CN113616573A - I type collagen production promoter - Google Patents
I type collagen production promoter Download PDFInfo
- Publication number
- CN113616573A CN113616573A CN202111025328.5A CN202111025328A CN113616573A CN 113616573 A CN113616573 A CN 113616573A CN 202111025328 A CN202111025328 A CN 202111025328A CN 113616573 A CN113616573 A CN 113616573A
- Authority
- CN
- China
- Prior art keywords
- extract
- concentration
- collagen
- motherwort
- polygonatum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
Abstract
The present invention provides a collagen type I production promoter characterized by being at least one selected from the group consisting of a motherwort (Leonurus japonicus) extract and a Polygonatum odoratum (Polygonatum odoratum) extract.
Description
Technical Field
The present invention relates to novel type I collagen production promoters, the use of these novel type I collagen production promoters in cosmetics, and cosmetics containing these novel type I collagen production promoters.
Background
Beauty maintaining and aging resisting are the targets that people pursue all the time, and the company is also always dedicated to searching products with the efficacies of beauty maintaining and aging resisting. The presence of collagen in the dermis layer is a major factor affecting skin aging. If the collagen content in the dermis is reduced, the skin shows aging signs, loses elasticity, becomes loose day by day, and wrinkles are formed, and if the collagen content in the dermis is rich, the skin looks full of elasticity and is young and glossy, so that the dream of beautifying, maintaining beauty and resisting aging can be realized if a product for promoting the increase of the collagen content in the dermis can be found.
The prior art methods of collagen supplementation include, 1. eating more collagen-rich foods: the skin (such as pig skin and fish skin), tendon (such as pig tendon, cattle tendon and pig trotter), and cartilage (bone and meat connection and brittle bone) of animals for daily food are rich in collagen. However, after the collagen is digested by the stomach and the intestines, the collagen is decomposed into micromolecular amino acid and polypeptide by protease, the shape and the structure of the collagen do not exist any more, and the oral administration of the collagen has little effect on improving the skin. 2. Smearing collagen: many smearing type collagen products such as facial masks and the like exist in the market, but because the collagen has large molecular weight, the collagen is difficult to penetrate through a tight skin barrier and be absorbed by the skin, and the collagen is smeared, so that the collagen of the skin cannot be increased. 3. Injection water light needle: by means of vacuum negative pressure technology, hyaluronic acid solution is injected into corium layer to make skin absorb 100% of nutrients. The effects of smoothing wrinkles, lightening melanin, whitening and tendering skin are achieved. However, the effect is short-lived by means of a percutaneous injection, the risks involved are also non-trivial and need to be very careful. 4. The collagen health-care product can be taken as oral liquid or powder; some are called collagen, some are called "collagen peptide", and some are called "hydrolyzed collagen".
The present inventors have determined that the above prior art methods of increasing collagen do not adequately address the needs of the individual and that such methods may be effective but are relatively slow to take effect. If a component capable of promoting the production of collagen can be directly added into the cosmetic, the component can penetrate into the dermal layer of the skin to directly promote the production of collagen in the dermal layer while the cosmetic is applied, and the method is more effective than the method for directly supplementing collagen in the prior art. Accordingly, the present inventors have made extensive and intensive studies to find a product capable of promoting the production of collagen in the dermal layer.
Motherwort herb, the name of traditional Chinese medicine. Is fresh or dried aerial part of Leonurus japonicus Houtt. of Labiatae. Harvesting fresh products from a spring seedling stage to an early summer flower stage; collecting the dried product in summer when stem and leaf flourish, flower do not bloom or bloom, sun-drying, or cutting and sun-drying. Widely distributed all over the country. The effects and actions of motherwort include: 1. the motherwort herb has the functions of obviously increasing coronary flow and quite obviously slowing down heart rate for isolated guinea pig heart caused by isoproterenol, and the motherwort herb preparation for intravenous injection can obviously increase coronary flow, reduce coronary resistance, slow down heart rate and reduce output and left ventricle work of anesthetized dogs. 2. The motherwort extract and decoction have strong and lasting excitation effect on uterus, can enhance the contraction of the uterus and simultaneously improve the tension and the contraction rate of the uterus. It is also indicated for traumatic injuries with blood stasis, for example, Yi mu Cao Gao recorded in Wai Tai Mian Yao. 3. And color spots are faded, if the whitening and freckle-removing cream added with the motherwort is used for massaging and washing the face continuously and the motherwort decoction is eaten for 2 to 3 times in one week, most of the color spots can be effectively faded, and more serious color spots can be faded. 4. For dysmenorrhea, it is usually combined with blood-enriching and blood-nourishing, qi-moving and pain-relieving herbs such as Yan Hu Suo, Dang Gui, Bai Shao, Xiang Fu and Chuan Xiu xi, etc. to relieve dysmenorrhea, and it is usually used in an amount of about 30 g, most of them have obvious effect. 5. For postpartum diseases such as postpartum hemorrhage or lochiorrhea, abdominal distending pain, small amount of bleeding, or blood clot complicated with uterine atony, it is often combined with Dang Gui, Jiu Bai Shao, ai Ye, Chuan Xiong, charred Haw thorn, and for cold patients, it is added with Pao Jiang, Tai Wu Yao, etc. with ideal effect. 6. The motherwort is compatible with other medicines to achieve the aim of treating kidney stones, acute nephritic edema, hematuria and other kidney diseases. 7. Can be used for inhibiting cancer, and herba Leonuri contains various trace elements. Selenium has the functions of enhancing the activity of immune cells, alleviating the occurrence of atherosclerosis and improving the disease defense system of the body; manganese has antioxidant, antiaging, antifatigue and cancer cell proliferation inhibiting effects. 8. Improving eyesight and replenishing vital essence, and motherwort has the function of treating dizziness and improving eyesight, and is used for treating dizziness and blurred vision caused by upward water dampness due to asthenia.
CN108670943A discloses a moisturizing, whitening and spot-lightening plant extract composition, which contains a plurality of plant extracts in an aqueous phase part, wherein the plant extracts contain motherwort herb extracts.
CN12516046A discloses an extract of motherwort herb acne removing cream and a preparation method thereof.
CN109758393A discloses a skin care product with whitening, moisturizing and anti-aging effects, and a preparation method and application thereof, wherein the skin care product contains 5-10 parts by weight of ginseng extract, 1-5 parts by weight of gentian extract, 5-10 parts by weight of pyracantha fortuneana extract, 1-5 parts by weight of motherwort extract and 3-5 parts by weight of polygonatum odoratum extract. Wherein, the paragraph 0039 of the specification states that the motherwort has the functions of promoting blood circulation, regulating menstruation, inducing diuresis, reducing edema, clearing heat and removing toxicity.
Yuzhu is called the superior product in Ben Cao gang mu, and has very high nutritive value and pharmacological action. The efficacy and effects of Yuzhu include: 1. nourishing yin, moistening dryness, promoting fluid production, quenching thirst, protecting health, and resisting aging. The polysaccharide, vitamin A and nicotinic acid in the polygonatum odoratum can enhance the disease resistance of a human body and delay aging. 2. Tranquilizing nerve and strengthening heart, and modern medicine considers that it contains cardiac glycoside, alkaloid, vitamin A matter and mucopolysaccharide, fragrant solomonseal rhizome fructan, etc. and has the functions of improving myocardial anoxia and adrenocorticoid hormone-like. Yuzhu contains steroid glycoside, and has effect on myocardium similar to that of herba Convallariae preparation. The rhizoma Polygonati Odorati glycoside has cardiotonic effect on isolated frog heart, and the effect of rhizoma Polygonati Odorati decoction is similar to rhizoma Polygonati Odorati glycoside. Meanwhile, polygonatum odoratum has certain curative effect on heart failure caused by palpitation, angina pectoris, rheumatic heart disease, coronary atherosclerotic heart disease and pulmonary heart disease. 3. The modern pharmacological research shows that more than 40 traditional Chinese medicines with the lipid regulating effect are classified into tonifying lipid-lowering medicines, blood-activating lipid-lowering medicines, phlegm-resolving dampness-removing lipid-lowering medicines and the like according to the properties of the traditional Chinese medicines. The methanol extract of polygonatum odoratum has an obvious effect of reducing the blood sugar value of a mouse with hyperglycemia caused by epinephrine. And show a tendency to improve the glucose tolerance function. One of the mechanisms of action is the inhibition of the hepatic glycolysis system. It has also been reported that rats gavaged with Yuzhu have an inhibitory effect on the increase in blood glucose in rats caused by glucose and alloxan. Wherein the polygonatum medicine has the function of reducing low-density lipoprotein and the function of reducing blood sugar, thereby effectively preventing the occurrence of diabetes. 4. Treating oral diseases caused by excessive internal heat and heart-fire rising. Traditional Chinese medicine reminds people of needing to control emotion, reduce tension and avoid worrying about trouble. Especially reduces the worries of delay, complex treatment and related to numerous interpersonal relationships so as to avoid heart-fire exuberance from inducing heart-brain diseases. The sweet soup cooked with rhizoma Polygonati Odorati has effects of clearing heart fire, nourishing yin, and lowering heart fire. 5. The polygonatum odoratum is sweet and greasy in taste, soft and moist in quality, and is a good medicine for nourishing yin and promoting the production of body fluid. The rhizoma Polygonati Odorati is rich in vitamin A substances and mucus, and vitamin A has effects in improving chapped and rough skin, and softening and lubricating skin. Thereby playing the role of beautifying and protecting skin. Modern medical research proves that the traditional Chinese medicine also has the effects of moistening the skin, dissipating chronic inflammation of the skin and treating traumatic injury and sprain. 6. For preventing dim eyesight, Yuzhu is the main herb, and is decocted with mint, ginger and honey for treating dim eyesight. 7. The plum-shizhen can be regarded as the superior products when the sleep is improved and the plum-shizhen is in the Ming dynasty, the polygonatum and the root of straight ladybell are boiled together, so that the effects of nourishing yin, moistening dryness, promoting the production of body fluid and quenching thirst can be achieved, and the effects of nourishing and nourishing yin are great when the two medicines are used together. Although polygonatum odoratum and adenophora stricta belong to the Chinese medicine, the polygonatum odoratum and adenophora stricta are decocted to have no medicinal flavor, can calm the nerves and fall asleep, and are particularly helpful for sleeping. 8. Has effects in improving immunity, regulating immunity, and inhibiting tumor growth. Long-term administration of rhizoma Polygonati Odorati can improve physical fitness and prevent various diseases.
CN109758393A discloses a skin care product with whitening, moisturizing and anti-aging effects, and a preparation method and application thereof, wherein the skin care product contains 5-10 parts by weight of ginseng extract, 1-5 parts by weight of gentian extract, 5-10 parts by weight of pyracantha fortuneana extract, 1-5 parts by weight of motherwort extract and 3-5 parts by weight of polygonatum odoratum extract. Wherein, the paragraph 0039 describes dried rhizome of Polygonatum odoratum (Mill.) Druce of Liliaceae, which mainly contains active ingredients such as steroidal saponins, polysaccharides, chelidonic acid, etc., wherein the Polygonatum odoratum mucopolysaccharide has good hygroscopicity, can effectively increase skin moisture content, and can scavenge free radicals in vivo, inhibit melanin formation, and exert skin caring and speckle removing effects.
As described above, although there are many descriptions about the effects and actions of leonurus japonicus and polygonatum odoratum in the prior art, it has never been disclosed or suggested that leonurus japonicus extract and polygonatum odoratum extract have collagen type I production promoting action. The present inventors have conducted extensive studies for the first time to find that leonurus extract, polygonatum extract, and the like have collagen type I production promoting effects, and if these extracts are added to cosmetics, the skin application of these extracts to the skin can promote the production of collagen type I in the dermis layer of the skin, increase the skin elasticity, and achieve the anti-aging effect, thereby completing the present invention.
Disclosure of Invention
The present invention provides, as a 1 st aspect, a collagen type I production promoter characterized by being at least one selected from the group consisting of an extract of Leonurus japonicus (Leonurus japonicus), and an extract of Polygonatum odoratum (Polygonatum odoratum).
The present invention provides, as a 2 nd aspect, the type I collagen production promoter according to the 1 st aspect, wherein the motherwort herb extract contains 0.0015 wt% or more of an active ingredient.
The present invention provides, as a 3 rd aspect, the type I collagen production promoter according to the 2 nd aspect, wherein the motherwort herb extract has an active ingredient content of 0.00185 wt% or more.
The present invention provides, as a 4 th aspect, the type I collagen production promoter according to the 1 st aspect, wherein the polygonatum odoratum extract contains 0.00025 wt% or more of an active ingredient.
The present invention provides, as a 5 th aspect, the type I collagen production promoter according to the 4 th aspect, wherein the polygonatum odoratum extract contains 0.0004 wt% or more of an active ingredient.
The present invention provides, as aspect 6, the type I collagen production promoter according to aspect 1, wherein the motherwort herb extract is obtained by extracting motherwort herb powder with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the extract with a mixture of water and a polyhydric alcohol having 3 to 6 carbon atoms.
The present invention provides, as a 7 th aspect, the type I collagen production promoter according to the 6 th aspect, wherein the motherwort herb extract is an extract obtained by ultrasonically stirring motherwort herb powder with ethanol and deionized water, heating and extracting, concentrating and drying, and then decoloring and deodorizing with water and butanediol.
The present invention provides, as an 8 th aspect, the type I collagen production promoter according to the 1 st aspect, wherein the polygonatum odoratum extract is an extract obtained by extracting a polygonatum odoratum powder with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the polygonatum odoratum powder with a mixture of water and a polyhydric alcohol having 3 to 6 carbon atoms.
The present invention provides, as a 9 th aspect, the type I collagen production promoter according to the 8 th aspect, wherein the polygonatum extract is an extract obtained by subjecting polygonatum powder, ethanol and deionized water to ultrasonic stirring, heating for extraction, concentration and drying, and then subjecting the extract to decoloring and deodorizing treatment with water and butanediol.
The present invention provides, as a 10 th aspect, use of the type I collagen production promoter described in the 1 st aspect for producing a cosmetic, characterized in that it is at least one selected from the group consisting of an extract of Leonurus japonicus (Leonurus japonicus), and an extract of Polygonatum odoratum (Polygonatum odoratum).
Detailed Description
The motherwort extract in the invention is an extract obtained by extracting motherwort powder with a mixed solution of water and alcohol. More preferably, the extract is obtained by extracting the extract with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the extract with a mixture of water and a lower polyol having 3 to 6 carbon atoms. More preferably, the extract is obtained by extracting with a mixture of water and a lower alcohol having 1 to 3 carbon atoms and then further extracting with a mixture of water and a lower polyol having 3 to 5 carbon atoms. Most preferred is an extract obtained by further extracting with a mixture of water and butylene glycol after extracting with a mixture of water and ethanol.
Examples of the lower alcohol having 1 to 6 carbon atoms include methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, tert-butanol, n-pentanol, 2-pentanol, tert-pentanol, n-hexanol, 2-hexanol, and 3-hexanol.
Examples of the lower polyhydric alcohol having 3 to 6 carbon atoms include propylene glycol, glycerol, butylene glycol, butanetriol, butanetetraol, pentanediol, pentanetriol, pentaerythritol, hexanediol, hexanetriol, and hexantetraol.
The mixing ratio of the mixed solution of water and a lower alcohol used in the first extraction is not particularly limited, but is preferably 3 to 7:7 to 3, more preferably 4 to 6:6 to 4, and most preferably 1: 1.
The mixing ratio of the mixed solution of water and a lower polyhydric alcohol used in the second extraction step is not particularly limited, but is preferably 2 to 6:8 to 4, more preferably 3 to 6:7 to 4, and most preferably 6: 4.
When the extraction is carried out with a mixed solution of water and a lower alcohol, the heating temperature is not particularly limited, and is preferably 70 to 90 ℃, more preferably 75 to 80 ℃. The heating time is also not particularly limited, but is preferably 5 to 10 hours, and more preferably 5 to 8 hours.
The polygonatum odoratum extract is obtained by extracting polygonatum odoratum powder with a mixed solution of water and alcohol. More preferably, the extract is obtained by extracting the extract with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the extract with a mixture of water and a lower polyol having 3 to 6 carbon atoms. More preferably, the extract is obtained by extracting with a mixture of water and a lower alcohol having 1 to 3 carbon atoms and then further extracting with a mixture of water and a lower polyol having 3 to 5 carbon atoms. Most preferred is an extract obtained by further extracting with a mixture of water and butylene glycol after extracting with a mixture of water and ethanol.
Examples of the lower alcohol having 1 to 6 carbon atoms include methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, tert-butanol, n-pentanol, 2-pentanol, tert-pentanol, n-hexanol, 2-hexanol, and 3-hexanol.
Examples of the lower polyhydric alcohol having 3 to 6 carbon atoms include propylene glycol, glycerol, butylene glycol, butanetriol, butanetetraol, pentanediol, pentanetriol, pentaerythritol, hexanediol, hexanetriol, and hexantetraol.
The mixing ratio of the mixed solution of water and a lower alcohol used in the first extraction is not particularly limited, but is preferably 3 to 7:7 to 3, more preferably 4 to 6:6 to 4, and most preferably 1: 1.
The mixing ratio of the mixed solution of water and the lower polyhydric alcohol used in the second extraction step is not particularly limited, but is preferably 5 to 10:5 to 1, more preferably 7 to 10:3 to 1, and most preferably 10: 1.
When the extraction is carried out with a mixed solution of water and a lower alcohol, the heating temperature is not particularly limited, and is preferably 70 to 90 ℃, more preferably 75 to 80 ℃. The heating time is also not particularly limited, but is preferably 5 to 10 hours, and more preferably 5 to 8 hours.
The present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.
Preparation example 1: preparation method of herba Leonuri (Leonurus japonicus) extract
Adding 5-10 parts by weight of a 1:1 mixed solution of deionized water and ethanol into 1 part by weight of motherwort powder according to the size of a reaction vessel, performing ultrasonic stirring under the condition that the motherwort powder is completely immersed in a solvent, then performing heating extraction at 75-80 ℃ for 5-8 hours, and then concentrating and drying to obtain a motherwort extract.
Adding 19 parts by weight of a 6:4 mixed solution of water and butanediol into 1 part by weight of the obtained motherwort extract, dissolving, carrying out decoloring and deodorizing treatment, standing at the low temperature of 6-8 ℃ for 48 hours after treatment, filtering to remove insoluble substances, adding 1, 2-hexanediol and p-hydroxyacetophenone serving as preservatives to enable the concentrations of the 1, 2-hexanediol and the p-hydroxyacetophenone to reach 0.4 wt% -0.6 wt% respectively, thus obtaining a semi-finished product of the motherwort extract, detecting in a quality control department, detecting that a product with the stachydrine content of more than 0.1% is a qualified product, and carrying out GMP packaging to obtain a finished product of the motherwort extract. The obtained motherwort herb extract finished product is a transparent liquid with the density of 0.95-1.05 g/cm from brown yellow to brown red3The refractive index is 1.340-1.400, and the concentration of active ingredients is 3-5 wt%.
Example 1 anti-aging efficacy test experiment of motherwort extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500 ml); trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell assay plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 5810R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) Test implementation
1. The extract solution of motherwort (Leonurus Japonicus extract using an extract solution having an active ingredient concentration of 3 wt%) obtained in the above preparation example was stored at room temperature for later use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90% for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the motherwort extract by using a DMEM medium according to a specified concentration after 24 hours to ensure that the concentration of the motherwort extract reaches 0.10%, 0.05% and 0.01% (namely the concentration of active ingredients of the motherwort extract reaches 0.003 wt%, 0.0015 wt% and 0.0003 wt% respectively), adding the diluted motherwort extract into an E-plate16 plate, continuously culturing for 24 hours, observing, and selecting the optimal concentration for subsequent experiments.
(5) Test results and discussion:
promoting collagen I production
TABLE 1
From the data in the above table, it can be seen that, in the predetermined concentration range, when the concentration of the motherwort extract is 0.10%, i.e., the concentration of the active components in motherwort is 0.003 wt%, the collagen content in the skin cell model is 0.6652 μ g/mL, when the concentration of the motherwort extract is 0.05%, i.e., the concentration of the active components in motherwort is 0.0015 wt%, the collagen content is 0.5455 μ g/mL, when the concentration of the motherwort extract is 0.01%, i.e., the concentration of the active components in motherwort is 0.0003 wt%, the collagen content is 0.2822 μ g/mL, and the collagen content in the negative control DMEM is 0.5083 μ g/mL. The results show that the leonurus extract with the leonurus extract concentration of 0.01% (namely when the leonurus active ingredient concentration is 0.0003 wt%) has no obvious collagen synthesis promoting capability, and the leonurus extract with the leonurus extract concentration of 0.1% and 0.05% (namely when the leonurus active ingredient concentration is 0.003 wt% and 0.0015 wt%) has a promoting effect on the generation of type I collagen in skin cells, and the results show that the leonurus extract with the leonurus extract concentration of 0.05% and 0.1% (namely when the leonurus active ingredient concentration is 0.003 wt% and 0.0015 wt%) has a certain anti-aging capability on human skin cells.
And (4) conclusion:
when the concentration of the active ingredients of the motherwort herb extract reaches more than 0.0015 wt%, the generation of type I collagen in a human skin fibroblast model can be promoted, and the anti-aging effect is excellent.
Example 2 anti-aging efficacy test experiment of motherwort extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500 ml); trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell assay plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 5810R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) Test implementation
1. The extract solution of Leonurus Japonicus (Leonurus japonica extract using an extract solution having an active ingredient concentration of 3.7 wt%) obtained according to the above preparation example was stored at room temperature for later use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90 percent for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the motherwort extract by using a DMEM medium according to the specified concentration after 24 hours to ensure that the concentration of the motherwort extract reaches 0.10 percent, 0.05 percent and 0.01 percent respectively (namely the concentration of the active ingredients of the motherwort extract reaches 0.0037 percent, 0.00185 percent and 0.00037 percent respectively), adding the diluted motherwort extract into an E-plate16 plate, continuously culturing for 24 hours, observing, and selecting the optimal concentration for subsequent experiments.
(5) Test results and discussion:
promoting collagen I production
TABLE 2
From the data in the above table, it can be seen that, in the predetermined concentration range, when the concentration of the motherwort extract is 0.10%, i.e., the concentration of the active components in motherwort is 0.0037 wt%, the collagen content in the skin cell model is 0.6669 μ g/mL, when the concentration of the motherwort extract is 0.05%, i.e., the concentration of the active components in motherwort is 0.00185 wt%, the collagen content is 0.5757 μ g/mL, when the concentration of the motherwort extract is 0.01%, i.e., the concentration of the active components in motherwort is 0.00037 wt%, the collagen content in the motherwort extract is 0.2848 μ g/mL, and the collagen content in the negative control DMEM is 0.5083 μ g/mL. The results show that the leonurus extract with the leonurus extract concentration of 0.01% (namely when the leonurus active ingredient concentration is 0.00037 wt%) has no obvious collagen synthesis promoting capability, and the leonurus extract with the leonurus extract concentration of 0.1% and 0.05% (namely when the leonurus active ingredient concentration is 0.0037 wt% and 0.00185 wt%) has a promoting effect on the generation of type I collagen in skin cells, and the results show that the leonurus extract with the leonurus extract concentration of 0.05% and 0.1% (namely when the leonurus active ingredient concentration is 0.0037 wt% and 0.00185 wt%) has a certain anti-aging capability on human skin cells.
And (4) conclusion:
when the concentration of the active ingredients of the motherwort herb extract reaches above 0.00185 wt%, the formation of type I collagen in a human skin fibroblast model can be promoted, and the anti-aging effect is excellent.
Example 3 anti-aging efficacy test experiment of motherwort extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500 ml); trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell assay plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 5810R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) Test implementation
1. The extract solution of Leonurus Japonicus (Leonurus japonica extract using an extract solution having an active ingredient concentration of 5.0 wt%) obtained according to the above preparation example was stored at room temperature for later use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90% for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the motherwort extract by using a DMEM medium according to a specified concentration after 24 hours to ensure that the concentration of the motherwort extract reaches 0.10%, 0.05% and 0.01% (namely the concentration of active ingredients of the motherwort extract reaches 0.005 wt%, 0.0025 wt% and 0.0005% respectively), adding the diluted motherwort extract into an E-plate16 plate for continuous culture for 24 hours, observing, and selecting the optimal concentration for subsequent experiments.
(5) Test results and discussion:
promoting collagen I production
TABLE 3
From the data in the above table, it can be seen that, in the predetermined concentration range, when the concentration of the motherwort extract is 0.10%, i.e., the concentration of the active components in motherwort is 0.005 wt%, the collagen content in the skin cell model is 0.7888 μ g/mL, when the concentration of the motherwort extract is 0.05%, i.e., the concentration of the active components in motherwort is 0.0025 wt%, the collagen content is 0.6122 μ g/mL, when the concentration of the motherwort extract is 0.01%, i.e., the concentration of the active components in motherwort is 0.0005 wt%, the collagen content is 0.2745 μ g/mL, and the collagen content in the negative control DMEM is 0.5083 μ g/mL. The results show that the leonurus extract with the leonurus extract concentration of 0.01% (i.e. when the leonurus active ingredient concentration is 0.0005 wt%) has no obvious collagen synthesis promoting capability, and the leonurus extract with the leonurus extract concentration of 0.1% and 0.05% (i.e. when the leonurus active ingredient concentration is 0.005 wt% and 0.0025 wt%) has a promoting effect on the generation of type I collagen in skin cells, and the results show that the leonurus extract with the leonurus extract concentration of 0.05% and 0.1% (i.e. when the leonurus active ingredient concentration is 0.005 wt% and 0.0025 wt%) has a certain anti-aging capability on human skin cells.
And (4) conclusion:
when the concentration of the active ingredients of the motherwort herb extract reaches more than 0.0025 wt%, the generation of type I collagen in a human skin fibroblast model can be promoted, and the anti-aging effect is excellent.
Preparation example 2 preparation of Polygonatum odoratum extract
Adding 5-10 parts by weight of a 1:1 mixed solution of deionized water and ethanol into 1 part by weight of Polygonatum odoratum powder (Polygonatum odoratum) according to the size of a reaction vessel, performing ultrasonic stirring under the condition that the Polygonatum odoratum powder is completely immersed in a solvent, heating and extracting at 75-80 ℃ for 5-8 hours, and then concentrating and drying to obtain the Polygonatum odoratum extract.
Adding 99 parts by weight of a 10:1 mixed solution of water and butanediol to 1 part by weight of the obtained polygonatum extract, dissolving, decoloring and deodorizing, standing at a low temperature of 6-8 ℃ for 48 hours after treatment, filtering to remove insoluble substances, and adding 1, 2-hexanediol and p-hydroxyacetophenone serving as preservatives to enable the concentrations of the 1, 2-hexanediol and the p-hydroxyacetophenone to reach 0.3-0.7 wt% respectively to obtain a semi-finished product of the polygonatum extract. Detecting the semi-finished product in quality control department, wherein the product with the content of polygonatum odoratum polysaccharide more than or equal to 0.2 wt% is a qualified product, and obtaining a finished product of polygonatum odoratum extract by GMP packaging. The obtained polygonatum odoratum extract finished product is light yellow to dark yellow transparent liquid with the density of 1.00-1.05 g/cm3The refractive index is 1.310-1.390, and the concentration of active ingredients is 0.5 wt% -0.9 wt%.
Example 4 anti-aging efficacy test report of Yuzhu extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500 ml); trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell assay plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 58)10R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) And (3) test implementation:
1. storing the EXTRACT of rhizoma Polygonati Odorati (POLYGONATUM ODORATUM EXTRACT, using EXTRACT with active ingredient concentration of 0.5 wt%) at room temperature for use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90 percent for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the polygonatum odoratum extract by using a DMEM medium according to a specified concentration after 24 hours so that the concentration of the polygonatum odoratum extract reaches 0.10 percent, 0.05 percent and 0.01 percent respectively (namely the concentration of active ingredients of the polygonatum odoratum extract reaches 0.0005wt percent, 0.00025wt percent and 0.00005wt percent respectively), adding the diluted polygonatum odoratum extract into an E-plate16 plate, continuously culturing for 24 hours, observing, and selecting an optimal concentration for subsequent experiments.
(5) Test results and discussion:
promoting collagen I production
TABLE 4
From the data in the above table, it can be seen that, in the test results of type i collagen production of Yuzhu extract within the predetermined concentration range, when the concentration of Yuzhu extract was 0.10%, i.e., the concentration of Yuzhu active ingredient was 0.0005 wt%, the collagen content was 0.6424 μ g/mL, when the concentration of Yuzhu extract was 0.05%, i.e., the concentration of Yuzhu active ingredient was 0.00025 wt%, the collagen content was 0.6055 μ g/mL, when the concentration of Yuzhu extract was 0.01%, i.e., the concentration of Yuzhu active ingredient was 0.00005 wt%, the collagen content was 0.2987 μ g/mL, and the collagen content of DMEM as a negative control was 0.5083 μ g/mL. The polygonatum extract with the concentration of 0.01 percent of the polygonatum extract, namely the concentration of the active ingredient of the polygonatum is 0.00005wt percent, has no obvious collagen promotion capability, the polygonatum extract with the concentration of 0.05 percent of the polygonatum extract, namely the concentration of the active ingredient of the polygonatum is 0.00025wt percent, and the polygonatum extract with the concentration of 0.1 percent of the polygonatum extract, namely the concentration of the active ingredient of the polygonatum is 0.0005wt percent have very good collagen generation promotion effects, and the polygonatum extract with the concentration of 0.05 percent of the polygonatum extract, namely the concentration of the active ingredient of the polygonatum is 0.00025wt percent, and the polygonatum extract with the concentration of 0.1 percent of the polygonatum extract, namely the concentration of the active ingredient of the polygonatum is 0.0005wt percent, have the anti-aging effect.
And (4) conclusion:
when the concentration of the active ingredients of the polygonatum odoratum extract reaches more than 0.00025 wt%, the generation of type I collagen in a human skin fibroblast model can be promoted, and the polygonatum odoratum extract has excellent anti-aging effect.
Example 5 anti-aging efficacy test report of Yuzhu extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500 ml); trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell detection plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 5810R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) And (3) test implementation:
1. storing the extractive solution (POLYGONATUM ODORATUM EXTRACT, extractive solution with active ingredient concentration of 0.8 wt%) at room temperature for use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90 percent for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the polygonatum odoratum extract by using a DMEM medium according to a specified concentration after 24 hours so that the concentration of the polygonatum odoratum extract reaches 0.10 percent, 0.05 percent and 0.01 percent respectively (namely the concentration of active ingredients of the polygonatum odoratum extract reaches 0.0008 percent, 0.0004 percent and 0.00008 percent respectively), adding the diluted polygonatum odoratum extract into an E-plate16 plate, continuously culturing for 24 hours, observing, and selecting the optimal concentration for subsequent experiments.
(5) Test results and discussion:
promoting collagen I production
TABLE 5
From the data in the above table, it can be seen that, in the test results of collagen production promotion of type i collagen of polygonatum extract within the predetermined concentration range, when the concentration of polygonatum extract is 0.10%, i.e., when the concentration of polygonatum active ingredient is 0.0008 wt%, the collagen content is 0.7015 μ g/mL, when the concentration of polygonatum extract is 0.05%, i.e., when the concentration of polygonatum active ingredient is 0.0004 wt%, the collagen content is 0.8638 μ g/mL, when the concentration of polygonatum extract is 0.01%, i.e., when the concentration of polygonatum active ingredient is 0.00008 wt%, the collagen content is 0.3275 μ g/mL, and the collagen content of DMEM as a negative control is 0.5083 μ g/mL. The polygonatum extract with the concentration of 0.01 percent of polygonatum extract, namely the concentration of 0.00008 percent of polygonatum active ingredient, has no obvious collagen promotion capability, the polygonatum extract with the concentration of 0.05 percent of polygonatum extract, namely the concentration of 0.0004 percent of polygonatum extract, and the polygonatum extract with the concentration of 0.1 percent of polygonatum extract, namely the concentration of 0.0008 percent of polygonatum active ingredient, both have very good collagen generation promotion effects, and the polygonatum extract with the concentration of 0.05 percent of polygonatum extract, namely the concentration of 0.0004 percent of polygonatum extract and the polygonatum extract with the concentration of 0.1 percent of polygonatum extract, namely the concentration of 0.0008 percent of polygonatum extract have anti-aging effects.
And (4) conclusion:
when the concentration of the active ingredients of the polygonatum odoratum extract reaches more than 0.0004 wt%, the generation of type I collagen in a human skin fibroblast model can be promoted, and the polygonatum odoratum extract has an excellent anti-aging effect.
Example 6 anti-aging efficacy test report of Yuzhu extract
(1) Description of the principle of the test method:
human type i collagen (Col i) collagen test principle: coating a microporous plate with a purified antibody to prepare a solid phase carrier, sequentially adding a specimen or a standard substance, a biotinylated anti-Col I antibody and HRP-labeled avidin into the micropore coated with the anti-Col I antibody, and developing with a substrate TMB after thorough washing. TMB is converted to blue by the catalysis of peroxidase and to the final yellow by the action of an acid. The shade of the color is positively correlated with ColI in the sample. The absorbance (OD value) was measured at a wavelength of 450nm with a microplate reader, and the sample concentration was calculated.
(2) Test drugs, materials, instruments and preparation: fibroblasts (HFF-1); human skin keratinocytes (HaCaT); DMEM medium (GIBCO, 500 ml); fetal bovine serum (GIBCO, 500ml)) (ii) a trypsin-EDTA (GIBCO, 100 ml); phosphate buffer (CORNING, 500ml), active oxygen detection kit (bi yun tian); human type I collagen (Col I) enzyme linked immunosorbent assay kit (Wuhan Huamei), RTCA instrument (Aisen); cell assay plate (E-plate 16); inverted microscope (OLYMPUS IX 73); a multifunctional microplate reader (TECAN, Infinite M200 Pro); incubator (Thermoscientific, DIRECT HEAT CO)2Incubator); centrifuge (eppendorf, Centrifuge 5810R); t25 cell culture flasks (COSTAR, 430168); cell culture plates (96 wells) (Costar, 3599); cell counter (Life, counter II FL).
(3) Test implementation conditions: sterile, ultra clean, cell requires 5% CO maintenance2And cultured at 37 ℃.
(4) And (3) test implementation:
1. storing the extractive solution (POLYGONATUM ODORATUM EXTRACT, extractive solution with active ingredient concentration of 0.9 wt%) at room temperature for use. When in use, the sample is taken out and shaken evenly and is diluted by serum-free culture medium in a gradient way.
2. And (5) culturing the cells. Selecting HFF-1 of logarithmic growth phase, digesting with 0.25% trypsin, subculturing with DMEM medium containing 10% fetal calf serum, placing in incubator at 37 deg.C and 5% CO2Culturing is carried out in the environment.
3. Selecting HFF-1 with the growth density of about 90 percent for experiment, spreading the HFF-1 in an E-plate16 plate for culture, diluting the polygonatum odoratum extract by using a DMEM medium according to a specified concentration after 24 hours so that the concentration of the polygonatum odoratum extract reaches 0.10 percent, 0.05 percent and 0.01 percent respectively (namely the concentration of active ingredients of the polygonatum odoratum extract reaches 0.0009 percent, 0.00045 percent and 0.00009 percent respectively), adding the diluted polygonatum odoratum extract into an E-plate16 plate, continuously culturing for 24 hours, observing, and selecting the optimal concentration for subsequent experiments.
(5) Test results and discussion:
TABLE 6
From the data in the above table, it can be seen that, in the predetermined concentration range, the collagen content of the Yuzhu extract is 0.7308 μ g/mL when the Yuzhu extract solution concentration is 0.10%, i.e., the Yuzhu active ingredient concentration is 0.0009 wt%, the collagen content is 0.6812 μ g/mL when the Yuzhu extract solution concentration is 0.05%, i.e., the Yuzhu active ingredient concentration is 0.00045 wt%, the collagen content is 0.3321 μ g/mL when the Yuzhu extract solution concentration is 0.01%, i.e., the Yuzhu active ingredient concentration is 0.00009 wt%, and the collagen content of the negative control DMEM is 0.5083 μ g/mL. The polygonatum extract with the concentration of 0.01 percent of polygonatum extract, namely the concentration of 0.00009 percent of polygonatum active ingredient, has no obvious collagen promotion capability, the polygonatum extract with the concentration of 0.05 percent of polygonatum extract, namely the concentration of 0.00045 percent of polygonatum extract and the polygonatum extract with the concentration of 0.1 percent of polygonatum extract, namely the concentration of 0.0009 percent of polygonatum active ingredient both have good collagen generation promotion effects, and the polygonatum extract with the concentration of 0.05 percent of polygonatum extract, namely the concentration of 0.00045 percent of polygonatum active ingredient and the polygonatum extract with the concentration of 0.1 percent of polygonatum extract, namely the concentration of 0.0009 percent of polygonatum active ingredient have anti-aging effects.
And (4) conclusion:
when the concentration of the active ingredients of the polygonatum odoratum extract reaches more than 0.00045 wt%, the generation of type I collagen in a human skin fibroblast model can be promoted, and the polygonatum odoratum extract has excellent anti-aging effect.
Claims (10)
1. A collagen I production promoter is characterized by being at least one selected from the group consisting of an extract of motherwort (Leonurus japonicus), and an extract of Polygonatum odoratum.
2. The type I collagen production-promoting agent according to claim 1, wherein the concentration of the active ingredient of said motherwort herb extract is 0.0015% by weight or more.
3. The type I collagen production promoter according to claim 2, wherein the concentration of the active ingredient of said motherwort extract is 0.00185 wt% or more.
4. The type I collagen production promoter according to claim 1, wherein the concentration of the active ingredient of Yuzhu extract is 0.00025 wt% or more.
5. The type I collagen production promoter according to claim 4, wherein the concentration of the active ingredient of Yuzhu extract is 0.0004 wt% or more.
6. The type I collagen production promoter according to claim 1, wherein the motherwort herb extract is obtained by extracting motherwort herb powder with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the extract with a mixture of water and a polyhydric alcohol having 3 to 6 carbon atoms.
7. The type I collagen production promoter according to claim 6, wherein the motherwort extract is an extract obtained by subjecting motherwort powder to ultrasonic agitation together with ethanol and deionized water, heating extraction, concentration drying, and decoloring and deodorizing with water and butanediol.
8. The type I collagen production promoter according to claim 1, wherein the polygonatum odoratum extract is obtained by extracting a polygonatum odoratum powder with a mixture of water and a lower alcohol having 1 to 6 carbon atoms and then further extracting the polygonatum odoratum powder with a mixture of water and a polyhydric alcohol having 3 to 6 carbon atoms.
9. The type I collagen production promoter according to claim 7, wherein the Polygonatum odoratum extract is obtained by subjecting Polygonatum odoratum powder to ultrasonic agitation with ethanol and deionized water, heat extraction, concentration drying, and decoloring and deodorizing with water and butanediol.
10. Use of the type I collagen production promoter according to claim 1 for the preparation of cosmetics, characterized by being at least one selected from the group consisting of motherwort (Leonurus japonicus) extract, Polygonatum odoratum (Polygonatum odoratum) extract.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111025328.5A CN113616573A (en) | 2021-09-02 | 2021-09-02 | I type collagen production promoter |
| PCT/CN2022/113589 WO2023030041A1 (en) | 2021-09-02 | 2022-08-19 | Type i collagen production accelerant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111025328.5A CN113616573A (en) | 2021-09-02 | 2021-09-02 | I type collagen production promoter |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113616573A true CN113616573A (en) | 2021-11-09 |
Family
ID=78388844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111025328.5A Pending CN113616573A (en) | 2021-09-02 | 2021-09-02 | I type collagen production promoter |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN113616573A (en) |
| WO (1) | WO2023030041A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023030041A1 (en) * | 2021-09-02 | 2023-03-09 | 株式会社资生堂 | Type i collagen production accelerant |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108498445A (en) * | 2018-06-26 | 2018-09-07 | 四川自然菲科技有限公司 | A kind of novel anti-aging type Essence |
| CN109730957A (en) * | 2019-03-14 | 2019-05-10 | 陕西中医药大学附属医院 | Compound cold cream and its preparation method with performance of keeping humidity |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103948529B (en) * | 2014-01-26 | 2016-09-07 | 广州雷诺生物科技有限公司 | A kind of cosmetic composition containing tomato pulp extract |
| CN109758393A (en) * | 2019-01-30 | 2019-05-17 | 浙江更土电子商务有限公司 | It is a kind of with whitening, moisturizing, the skin care item of anti-aging effects and its preparation method and application |
| CN109730942A (en) * | 2019-01-30 | 2019-05-10 | 浙江更土电子商务有限公司 | One kind containing collagen hydrolysate, the skin care item and its preparation method and application of moisturizing moisturizing |
| CN109700726B (en) * | 2019-03-19 | 2021-11-05 | 广东芭薇生物科技股份有限公司 | A skin external composition and cosmetic for improving wrinkle |
| CN114929195B (en) * | 2020-01-02 | 2024-08-06 | 株式会社Lg生活健康 | Composition containing plant extract |
| CN113616573A (en) * | 2021-09-02 | 2021-11-09 | 株式会社资生堂 | I type collagen production promoter |
-
2021
- 2021-09-02 CN CN202111025328.5A patent/CN113616573A/en active Pending
-
2022
- 2022-08-19 WO PCT/CN2022/113589 patent/WO2023030041A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108498445A (en) * | 2018-06-26 | 2018-09-07 | 四川自然菲科技有限公司 | A kind of novel anti-aging type Essence |
| CN109730957A (en) * | 2019-03-14 | 2019-05-10 | 陕西中医药大学附属医院 | Compound cold cream and its preparation method with performance of keeping humidity |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023030041A1 (en) * | 2021-09-02 | 2023-03-09 | 株式会社资生堂 | Type i collagen production accelerant |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2023030041A1 (en) | 2023-03-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101059471B1 (en) | Cosmetic composition for skin aging | |
| CN103585097B (en) | Epidermal growth factor-loaded bletilla rhizome polysaccharide compound, and preparation method and application thereof | |
| JP2011505353A (en) | Skin external preparation composition containing a compound herbal extract as an active ingredient | |
| CN105902454B (en) | One kind being used for the regenerated Moringa compound Essence of cell repair | |
| CN102258442A (en) | Compound traditional Chinese medicine extract and application thereof to whitening, moisturizing and anti-aging skin care product | |
| CN102579315B (en) | Plant composition applied to cosmetics and application method for plant composition | |
| CN110772454B (en) | Skin-brightening, moisturizing, soothing and anti-aging compound essential oil, and preparation method and application thereof | |
| CN103484308A (en) | Health-care wine and preparation method thereof | |
| KR100611796B1 (en) | Cosmetic composition containing medicinal herbs called sipgeondaebodan and method for preparing the same | |
| KR100780180B1 (en) | Chinese composition preventing loss of hair and promoting growth of hair and method for preparing the same | |
| CN109464354B (en) | Skin care composition with effects of reducing subcutaneous fat content and improving skin firmness | |
| CN113616573A (en) | I type collagen production promoter | |
| KR20140145278A (en) | Cosmetic composition containing Cimicifuga heracleifolia extract, Cornus officinalis extract and Geranium nepalense extract for skin convergence and elasticity effect | |
| KR20100124132A (en) | Anti-wrinkle cosmetic composition containing oriental herb extract treated by enzyme and its extraction method | |
| CN114831918A (en) | Traditional Chinese medicine composition fermentation liquor, preparation method and application thereof | |
| CN115137675A (en) | Compound plant extract with moisturizing effect and application thereof | |
| CN109394801A (en) | The composition of the decomposition of the generation and promotion glycosylation end products for inhibiting glycosylation end products containing chestnut Herba Visci extract | |
| CN107823114A (en) | A kind of anti-acne skin care item containing sealwort stem cell extract | |
| CN103301357A (en) | Formula and preparation method for traditional Chinese medicine preparation for promoting hair blacking | |
| CN113413419A (en) | Ovary care composition containing jujube exosome and preparation method thereof | |
| CN113908087A (en) | Natural moisturizing combined extract, preparation method and application thereof | |
| CN106942561A (en) | A kind of beverage composition for treating dental erosion and preparation method and application | |
| KR20160057183A (en) | Cosmetic composition containing the fermented extract of allium monanthum, allium scorodorpasum, allium tuberosum, allium fistulosum and scilla scilloides | |
| CN108524850A (en) | A kind of liquor preparation for enhancing immune prevention cardiovascular and cerebrovascular disease | |
| CN114788796B (en) | Whitening and antibacterial cosmetic containing Fuding white tea extract and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |