CN115105448B

CN115105448B - Preparation method and application of skin care product raw materials for preventing radiation, promoting blood circulation, nourishing face and resisting aging

Info

Publication number: CN115105448B
Application number: CN202210827829.3A
Authority: CN
Inventors: 贺英
Original assignee: Qinhuangdao Purun High Tech Development Co ltd
Current assignee: Qinhuangdao Taizhi Medical Technology Co ltd
Priority date: 2022-07-14
Filing date: 2022-07-14
Publication date: 2023-07-18
Anticipated expiration: 2042-07-14
Also published as: CN116270345A; CN116270345B; CN115105448A

Abstract

The application relates to the technical field of radiation-proof cosmetics, in particular to a preparation method and application of a skin care product raw material for radiation protection, blood circulation activating, skin care and aging resistance. The skin care product can be in the form of emulsion, is prepared by using a composition consisting of the tobacco extract and the rose extract, has the effect of radiation protection on skin tissues, has the effects of activating blood circulation, nourishing skin and resisting aging compared with the skin care emulsion which does not have the composition generally, and has wide application prospect.

Description

Preparation method and application of skin care product raw materials for preventing radiation, promoting blood circulation, nourishing face and resisting aging

Technical Field

The application relates to the technical field of radiation-proof cosmetics, in particular to a preparation method and application of a skin care product raw material for resisting radiation, activating blood, nourishing face and resisting aging.

Background

In daily life, people are exposed to solar radiation and electromagnetic radiation. The light aging caused by ultraviolet rays in sunlight is a damage accumulation process, and the ultraviolet rays can promote skin dryness, quicken skin aging, generate or deepen black spots and freckles, or cause dermatitis and possibly even skin cancer. Most of the skin care products with the radiation protection function sold in the prior art are added with heavy metals such as lead and the like, which cause harm to human health. Therefore, it is very necessary to develop a natural and harmless skin care product with the function of radiation protection.

Disclosure of Invention

In view of this, the purpose of the present application is to avoid selecting heavy metal elements as raw materials of skin care products, such as natural herbaceous crops, so as to reduce the harm of skin care products themselves to skin.

In a first aspect, embodiments of the present application disclose a composition for use in a cosmetic comprising a tobacco extract and a rose extract, the cosmetic comprising the composition and other ingredients acceptable in the cosmetic; wherein, based on the total weight of the skin care emulsion, the tobacco extract accounts for 0.001 to 3.5 weight percent, and the rose extract accounts for 0.001 to 15 weight percent.

In an embodiment of the present application, the tobacco extract comprises anthocyanin, procyanidine A1, procyanidine C, 3-hydroxyflavone, chalcone, aurone, isorhamnetin-3, 7-O-diglucoside, isorhamnetin-3-O-glucoside, kaempferol-7-O-beta-D-glucoside, kaempferol-3-O-rutinoside, quercetin 3,4' -diglucoside, and quercetin 3-O- (6 "-galloyl) -beta-D-glucoside.

In an embodiment of the present application, the tobacco extract is extracted with a mixed reagent comprising water, acetone, methanol, formic acid, trifluoroacetic acid and ethyl acetate.

In embodiments of the present application, the rose extract comprises phenethyl alcohol, 1-nonanol, linalool, geraniol, 2-heptanol, acetic acid, benzyl alcohol, isopentenyl alcohol, 2-methylbutanol, 2-propanol, 1-octanol, nerol, eugenol, methyl eugenol, 2, 6-bis (1, 1-dimethylethyl) -4-methylphenol, allyl nodular ether, benzaldehyde, neral, nonanal, decanal, geranial, undecanal, citronellal, methylheptenone, 2-nonanone, 2-tridecanone, 2-undecanone, 2-heptanone, geranylacetone, terpinene, alpha-cedrene, thunberene, myrcene, alpha-travelling alkene, dipentene, alpha-farnesene, neryl formate, citronellyl acetate, geranyl acetate, benzyl acetate, methyl eicosanoate, pear alcohol ester, n-tridecane, 3-methylene-1, 1-dimethyl-alpha-vinylcyclohexane, nitrone hydrochloride, and amine salts of methyl-isopropyl alcohol hydrochloride.

In embodiments of the present application, the additional acceptable ingredients are selected from one or more of emollients, film forming agents, emulsifiers, humectants, stabilizers, and solvents.

In the embodiment of the application, the product form of the cosmetic can be one or more of foundation liquid, foundation cream, barrier cream, pre-makeup cream, face-cleansing cream, skin-care emulsion, concealer liquid and face-repair cream.

In a second aspect, embodiments of the present application disclose a skin care lotion comprising, by weight, 0.02 to 3.2% of a tobacco extract, 0.001 to 8.5% of a rose extract, and other ingredients acceptable in cosmetics.

In the embodiment of the application, the skin care cream comprises 1.8-3.0 wt% of tobacco extract, 3-7 wt% of rose extract, 0.01-2.3 wt% of casein phosphopeptide, 3-8 wt% of PEG-12 polydimethylsiloxane, 8-15 wt% of propylene glycol, 0.1-0.5 wt% of magnesium stearate and the balance of water.

In the examples herein, 1.8 to 2.0wt% of tobacco extract, 5 to 6.5wt% of rose extract, 0.01 to 1.65wt% of fucoidin, 3 to 8wt% of sorbitan olive oleate, 8 to 15wt% of 1, 2-pentanediol and 0.1 to 0.5wt% of magnesium stearate, and the balance being water.

In a third aspect, the application discloses application of the composition of the first aspect or the skin care emulsion of the second aspect in preparing skin-care preparations for promoting blood circulation and beautifying.

In the embodiments of the present application,

compared with the prior art, the application has the following beneficial effects:

the skin care emulsion is prepared from the composition consisting of the tobacco extract and the rose extract, has the effect of radiation protection on skin tissues, has the effects of activating blood circulation, nourishing skin and resisting aging, and has wide application prospect compared with the skin care emulsion which does not have the composition generally.

Drawings

Fig. 1 is an electron microscope image of the skin tissue of a mouse provided by the normal group of animal experiments in the present application.

Fig. 2 is an electron microscope image of the skin tissue of the mice provided by the animal experiment model group.

Fig. 3 is an electron microscope image of the skin tissue of the mice provided by the positive control group of the animal experiment in the application.

Fig. 4 is an electron microscope image of the skin tissue of the mice provided by the experimental animal group (example 2) of the present application.

Fig. 5 is an electron microscope image of the skin tissue of the mice provided by the experimental animal group (example 3) of the present application.

FIG. 6 is an electron micrograph of the skin tissue of a mouse provided by the experimental animal group (comparative example 1) of the present application.

Fig. 7 is an electron micrograph of the skin tissue of a mouse provided by the experimental animal group (comparative example 2) of the present application.

Fig. 8 is an electron micrograph of the skin tissue of a mouse provided by the experimental animal group (comparative example 3) of the present application.

Fig. 9 is an electron micrograph of the skin tissue of a mouse provided by the experimental animal group (comparative example 4) of the present application.

Detailed Description

In order to make the objects, technical solutions and advantages of the present application more apparent, the present application will be further described in detail with reference to examples. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the present application.

The inventor creatively discovers that the tobacco extract and the rose extract are subjected to homogenization treatment to prepare the water-in-water type multi-suspension liposome skin care emulsion which has liposome results, is easier to be absorbed through skin penetration, can form a skin cell tissue barrier, has the function of preventing radiation and aging by applying skin care products, and has stable multiphase structure. In addition, the inventor prepares the skin care product into a skin care product, which not only has the radiation-proof effect on skin tissues, but also has the effects of activating blood circulation, nourishing skin and resisting aging. The following examples are provided to illustrate the reagents and apparatus which are not specifically described in the following examples, and the experimental methods which are not specifically described are all conventional experimental methods.

Tobacco extract

Tobacco flower (academic name: nicotiana alata Link et Otto) is a perennial herb of the family Solanaceae, the genus Nicotiana is limited, up to 1.5 meters, and the whole is adhered with hair. The basal part of the blade slightly holds the stem or has a wing-shaped handle, and forms an oval or oval oblong shape upwards, and the basal part is near to the stem or has no ear, and the basal part is near to the inflorescence to form a needle shape. Pseudo-overall inflorescence type, for evacuating several flowers; calyx cup-shaped or bell-shaped, split drill-shaped needle-shaped, unequal in length; the corolla is light green, stamens are not equal in length, capsules are egg-shaped, and seeds are grey brown. Argentina and Brazil are original products. There are introduced cultivation in the cities of Harbin, beijing, nanjing and the like. The flower tobacco is mainly used as the cultivated flower, has bright color and rich color, and has good ornamental value. The embodiment of the application carries out the relevant extraction steps to obtain the tobacco extract, and the tobacco extract is applied to skin care products. The process for extracting the tobacco flower is as follows:

1. materials and methods

1. Flower tobacco and primary material for extraction

The flower tobacco is purchased in the Beijing flower market. Collecting flowers in the period of mucous secretion of the stigmas in the full-bloom stage in sunny days, taking colored petal parts, and sealing and preserving.

2. Extraction process

Grinding the above materials with liquid nitrogen to powder, extracting with 10 mL/g of extracting agent, mixing extractive solutions, sealing during each extraction process for 24 hr or more, rotary evaporating, concentrating, and drying to obtain tobacco extract.

Specifically, the extraction reagent in example 1 is a mixed solution of water, acetone, methanol, formic acid, trifluoroacetic acid and ethyl acetate, and the volume ratio of the formulation is 65:65:80:2:1:1. The extraction reagent of comparative example 1 is a mixed solution of water, methanol, formic acid and trifluoroacetic acid, and the volume ratio of the formulation is 27:70:2:1.

3. Determination of the content of anthocyanidin-related Components

The extracts of example 1 and comparative example 1 were dissolved with the corresponding extraction reagents, and then sequentially filtered through filter paper and 0.22 μm filter paper, and the filtrate was used as a sample.

The standard substance is as follows: anthocyanin (Beijing carboline technologies Co., ltd.), procyanidin A1 (Shanghai Yuan Ye Biotechnology Co., ltd.) and procyanidin C (Shanghai Yuan Ye Biotechnology Co., ltd.).

The HPLC-TOF-MS method is adopted to measure the content of the related anthocyanin in the extract, and the chromatographic conditions are as follows: the column was an Alltima HP HILIC 86466 (250 mm. Times.4.6 mm, beijing and Yu Wei technology Co., ltd.); the workstation is EZchrom Elite, version 3.2.1; a detector UV6000; the detection wavelength is 290nm; column temperature is 25 ℃; sample injection amount is 10 mu L;

the flow rate was 0.8mL/min.

The elution conditions were: the mobile phase adopts phase A (acetonitrile) and phase B (methanol: water volume ratio is 98.5:1.5) for gradient elution, and the elution procedure is 0-3 min (100% A); 3 to 20min (100 to 70 percent of A); 20 to 30min (70% → 0% A); 30 to 32min (0% → 100% A); 32.fwdarw.35 min (100% A).

4. Determination of the content of flavonols and their sugar-generic compounds

Detecting a sample: the extracts of example 1 and comparative example 1 were dissolved with the corresponding extraction reagents, and then sequentially filtered through filter paper and 0.22 μm filter paper, and the filtrate was used as a sample. .

The standard substance is as follows: kaempferol-7-O- β -D-glucoside (CAS: 16290-07-6, ai Ke reagent, chromatographically pure), kaempferol-3-O-rutinoside (CAS: 17650-84-9, abisma (Shanghai) Biotechnology Co., ltd., purity 98%), quercetin 3,4' -diglucoside (CAS: 29125-80-2, shanghai-derived leaf Biotechnology Co., ltd., HPLC. Gtoreq.98.5%), quercetin 3-O- (6 "-galloyl) -beta-D-glucoside (CAS: 56316-75-7, HPLC. Gtoreq.98.5%, shanghai-associated Mich Biotechnology Co., ltd.), 3-hydroxyflavone (CAS: 577-85-5, beijing-BAILING Co., purity 98.5%), chalcone (CAS: 94-41-7, beijing-BAILING Co., purity 98.0%), and hesperidone (CAS: 38216-53-4, beijing NOKOKatsumami Co., ltd., purity 98%), isorhamnetin-3, 7-O-diglucoside (CAS: 6758-51-6, sesaminide), isorhamnetin-3-O-3 (CAS: 94-41-7, biotechnology Co., ltd., purity 50%), 50%)

HPLC-DAD analysis conditions: triple quadrupole liquid chromatograph-mass spectrometer (zemoeid), DAD detector, wavelength 350nm; mobile phase: phase A is acetonitrile and phase B is 80.1% formic acid aqueous solution; the column was Angilent Eclipse plus C (150X 2.1mm,5 μm); the flow rate is 0.3mL/min; the column temperature is 35 ℃; the sample injection amount is 5 mu L; gradient elution.

And (3) preparing a standard curve according to peak areas corresponding to different concentrations of each standard substance, calculating the concentration of related active ingredients in the sample according to the standard curve and the peak areas corresponding to the sample, and calculating the content of each active ingredient in the extract according to the concentration.

2. Results

TABLE 1

Table 1 shows the anthocyanin-related component, flavonols and glycoside component contents of the tobacco extract obtained in example 1 and comparative example 1, respectively, wherein "-" indicates undetected. As is clear from Table 1, the tobacco extracts prepared in example 1 and comparative example 1 each contained anthocyanin, procyanidin A1 and procyanidin C, but the extracts prepared in example 1 contained low anthocyanin content, whereas procyanidin A1 and procyanidin C contained high anthocyanin content, whereas comparative example 1 contained the opposite anthocyanin. The extracts obtained in example 1 and comparative example 1 each contained 3-hydroxyflavone, chalcone, orange ketone, kaempferol-7-O-beta-D-glucoside, kaempferol-3-O-rutinoside, quercetin 3,4 '-diglucoside and quercetin 3-O- (6' -galloyl) -beta-D-glucoside in a small amount. The extract obtained in comparative example 1 did not contain isorhamnetin-3, 7-O-diglucoside and isorhamnetin-3-O-glucoside.

Rose extract

The flos Rosae Rugosae is of Rosa of Rosaceae, has edible fruit, no sugar, and is rich in vitamin C, and is commonly used in herb tea, jam, jelly, fruit juice, bread, etc.; the fresh flower can be steamed into aromatic oil, the main component of the oil is levo aromatic alcohol, the content of the oil can reach six thousandths, the fresh flower can be used for eating and cosmetics, the petals can be used for preparing cake stuffing, rose wine and rose syrup, and after the fresh flower is dried, tea can be made, and the flower buds can be used as medicines for treating liver, stomach ache, chest and abdominal distension and irregular menstruation.

The embodiment of the application carries out relevant extraction steps to obtain the rose extract, and the rose extract is applied to skin care products. The extraction process for roses is as follows:

1. rose extraction process

The roses referred to in this application are purchased from the Beijing flower market. The extraction process is as follows:

accurately weighing 200g of dried rose petal powder, placing into 500mL round bottom flask, adding 200mL distilled water, mixing, treating with microwave hydrothermal synthesis instrument (XH-800 SP, beijing Xiangnin) for 20min and 27kW, transferring to connected steam distillation device for extraction, adding NaCl into distillate, standing, extracting with 50mL diethyl ether for 3 times, mixing the extractive solutions, and anhydrous Na ₂ SO ₄ Drying, filtering, rotary evaporating to recover diethyl ether, and vacuum drying at 40deg.C for 1 hr to obtain flos Rosae Rugosae extract.

2. Related component detection

And (3) analyzing relevant components in the rose extract by adopting a gas chromatography-mass spectrometry (GC-MS). The chromatographic column is HP-5MS (Agilent, 19091S-433-INT) with a specification of 30m×0.25mm×0.25 μm; the carrier gas is nitrogen, and the air inflow is 1.0mL/min; the temperature of the sample inlet is 260 ℃; heating to 250 ℃ at a speed of 50 ℃/min; 1 sample injection amount, no split flow; EI ion source, 70eV, scanning range 50-650 amu, transmission line temperature 250 ℃. And (3) automatically searching through a WILEY and MAINLIB spectrum library, and simultaneously adopting a retention index and a reference document to assist mass spectrum searching qualitative, and calculating the relative content of each component according to a peak area normalization method of a total ion flow chromatographic peak by a quantitative analysis result.

3. Results

TABLE 2 relative content of rose extract components (%)

Table 2 shows the relative amounts of alcohols, phenols, aldehydes, ketones, alkenes, esters and ingredients in the rose extract, for a total of 48 ingredients, with higher amounts of alcohols.

Composition and skin care product

The present inventors found that a composition which can be used in cosmetics is obtained by mixing the tobacco extract and the rose extract obtained as described above. The cosmetic comprises the composition and other acceptable ingredients in the cosmetic; wherein, based on the total weight of the skin care emulsion, the tobacco extract accounts for 0.001 to 3.5 weight percent, and the rose extract accounts for 0.001 to 15 weight percent. The composition is applied to cosmetics, has the effect of radiation protection on skin tissues, and also has the effects of activating blood circulation, maintaining beauty and resisting aging.

Wherein the tobacco extract is prepared from example 1, specifically, the tobacco extract comprises anthocyanin, procyanidine A1, procyanidine C, 3-hydroxyflavone, chalcone, aurone, isorhamnetin-3, 7-O-diglucoside, isorhamnetin-3-O-glucoside, kaempferol-7-O-beta-D-glucoside, kaempferol-3-O-rutinoside, quercetin 3,4 '-diglucoside and quercetin 3-O- (6' -galloyl) -beta-D-glucoside.

Wherein the rose extract is prepared from the above embodiments. In particular, the rose extract comprises phenethyl alcohol, 1-nonanol, linalool, geraniol, 2-heptanol, acetic acid, benzyl alcohol, isopentenyl alcohol, 2-methylbutanol, 2-propanol, 1-octanol, nerol, eugenol, methyl eugenol, 2, 6-bis (1, 1-dimethylethyl) -4-methylphenol, allyl nodal ether, benzaldehyde, neral, nonanal, decanal, geranial, undecanal, citronellal, methylheptenone, 2-nonanone, 2-tridecanone, 2-undecanone, 2-heptanone, geranylacetone, terpinene, alpha-cedrene, thunber, myrcene, alpha-farnesene, neryl formate, citronellyl acetate, geranyl acetate, benzyl acetate, methyl eicosanoate, sorbitol ester, n-tridecane, 3-methylene-1, 1-dimethyl-alpha-vinylcyclohexane, nitrone hydrochloride, and nitrone hydrochloride.

In the cosmetic prepared using the composition, the product form of the cosmetic may be one or more of foundation, make-up cream, pre-make-up cream, facial cleanser, skin cream, skin lotion, concealer, and make-up cream, but is not limited thereto, and skin cream is preferred.

In particular embodiments, the other acceptable ingredients are selected from one or more of emollients, film formers, emulsifiers, moisturizers, stabilizers, and solvents, all in amounts conventional for cosmetic products.

In embodiments of the present application, the emollient is selected from one or more of ethoxy-modified silicone, cyclopentadimethicone, phenyl trimethicone, tocopheryl acetate, pentaerythritol tetraisostearate, butylene glycol dicaprylate/dicaprate, isododecane, diisostearyl malate octyl polymethylsiloxane, long chain alcohol benzoate, oil soluble lanolin, bean myristate, octyl octanoate, oleyl oleate, hexyl laurate, isopropyl palmitate, PEG-6 caprylate/caprate triglyceride, PEG-60 corn oil glycerol mixed ester, PEG-7 olive oleate, glycerol tricaprylate, glycerol tricaprate, and isotridecyl alcohol isononanoate. However, without limitation, emollients commonly used in the art may be used, and the present application is preferably PEG-6 caprylic/capric triglyceride, oil-soluble lanolin, and long chain alcohol benzoate.

In the embodiments of the present application, the film forming agent is selected from one or more of protein film forming agents, acrylic resin film forming agents, polyethylene copolymers, copolymers of various hydrocarbons, silicone polymers and silicone acrylates, but is not limited thereto, and film forming agents commonly used in the art may be used. Preferred film formers for embodiments of the present application are protein film formers such as casein, fucoidan, and the like.

In an embodiment of the present application, the emulsifier is selected from one or more of the group consisting of PEG-12 polydimethylsiloxane, PEG-12 polydimethylsiloxane ethyl polydimethylsiloxane, cetyl poly PEG-10/PPG-10/1 polydimethylsiloxane, sorbitan olive oleate, and PEG-10/PPG-18/18 polydimethylsiloxane. But not limited thereto, an emulsifier commonly used in the art may be used.

In embodiments herein, the humectant is selected from one or more of glycerin, butylene glycol, 1, 2-pentanediol, sodium hyaluronate, and propylene glycol.

In an embodiment of the present application, the stabilizer is selected from one or more of sodium hydrogen phosphate, sodium stearate, and magnesium stearate.

In embodiments of the present application, the solvent is selected from water and/or ethanol. However, the solvent is not limited thereto, and solvents commonly used in the art may be used, and water and ethanol are preferred herein, and the water is deionized water, purified water, distilled water, or the like.

Specifically, the embodiment of the application provides skin care cream which comprises 0.02-3.2 wt% of tobacco extract, 0.001-8.5 wt% of rose extract and other acceptable components in cosmetics.

Preferably, the skin care cream comprises 1.8 to 3.0wt% of tobacco extract, 3 to 7wt% of rose extract, 0.01 to 2.3wt% of casein phosphopeptide (Shaanxi Lloyd Biotechnology Co., ltd.), 3 to 8wt% of PEG-12 polydimethylsiloxane, 8 to 15wt% of propylene glycol and 0.1 to 0.5wt% of magnesium stearate, and the balance of water.

Preferably, the skin care cream comprises 1.4 to 2.0wt% of tobacco extract, 5 to 6.5wt% of rose extract, 0.01 to 1.65wt% of fucoidan (cistanin, purity 98%), 5 to 9.5wt% of sorbitan olive oleate, 10 to 18wt% of 1, 2-pentanediol and 0.1 to 0.5wt% of magnesium stearate, and the balance being water.

The formulation of the skin care cream provided in the present application is shown in table 3.

Table 3 skin care cream formulation

The preparation methods of examples 2 to 7 and comparative examples 1 to 2 specifically include:

1) Heating and fully mixing the components of the formula A to realize homogenization;

2) Dissolving the phase B component in water to obtain an aqueous phase solution;

3) Adding phase A and phase B into homogenizing emulsifying machine, homogenizing, performing microfluid pressurizing treatment for multiple times, and filtering to obtain skin care emulsion.

The skin care cream of comparative example 3 was prepared by:

1) Weighing 50g of rheum officinale and 50g of phellodendron, cleaning, mixing, decocting with water twice, and combining the decoctions. Precipitating with ethanol for 48 hours, filtering to obtain filtrate, and concentrating the filtrate under reduced pressure to obtain concentrated solution, 200mL;

2) 15g of tragacanth (content 99%, xian Zhengzhu pharmaceutical excipients Co., ltd.), 1.0g of propionic acid innocuous pill ketone and 500mL of clear cod liver oil are weighed and mixed to be used as an oil phase;

3) 1.0g of hydrocortisone, 10.0g of tetracaine hydrochloride, 250g of honey and 0.8g of nipagin are weighed according to the prescription, and 500mL of distilled water is added to dissolve various components to be used as a water phase. Under strong stirring, slowly pouring the water phase concentrated solution and the Chinese medicinal liquid into the oil phase solution in a fine flow state for emulsification, and obtaining the skin care emulsion after the emulsification is completed.

Characterization of skin care milk Performance

1. Stability investigation

The prepared nanometer skin care emulsion is placed for 30d at the temperature of 4 ℃,25 ℃ and 60 ℃ respectively, and the average particle size and Zeta potential value of the skin care emulsion are measured for 1 time per Sd.

And diluting a proper amount of nano skin care emulsion to be detected by deionized water for 1 000 times, shaking uniformly, and respectively measuring the average particle size and Zeta potential of the prepared nano skin care emulsion by using a ZS90 type nano particle size and Zeta potential analyzer of Malvern company in UK.

2. Results

TABLE 4 average particle size (nm)

TABLE 5 Zeta (mV)

The average particle diameters of the nano skin care emulsions provided in examples 2 to 7 and comparative examples 1 to 4 after being stored at 4 ℃,25 ℃ and 50 ℃ for 30 days, respectively, are shown in table 4, and as a result, the average particle diameters of the nano skin care emulsions provided in examples 2 to 7 and comparative examples 1 to 2 were found to be less changed within 30 days under the three temperature environments. The average particle diameter of the nano skin care emulsion provided in comparative example 3 increases more when the emulsion is stored for 0 to 30 days. The initial average particle size of the nano skin care emulsion provided in comparative example 4 exceeds 200nm, and the nano skin care emulsion can basically undergo precipitation and polymerization at 50 ℃ and has a nano configuration damaged.

The Zeta potentials of the nano skin care emulsions provided in examples 2 to 7 and comparative examples 1 to 4 after 30 days of storage at 4 ℃,25 ℃ and 50 ℃ respectively are shown in table 5, and as a result, the nano skin care emulsions provided in examples 2 to 7 and comparative examples 1 to 2 were found to be lower in Zeta potentials in these three temperature environments and less in change within 30 days. The nano skin care emulsion provided by the comparative examples 3-4 has lower Zeta potential increase when stored for 0-30 days, and the nano configuration of the skin care emulsion provided by the two surfaces is seriously damaged, so that the stability is poor.

Ultraviolet radiation resistant activity of skin care milks

The application further verifies that the skin care cream provided by each example and comparative example has ultraviolet radiation resistant activity, and is specifically as follows:

1. UVA radiation human epidermis keratinocyte injury model

Human epidermal keratinocytes scientific, cat: 2110 Divided into 6 groups, normal cell control group and 5 UVA damaged groups. Culturing human epidermal keratinocyte with DMEM medium to fusion degree of 70-80%, placing into 96-well plate, adding 100 μl cell suspension into each well, 4 multiple wells, incubating for 24 hr, and performing ultraviolet irradiation on each UVA injury groupTreating, wherein the cell is 15cm away from an irradiation light source, and the irradiation intensity is 5.4W/m ² The irradiation time is respectively 15, 20, 30, 60 and 120min, and after the culture is continued for 24h, the MTT method is adopted for detection. When the UVA irradiation time was 30min, the cell death rate was around half the death value, which was the ideal dose for modeling, so the subsequent UVA damage building uv-radiation resistant active mode experiment of the paint seed oil nanoemulsion of table 2 was chosen for an irradiation time of 30 min.

2. In vitro anti-UVA radiation assay

Cells were randomly divided into control, model and experimental groups.

Human epidermal keratinocytes were inoculated into 96-well cell culture plates (6 000 cells/well) and placed in a volume fraction of 5% CO ₂ Culturing in an incubator at 37 ℃ for 24 hours; firstly, receiving UVA radiation by a model group and an experimental group according to a damage model result, wherein UVA radiation is not performed by a normal group; then, nanometer skin care emulsion with serial mass concentration (5 mg/L, 1mg/L and 0.1 mg/L) is respectively added into cell fluid of an experiment group and a control group, the same amount of DMEM culture medium is added into a model group, and after incubation is carried out for 24 hours, the detection is carried out by adopting an MTT method.

3. MTT assay

After adding 20 mu L of MTT solution with the mass concentration of 5g/L into each hole of the cell culture plate and continuously culturing for 4 hours, removing supernatant, adding 100 mu L/hole of DMSO, and carrying out light-proof oscillation for 15min to detect an Optical Density (OD) value by using a microplate reader, detecting a light absorption value at a wavelength of 570nm, and calculating the cell survival rate. Cell viability = experimental absorbance at/blank absorbance x 100%.

5. Data processing

All experimental data are expressed as mean and standard deviation, and data were processed using SPSS13.0 software and subjected to multiple comparisons and significance differential labeling.

6. Results

TABLE 6 cell viability (%)

Table 6 shows the results of in vitro anti-UVA experiments on human epidermal keratinocytes by the skin care milk provided herein.

As shown in Table 6, the cell activities of the human epidermal keratinocytes of the model group were 51.6% after irradiation with ultraviolet rays, which indicates that the modeling was successful. The skin care milk provided by the embodiment of the present application may have the effect of promoting the proliferation of human epidermal keratinocytes, as the cell activity of the skin care milk provided by the embodiment of the present application is rather increased in the skin care milk culture provided by the embodiment 2 of different concentrations of the control group.

In the experimental group, the skin care milk provided in examples 2 to 7 has an accelerating effect on the activity of human epidermal keratinocytes under ultraviolet irradiation. Whereas the skin care milks provided in comparative examples 1,2 and 4 had activities against human epidermal keratinocytes under ultraviolet irradiation, the skin care milk provided in comparative example 4 even resulted in a decrease in activities of human epidermal keratinocytes. The skin care cream provided in comparative example 3 also has an accelerating effect on the activity of human epidermal keratinocytes under ultraviolet irradiation and an ultraviolet irradiation resisting effect, but the stability analysis of the skin care cream provided by the skin care cream is combined, so that the stability is poor, and the skin care cream has no practical storage and application value.

Animal experiment

1. Experimental animal

SPF-class SD female rats (medical laboratory animal center, guangdong province) including naturally aged rats with aged teeth, body weight (490+ -20) g; the weight of the young rats (170+/-20 g), the temperature (25+/-1) DEG C and the relative humidity (70%).

2. Test article

The skin care creams provided in examples 2 to 7 and comparative examples 1 to 4, commercially available repair creams (appropriately herbal) served as positive controls. The Kema depilatory cream was purchased from manting cosmetic limited, guangzhou.

3. Experiment grouping and experiments

Taking naturally-aged SPF-class SD female aged rats, and randomly dividing the SPF-class SD female aged rats into an experimental group, a positive control group and a model group 3, wherein 10 rats are selected from each group; 10 SPF-class SD female cyan-aged rats were also taken as normal control groups.

The rats of each group were placed under the same environment and fed with the same feed, and the back of the rats was dehaired with the Kema dehaired cream 24 hours before the experiment, and the dehaired area was 2.5cm×2.5cm.

After observing the skin for no irritation, the following operations were performed on the dehaired skin areas, respectively: the experimental groups smeared 100mg of the skin care milk provided in examples 2 to 7 and comparative examples 1 to 4; the positive control group was coated with 100mg of commercial repair cream; the model control group was not treated.

After the skin to be tested is lightly massaged until the medicine is safely absorbed, each group is covered by a preservative film and fixed and protected by an adhesive tape for 1 hour. 2 times daily for 30 days, 3 treatment courses.

4. On day 31, all rats were sacrificed by cervical dislocation, and 1cm of the dehairing sites of each rat was cut with small ophthalmic scissors ² After removing adhered fat, 100mg of skin to be tested is weighed on an electronic precision balance, and digested with 2ml of 6M HCL solution, and the digested solutions are centrifuged, and then supernatant is taken as a detection sample.

5. Transmission electron microscope detection

After digestion, the cells of each group after drug drying were washed 1 time with precooled PBS (pH 7.4), centrifuged at 1500r/min for 5min, the supernatant was discarded, 1ml of 2.5% glutaraldehyde solution was added, fixed at 4℃for 24h, stepwise ethanol dehydration was performed, and the resin-embedded sections were observed by transmission electron microscopy.

6. DNA global methylation level detection

Extracting DNA from each group of samples after drug is dried according to the instruction of the kit, adding the sample according to the instruction, ensuring that the liquid in each hole is at the bottom, placing the liquid at 37 ℃ for incubation for 60min, washing the liquid on a plate washer for 3 times, adding 5-me Detection Complex Solution, incubating for 50min at room temperature, adding DS, placing the liquid at room temperature for 5min, stopping the reaction when 50% of PC holes turn into deep blue, and reading the OD value at 450nm of an enzyme-labeling instrument when the color change is complete.

6. Data processing

7. Results

As shown in figure 1, the skin fibroblasts of the normal group mice have plump cell morphology, uniform electron density, complete nuclear membrane, visible nucleoli, abundant cytoplasm, complete organelle structure, regular mitochondrial morphology, complete body purification and abundant matrix. As shown in fig. 2, the cell morphology of the model group mice is obviously changed, the cell electron density is reduced, the nuclear membrane and nucleolus are unclear, the cytoplasmic components are obviously reduced, a large amount of vacuoles are formed in cytoplasm, mitochondria are obviously swelled and deformed, and the morphology is irregular. As shown in fig. 3 to 5, the cells of the experimental group (examples 2 to 3) and the positive control group had substantially complete cell morphology, uneven electron density, complete nuclear membrane, visible nucleoli, reduced cytoplasmic components, and reduced number of organelles with clear structures in the cytoplasm; more bilayer or unilateral closed membrane structures are seen, endocytosing cytoplasmic components of mitochondria or other organelles, suggesting autophagosome formation. As shown in fig. 6 to 9, the cell morphology of the mice of the experimental group (comparative examples 1 to 4) was significantly covered, the electron density was lowered, the nuclear membrane and nucleolus were unclear, the cytoplasm was decreased, the cell damage was serious, and especially comparative example 4, thereby indicating that the skin lotions provided in comparative examples 1 to 4 could not exert an anti-aging effect on the skin cells of the mice of the model group.

TABLE 7 OD450

As can be seen from Table 7, the overall methylation level of the DNA was significantly reduced in the model group compared to the normal group, while that of the positive control group. In the experimental group, the skin care cream provided in examples 2 to 7 showed a significant increase in overall methylation water of DNA after intervention in model mice, whereas comparative examples 1 to 4 did not have a significant promoting effect.

From the above results, it was found that the destruction of cell organelles, especially mitochondria, in human skin fibroblasts was significantly reduced in the large and medium dose group, and the cell morphology was also significantly improved, as compared with the model group. The overall methylation level of the cell DNA is obviously reduced, and the cell ultrastructure has obvious aging changes such as mitochondrial swelling, nuclear membrane inward folding and the like. After the skin care emulsion provided by the embodiment of the application intervenes in the aging mice, the DNA methylation level is higher, so that the skin care emulsion provided by the embodiment of the application has the effect of resisting the aging process of the mice, plays a role in skin care and beautifying, reduces damage to skin organelles of the mice, particularly mitochondria, and improves cell morphology.

In summary, the skin care emulsion is prepared by using the composition composed of the tobacco extract and the rose extract, has the effect of radiation protection on skin tissues, has the effects of activating blood circulation, maintaining beauty and resisting aging, and has wide application prospect compared with the skin care emulsion which does not have the composition generally.

The foregoing is merely a preferred embodiment of the present application, but the scope of the present application is not limited thereto, and any changes or substitutions easily contemplated by those skilled in the art within the technical scope of the present application should be covered by the scope of the present application.

Claims

1. A composition for use in a cosmetic comprising a tobacco extract and a rose extract, the cosmetic comprising the composition and other ingredients acceptable in the cosmetic; wherein, based on the total weight of the cosmetic, the tobacco extract accounts for 0.001-3.5wt% and the rose extract accounts for 0.001-15wt%;

the tobacco extract is obtained by extracting with a mixed reagent, wherein the mixed reagent is a mixed solution of water, acetone, methanol, formic acid, trifluoroacetic acid and ethyl acetate, and the formula volume ratio is 65:65:80:2:1:1.

2. The composition of claim 1, wherein the tobacco flower extract comprises anthocyanin, procyanidin A1, procyanidin C, 3-hydroxyflavone, chalcone, hesperidone, isorhamnetin-3, 7-o-diglucoside, isorhamnetin-3-o-glucoside, kaempferol-7-o- β -D-glucoside, kaempferol-3-o-rutinoside, quercetin 3,4' -diglucoside, and quercetin 3-o- (6 "-galloyl) - β -D-glucoside.

3. The composition of claim 1, wherein the rose extract comprises phenethyl alcohol, 1-nonanol, linalool, geraniol, 2-heptanol, acetic acid, benzyl alcohol, isopentenyl alcohol, 2-methylbutanol, 2-propanol, 1-octanol, nerol, eugenol, methyl eugenol, 2, 6-bis (1, 1-dimethylethyl) -4-methylphenol, allyl butyl ether, benzaldehyde, neral, nonanal, decanal, geranial, undecanal, citronellal, methylheptenone, 2-nonanone, 2-tridecanone, 2-undecanone, 2-heptanone, geranylacetone, terpinene, α -cedrene, thurene, myrcene, α -hotel, dipentene, α -farnesene, neryl formate, citronellyl acetate, geranyl acetate, methyl eicosanoate, pear alcohol ester, n-tridecane, 3-methylene-1, 1-dimethyl- α -vinylcyclohexane, p-toluidine hydrochloride, and amine salts thereof.

4. The composition of claim 1, wherein the acceptable additional ingredients are selected from one or more of emollients, film forming agents, emulsifiers, humectants, stabilizers, and solvents.

5. The composition of claim 1, wherein the cosmetic product is in the form of one or more of a foundation, a barrier, a pre-make-up, a face wash, a skin care lotion, a concealer, and a make-up cream.