CN116492251A - Antioxidant and saccharification inhibitor and application thereof - Google Patents
Antioxidant and saccharification inhibitor and application thereof Download PDFInfo
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- CN116492251A CN116492251A CN202310268215.0A CN202310268215A CN116492251A CN 116492251 A CN116492251 A CN 116492251A CN 202310268215 A CN202310268215 A CN 202310268215A CN 116492251 A CN116492251 A CN 116492251A
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- Prior art keywords
- antioxidant
- plant
- butanediol
- extract
- water
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- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 229940119170 jojoba wax Drugs 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229940017219 methyl propionate Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000004200 microcrystalline wax Substances 0.000 description 1
- 235000019808 microcrystalline wax Nutrition 0.000 description 1
- 229940114937 microcrystalline wax Drugs 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 description 1
- GOQYKNQRPGWPLP-UHFFFAOYSA-N n-heptadecyl alcohol Natural products CCCCCCCCCCCCCCCCCO GOQYKNQRPGWPLP-UHFFFAOYSA-N 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 229940055577 oleyl alcohol Drugs 0.000 description 1
- XMLQWXUVTXCDDL-UHFFFAOYSA-N oleyl alcohol Natural products CCCCCCC=CCCCCCCCCCCO XMLQWXUVTXCDDL-UHFFFAOYSA-N 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000011814 protection agent Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 229940032094 squalane Drugs 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003760 tallow Substances 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229940099259 vaseline Drugs 0.000 description 1
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
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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 invention discloses an antioxidant and saccharification inhibitor, which is prepared by the following steps of pretreatment; extracting and concentrating. The antioxidant and saccharification inhibitor is kava extract, has the effects of resisting oxidation and inhibiting protein saccharification, can inhibit collagenase activity, inhibit synthesis secretion of cell factor (IL-12) (inhibit cell aging), and improve skin relaxation. Can be used in various cosmetics, and has good compatibility with various cosmetic components.
Description
Technical Field
The invention relates to an antioxidant and saccharification inhibitor and application thereof, relates to A61K, and in particular relates to the field of cosmetic preparations.
Background
Since skin aging resistance has been pursued by people who are loving beauty since ancient times, research on skin aging has progressed to a cellular level with progress of technology, and it is known that skin cell aging is caused by oxidative damage of cells or oxidative damage of proteins to induce inflammatory cytokines to peripheral cells, it is necessary for skin external preparations to inhibit oxidative damage of cells, inhibit protein glycation, and inhibit induction of secretion of aging factors in order to inhibit skin aging. The kava is a plant of the genus Piperaceae, and has an effect of inhibiting fat synthesis and a moisturizing effect when applied to cosmetics in the conventional process, but no effect of inhibiting the induction of aging factor synthesis is disclosed with respect to an antioxidant effect, an anti-protein glycation effect. And the extract in the present application also has the effects of inhibiting collagenase activity and improving skin relaxation.
Disclosure of Invention
In order to increase the extraction rate of the effective components of the kava, so that the kava has the effects of resisting oxidation, inhibiting saccharification, inhibiting collagenase activity, inhibiting the secretion of induced aging factors and improving relaxation, the first aspect of the invention provides an antioxidant and saccharification inhibitor, which is prepared by the following steps:
(1) Pre-washing the plant to be extracted with water, removing surface foreign matters, drying, and crushing;
(2) Mixing crushed plant to be extracted with an extractant, extracting, and filtering to obtain crude plant extract;
(3) Concentrating the crude plant extract under reduced pressure to obtain plant concentrate, and adding water and antiseptic into the plant concentrate to obtain plant extract.
As a preferred embodiment, the plant to be extracted is a plant of the genus piperaceae, the plant of the genus piperaceae is kava (Macropiper excelsum), and the extraction site of the kava is a leaf.
As a preferred embodiment, the mass ratio of the slip blades to the extracting agent is 1: (4-6), and more preferably, the mass ratio of the slip blades to the extracting agent is 1:5.
as a preferred embodiment, the extractant is selected from one or a combination of several of water, lower alcohol, polyhydric alcohol, ester extractant, ketone extractant, ether extractant, hydrocarbon extractant.
As a preferred embodiment, the lower alcohol is selected from one or a combination of several of methanol, ethanol and propanol, and the polyhydric alcohol is selected from one or a combination of several of ethylene glycol, propylene glycol, butylene glycol and glycerin.
As a preferred embodiment, the ester extractant is selected from one or a combination of more of ethyl acetate, butyl acetate and methyl propionate; the ketone extractant is selected from one or a combination of more of acetone and methyl ethyl ketone; the ether extractant is selected from one or a combination of more of diethyl ether and isopropyl ether; the hydrocarbon extractant is n-hexane.
As a preferred embodiment, the extractant is a mixed solution of ethanol and water or a mixed solution of butanediol and water, and the butanediol is selected from one or a combination of more than one of 1, 2-butanediol, 1, 3-butanediol and 1, 4-butanediol.
As a preferred embodiment, the mass fraction of the ethanol in the mixed solution of the ethanol and the water is 10-60%, and the mass fraction of the butanediol in the mixed solution of the butanediol and the water is 10-70%. Further preferably, the butanediol is 1, 3-butanediol.
As a preferred embodiment, the mass fraction of ethanol in the mixed solution of ethanol and water is 45-55%. Further preferably, the mass fraction of ethanol in the mixed solution of ethanol and water is 50%.
As a preferred embodiment, the parameters extracted in the step 2 are: extracting at 0-80deg.C for 1-168 hr; the pH of the plant crude extract is 3-9.
As a preferred embodiment, the parameters extracted in the step 2 are: the extraction temperature is 30-50deg.C, and the extraction time is 1.5-3h; the pH of the plant crude extract is 4-8.
As a preferred embodiment, the preservative in the step 3 is 1, 3-butanediol, and the total addition mass fraction of the 1, 3-butanediol is 35-55%.
As a preferred embodiment, the pH is adjusted by adding an acid-base modifier selected from one or a combination of several of sodium hydroxide, sodium carbonate, potassium hydroxide, sodium citrate, hydrochloric acid, phosphoric acid, sulfuric acid.
The second aspect of the invention provides an application of an antioxidant and saccharification inhibitor, which is applied to daily chemical products, wherein the daily chemical products are selected from one of emulsion, cream, toning lotion, essence, peel-off facial mask, lipstick, foundation, sheet facial mask, foundation liquid, cosmetic pressed powder, blush, powdery mildew, facial cleanser, body shampoo, hair shampoo, soap, hair tonic and bath agent.
In a preferred embodiment, the daily chemical product comprises one or more of hydrocarbon auxiliary agents, ester auxiliary agents, triglyceride auxiliary agents, fatty acids, higher alcohols, surfactants, polyols, thickening gelling agents, antioxidants, ultraviolet ray protection agents, color agents, preservatives, pH adjusters, moisturizers, skin conditioning agents, plant components and microorganism components.
As a preferred embodiment, the hydrocarbon auxiliary is selected from one or a combination of squalane, vaseline and microcrystalline wax; the ester auxiliary agent is one or a combination of jojoba oil, carnauba wax and octyl dodecyl oleate; the triglyceride auxiliary agent is selected from one or a combination of olive oil, beef tallow and coconut oil; the fatty acid is selected from one or a combination of stearic acid, oleic acid and retinoic acid; the higher alcohol is selected from one or a combination of oleyl alcohol, stearyl alcohol and octyl dodecanol; the surfactant is selected from one or a combination of sulfosuccinic acid ester, polyoxyethylene alkyl sodium sulfate anionic surfactant, alkyl betaine salt amphoteric surfactant, dialkyl ammonium salt cationic surfactant, sorbitan fatty acid ester polyoxyethylene adduct, fatty acid monoglyceride polyoxyethylene adduct, polyoxyethylene alkyl ether and polyoxyethylene fatty acid ester nonionic surfactant; the polyalcohol is selected from one or a combination of polyethylene glycol, glycerol and 1, 3-butanediol; the skin conditioning agent is selected from one or a combination of dipotassium glycyrrhizinate, hyaluronic acid, hyaluronate and hydroxy pinacolone retinoate; the plant component is selected from, but not limited to, camellia extract; the microbial component is selected from, but not limited to, a bifidobacterium culture medium component.
As a preferred embodiment, the material is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 0.5-2%,1, 3-butanediol 1-5%, nipagin ester 0.2-0.3%, purified water was supplemented to 100%.
As a preferred embodiment, the material is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: 0.5-2% of the extract of example 1, 0.5-3% of beta-glucan, 0.2-6% of beta-glucan, 1-5% of 1, 3-butanediol, 0.05-0.1% of sodium citrate and the purified water is supplemented to 100%.
As a preferred embodiment, the material is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 0.5-2%, hydroxy pinacolone retinoate 0.05-1%, bifidobacterium culture solution 1-3%,1, 3-butanediol 0.1-5%, phenoxyethanol 0.5-0.8%, sodium citrate 0.05-0.1%, purified water up to 100%.
As a preferred embodiment, the material is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 0.5-2%, centella asiatica extract hydroxy pinacolone retinoate 0.2-2%, sodium hyaluronate 0.8-2%,1, 3-butylene glycol 0.1-5%, phenoxyethanol 0.5-0.8%, sodium citrate 0.05-0.1%, purified water up to 100%.
Compared with the prior art, the invention has the following beneficial effects:
(1) According to the invention, the antioxidation and saccharification inhibitor is kava extract, and the ethanol water solution with the mass fraction of 50% is adopted as an extractant, so that the dissolution rate of active ingredients can be increased, and the complexity of the subsequent separation step is reduced.
(2) According to the invention, the antioxidant and saccharification inhibitor is a kava extract, and the 1, 3-butanediol with the mass fraction of 30% is added into the kava concentrated solution, so that the stability of the kava extract can be improved, and the solubility of active ingredients can be increased.
(3) The anti-oxidation and saccharification inhibitor is kava extract, and the extraction yield of the extract can be improved and the structural stability of the active ingredients can be maintained by adopting the extraction temperature of 40 ℃ and the extraction time of 2 hours.
(4) The antioxidant and saccharification inhibitor is a kava extract, and the feed liquid ratio of a kava crushing blade to an extractant is 1: (4-6) can reduce the extraction cost while increasing the extraction rate of the active ingredient in the kava extract.
(5) The antioxidant and saccharification inhibitor is kava extract, has the effects of resisting oxidation and inhibiting protein saccharification, can inhibit collagenase activity, inhibit synthesis secretion of cell factor (IL-12) (inhibit cell aging), and improve skin relaxation.
(6) The antioxidant and saccharification inhibitor is a kava extract, can be used in various cosmetics, and has good compatibility with various cosmetic components.
Drawings
FIG. 1 is a graph showing skin elasticity change for four weeks of skin without applying lotion;
FIG. 2 is a graph showing skin elasticity change for four weeks with the lotion of comparative example 1;
fig. 3 is a graph showing skin elasticity change for four weeks of skin using the lotion of application example 1.
Detailed Description
Example 1
A preparation method of an antioxidant and saccharification inhibitor comprises the following steps:
(1) Pre-washing the plant to be extracted with water, removing surface foreign matters, drying, and crushing;
(2) Mixing crushed plant to be extracted with an extractant, extracting, and filtering to obtain crude plant extract;
(3) Concentrating the crude plant extract under reduced pressure to obtain plant concentrate, and adding water and antiseptic into the plant concentrate to obtain plant extract.
The plant to be extracted is a blade of a slips; the mass ratio of the slip blades to the feed liquid of the extractant is 1:5. 100g of slips blade and 500g of extractant.
The extractant is a mixed solution of ethanol and water, and the mass fraction of the ethanol in the mixed solution of ethanol and water is 50%.
The extraction temperature is 40 ℃, the extraction time is 2 hours, the pH of extracted acid and alkali is 6.5, the acid and alkali regulator is sodium citrate, and the mass fraction of the whole added acid and alkali regulator is 0.05%.
The preservative in the step 3 is 1, 3-butanediol, and the mass fraction of the integral addition of the 1, 3-butanediol is 30%.
The final plant extract is yellow solution with a mass of 4.0kg and a solid content of 0.28%.
Example 2
A preparation method of an antioxidant and saccharification inhibitor comprises the same steps as in example 1, wherein the extraction agent is a mixed solution of ethanol and water, and the mass fraction of ethanol in the mixed solution of ethanol and water is 20%.
The final plant extract was a yellow solution with a mass of 4.2kg and a solid content of 0.22%.
Performance testing
1. Evaluation test of antioxidant Effect (DPPH radical scavenging Effect)
DPPH 2.4 parts was dissolved in 20 parts of ethanol, and 20 parts of purified water was added thereto to prepare a DPPH solution. To 22.4 parts of this DPPH solution, 19.2 parts of an 18v/v% ethanol solution and 4.8 parts of a 2M acetic acid-sodium acetate buffer (pH 5.5) were added to prepare a DPPH additive solution. In order to eliminate the influence of the color tone of the extract itself on the test, a 50v/v% ethanol solution was used instead of the DPPH solution, and a solution in which 18v/v% ethanol solution and 2M acetic acid-sodium acetate buffer were mixed was used as a control solution. The kava extract of the present invention (extract solution of example 1) was then diluted with purified water to prepare sample solutions having final mass concentrations of 0.5%, 1.0% and 2.0%, respectively. The sample solution and the DPPH additive solution were mixed in a ratio of 1:3, and after standing at room temperature for 10 minutes, the difference between the absorbance at 550nm when each test solution was mixed with the DPPH additive solution and the absorbance at 550nm when each test solution was mixed with the control solution was measured, and the residual amount of DPPH radical was confirmed. The same procedure as described above was carried out using a 30% by volume aqueous solution of 1, 3-butanediol [ control (1.3-BG) ] (2.0 wt%) as a control instead of the extract solution of example 1, and the relative value of the residual DPPH radical rate at the time of adding each sample to the residual DPPH radical rate obtained here was determined. The test results are shown in Table 1.
TABLE 1
As shown in Table 1, it was confirmed that the extract of the present invention had excellent active oxygen (DPPH radical) eliminating effect. Therefore, the kava extract provided by the invention can inhibit oxidative damage of cells and inhibit aging of the cells.
2. Evaluation test of protein glycation inhibition effect
AGE production inhibition effect, i.e., protein glycation inhibition effect was evaluated by glyceraldehyde-mediated fluorescence production and color development of BSA (bovine serum albumin). First, 50. Mu.L of the kava extract of the present invention (extract solution of example 1), 50. Mu.L of a BSA aqueous solution of 50mg/mL, 50. Mu.L of an 8mM glyceraldehyde aqueous solution and 50. Mu.L of a PBS solution were mixed and stirred to prepare a sample solution. The concentrations of the sample solutions were adjusted so that the final mass concentrations of the extract solutions of example 1 were 0.25%, 0.5% and 1.0%, respectively. The sample solutions were allowed to stand at 37℃for 24 hours, and then fluorescence generation (excitation: 355nm, emission: 460nm: fluorescence microplate reader (manufactured by Fluoscan Acent, thermo Fisher Scientific Co.) and absorbance (wavelength 405nm: microplate reader (manufactured by Multiskan FC, thermo Scientific Co.)) were measured for each sample solution. The same procedure was carried out using 30% by volume of 1, 3-butanediol [ control group (1.3-BG) ] (1.0%) as a control instead of the extract of example 1, and the fluorescence measurement value obtained here, and the relative value and percentage (%) of the absorbance in each sample solution were obtained as protein glycation degree (%). The same test was performed with the extract of example 1 replaced by Aminoguanidine (AG) added at 1mM as a positive control. The absorbance calculation results are shown in Table 2, and the fluorescence values are shown in Table 3.
TABLE 2
TABLE 3 Table 3
As shown in tables 2 and 3, the kava extract of the present invention has a protein glycation inhibition effect, and thus it can be seen that the kava extract of the present invention inhibits glycation of cells and suppresses aging of cells.
Evaluation test of Activity inhibitory Effect of SA-. Beta.Gal
SA-. Beta. Gal (senescence associated beta galactosidase) is used as an index for evaluating aging in a cell unit because it is activated in aging cells. Cell aging was evaluated by SA-. Beta.Gal index.
Step (1) saccharification of BSA
50mg/mL of aqueous BSA was mixed with 8mM glyceraldehyde solution and incubated at 37℃for 7 days to induce saccharification of BSA. The sample addition zone was set, and the kava extract (extract solution of example 1) of the present invention was added to the culture solution for inducing BSA saccharification as a sample solution. The concentrations of the sample solutions were adjusted so that the final mass concentrations of the extract solutions of production example 1 were 0.125%, 0.25% and 0.5%, respectively. On the other hand, the same procedure was carried out by setting a control group to which 30% by volume of 1, 3-butanediol [1.3-BG ] (0.5% by weight) was added instead of the extract solution of example 1.
Step (2) Activity measurement of SA-. Beta.Gal
NHDF (fibroblast from 36 year old donor) was used at 9.0X10 4 The wells were inoculated in 24-well plates and incubated on Eagle minimal essential medium containing 0.5% new born calf serum for 24 hours. Then, the Eagle minimum essential medium containing 0.5% of the newborn calf serum was replaced with a serum-free medium, and the culture solution of the test zone or the culture solution of the control zone containing the glycosylated BSA prepared in the above step (1) was added, respectively, and the culture was continued for another 6 days. In the comparative zone (1), a test zone was set in which a culture solution containing fresh BSA at the same concentration (i.e., BSA not inducing saccharification) was added to an NHDF-cultured well plate instead of the culture solution containing the above-mentioned saccharified BSA, and the culture was similarly carried out for 6 days. After incubation, the medium was removed, SPiDER-. Beta.Gal (SG 02: tonic chemistry) was added and incubated at 37℃for 30 minutes. Then, the cells were peeled off with trypsin, and the amount of fluorescence of each cell was measured with a flow cytometer (ex=488/em=538 nm). The fluorescence values of the added sample region and the comparison region were calculated when the fluorescence value of the control region was 100, and the result was used as the activity rate of SA-. Beta.Gal. The results are shown in Table 4.
TABLE 4 Table 4
As shown in Table 4, the kava extract of the present invention inhibited the activity of SA-. Beta.Gal. The kava extract of the present invention inhibits cell aging.
Synthesis inhibition of SASP factor (IL-12)
Step (1) saccharification of BSA
50mg/mL of aqueous BSA was mixed with 8mM glyceraldehyde solution and incubated at 37℃for 7 days to induce saccharification of BSA. The sample addition zone was set, and the kava extract (extract solution of example 1) of the present invention was added to the culture solution for inducing BSA saccharification as a sample solution. The same procedure was carried out by setting the comparative zone (1) to which 30% by volume of 1, 3-butanediol [1.3-BG ] (0.5% by weight) was added in place of the extract solution of example 1.
Step (2) determination of the amount of SASP factor (IL-12) synthesized
NHDF (fibroblast from 36 year old donor) was used at 9.0X10 4 The wells were inoculated in 24-well plates and incubated on Eagle minimal essential medium containing 0.5% new born calf serum for 24 hours. Then, the Eagle minimum essential medium containing 5% of fresh calf serum was replaced with a serum-free medium, and the culture solutions of the test areas containing the glycosylated BSA prepared in the above step (1) were added, respectively, and the culture was continued for another 6 days. Control group: the NHDF was replaced with a well plate of fibroblasts (NB 1 RGB) derived from neonates, and a test zone was set for culturing without adding the glycosylated BSA; comparison zone (2): the test area in which the above-mentioned glycosylated BSA was not added to the well plate on which NHDF was set was cultured, and the culture was performed for 6 days in the same manner. After the culture, the culture supernatant was collected and analyzed with Human Cytokine Antibody Array (human cytokine antibody chip) (42 target 8 membrane) ab133997 (Abcam). The activity of SA-. Beta.Gal is expressed as a relative value when the culture supernatants of the sample addition region, the comparison regions (1) and (2) were analyzed, wherein the value when the culture supernatant of fibroblasts from the neonate was analyzed was set to 100. The results are shown in Table 5.
TABLE 5
Sample preparation | IL-12 Synthesis Rate (%) |
Control zone | 100.0 |
Example 1 | 106.5 |
Comparison area (1) | 130.6 |
Comparison area (2) | 122.6 |
As shown in Table 5, the kava extract of the present invention has an effect of inhibiting cytokine (IL-12), which is one of SASP factors. Thus, the kava extract of the present invention has an effect of inhibiting the induction of cell aging.
5. Evaluation test for inhibition of collagenase Activity
Step (1) saccharification of BSA
50mg/mL of aqueous BSA was mixed with 8mM glyceraldehyde solution and incubated at 37℃for 7 days to induce saccharification of BSA. The sample addition zone was set, and the kava extract (extract solution of example 1) of the present invention was added to the culture solution for inducing BSA saccharification as a sample solution. The same procedure was carried out by setting the comparative zone (1) to which 30% by volume of 1, 3-butanediol [1.3-BG ] (0.5% by weight) was added in place of the extract solution of example 1.
Measurement of collagenase Activity value in step (2)
NHDF (fibroblast from 36 year old donor) was used at 9.0X10 4 The wells were inoculated in 24-well plates and incubated on Eagle minimal essential medium containing 0.5% new born calf serum for 24 hours. Then, the Eagle minimum essential medium containing 5% of newborn calf serum was replaced with a serum-free medium, and the culture solutions containing the sample-added regions of the saccharified BSA prepared in the above step (1) were added, respectively, and the culture was continued for another 6 days. Control zone: set up 9.0X10 inoculation in 24 well plates 4 Well, the same was carried out for 6 days without adding the above-mentioned saccharified BSA to the test area for culture. After the cultivation, a culture supernatant was collected, and the culture supernatant was used as a collagenase enzyme solution. The collagenase activity was measured using a collagenase measurement kit from primaricell corporation. To the microtubules were added 50. Mu.L of the collagenase solution described above and 50. Mu.L of FITC-labeled type I collagen matrix solution. After reaction at 37℃for 30 minutes, 300. Mu.L of a reaction termination solution was added, and unreacted collagen matrix was precipitated by centrifugation, and the fluorescence intensity of the supernatant was measured (Ex=355, em=460). Measurement results with no saccharification BSA addedThe relative value of the culture supernatant of the cultured NHDF is 100. The results are shown in Table 6.
TABLE 6
Sample preparation | Collagenase activity rate (%) |
Control zone | 100.0 |
Example 1 | 83.8 |
Comparison area (1) | 165.5 |
As shown in table 6, the kava extract of the present invention has an effect of inhibiting collagenase activity. Therefore, the kava extract of the invention has the effects of inhibiting the decomposition of dermal collagen and improving skin elasticity.
Application example 1
The application of the antioxidant and saccharification inhibitor is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 1%,1, 3-butanediol 5%, nipagin ester 0.2%, purified water was supplemented to 100%.
Application example 2
The application of the antioxidant and saccharification inhibitor is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 0.5%, beta-glucan 0.2%,1, 3-butanediol 1%, sodium citrate 0.05%, purified water was supplemented to 100%.
Application example 3
The application of the antioxidant and saccharification inhibitor is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 2%, hydroxy pinacolone retinoate 0.05%, bifidobacterium broth 1%,1, 3-butanediol 0.1%, phenoxyethanol 0.5%, sodium citrate 0.05%, purified water make up to 100%.
Application example 4
The application of the antioxidant and saccharification inhibitor is applied to daily chemical products, and the daily chemical products are prepared from the following raw materials in percentage by weight: example 1 extract 2%, centella asiatica extract hydroxy pinacolone retinoate 0.2%, sodium hyaluronate 0.8%,1, 3-butylene glycol 0.1%, phenoxyethanol 0.5%, sodium citrate 0.05%, purified water make up to 100%.
Comparative example 1 was used
The application of the antioxidant and saccharification inhibitor is applied to daily chemical products, and the specific raw materials of the daily chemical products are the same as those of application example 1, wherein the specific raw materials are as follows: the extract of example 1 was replaced with 30% by mass of aqueous 1, 3-butanediol.
Performance testing
Skin elasticity evaluation test
3 test sites were set on the inner side of the forearm of 5 subjects (women aged 20 to 30 years) as the non-applied region, the comparative control lotion application region of comparative formulation application example 1, and the lotion application region containing application example 1 of the present invention. First, elasticity (R2) of the skin before applying the lotion was measured with a curemeter (courage+khazaka, germany) as an initial value. Each lotion was then applied to the test site twice daily for 4 weeks. After the completion of the test, skin elasticity (R2) was measured using a curemeter similarly to the start of the test, and the average value of the measured values of 5 persons was calculated to calculate the average change rate (%).
The test results are shown in FIGS. 1 to 3.
In the areas where the sample was not applied, as shown in fig. 1 and 2, the skin elasticity was reduced by 4.5%, while the application of the lotion of comparative example 1 was reduced by 2.8%, while in the areas where the lotion of application example 1 of the present invention was applied, the skin elasticity was improved by 0.6%. This indicates that the extract of the present invention has an effect of improving skin relaxation.
Claims (10)
1. An antioxidant and saccharification inhibitor is characterized in that the preparation method comprises the following steps:
(1) Pre-washing the plant to be extracted with water, removing surface foreign matters, drying, and crushing;
(2) Mixing crushed plant to be extracted with an extractant, extracting, and filtering to obtain crude plant extract;
(3) Concentrating the crude plant extract under reduced pressure to obtain plant concentrate, and adding water and antiseptic into the plant concentrate to obtain plant extract.
2. The antioxidant, glycation inhibiting agent of claim 1, wherein the plant to be extracted is a plant of the genus piperaceae, the plant of the genus piperaceae is kava, and the extraction site of the kava is a leaf.
3. The antioxidant, glycation preventing agent of claim 1, wherein the extractant is selected from one or more of water, lower alcohols, polyols, ester extractants, ketone extractants, ether extractants, hydrocarbon extractants.
4. The antioxidant, glycation preventing agent of claim 3, wherein the lower alcohol is selected from one or more of methanol, ethanol, propanol, and the polyhydric alcohol is selected from one or more of ethylene glycol, propylene glycol, butylene glycol, glycerin.
5. The antioxidant and saccharification inhibitor of claim 4, wherein the extractant is a mixed solution of ethanol and water or a mixed solution of butanediol and water, and the butanediol is selected from one or a combination of several of 1, 2-butanediol, 1, 3-butanediol, and 1, 4-butanediol.
6. The antioxidant and saccharification inhibitor of claim 5, wherein the mass fraction of ethanol in the mixed solution of ethanol and water is 10-60% and the mass fraction of butanediol in the mixed solution of butanediol and water is 10-70%.
7. The antioxidant, glycation inhibiting agent of claim 1, wherein the parameters extracted in step 2 are: extracting at 0-80deg.C for 1-168 hr; the pH of the plant crude extract is 3-9.
8. Use of an antioxidant, glycation inhibiting agent according to any one of claims 1-7, in a daily chemical product.
9. The use of an antioxidant, glycation preventing agent according to claim 8, wherein said daily chemical product is selected from one of an emulsion, a cream, a lotion, an essence, a peel-off mask, a lipstick, a foundation, a spread mask, a foundation, a pressed compact, a blush, a powder, a facial cleanser, a body shampoo, a hair shampoo, a soap, a hair tonic, a bath agent.
10. The use of antioxidant and glycation preventing agent according to claim 8, wherein the daily chemical product comprises one or more of hydrocarbon auxiliary agent, ester auxiliary agent, triglyceride auxiliary agent, fatty acid, higher alcohol, surfactant, polyalcohol, thickening gel, antioxidant, ultraviolet defending agent, colorant, preservative, pH regulator, humectant, skin conditioner, plant component, and microorganism component.
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