CN112315831A

CN112315831A - Anti-radiation whitening and moisturizing face cream and preparation method thereof

Info

Publication number: CN112315831A
Application number: CN202011294826.5A
Authority: CN
Inventors: 熊林
Original assignee: 熊林
Current assignee: Guangzhou Wanqian vermicelli Cosmetics Co.,Ltd.
Priority date: 2020-11-18
Filing date: 2020-11-18
Publication date: 2021-02-05
Anticipated expiration: 2040-11-18
Also published as: CN113599293A; CN112315831B

Abstract

The invention discloses an anti-radiation whitening and moisturizing cream and a preparation method thereof. The melanin can protect the organism from damage caused by sunlight, ultraviolet light, nuclear radiation, ionizing radiation, X radiation, gamma rays and the like, and the curdlan can firmly aggregate the melanin together to protect the skin. The curdlan can be converted into irreversible high-strength gel by heating above 80 deg.C, so as to firmly lock water and melanin, prevent melanin from entering into skin, and prevent other skin care components from permeating into skin. The verbascoside has super-strong antioxidant activity, and is expressed in large amount in rehmannia rhizome by using transgenic technology. The rehmannia root-like root culture has the advantages of fast growth, stable yield and no influence of seasons compared with a simple plant body.

Description

Anti-radiation whitening and moisturizing face cream and preparation method thereof

Technical Field

The invention relates to the technical field of cosmetics, in particular to an anti-radiation whitening and moisturizing cream and a preparation method thereof.

Background

With the wide application of radiation technology in production, scientific research, medical treatment, military and other aspects, the chances that a human body is irradiated by radioactive rays are more and more, and the harm of radiation to the human body is more and more obvious and serious.

Radiation is an important oxidative stressor. Ionizing radiation can act on water molecules in a body, the water molecules are ionized and excited to generate a large number of free radicals with strong oxidizing performance, and the free radicals and biological macro molecules act, so that tissue cell degeneration and necrosis are indirectly caused, and radiation damage to the body is caused. Therefore, the search for effective radioprotectors from the perspective of resisting oxidation and scavenging free radicals has become an important idea for modern medicine development.

The skin is used as the first line of defense of the body, and can cause the problems of cell damage, skin melanin accumulation, skin aging and the like after being radiated.

Therefore, it is necessary to design an anti-radiation whitening moisturizing cream with good anti-radiation and moisturizing effects and a preparation method thereof.

Disclosure of Invention

The invention aims to provide an anti-radiation whitening and moisturizing cream and a preparation method thereof, so as to solve the problems in the background technology.

In order to solve the technical problems, the invention provides the following technical scheme: an anti-radiation whitening and moisturizing cream comprises the following components in parts by weight: 3-10 parts of verbascoside, 2-7 parts of melanin, 5-10 parts of curdlan, 4-6 parts of glycerol, 3-10 parts of hyaluronic acid, 0.8-3 parts of glutathione, 3-5 parts of an emulsifier, 3-10 parts of pearl powder, 0.6-1.5 parts of vitamin E and 8-20 parts of water.

A preparation method of an anti-radiation whitening and moisturizing cream comprises the following specific steps:

the method comprises the following steps: stirring melanin, 1/2 water and curdlan fermentation liquor at 90 deg.C under 60-120rpm for 4h to obtain phase A;

step two: mixing acteoside, hyaluronic acid, glycerol, Margarita powder, arginine, glutathione, vitamin E and 1/2 water at 70 deg.C under 100-;

step three: mixing emulsifier with phase A and phase B, stirring at 40 deg.C and 80-200rpm for 3 hr to obtain cream.

According to the technical scheme, the melanin is extracted from lachnum tricholobus fermentation liquor or black sesame.

According to the technical scheme, the curdlan fermentation liquor is obtained by utilizing pseudomonas fermentation: 30g/L of sucrose, 8g/L of yeast powder, 3g/L of calcium carbonate, 7.5-8.5 of initial pH value of fermentation liquor, 3-5 days of fermentation period and 30-35 ℃ of fermentation temperature; taking the fermented fermentation liquor, centrifuging for 30 minutes at 8000rpm of 5000-.

According to the above technical scheme, the acteoside is extracted from rehmannia glutinosa hairy root.

According to the technical scheme, the rehmannia root hairy roots are infected with the agrobacterium rhizogenes A4, and the explant age is 7-14D.

According to the technical scheme, the rehmannia glutinosa leaves (0.5 x 0.5cm) are infected by agrobacterium rhizogenes for 5-10min, then bacterial liquid is sucked on sterile filter paper, the leaves are placed in co-culture solid culture media upside down, 10 leaves are placed in each culture dish, co-culture is carried out at 26 ℃ under the dark condition, the co-culture medium is an MS medium, and 100 mu mol/L of AS is added; after co-culture for 2 days, washing with sterile water for 3 times, sucking the liquid with sterile filter paper, inoculating the leaf into a screening medium, and culturing at 26 deg.C in the dark, wherein the screening medium is MS + Cef (0.4g/L) + AS (100 μmol/L).

According to the technical scheme, the Ri plasmid of the agrobacterium rhizogenes contains an Arabidopsis MYB (21R1R2R3-MYB) gene, the MYB gene is amplified through PCR, and the MYB gene is connected to the Ri plasmid after double cutting by enzyme.

According to the technical scheme, the rehmannia glutinosa hairy roots surviving in the screening culture medium are subjected to PCR detection: according to the designed pair of MYB gene primers, the electrophoresis detection shows that the same band as the gene primers appears, but the band is not amplified by the unconverted rehmannia root hairy root, so that the recombination gene is proved to be integrated into the genome of the rehmannia root hairy root.

According to the technical scheme, the rehmannia root-like roots are transferred into a liquid culture medium for amplification and multiplication culture: culturing with YEB culture medium, beef extract 5g/L, yeast extract 1g/L, peptone 5g/L, magnesium sulfate heptahydrate 0.5g/L, and agar 15g/L for 20D, adding SA with final concentration of 25 μmol/L, and collecting rehmannia root after 12 hr.

Melanin can protect the body from damage caused by sunlight, ultraviolet light, nuclear radiation, ionizing radiation, X-radiation, gamma rays and the like, and is an endogenous anti-radiation agent. The melanin can remove OH and O₂ ^ˉAnd free radicals such as DPPH, chelated Fe²⁺And the lipid peroxidation prevention function and the like, and the stability under the conditions of temperature, illumination, pH and the like is good.

The percentages of lachnum intracellular melanin C, H, N, O and S were 60.71%, 7.61%, 6.19%, 24.94% and 0.55%, respectively, with the presence of carbonyl and-NH 2 or-OH; the silanized derivative of the degradation product of the black sesame melanin is analyzed by gas chromatography-mass spectrometry to find that the degradation product contains catechuic acid, catechol and hydroquinone, and the result shows that the black sesame melanin belongs to catechol melanin.

The curdlan is extracted from pseudomonas fermentation exoproducts, and the purer curdlan is white powdery solid and can be dissolved in water. The molecular weight of curdlan is 618KDa, the curdlan is straight-chain beta-1, 3-glucan, the surface is porous, the specific surface area is large, and the curdlan is a microbial gel and a flocculating agent.

Because of the structural characteristics of curdlan and melanin, curdlan can firmly coagulate melanin together to resist external radiation damage, thereby achieving the effect of protecting skin; can also prevent melanin from entering the skin, and increase the melanin content of the skin.

The gel polysaccharide has the characteristics that the gel polysaccharide can be converted into irreversible high-strength gel by heating to more than 80 ℃, can firmly lock water, and does not influence the permeation of verbascoside, pearl powder, glycerol, pearl powder, arginine, glutathione and vitamin E to the inside of the skin.

Thalli in the pseudomonas fermentation liquor are removed through centrifugation, residual thalli can be killed through high temperature, and residual cane sugar, yeast powder and calcium carbonate in the fermentation liquor can be coagulated and remained in the gel polysaccharide colloid, so that the extraction step is saved; and the yeast powder also has whitening effect.

Acteoside belongs to diglycoside, is p-phenylethanoid glycoside, contains multiple conjugated unsaturated systems and phenolic hydroxyl structural units, and has strong antioxidant activity. Can combine free radicals in vivo, avoid the accumulation of free radicals in vivo, promote the synthesis of DNA and RNA, and effectively resist the biological effect caused by ionizing radiation. Besides, the acteoside has small steric hindrance, is beneficial to approaching to free radicals, and also contains catechol hydroxyl in the structure, so that the acteoside is an important group with antioxidant activity.

An Arabidopsis MYB (21R1R2R3-MYB) gene is a regulation gene for promoting expression of verbascoside polysaccharide, and rehmannia root hairy roots with high verbascoside expression can be obtained by infecting a rehmannia root explant by using agrobacterium rhizogenes. The rehmannia root-like root culture has the advantages of fast growth, stable yield and no influence of seasons compared with a simple plant body. Utilizes the characteristic that Salicylic Acid (SA) can promote the expression of verbascoside, and stimulates the rehmannia glutinosa hairy roots for 12h by using SA to obtain high-concentration verbascoside.

Compared with the prior art, the invention has the following beneficial effects: in the invention, the raw materials are mixed,

(1) the melanin can protect organisms from being damaged by sunlight, ultraviolet light, nuclear radiation, ionizing radiation, X radiation, gamma rays and the like, and the gel polysaccharide can firmly aggregate the melanin together to resist the external radiation damage, so that the skin is protected; can also prevent melanin from entering the inside of the skin and increase the melanin content of the skin;

(2) the curdlan can be converted into irreversible high-strength gel by heating above 80 deg.C, and can firmly lock water content without affecting permeation of verbascoside, hyaluronic acid, Margarita powder, glycerol, Margarita powder, arginine, glutathione, and vitamin E into skin; residual thalli can be killed through high temperature, and residual cane sugar, yeast powder and calcium carbonate in the fermentation liquor can be coagulated and remained in the gel polysaccharide colloid, so that the extraction step is saved; the residual yeast powder also has whitening effect;

(3) the verbascoside has super-strong antioxidant activity, and promotes the large-scale expression of the verbascoside in the rehmannia glutinosa hairy roots by utilizing a transgenic technology; the rehmannia root hairy root culture has the advantages of fast growth, stable yield and no influence by seasons compared with a simple plant body; utilizes the characteristic that Salicylic Acid (SA) can promote the expression of verbascoside, and stimulates hairy roots of rehmannia for 12h by using SA to obtain high-concentration verbascoside.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The invention provides the technical scheme that: an anti-radiation whitening and moisturizing cream comprises the following components in parts by weight: 3-10 parts of verbascoside, 2-7 parts of melanin, 5-10 parts of curdlan, 4-6 parts of glycerol, 3-10 parts of hyaluronic acid, 0.8-3 parts of glutathione, 3-5 parts of emulsifier, 3-10 parts of pearl powder, 0.6-1.5 parts of vitamin E and 8-20 parts of water.

Example 1

An anti-radiation whitening and moisturizing cream comprises the following components in parts by weight: 5 parts of verbascoside, 4 parts of melanin, 8 parts of curdlan, 5 parts of glycerol, 6 parts of hyaluronic acid, 1.8 parts of glutathione, 4 parts of emulsifier, 9 parts of pearl powder, 0.9 part of vitamin E and 15 parts of water.

the method comprises the following steps: stirring melanin, 1/2 water and curdlan fermentation liquor at 90 ℃ and 100rpm for 4h to obtain phase A;

step two: mixing acteoside, hyaluronic acid, glycerol, Margarita powder, arginine, glutathione, vitamin E and 1/2 water at 70 deg.C and 120rpm for 1h, and cooling to room temperature to obtain phase B;

step three: mixing emulsifier with phase A and phase B, stirring at 40 deg.C and 160rpm for 3 hr to obtain cream.

According to the technical scheme, the curdlan fermentation liquor is obtained by utilizing pseudomonas fermentation: 30g/L of sucrose, 8g/L of yeast powder, 3g/L of calcium carbonate, 7.5-8.5 of initial pH value of fermentation liquor, 3-5 days of fermentation period and 30-35 ℃ of fermentation temperature; taking the fermented fermentation liquor, centrifuging at 8000rpm for 30 minutes, taking the supernatant, and evaporating and concentrating at low temperature to obtain the curdlan fermentation liquor.

Example 2

An anti-radiation whitening and moisturizing cream comprises the following components in parts by weight: 4 parts of verbascoside, 5 parts of melanin, 6 parts of curdlan, 4 parts of glycerol, 4 parts of hyaluronic acid, 1 part of glutathione, 3 parts of an emulsifier, 6 parts of pearl powder, 1.2 parts of vitamin E and 10 parts of water.

the method comprises the following steps: stirring melanin, 1/2 water and curdlan fermentation liquor at 90 ℃ and 60rpm for 4h to obtain phase A;

step two: mixing acteoside, hyaluronic acid, glycerol, Margarita powder, arginine, glutathione, vitamin E and 1/2 water at 70 deg.C under stirring at 180rpm for 1h, and cooling to room temperature to obtain phase B;

step three: mixing emulsifier with phase A and phase B, stirring at 40 deg.C and 120rpm for 3 hr to obtain cream.

According to the technical scheme, the curdlan fermentation liquor is obtained by utilizing pseudomonas fermentation: 30g/L of sucrose, 8g/L of yeast powder, 3g/L of calcium carbonate, 7.5-8.5 of initial pH value of fermentation liquor, 3-5 days of fermentation period and 30-35 ℃ of fermentation temperature; taking the fermented fermentation liquor, centrifuging at 6000rpm for 30 minutes, taking the supernatant, and evaporating and concentrating at low temperature to obtain the gel polysaccharide fermentation liquor.

Test 1 mullein glycoside production test

Test samples: inoculating transgenic and non-transgenic hairy roots with length of 2-3cm into MS liquid culture medium, respectively, shake culturing at 26 deg.C and 130r/min in dark for 72h, and cleaning.

And (3) chromatography: the chromatographic column is Athena C18, (4.6X 250mm,5 μm); acetonitrile-0.1% acetic acid (16:84) is used as a mobile phase; the detection wavelength is 334 nm; the flow rate is 1.0mL min < -1 >; the column temperature is 30 ℃; the sample injection amount is 20 mu L; the theoretical plate number is not less than 5000 calculated according to acteoside chromatographic peak; under the above conditions, the separation degree between chromatographic peaks was good.

Preparation of a test solution: precisely weighing 0.4g of rehmannia hairy root powder, placing the powder in a conical flask with a plug, precisely adding 25ml of methanol, weighing the weight, performing reflux extraction at 65 ℃ for 1.5h, cooling to room temperature, and complementing weight loss; shaking up, filtering, precisely measuring 10ml of subsequent filtrate, and concentrating to near dryness; dissolving the residue with mobile phase, transferring into 5ml measuring flask, fixing volume with mobile phase, shaking, and filtering with 0.22 μm microporous membrane.

The reference solution is prepared by precisely weighing herba Thesii glucoside standard 0.08mg, placing in 5ml volumetric flask, and diluting with mobile phase to constant volume to obtain 0.016mg/ml reference solution.

Weighing two groups of hairy roots 85g in parallel, 6 parts of each hairy root, preparing a test solution, injecting samples according to the chromatographic conditions, respectively measuring the content of verbascoside, and calculating the RSD value.

Sampling control solution repeatedly for 6 times according to the above chromatographic conditions, measuring peak area of acteoside, and calculating RSD value.

Acteoside content	1	2	3	4	5	6	RSD％
								Transgenic genome	0.65	0.64	0.68	0.67	0.65	0.69	3.14
Non-transgenic group	0.47	0.48	0.46	0.49	0.48	0.49	2.35

TABLE 1RSD value calculation results

As can be seen from Table 1, the transgenic plants produced more acteoside than the non-transgenic plants, resulting in a 34% increase in yield.

Test 2. radiation protection effect test:

cell culture: DMEM medium containing 10% fetal bovine serum in 5% CO₂And culturing Human Skin Fibroblasts (HSF) in an incubator at 37 ℃.

Establishing an HSF radiation damage model: a radiation damage model is made by a linear accelerator. HSF was collected during logarithmic growth phase and cell suspension concentration was adjusted to 2.5X 10⁴and/mL. The experiments were divided into 3 groups: negative control group of non-radiation 1 group, 16Gy radiation group and 16Gy addition whitening and moisturizing cream protection group, collecting HSF in logarithmic growth phase, and adjusting cell suspension concentration to 2.5 × 10⁴Perml was inoculated in 96-well plates. After 5% CO2 and incubation at 37 ℃ for 48h, 6MV X-ray is used for one-time radiation except for a negative control group, the radiation dose rate is 3Gy/min, the radiation field size is 25cm multiplied by 25cm, and the distance between a radiation source and the center of a target is 100 cm. During radiation, an equivalent scattering plate with the thickness of 1cm is placed on the 96-well plate, and an equivalent scattering plate with the thickness of 3cm is placed below the 96-well plate. The OD of each group of cells was measured by MTT method at 2, 12, 24, 48 after irradiation, respectively₅₇₀Each group of 6 multiple holes.

OD₅₇₀	2	12	24	48
					Negative control group	0.815	0.853	0.919	1.026
Radiation group	0.786	0.794	0.812	0.833
					Protection group	0.823	0.871	0.936	1.102

TABLE 2 OD of the test cells in each group_{570 value}

Group of	Growth rate
		Negative control group	25.89％
Radiation group	5.98％
		Protection group	33.9％

TABLE 3 human skin fibroblast growth rate

As can be seen from Table 3, the growth rate of human skin fibroblasts added with the whitening and moisturizing cream is greatly improved compared with that of a non-protective radiation group, which shows that the anti-radiation whitening and moisturizing cream has strong anti-radiation capability.

Compared with a non-radiation negative control group, the human skin fibroblasts added with the whitening and moisturizing cream are also improved, and the result shows that the anti-radiation whitening and moisturizing cream disclosed by the invention has the function of promoting the human skin fibroblasts.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An anti-radiation whitening and moisturizing cream comprises the following components in parts by weight: 3-10 parts of verbascoside, 2-7 parts of melanin, 5-10 parts of curdlan, 4-6 parts of glycerol, 3-10 parts of hyaluronic acid, 0.8-3 parts of glutathione, 3-5 parts of emulsifier, 3-10 parts of pearl powder, 0.6-1.5 parts of vitamin E and 8-20 parts of water.

2. A preparation method of an anti-radiation whitening and moisturizing cream comprises the following specific steps:

3. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 2, characterized in that: the melanin is extracted from Lachnum micranthum fermentation liquor or semen Sesami Niger.

4. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 2, characterized in that: the curdlan fermentation liquor is obtained by utilizing pseudomonas fermentation: 30g/L of sucrose, 8g/L of yeast powder, 3g/L of calcium carbonate, 7.5-8.5 of initial pH value of fermentation liquor, 3-5 days of fermentation period and 30-35 ℃ of fermentation temperature; taking the fermented fermentation liquid, centrifuging for 30 minutes at 8000rpm of 5000-.

5. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 2, characterized in that: the verbascoside is extracted from rehmannia glutinosa hairy root.

6. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 5, characterized in that: the rehmannia root hairy roots are infected with the agrobacterium rhizogenes A4, and the explant age is 7-14D.

7. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 6, characterized in that: infecting rehmannia leaves (0.5 x 0.5cm) by using agrobacterium rhizogenes for 5-10min, then sucking dry bacterial liquid on sterile filter paper, inversely placing the leaves in co-culture solid culture media, placing 10 leaves in each culture dish, co-culturing at 26 ℃ under a dark condition, wherein the co-culture media are MS media, and adding 100 mu mol/L AS; after co-culture for 2 days, washing with sterile water for 3 times, sucking the liquid with sterile filter paper, inoculating the leaf into a screening medium, and performing dark culture at 26 ℃ in the presence of MS + Cef (0.4g/L) + AS (100 μmol/L).

8. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 7, characterized in that: the Ri plasmid of the agrobacterium rhizogenes contains an Arabidopsis MYB (21R1R2R3-MYB) gene, the MYB gene is amplified through PCR, and the MYB gene is connected to the Ri plasmid after double cutting by enzyme.

9. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 8, characterized in that: and (3) performing PCR detection on the viable rehmannia glutinosa hairy roots of the screening culture medium: according to the designed pair of MYB gene primers, a band which is the same as the gene primer appears in the electrophoresis detection, but the band is not amplified in the unconverted rehmannia root hairy root, so that the recombinant gene is proved to be integrated into the genome of the rehmannia root hairy root.

10. The preparation method of the anti-radiation whitening and moisturizing cream according to claim 9, characterized in that: the rehmannia root-like roots are transferred into a liquid culture medium for amplification and multiplication culture: culturing with YEB culture medium, beef extract 5g/L, yeast extract 1g/L, peptone 5g/L, magnesium sulfate heptahydrate 0.5g/L, and agar 15g/L for 20D, adding SA with final concentration of 25 μmol/L, and collecting rehmanniae radix hairy root after 12 hr.