CN117547497B - Whitening and anti-aging composition for inhibiting tyrosinase activity and application thereof - Google Patents
Whitening and anti-aging composition for inhibiting tyrosinase activity and application thereof Download PDFInfo
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- CN117547497B CN117547497B CN202311860256.5A CN202311860256A CN117547497B CN 117547497 B CN117547497 B CN 117547497B CN 202311860256 A CN202311860256 A CN 202311860256A CN 117547497 B CN117547497 B CN 117547497B
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Abstract
A whitening and anti-aging composition for inhibiting tyrosinase activity and application thereof belong to the technical fields of health foods, functional foods, health medicines and cosmetics, and the composition comprises the following components in parts by mass: 5 to 10 parts of hedyotis diffusa extract, 0.5 to 1.5 parts of sugarcane polyphenol, 5 to 10 parts of malus hupehensis leaf extract and 1 to 3 parts of coumarin derivative. The white tongue herba hedyotis extract, the Hupeh begonia leaf extract and the coumarin derivative are matched for use, so that the whitening and anti-aging effects can be greatly improved; the oxidation resistance can be further improved by adding the sugarcane polyphenol, and the anti-aging effect is improved; the tyrosinase activity inhibition components are matched for use, so that the whitening effect is more obvious, the effect is quick, and the use experience is improved.
Description
Technical Field
The invention belongs to the technical field of health-care foods, functional foods, health-care medicines and cosmetics, and particularly relates to a whitening and anti-aging composition for inhibiting tyrosinase activity and application thereof.
Background
Melanin is an indispensable pigment in healthy human skin and hair, and the existence of melanin can enable the skin to have different degrees of color, can absorb heat energy in sunlight, maintain body temperature, can eliminate free radicals formed by ultraviolet irradiation, and can eliminate short wave ultraviolet injury. When the melanocytes are irradiated by sunlight for a long time, in order to resist ultraviolet injury, more melanin can be generated after the melanocytes are irradiated by sunlight for a long time, so that the daily life of people is affected, namely, a part of generated melanin enters capillary vessels and is finally metabolized by kidneys; the other part is transferred to the stratum corneum for deposition, so that the exposed skin is blackened, and facial chloasma can also be generated, and the facial skin appears to be aged and affects the image; thus, oral or topical medications are required to whiten the face against aging.
Currently, inhibition of melanin production and transport is a major approach to whitening, i.e., most of the whitening ingredients achieve the effect of preventing darkening by inhibiting, blocking or affecting melanin production and transfer. In skin melanin biosynthesis, tyrosinase catalyzes the conversion of dopa into dopaquinone, which spontaneously undergoes a series of reactions to finally form melanin. Tyrosinase is thus a key enzyme in melanin synthesis, and its activity directly influences the rate and quantity of melanin synthesis. The substance with tyrosinase activity inhibiting effect can reduce conversion of dopa into dopaquinone, and prevent melanin from being generated in large quantity, so as to achieve whitening effect.
In addition, some products are used for accelerating the metabolism of melanin and separating from the stratum corneum to achieve the whitening effect.
However, at present, the whitening and anti-aging medicine or cosmetic products on the market are various, the whitening effect of the components is also uneven, the effect is slow, and the products with unobvious effects are numerous, so that better product upgrading can be realized, and the achievement with higher speed and high efficiency is always the aim pursued in the field of whitening and anti-aging.
Disclosure of Invention
Aiming at the problems of good and uneven effects, slow onset of action, obvious effects, most products and poor use experience of the existing whitening anti-aging products. The invention provides a whitening and anti-aging composition for inhibiting tyrosinase activity and application thereof, wherein the whitening and anti-aging effects can be greatly improved by adopting a white tongue snake grass extract, a Hubei begonia leaf extract and coumarin derivatives to be matched; the oxidation resistance can be further improved by adding the sugarcane polyphenol, and the anti-aging effect is improved; the tyrosinase activity inhibition components are matched for use, so that the whitening effect is more obvious, the effect is quick, and the use experience is improved. The specific technical scheme is as follows:
a whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 5 to 10 parts of hedyotis diffusa extract, 0.5 to 1.5 parts of sugarcane polyphenol, 5 to 10 parts of malus hupehensis leaf extract and 1 to 3 parts of coumarin derivative.
In the technical scheme, the preparation method of the hedyotis diffusa extract comprises the steps of smashing hedyotis diffusa, percolating with 5-6 times of methanol, and filtering to obtain an extract; distilling to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of (8-10) of purified water=1, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting by using an ethanol aqueous solution to obtain an eluent, then adding active carbon to decolorize, filtering, distilling and concentrating, and drying to obtain the hedyotis cinerea extract powder.
The volume concentration of the ethanol water solution is 70-75%.
The distillation temperature of the methanol is 90-95 ℃.
The temperature of distillation concentration is 120-130 ℃.
In the technical scheme, the preparation method of the malus hupehensis leaf extract comprises the steps of crushing malus hupehensis leaves, percolating with methanol 5-6 times of the mass of the malus hupehensis leaves, and filtering to obtain an extract; distilling to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of (8-10) of purified water=1, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting by using an ethanol aqueous solution to obtain an eluent, then adding active carbon to decolorize, filtering, distilling and concentrating, and drying to obtain the extract powder of the malus hupehensis leaves.
The volume concentration of the ethanol water solution is 70-75%.
The distillation temperature of the methanol is 90-95 ℃.
The temperature of distillation concentration is 120-130 ℃.
In the technical scheme, the mixing method of the composition comprises the steps of firstly mixing the hedyotis diffusa extract, the malus hupehensis leaf extract and the sugarcane polyphenol, then adding the coumarin derivative, and mixing for 1-3 min at the rotating speed of 10-14 rpm to obtain the composition.
In the above technical scheme, the coumarin derivative has a molecular structural formula as follows:
wherein R1 is OH; and R2 is H.
The whitening and anti-aging composition for inhibiting tyrosinase activity is applied to health food, functional food, health medicine and cosmetics with whitening and anti-aging effects.
The whitening and anti-aging composition for inhibiting tyrosinase activity and the application thereof have the beneficial effects that compared with the prior art, the whitening and anti-aging composition for inhibiting tyrosinase activity has the following beneficial effects:
1. the hedyotis diffusa contains tens of compounds such as iridoid, acid-containing compounds, anthraquinone compounds, volatile components, phenylpropanoids, coumarin, polysaccharides and the like, and also contains various trace elements, and has various pharmacological activities such as anti-tumor, anti-inflammatory, antibacterial, antioxidant, immunity enhancing and the like. The herba Hedyotidis Diffusae can be separated to obtain 171 kinds of compounds including 32 kinds of iridoid compounds, 26 kinds of flavonoids, 24 kinds of anthraquinones, 26 kinds of phenols and their derivatives, and 50 kinds of volatile oil compounds, and these phytochemicals and extracts have various pharmacological activities such as anti-tumor, antioxidant, antiinflammatory, fibroblast proliferation inhibiting, immunity regulating, and nervous system protecting effects.
The invention adopts the processes of methanol extraction, resin adsorption, ethanol elution, active carbon decoloration and the like to obtain the hedyotis cinerea extract, and the whitening and anti-aging composition is added to inhibit the activity of tyrosinase, so that the effects of preventing melanin from generating, promoting melanin metabolism, whitening and resisting oxidation, enhancing immunity and resisting aging are achieved.
2. The sugarcane polyphenol is a powder substance prepared by taking sugarcane as a raw material, squeezing, filtering, extracting, removing excessive sugar and salt to obtain sugarcane molasses, taking water and ethanol as extraction solvents, filtering, vacuum concentrating, ion exchange, spray drying and the like. The main characteristic component is total polyphenol, and has extremely strong oxidation resistance; can inhibit the activity of alpha-glucosidase and alpha-amylase, and reduce the rise of blood sugar into an index; the invention has the effects of inhibiting the activity of proinflammatory cytokines and resisting inflammation, and the invention adds the sugarcane polyphenol to improve the anti-aging effect of the whitening anti-aging composition.
3. The folium Mali Hupehensis is rich in flavonoids, and also contains polyphenol and phlorizin, and the content of phlorizin in tender leaf of Malus Hupehensis in high altitude area is higher than that in low altitude area. The total flavone of the malus hupehensis can obviously increase proliferation and differentiation of osteoblasts, the active ingredient phlorizin of the total flavone of the malus hupehensis can maintain dynamic balance of bone metabolism by enhancing differentiation of osteoblasts and reducing activity of osteoclasts, and the flavone ingredients such as phlorizin in the total flavone of the malus hupehensis have estrogen-like structures, can be combined with estrogen receptors, play the same role of estrogen and improve the anti-aging effect. The folium Mali Hupehensis also effective in nourishing liver and regulating stomach, promoting salivation, quenching thirst, resolving food stagnation, and removing food stagnation, and can be used for treating acute and chronic liver injury, chronic hepatitis, and fatty liver, and reducing blood sugar.
The invention adopts the processes of methanol extraction, resin adsorption, ethanol elution, active carbon decoloration and the like to obtain the extract of the malus hupehensis leaves, and the whitening and anti-aging composition is added to inhibit the activity of tyrosinase, so that the effects of preventing melanin from generating, promoting melanin metabolism, whitening and resisting aging are achieved.
4. The coumarin derivative is a novel tyrosinase inhibitor, has reversible and mixed inhibition effect on tyrosinase, and can obviously inhibit the activity of tyrosinase, so that the synthesis of melanin is effectively reduced. Compared with hydroquinone, the coumarin derivative has stronger performance of inhibiting tyrosinase in cells and no toxicity. Can be complemented with hydroquinone products during pigmentation treatment to reduce the risk of rebound pigmentation associated with hydroquinone and the initiation of exogenous chloasma. The invention adopts coumarin derivatives and the malus hupehensis leaf extract for composite use, can greatly improve the whitening effect, and can inhibit the tyrosinase activity with high efficiency (the inhibition rate is improved by more than 30%).
The whitening and anti-aging composition provided by the invention has the advantages that the generation of new melanin is inhibited and the existing melanin is decomposed, so that the whitening speed can be greatly improved, compared with the two mechanisms which are independently used, the whitening and anti-aging composition has the advantages of quicker onset of action, obvious whitening and repairing skin effect within 7 days at the shortest, and the use experience is improved.
Detailed Description
The invention will be further illustrated with reference to specific examples, but the invention is not limited to these examples.
Example 1
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 8 parts of hedyotis diffusa extract, 1 part of sugarcane polyphenol, 8 parts of malus hupehensis leaf extract and 2 parts of coumarin derivatives. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 12rpm for 2 min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 5.5 times of methanol, and filtering to obtain extractive solution; distilling at 92 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:9, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 72% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 125 ℃, and drying to obtain hedyotis diffusa extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 5.5 times of methanol, and filtering to obtain extractive solution; distilling at 92 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:9, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 72% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 125 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 82 |
| Comparative example 1 | 76 |
| Comparative example 2 | 73 |
| Comparative example 3 | 75 |
| Comparative example 4 | 52 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 30% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 8 parts of hedyotis diffusa extract, 1 part of sugarcane polyphenol, 8 parts of malus hupehensis leaf extract, 2 parts of coumarin derivatives, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 8 parts of purified water) to be matched for use, 30 subjects are selected, and the composition is taken orally twice a day, and externally applied for uniformly applying faces once a day in the morning and evening, takes effect in 7 days, and has good user experience; after 1 month of use, the skin becomes obviously white and young.
Example 2
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 5 parts of hedyotis diffusa extract, 1.5 parts of sugarcane polyphenol, 5 parts of malus hupehensis leaf extract and 3 parts of coumarin derivatives. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 14rpm for 1 min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 90deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 70% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 130 ℃, and drying to obtain hedyotis diffusa extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 90deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 70% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 130 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 83 |
| Comparative example 1 | 75 |
| Comparative example 2 | 72 |
| Comparative example 3 | 76 |
| Comparative example 4 | 51 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 32% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 5 parts of hedyotis diffusa extract, 1.5 parts of sugarcane polyphenol, 5 parts of malus hupehensis leaf extract, 3 parts of coumarin derivatives, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 5 parts of purified water) to be matched for use, 30 subjects are selected, the oral administration is carried out twice a day, two granules are taken once, the external application is carried out once a day for each day in the morning and evening, 7 days, the effects are achieved, and the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Example 3
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 10 parts of hedyotis diffusa extract, 0.5 part of sugarcane polyphenol, 10 parts of malus hupehensis leaf extract and 1 part of coumarin derivative. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 10rpm for 3min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 5 times of methanol, and filtering to obtain extractive solution; distilling at 95deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 75% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 120 ℃, and drying to obtain the hedyotis cinerea extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 5 times of methanol, and filtering to obtain extractive solution; distilling at 95deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 75% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 120 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
from the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 31% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 10 parts of hedyotis diffusa extract, 0.5 part of sugarcane polyphenol, 10 parts of malus hupehensis leaf extract, 1 part of coumarin derivative, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 10 parts of purified water) to be matched for use, 30 subjects are selected, the oral administration is carried out twice a day, two granules are taken once, the external application is carried out once a day for each day in the morning and evening, 7 days, the effects are achieved, and the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Example 4
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 5 parts of hedyotis diffusa extract, 0.5 part of sugarcane polyphenol, 5 parts of malus hupehensis leaf extract and 1 part of coumarin derivative. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 10rpm for 1 min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 5 times of methanol, and filtering to obtain extractive solution; distilling at 90deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 70% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 120 ℃, and drying to obtain hedyotis diffusa extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 5 times of methanol, and filtering to obtain extractive solution; distilling at 90deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 70% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 120 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 82 |
| Comparative example 1 | 75 |
| Comparative example 2 | 71 |
| Comparative example 3 | 75 |
| Comparative example 4 | 51 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 31% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 5 parts of hedyotis diffusa extract, 0.5 part of sugarcane polyphenol, 5 parts of malus hupehensis leaf extract, 1 part of coumarin derivative, 2 parts of poloxamer, 1 part of alkyl glycoside, 0.8 part of carbomer and 5 parts of purified water), 30 subjects are selected for use in a matched mode, the oral administration is carried out twice a day, two tablets are taken once, the external application is carried out for each time in the morning and evening, the effects are achieved in 7 days, and the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Example 5
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 10 parts of hedyotis diffusa extract, 1.5 parts of sugarcane polyphenol, 10 parts of malus hupehensis leaf extract and 3 parts of coumarin derivatives. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 14rpm for 3min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 95deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 75% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 130 ℃, and drying to obtain the hedyotis cinerea extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 95deg.C to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 75% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 130 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 82 |
| Comparative example 1 | 75 |
| Comparative example 2 | 72 |
| Comparative example 3 | 74 |
| Comparative example 4 | 51 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 31% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 10 parts of hedyotis diffusa extract, 1.5 parts of sugarcane polyphenol, 10 parts of malus hupehensis leaf extract, 3 parts of coumarin derivatives, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 10 parts of purified water) to be matched for use, 30 subjects are selected, the oral administration is carried out twice a day, two granules are taken once, the external application is carried out once a day for each day in the morning and evening, 7 days, the effects are achieved, and the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Example 6
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 6 parts of hedyotis diffusa extract, 0.8 part of sugarcane polyphenol, 6 parts of malus hupehensis leaf extract and 1.5 parts of coumarin derivatives. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 12rpm for 1 min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 5.2 times of methanol, and filtering to obtain extractive solution; distilling at 91 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8.5, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 72% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 122 ℃, and drying to obtain hedyotis diffusa extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 5.2 times of methanol, and filtering to obtain extractive solution; distilling at 91 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:8.5, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with 72% ethanol water solution to obtain eluent, adding active carbon to decolorize, filtering, concentrating by distillation at 122 ℃, and drying to obtain the malus hupehensis leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 81 |
| Comparative example 1 | 74 |
| Comparative example 2 | 70 |
| Comparative example 3 | 75 |
| Comparative example 4 | 49 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 32% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 6 parts of hedyotis diffusa extract, 0.8 part of sugarcane polyphenol, 6 parts of malus hupehensis leaf extract, 1.5 parts of coumarin derivatives, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 8 parts of purified water) to be matched for use, 30 subjects are selected, and the composition is taken orally twice a day, two tablets are taken orally, and the composition is applied on the face for each time uniformly in the morning and evening every day, and takes effect for 7 days, so that the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Example 7
A whitening and anti-aging composition for inhibiting tyrosinase activity comprises the following components in parts by mass: 9 parts of hedyotis diffusa extract, 1.2 parts of sugarcane polyphenol, 9 parts of malus hupehensis leaf extract and 2.5 parts of coumarin derivatives. The composition is prepared by mixing herba Hedyotidis Diffusae extract, folium Mali Hupehensis extract and sugarcane polyphenol, adding coumarin derivative, and mixing at 14rpm for 3min.
The preparation method of herba Hedyotidis Diffusae extract comprises pulverizing herba Hedyotidis Diffusae, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 94 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 74% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 128 ℃, and drying to obtain the hedyotis cinerea extract powder.
The preparation method of the Malus hupehensis leaf extract comprises pulverizing Malus hupehensis leaf, percolating with 6 times of methanol, and filtering to obtain extractive solution; distilling at 94 ℃ to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of purified water=1:10, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting with an ethanol water solution with the volume concentration of 74% to obtain an eluent, then adding activated carbon to decolorize, filtering, concentrating by distillation at 128 ℃, and drying to obtain the Hupeh crab leaf extract powder.
The composition of the embodiment is applied to health-care food, functional food, health-care medicine and cosmetics with whitening and anti-aging effects.
Comparative example 1 was prepared: the quality of the components of the hedyotis diffusa extract is replaced by starch without adding the hedyotis diffusa extract, and other parameters are the same as those of the embodiment.
Comparative example 2 was prepared: the components of the extract of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
Comparative example 3 was prepared: the component mass of coumarin derivatives is replaced by starch without coumarin derivatives, and other parameters are the same as those of the embodiment.
Comparative example 4 was prepared: the components of the extract and coumarin derivatives of the malus hupehensis are replaced by starch, and other parameters are the same as those of the embodiment.
The raw materials of the composition in the embodiment are food, medicine and cosmetic allowable added raw materials.
Tyrosinase activity inhibition assay: tyrosinase activity inhibition rate measurement was performed by using the compositions of this example and the compositions of comparative examples 1 to 4, respectively. Firstly preparing tyrosinase solution with pH of 6.8 and 50mmol/LPBS buffer solution, and then preparing 10g/L ethanol water solution (volume concentration of 40%) composition solution; in a 96-well plate, 80 mu L of a tyrosine 40 mu L, PBS buffer solution, 60 mu L of a composition solution and 50 mu L of a tyrosinase solution of 60U/mL are sequentially added, incubated for 30min at 37 ℃, detected at 475nm by an enzyme-labeled instrument, PBS buffer solution is taken as a blank group, and the inhibition rate of tyrosinase is calculated as follows:
inhibition ratio (%) = [ (A-B) - (C-D) ]/(A-B) ×100%
Wherein A is the absorbance after incubation of the blank solution, B is the absorbance before incubation of the blank solution, C is the absorbance after incubation of the composition solution, and D is the absorbance before incubation of the composition solution.
The results are shown in the following table:
| composition and method for producing the same | Tyrosinase activity inhibition rate% |
| This embodiment | 82 |
| Comparative example 1 | 75 |
| Comparative example 2 | 71 |
| Comparative example 3 | 74 |
| Comparative example 4 | 50 |
From the inhibition rate results, the hedyotis diffusa extract and the malus hupehensis leaf extract have good inhibition effects, wherein the inhibition rate can be greatly improved by 32% when the coumarin derivative and the malus hupehensis leaf extract are used together.
The composition of the embodiment is prepared into capsules and external application products (the external application product formula comprises 9 parts of hedyotis diffusa extract, 1.2 parts of sugarcane polyphenol, 9 parts of malus hupehensis leaf extract, 2.5 parts of coumarin derivatives, 2 parts of poloxamer, 1 part of alkyl glycoside, 1 part of carbomer and 10 parts of purified water) to be matched for use, 30 subjects are selected, and the composition is taken orally twice a day, two tablets are taken orally, and the composition is applied on the face for each time uniformly in the morning and evening every day, and takes effect for 7 days, so that the user experience is good; after 1 month of use, the skin becomes obviously white and young.
Claims (6)
1. A whitening and anti-aging composition for inhibiting tyrosinase activity is characterized by comprising the following components in parts by weight: 5 to 10 parts of hedyotis diffusa extract, 0.5 to 1.5 parts of sugarcane polyphenol, 5 to 10 parts of malus hupehensis leaf extract and 1 to 3 parts of coumarin derivative;
the preparation method of the hedyotis diffusa extract comprises the steps of smashing hedyotis diffusa, percolating with methanol 5-6 times of mass, and filtering to obtain an extract; distilling to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of (8-10) of purified water=1, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting by using an ethanol aqueous solution to obtain an eluent, then adding active carbon to decolorize, filtering, distilling and concentrating, and drying to obtain hedyotis diffusa extract powder;
the preparation method of the malus hupehensis leaf extract comprises the steps of smashing malus hupehensis leaves, percolating the crushed malus hupehensis leaves with methanol of which the mass is 5-6 times, and filtering the percolate to obtain an extracting solution; distilling to remove methanol to obtain extract; diluting and dispersing the extract according to the mass ratio of (8-10) of purified water=1, then adopting a macroporous structure polymer adsorption resin column to adsorb, eluting by using an ethanol aqueous solution to obtain an eluent, then adding active carbon to decolorize, filtering, distilling and concentrating, and drying to obtain the extract powder of the malus hupehensis leaves.
2. The whitening and anti-aging composition for inhibiting tyrosinase activity according to claim 1, wherein the volume concentration of the aqueous ethanol solution is 70-75%.
3. The whitening and anti-aging composition for inhibiting tyrosinase activity according to claim 1, wherein the distillation temperature of methanol is 90 ℃ to 95 ℃; the temperature of distillation concentration is 120-130 ℃.
4. The whitening and anti-aging composition for inhibiting tyrosinase activity according to claim 1, wherein the composition is prepared by mixing the extract of hedyotis diffusa, the extract of begonia hupehensis leaf and the polyphenol of sugarcane, adding coumarin derivatives, and mixing at a slow speed.
5. The whitening and anti-aging composition for inhibiting tyrosinase activity according to claim 4, wherein the slow mixing speed is 10rpm to 14rpm for 1min to 3min.
6. The use of a whitening and anti-aging composition for inhibiting tyrosinase activity according to claim 1, wherein the composition is used for preparing a medicine or cosmetic having whitening and anti-aging effects.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020071228A (en) * | 2001-03-05 | 2002-09-12 | 연규동 | Extract of Kalopanax pictus, method for preparation and use thereof |
| CN104435630A (en) * | 2013-09-13 | 2015-03-25 | 广东轻工职业技术学院 | Traditional Chinese medicine composition with whitening and anti-aging effects and preparation method and application of extract thereof |
| KR20150129467A (en) * | 2014-05-12 | 2015-11-20 | 한국콜마주식회사 | Skin whitening coumarin compound having inhibitory ability of melanin synthesis, composition for whitening skin and purification method of cumarin compound |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020071228A (en) * | 2001-03-05 | 2002-09-12 | 연규동 | Extract of Kalopanax pictus, method for preparation and use thereof |
| CN104435630A (en) * | 2013-09-13 | 2015-03-25 | 广东轻工职业技术学院 | Traditional Chinese medicine composition with whitening and anti-aging effects and preparation method and application of extract thereof |
| KR20150129467A (en) * | 2014-05-12 | 2015-11-20 | 한국콜마주식회사 | Skin whitening coumarin compound having inhibitory ability of melanin synthesis, composition for whitening skin and purification method of cumarin compound |
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| Title |
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| 湖北海棠的化学成分和药理活性研究进展;李艳 等;中国实验方剂学杂志;20160131;第22卷(第2卷);第226-229页 * |
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