt microcapsule can resist the aging increase caused by blue light, and has obvious effect. See figure 10 for details.

7. Gel film human body efficacy verification

Firstly, the eye mask has the function of treating the periocular problems

Selecting 60 25-40 years old pigmented black eye female subjects, randomly dividing into three groups, namely a blank group: without any periocular care product; control group: the microcapsule gel not containing the loaded lutein/sodium copper chlorophyllin prepared as in example 5 using the blank gum; experimental groups: the lutein/sodium copper chlorophyllin loaded microcapsule gel prepared as in example 5 was used.

The treatment is carried out for 120 days, once in the morning and at night, 10 minutes each time. In the experiment, the influence of other factors on the test result is reduced as much as possible (for example, the test subject is required to keep the sleep time for about 8 hours every day)

Through tests, a subject takes facial photos under the same environment of 60 days and 120 days before use, and after the color is uniformly corrected by Adobe photoshop CC, ImageJ software obtains CIEL a b chromaticity space color data (CIE International Commission on illumination) of a black eye area of a lower eyelid to evaluate the curative effect of treating the periocular problems by using the gel patch, and the result shows that the lutein/sodium copper chlorophyllin microcapsule gel is effective in improving the black eye and has no irritation. See fig. 11, table 1.

TABLE 1 dark circles data Change before and after use of lutein/sodium copper chlorophyllin microcapsule gel

Subjective testing: by way of physician evaluation, physicians were asked to score questions, with each item being a maximum of 5 points, averaged per group, and scored once per week.

Table 2 subjective test change before and after eye use of lutein/sodium copper chlorophyllin microcapsule gel

② the facial mask acts on facial problems

Selecting 60 female subjects with dark skin and mild fine wrinkles of 35-60 years old, randomly dividing the female subjects into three groups, namely a blank group: without any periocular care product; control group: example 5 preparation of microcapsules without loaded lutein/sodium copper chlorophyllin using a blank gum; experimental groups: example 5 was used to prepare a lutein/sodium copper chlorophyllin loaded microcapsule gel.

Subjective testing: by way of physician evaluation, physicians were asked to score questions, with each item being a maximum of 5 points, averaged per group, and scored once per week.

Table 3 subjective test change before and after using lutein/sodium copper chlorophyllin loaded microcapsule gel on face

As can be seen from table 3: compared with the microcapsule gel without the lutein/sodium copper chlorophyllin and the eye nursing product, the lutein/sodium copper chlorophyllin microcapsule gel can obviously improve the facial darkness, reduce the wrinkle quantity and reduce the wrinkle depth.

In conclusion, the composition obtains excellent blue light resistance (400-450nm) by compounding the lutein/sodium copper chlorophyllin, so that the blue shift of the resistant wave band acts on the short-wave blue light (400-450nm) with larger harm degree. Meanwhile, cosmetics such as eye masks and facial masks prepared by the blue light resistant compound can protect blue light damage of skin around eyes and faces, reduce oxidative stress and pigmentation of corresponding parts, reduce black eye circles and fine lines, have strong adhesion, are not easy to fall off, can be conveniently used in activity time, and have practical popularization and application values.