CN116270399A - Preparation method of umbilical cord extract and whitening composition thereof - Google Patents
Preparation method of umbilical cord extract and whitening composition thereof Download PDFInfo
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- CN116270399A CN116270399A CN202310285852.9A CN202310285852A CN116270399A CN 116270399 A CN116270399 A CN 116270399A CN 202310285852 A CN202310285852 A CN 202310285852A CN 116270399 A CN116270399 A CN 116270399A
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
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Abstract
The invention belongs to the field of medical, dental or cosmetic preparations, and particularly relates to a preparation method of an umbilical cord extract and a whitening composition thereof. The umbilical cord extract is used in cosmetics, and comprises umbilical cord extract, fermentation product filtrate of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, calendula extract, flos Albiziae extract, and rhizoma Gastrodiae root extract. The whitening composition containing the umbilical cord extract has the effects of whitening, moisturizing, removing freckles, repairing, protecting skin barriers, resisting wrinkles and resisting aging.
Description
Technical Field
The invention belongs to the field of medical, dental or cosmetic preparations, and particularly relates to a preparation method of an umbilical cord extract and a whitening composition thereof.
Background
Skin aging refers to the aging and sexual damage of skin function, and the protective ability, regulation ability and the like of the skin on the body are reduced, so that the skin cannot adapt to the change of internal and external environments, and the overall appearance conditions such as color, luster, shape, texture and the like are changed. Skin aging has many causes, and is largely divided into endogenous aging and exogenous aging. Endogenous aging is an age factor, and as the age increases, the human body gradually ages, and skin also ages, loosens, wrinkles and the like, which is a natural phenomenon and is irreversible. Because people can continuously perform limb activities in life, faces such as smiles, laughs, frowns and the like can move along with the skins, and the muscles and the skins with higher use frequency are more easy to age. Exogenous aging is an environmental factor, which varies with the living environment and habits of different people. Some people are in the open air for a long time due to work or life, and are exposed to wind and sun, so that skin is more easily roughened, even blackened, long-spotted and the like.
Whereas, from a cellular standpoint, skin aging is divided into a decline in cell viability in the epidermis and dermis. The former decrease causes a decrease in skin resistance to the outside, a decrease in protective power, and an increase in metabolic disturbance and pigment accumulation due to ultraviolet rays. The latter decrease may result in decreased collagen and elastin secretion by fibroblasts therein. Collagen has the function of keeping water storage capacity, and the content is reduced, so that the skin is loosened and wrinkles are increased; the function of the elastin fibers is to provide a supportive effect and reduced levels can result in loss of skin elasticity.
Melanin is a protective pigment for living things, and is a generic name for phenols or indole biomacromolecule pigments with complex and diverse structures in animals. It provides structural strength to the organism or protects the organism from environmental stresses such as ultraviolet light, ionizing radiation, heavy gold contamination, low and high temperatures, etc. Melanin is a class of photoprotectors, radioprotectors, chelators, biological antioxidants and immune promoters. When the skin is irradiated by the line, the skin can self-protect, and the skin cells are protected by secreting the melanins to activate tyrosinase to oxidize dopa to form melanin. The melanin is moved due to the metabolic activity of the cells, and is transferred to the epidermis layer of the skin to form freckle, sunburn, black spot and other shapes.
Disclosure of Invention
The invention aims to provide a preparation method of an umbilical cord extract and a whitening composition containing the umbilical cord extract, wherein the whitening composition has the essence for removing freckles, whitening, moisturizing and repairing skin with good stability.
A method for preparing an umbilical cord extract, comprising the steps of:
(1) Washing mammal umbilical cord in buffer solution, and removing blood and blood vessel;
(2) The rest umbilical cord tissue is Wharton's jelly, after being cut into fragments, the Wharton's jelly is placed in a buffer solution for Wen Jiaodong;
(3) Centrifuging after stirring to obtain supernatant, and obtaining umbilical cord extract.
In particular, the buffer is a PBS buffer;
the length of the fragments in the step (2) is 0.5-3cm;
in the step (2), the temperature range is 3-8 ℃, and the stirring time is 10-30h;
in the step (2), the addition amount of the buffer solution is 1-5 times of the original mass of the umbilical cord;
in the step (3), the centrifugation time is 10-20min, and the rotating speed is 2500-5500rpm.
The invention also provides application of the umbilical cord extract in the field of cosmetics, specifically comprising the following steps:
the whitening composition comprises the following raw materials in parts by mass:
in particular, humectants, water, emulsifiers, thickeners and preservatives are also included.
In particular, the method comprises the following steps of:
in particular, the humectant is at least one of glycerin, hyaluronic acid, sodium polyglutamate or schizophyllan.
In particular, the preservative is at least one of 1, 2-hexanediol, or butanediol.
In particular, the emulsifier is at least one of PEG/PPG-17/6 copolymer, jojoba wax PEG-120 esters or jojoba oil PEG-150 esters.
The invention also provides a preparation method of the whitening composition, which comprises the following steps:
(1) Mixing water, emulsifier, thickener and humectant, and heating;
(2) Cooling, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, calendula extract, flos Albiziae extract, and rhizoma Gastrodiae (root extract;
(3) Continuously cooling, adding antiseptic, stirring, discharging to obtain whitening composition,
the heating temperature in the step (1) is 80-90 ℃;
the temperature of the step (2) is reduced to 40-50 ℃;
the continuous cooling in the step (3) is carried out until the temperature is reduced to 35-39 ℃;
the stirring time in the step (2) and the step (3) is 15-30min.
Umbilical cord extract: contains chondroitin sulfate, glycogen and lipid. And has a large amount of protein and collagen, and has good conditioning effect on skin. Firstly, by repairing endogenous cells, the aging and death are inhibited, the metabolism is accelerated, the skin barrier is repaired, and the aging is delayed; and then deeply nourishing the cells to enable the cells to be full of vigor again, so that the skin can be full of gloss, ruddy and fine.
Radix astragali Mongolici (Astragalus membranaceus) root extract: the active ingredient of the astragalus saponin contained in the composition can stimulate the proliferation of cortical cells and has the effect of conditioning skin. Has effects of resisting aging and preventing wrinkle.
Radix Saposhnikoviae (SAPOSHNIKOVIA DIVARICATA) root extract: has effects in scavenging various free radicals, and can be used as antioxidant; has certain inhibiting effect on melanin and good whitening effect; can shrink pores and be used as astringents; meanwhile, the product has antibacterial and anti-inflammatory effects.
Calendula (CALENDULA OFFICINALIS) flower extract: anti-inflammatory and antibacterial effects, the calendula extract has a proliferation effect on epidermal cells and dermal cells, inhibits elastase activity, combines its excellent antioxidant properties, and thus can enhance skin activity while having a moisturizing effect.
Albizia julibrissin (ALBIZIA JUUBRISSIN) flower extract: the characteristic active ingredient is albizia julibrissin which has antioxidant activity, anti-saccharification effect, and can prevent reducing sugar and protein from undergoing browning reaction, and simultaneously reverse glycosylated collagen, protect and restore the structure and activity of collagen, reduce skin wrinkles and improve pigmentation.
Gastrodia ELATA (Gastrodia ELATA) root extract: has good activation effect on superoxide dismutase, catalase and glutathione peroxidase, and can effectively eliminate free radicals to play an anti-aging role; the rhizoma Gastrodiae extract also has activating effect on melanocyte.
Sodium polyglutamate: the special molecular structure ensures that the skin moisturizing composition has extremely strong moisturizing capability, can effectively increase the moisturizing capability of skin and promote skin health, has super-strong moisturizing capability better than hyaluronic acid and collagen, and is a new-generation biotechnology moisturizing component. As whitening agents, has long-lasting anti-crease properties.
Schizophyllan: is an ideal component for cosmetics, and has the effects of resisting inflammation and enhancing immunity. Can effectively repair sunburn skin, has the functions of moisturizing and anti-wrinkle on the skin, and enables the skin to be tender and plump.
Compared with the prior art:
1. the whitening composition containing the umbilical cord extract has the effects of whitening, moisturizing, removing freckles, repairing, protecting skin barriers, resisting wrinkles and resisting aging.
2. The raw material components of the invention, namely the fermentation product filtrate of the saccharomyces cerevisiae, the lysate of the fermentation product of the saccharomyces cerevisiae and the umbilical cord extract, have the effects of reducing the biotoxicity, having high permeability, improving the activity of cells and accelerating the metabolism
3. The invention adds substances such as whitening, moisturizing, anti-allergic components and the like, and after the substances are absorbed by cells, the melanin synthesis is inhibited, the pigmentation is failed, the whitening effect is achieved, the substances for resisting inflammation are contained, the proliferation and migration of relevant cells which cause inflammation at the damaged part are effectively inhibited, and the cell regeneration capability is enhanced, so that the repairing and anti-aging effects are achieved.
4. The added polyglutamic acid is an amino acid anionic polymer with viscosity, also called gamma-PGA, has excellent skin care and maintenance effects, and promotes the synthesis of skin natural moisturizing factor PCA; protecting hyaluronic acid, and enhancing skin water holding capacity with synergistic effect of hyaluronic acid; inhibiting tyrosinase activity and reducing melanin synthesis; activating fibroblast to synthesize collagen and elastin, and making skin elastic, moist and transparent. The gamma-PGA can be used for constructing an embedded slow-release system, promoting skin absorption, forming a film on the surface of skin, isolating environmental stimulus and reducing skin inflammation.
Drawings
The invention will be further described with reference to the drawings and examples.
FIG. 1 is a drawing of the umbilical cord extract extraction process of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
In specific embodiments, steps, material selections, numerical parameters that are not described in detail are all routine selections in the art, or any prior art that is presently disclosed.
The umbilical cord extract adopted by the invention is derived from sheep umbilical cords which are legal sources in accordance with ethics.
Example 1
A method for preparing a whitening composition, comprising the steps of:
(1) Weighing the raw materials for standby, mixing water, an emulsifying agent, a thickening agent and a humectant, and heating to 85 ℃;
schizophyllan 1
(2) Cooling to 45deg.C, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, herba Sidae Rhombifoliae extract, flos Albiziae extract, and rhizoma Gastrodiae root extract, and stirring for 15min;
(3) Continuously cooling to 37 ℃, adding preservative, stirring for 15min, and discharging to obtain the whitening composition.
The preparation method of the umbilical cord extract comprises the following steps:
(1) Washing the umbilical cord of sheep in Phosphate Buffered Saline (PBS);
(2) Removing blood and blood vessels from umbilical cord, cutting the remaining umbilical cord tissue into Wharton's jelly, soaking in phosphate buffer solution with the mass of 3 times of that of umbilical cord, wherein the length of Wharton's jelly is 1 cm-2 cm.
(3) The Walker gum soaked in phosphate buffer was stirred for 24h at 4 ℃.
(4) Centrifuging at 4000rpm for 15min, collecting supernatant, and collecting supernatant as umbilical cord extract.
Example 2
A method for preparing a whitening composition, comprising the steps of:
(1) Weighing the raw materials for standby, mixing water, an emulsifying agent, a thickening agent and a humectant, and heating to 90 ℃;
umbilical cord extract 2
Saccharomyces cerevisiae fermentation product filtrate 2
Lysate 2 of fermentation product of Saccharomyces cerevisiae
Jojoba wax PEG-120 esters 3
Acrylamide dimethyl taurate VP copolymer 3
1.5 parts of butanediol.
(2) Cooling to 50deg.C, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, herba Sidae Rhombifoliae extract, flos Albiziae extract, and rhizoma Gastrodiae root extract, and stirring for 15min;
(3) Continuously cooling to 39 ℃, adding preservative, stirring for 30min, and discharging to obtain the whitening composition.
The preparation method of the umbilical cord extract comprises the following steps:
(1) Washing the umbilical cord of sheep in Phosphate Buffered Saline (PBS);
(2) Removing blood and blood vessels from umbilical cord, cutting the remaining umbilical cord tissue into Wharton's jelly, soaking in phosphate buffer solution with the mass of 5 times of that of umbilical cord, wherein the length of Wharton's jelly is 0.5 cm-2 cm.
(3) The Walker gum soaked in phosphate buffer was stirred for 30h at 3 ℃.
(4) Centrifuging at 2500rpm for 20min, collecting supernatant, and collecting supernatant as umbilical cord extract.
Example 3
A method for preparing a whitening composition, comprising the steps of:
(1) Weighing the raw materials for standby, mixing water, an emulsifying agent, a thickening agent and a humectant, and heating to 80 ℃;
umbilical cord extract 2
Saccharomyces cerevisiae fermentation product filtrate 2
Lysate 2 of fermentation product of Saccharomyces cerevisiae
Radix astragali Mongolici extract 1
Radix Saposhnikoviae root extract 1
Calendula extract 1
Albizia flower extract 1
Gastrodia elata root extract 1
Glycerol 2
Schizophyllan 3
Water 74.5
Jojoba wax PEG-120 esters 3
Acrylamide dimethyl taurate VP copolymer 3
1.5 parts of butanediol.
(2) Cooling to 40deg.C, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, herba Sidae Rhombifoliae extract, flos Albiziae extract, and rhizoma Gastrodiae root extract, and stirring for 30min;
(3) Continuously cooling to 35 ℃, adding preservative, stirring for 15min, and discharging to obtain the whitening composition.
The preparation method of the umbilical cord extract comprises the following steps:
(1) Washing the umbilical cord of sheep in Phosphate Buffered Saline (PBS);
(2) Removing blood and blood vessels from umbilical cord, cutting the remaining umbilical cord tissue into Wharton's jelly, soaking in phosphate buffer solution with the mass of 1 times of that of umbilical cord, wherein the length of Wharton's jelly is 1.5 cm-3 cm.
(3) The Walker gum soaked in phosphate buffer was stirred for 10h at 8 ℃.
(4) Centrifuging at 5500rpm for 10min, collecting supernatant, and collecting supernatant as umbilical cord extract.
Example 4
A method for preparing a whitening composition, comprising the steps of:
(1) Weighing the raw materials for standby, mixing water, an emulsifying agent, a thickening agent and a humectant, and heating to 85 ℃;
(2) Cooling to 45deg.C, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, herba Sidae Rhombifoliae extract, flos Albiziae extract, and rhizoma Gastrodiae root extract, and stirring for 15min;
(3) Continuously cooling to 37 ℃, adding preservative, stirring for 15min, and discharging to obtain the whitening composition.
The preparation method of the umbilical cord extract comprises the following steps:
(1) Washing the umbilical cord of sheep in Phosphate Buffered Saline (PBS);
(2) Removing blood and blood vessels from umbilical cord, cutting the remaining umbilical cord tissue into Wharton's jelly, soaking in phosphate buffer solution with the mass of 3 times of that of umbilical cord, wherein the length of Wharton's jelly is 1 cm-2 cm.
(3) The Walker gum soaked in phosphate buffer was stirred for 24h at 4 ℃.
(4) Centrifuging at 4000rpm for 15min, collecting supernatant, and collecting supernatant as umbilical cord extract.
Comparative example 1
The difference from example 1 is that the following components are included:
umbilical cord extract 2
Saccharomyces cerevisiae fermentation product filtrate 2
Lysate 2 of fermentation product of Saccharomyces cerevisiae
Glycerol 2
Schizophyllan 3
Water 77
Jojoba wax PEG-120 esters 3
Acrylamide dimethyl taurate VP copolymer 3
1.5 parts of butanediol.
The preparation method was the same as in example 1 except that the umbilical cord extract, the filtrate of the fermentation product of Saccharomyces cerevisiae, and the lysate of the fermentation product of Saccharomyces cerevisiae were not contained.
Comparative example 2
The difference from example 1 is that the following components are included:
the preparation method was the same as in example 1 except that the related steps of the astragalus root extract, the radix sileris extract, the calendula extract, the albizia flower extract and the gastrodia tuber root extract were not contained.
Sample stability test
1. Heat resistance test: the temperature of the incubator is adjusted to 40 ℃, three samples of the four prepared examples are taken and filled in transparent glass bottles, the sample filling amount is 20 ml/bottle, the samples are placed in the incubator after being sealed, and the samples are taken out after three months, are restored to room temperature, and observe the appearance change.
2. Cold resistance test: the temperature of the incubator is regulated to-10 ℃, three samples of the four prepared examples are taken and filled in transparent glass bottles, the sample filling amount is 20 ml/bottle, the samples are placed in the incubator after being sealed, taken out after three months, and the samples are recovered to room temperature, and the appearance change is observed.
3. And (3) normal temperature test: three samples of the four prepared examples were taken and placed in transparent glass bottles, the sample loading amount was 20 ml/bottle, and after the bottle was sealed and left at room temperature for 6 months, the appearance change of the essence was observed.
No phenomena such as precipitation and precipitation are observed in the heat resistance test, the cold resistance test and the normal temperature test, and the original appearance is maintained.
Human body skin patch test
Four examples obtained are referred to the human skin patch experiments in 2022 cosmetic safety Specification.
Skin reactions were observed as standard at 30min (after the disappearance of the indentations), 24h and 48h, respectively, and the observations were recorded, see table 1.
TABLE 1 human safety test results
Human subjective feeling test
90 women were recruited, aged 20-50 years, into 6 groups of 15 people each. The 6 groups of subjects respectively smeared with corresponding samples at night, the finished product of the 1 st group dressing example 1, the finished product of the 2 nd group dressing example 2, the finished product of the 3 rd group dressing example, the finished product of the 4 th group dressing example 4, the finished product of the 5 th group dressing comparative example 1 and the finished product of the 6 th group dressing comparative example 2, and the effects before and after use are shown in table 2.
TABLE 2 human subjective feeling test results
Note that: in table 2, for each subject, a questionnaire was issued, a score of 0 for no corresponding effect was considered, a score of 50 for a slight effect was considered, a score of 100 for a significant effect was considered, and finally the final average score was calculated.
Cell Activity test
(1) Cell inoculation: according to 2.2 x 10 4 Cell/well seeding Density cells were seeded into 96 well plates and incubated overnight in incubator (37 ℃ C., 5% CO) 2 )。
(2) Experimental grouping: the experiment sets a blank control group and an experiment group, 3 cell-free holes are additionally set as zeroing holes, 5 concentration gradients are set in the experiment group based on the previous embodiment of the sample, and 3 compound holes are set under each gradient concentration.
(3) Preparing liquid: according to the experimental design, test substances with different concentrations are prepared by using a basic culture medium.
TABLE 3 test concentration setting table
(4) Administration: taking out the 96-well plate after 24 hours, removing the old culture medium, adding 100 mu L of culture solution into each well of a blank control group, and adding 100 mu L of culture solution containing samples with corresponding concentrations into each well of a sample group; the zeroed group was inoculated without cells and only 100. Mu.L of cell culture medium was added. And after the administration is finished, the culture medium is returned to the incubator for continuous culture.
(5) And (3) detection: after cell culture for 24 hours, the supernatant was discarded and MTT working solution was added. Incubate at 37℃for 4h in the absence of light, after incubation, discard supernatant, add 100. Mu.L DMSO per well, read OD at 490 nm.
(6) Cell relative viability calculation: calculating according to a formula;
(7) Statistical analysis was performed using the SPSS 19.0 statistical software package to descriptive statistics of the measurements of the test area. The change in the analytical value was calculated and the difference between the control group and the sample group. If the test data are normally distributed, adopting an independent T test method to carry out statistical analysis; if the test data are in non-normal distribution, adopting a rank sum test method to carry out statistical analysis. The statistical methods all use a two-tailed test with a test level α=0.05.
The results are shown in Table 4.
TABLE 4MTT assay results
Human body instrument test
The effect of the examples was evaluated scientifically by instrumental measurements.
A total of 10 volunteers were enrolled, and skin individual type angle ITA ° values, melanin index (MI value), skin whiteness (L value), skin moisture content (MMV value), and skin water loss rate (TEWL value) indexes were collected by CK probes at 0W, 2W, 4W, and 6W, respectively, using the samples prepared in example 4 and the matrix samples at random on the left and right faces, respectively.
Experimental instrument:
skin color tester (colorimeter CL400; CK, germany): the XYZ value is obtained by using a three primary color method after correcting the reflected light quantity measured by a probe by using a special color matrix, and is calculated and converted into an Lxa.b.ITA degree value.
Skin melanin and heme tester (Mexameter MX 18): the measurement principle is based on the principle of spectral absorption (RGB), and the content of melanin in the skin is determined by measuring the reflection amount of light of a specific wavelength on the skin of a human body.
Skin moisture content tester (corneteter CM825, CK, germany): the moisture content of the human skin cuticle is measured by adopting a capacitance method.
Percutaneous moisture loss tester (Tewameter TM300; CK, germany): the water vapor pressure gradient at different points of the epidermis formed by the water dispersion of the angle layer is measured by two groups of sensors with different temperatures and humidity, and the water content evaporated through the epidermis is measured.
Facial image analyzer VISIA (CK, germany): the face image shooting is carried out by adopting 5 different light sources, so that the change of the face skin state can be visually observed.
Experimental samples:
the samples prepared in example 4, and the matrices shown in table 5 below;
table 5 matrix formulation
| Trade name | Name of the name | Additive amount |
| Deionized water | Water and its preparation method | 77.1 |
| EDTA-2NA | EDTA disodium salt | 0.05 |
| AC Yeast Beta-Glucan creeping glycans | Beta-glucan | 0.05 |
| ALLANTOIN (ALLANTOIN) | Allantoin | 0.2 |
| Glycerol | Glycerol | 5 |
| ULTREZ 21 POLYMER | Acrylic acid (esters) C10-30 alkanol acrylate cross-linked polymers | 0.15 |
| Low molecular weight sodium hyaluronate | Sodium hyaluronate | 0.05 |
| T.I.O | Glycerol tris (ethylhexanoate) | 2 |
| DM10 | Polydimethylsiloxane | 3 |
| Synthesis of Squalane Squalane Syn | Hydrogenated polyisobutene | 4 |
| ELDEW PS-203 | Phytosterol/octyldodecanol lauroyl glutamate | 0.15 |
| olivem 1000 | Cetylus spermacetiFatty alcohol olive oil acid ester | 1.6 |
| SEPINOV EMT10 | Hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer | 1 |
| Spectrastat PHL | Octanoyl hydroxamic acid | 1 |
| Hexanediol (Hexadiol) | 1, 2-hexanediol | 0.5 |
| 1.3 butanediol | Butanediol (butanediol) | 4 |
| Triethanolamine TEA | Triethanolamine salt | 0.15 |
The test method comprises the following steps:
(1) Volunteer requirements
The volunteer 10 persons were recruited, and five indexes of skin individual type angle ITA DEG value, melanin index (MI value), skin whiteness (L value), skin moisture content (MMV value) and percutaneous water loss rate (TEWL value) of four areas in total of each of the plaque area and the non-plaque area of the left and right faces of the volunteer were tested. The tested part is forbidden to use the preparation which influences the whitening and freckle removing test; subjects were prohibited from dropping, injecting, orally taking or otherwise ingesting formulations that affected the whitening and freckle-removing test; the subject is mainly subjected to indoor activities, and long-term exposure to light is avoided.
1.1 inclusion criteria
25-60 years old, healthy female or male;
at least one obvious color spot with the ITA degree difference of more than 10 degrees and the peripheral adjacent skin at the tested part, and the diameter of the color spot is not less than 3mm (the color spot cannot be freckle, pigmented nevus and the like which are difficult to improve by using an external preparation clinically);
no allergic diseases, no allergic history of cosmetics and other external preparations;
no medicine affecting the light sensitivity is used in the recent history of no light-sensitive disease;
the skin of the tested part should not have the phenomena of birthmarks, inflammations, scars, hair and the like;
the test procedure can be understood, voluntarily taking part in the test and signing a written informed consent.
1.2 exclusion criteria
a) Pregnant or lactating women, or those who have a pregnancy preparation plan recently;
b) Patients with skin history such as psoriasis, eczema, atopic dermatitis, severe acne, etc.; or suffering from other chronic systemic diseases;
c) For the last 1 month, anti-inflammatory drugs such as corticosteroids, etc. are orally taken or externally used;
d) For about 2 months, any skin color affecting product or drug (such as hydroquinone type preparation) such as percarboxylic acid, salicylic acid, etc.;
e) The test part uses the vitamin A acid preparation or the medical and aesthetic therapist who carries out chemical stripping, laser, pulse light and the like within 3 months;
f) Unavoidable long-term sun exposure;
g) Other clinical trials were enrolled in the last 2 months;
h) Other clinical evaluations were considered unsuitable for the participants.
1.3 subject restriction
a) During the test, the tested part must use the test product or the control product provided by the test mechanism, and any other product with the effects of removing spots and whitening or possibly influencing the test result cannot be used;
b) No insolation is possible during the test and the test site should be well-protected from sunlight.
(2) Test index
Individual skin type angle ITA ° value: the skin colorimeter or the reflectance spectrophotometer measures the skin Lxa xb xcolor space data to represent the parameters of the skin color of the human body, and the larger the ITA value is, the lighter the skin complexion is, and the darker is the reverse. The calculation formula is as follows:
melanin index (MI value) MI value characterizes melanin index, ranging from 0 to 999, with higher measured values indicating darker skin color.
Skin brightness (L-value): indicating a black to white balance, a larger L value indicates a whiter skin.
Skin moisture content (MMV value): MMV values characterize skin moisture, with greater MMV values indicating higher stratum corneum moisture content.
Percutaneous moisture loss (TEWL value): TEWL values are used to evaluate skin barrier function and are important parameters for examining skin barrier after use of the product, with smaller values leading to less skin loss of water through the skin.
(3) Evaluation method and data statistics method
The test adopts a double-blind method, the test period is 6 weeks, the skin data acquisition times are 4 times, and the skin data acquisition times are 0W, 2W, 4W and 6W respectively. Qualified subjects are screened according to requirements, and when volunteers visit for the first time, volunteer informed consent is signed, 0-week data are collected, test areas (including four areas of left and right face spots and non-spot areas) are delineated, and samples are issued. When volunteers visit each time, after the faces are cleaned by the designated face cleaning, the volunteers sit still for 20-30min in the test environment, and after sitting still, the face data are collected.
Statistical analysis of the data was performed using statistical analysis software. The metering data are expressed as: the mean value is +/-standard deviation, normal distribution inspection is carried out, normal distribution requirements are met, paired t inspection is adopted for comparison before and after the mean value is self, and otherwise, two related sample rank sum inspection is adopted; comparing the grade data before and after use, and adopting two related sample ranks and tests; comparison between test product and control group used independent sample t-test or rank-sum test. The above statistical analysis was a two-tailed test with a significance level of α=0.05.
(4) Test notice
4.1 test Environment
Laboratory temperature 21±1 ℃; relative humidity 50% + -10%
4.2 test sites
The total of four areas of the left and right facial speckle regions and the non-speckle region
4.3 sample application method
2 times per day, respectively and uniformly applying on both sides of face.
(5) Test results
5.1 individual type angle ITA deg. values as shown in table 6.
Table 6ITA value test results
Note "-": p >0.05, indicating no significant difference, "": p <0.05, representing significant differences, "x": p <0.01, ", x": p <0.001, representing a very significant difference.
5.2 melanin index MI values, as shown in table 7.
TABLE 7 melanin index MI test results
Note "-": p >0.05, indicating no significant difference, "": p <0.05, representing significant differences, "x": p <0.01, ", x": p <0.001, representing a very significant difference.
5.3 skin whiteness L values, as shown in table 8.
Table 8 skin whiteness L value test results
Note "-": p >0.05, indicating no significant difference, "": p <0.05, representing significant differences, "x": p <0.01, ", x": p <0.001, representing a very significant difference.
5.4 skin moisture content MMV values as shown in table 9.
TABLE 9 skin moisture content MMV value test results
Note "-": p >0.05, indicating no significant difference, "": p <0.05, representing significant differences, "x": p <0.01, ", x": p <0.001, representing a very significant difference.
5.5 skin trans-skin water loss TEWL values as shown in table 10.
TABLE 10 results of the test for the percutaneous Water loss rate TEWL value of skin
Note "-": p >0.05, indicating no significant difference, "": p <0.05, representing significant differences, "x": p <0.01, ", x": p <0.001, representing a very significant difference.
The foregoing detailed description is directed to one of the possible embodiments of the present invention, which is not intended to limit the scope of the invention, but is to be accorded the full scope of all such equivalents and modifications so as not to depart from the scope of the invention.
Claims (10)
1. The preparation method of the umbilical cord extract is characterized by comprising the following steps of:
(1) Washing mammal umbilical cord in buffer solution, and removing blood and blood vessel;
(2) The rest umbilical cord tissue is Wharton's jelly, after being cut into fragments, the Wharton's jelly is placed in a buffer solution for Wen Jiaodong;
(3) Centrifuging after stirring to obtain supernatant, and obtaining umbilical cord extract.
2. The method for producing an umbilical cord extract according to claim 1, wherein,
the buffer solution is PBS buffer solution;
the length of the fragments in the step (2) is 0.5-3cm;
in the step (2), the temperature range is 3-8 ℃, and the stirring time is 10-30h;
in the step (2), the addition amount of the buffer solution is 1-5 times of the original mass of the umbilical cord;
in the step (3), the centrifugation time is 10-20min, and the rotating speed is 2500-5500rpm.
3. Use of the umbilical cord extract of claim 1 in the cosmetic field.
4. The whitening composition is characterized by comprising the following raw materials in parts by mass:
5. the whitening composition according to claim 4, further comprising a humectant, water, an emulsifier, a thickener, and a preservative.
6. The whitening composition according to claim 5, further comprising the following raw materials in parts by mass:
7. the whitening composition according to claim 6, wherein the humectant is at least one of glycerin, hyaluronic acid, sodium polyglutamate or schizophyllan.
8. The whitening composition according to claim 6, wherein the preservative is at least one of 1, 2-hexanediol, or butanediol.
9. The whitening composition according to claim 6, wherein the emulsifier is at least one of PEG/PPG-17/6 copolymer, jojoba wax PEG-120 esters or jojoba oil PEG-150 esters.
10. A method for preparing the whitening composition according to any one of claims 5 to 9, comprising the steps of:
(1) Mixing water, emulsifier, thickener and humectant, and heating;
(2) Cooling, adding umbilical cord extract, filtrate of fermentation product of Saccharomyces cerevisiae, lysate of fermentation product of Saccharomyces cerevisiae, radix astragali extract, radix Saposhnikoviae root extract, calendula extract, flos Albiziae extract, and rhizoma Gastrodiae (root extract;
(3) Continuously cooling, adding antiseptic, stirring, discharging to obtain whitening composition,
the heating temperature in the step (1) is 80-90 ℃;
the temperature of the step (2) is reduced to 40-50 ℃;
the continuous cooling in the step (3) is carried out until the temperature is reduced to 35-39 ℃;
the stirring time in the step (2) and the step (3) is 15-30min.
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