CN112545945B - A method for preparing anti-aging extract of ground active components of Ginseng radix and its application - Google Patents
A method for preparing anti-aging extract of ground active components of Ginseng radix and its application Download PDFInfo
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- CN112545945B CN112545945B CN202011577356.3A CN202011577356A CN112545945B CN 112545945 B CN112545945 B CN 112545945B CN 202011577356 A CN202011577356 A CN 202011577356A CN 112545945 B CN112545945 B CN 112545945B
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- ginseng
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- 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]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- 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
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- 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
Abstract
The present invention relates to the field of cosmetics. Aims to solve the application problem of the ground part of the ginseng in cosmetics. In particular to a preparation method of an active ingredient extract of the overground part of ginseng and application of the active ingredient extract of the overground part of ginseng in cosmetics. The ginseng preparation disclosed by the invention is prepared by ultra-high pressure extraction technology of the overground part of ginseng, and the activity of the overground part of ginseng is researched; the invention provides an extract of the overground part of ginseng by establishing a cell radiation model and optimizing and screening effective concentration. The ground part extract of the ginseng prepared by detection and extraction has the functions of radiation resistance, oxidation resistance and aging resistance.
Description
Technical Field
The invention relates to the field of cosmetics, in particular to a preparation method of an anti-aging ginseng overground part active ingredient extract and application of the extract in preparation of cosmetics.
Background
Ginseng has many efficacies in cosmetics, and its anti-aging function is widely known.
The ginseng has good effects on resisting natural aging, resisting photoaging and resisting gene aging.
The common medicinal parts of the existing ginseng products are ginseng roots and rhizomes (underground parts), and the overground parts of ginseng, including ginseng stems, leaves and flowers, are usually ignored by people, and the report of the whole application is not reported.
With the improvement of the living standard of human beings, the protection consciousness of people against radiation is continuously strengthened, and the screening of a natural and effective radiation-resistant medicine becomes a research hotspot of many scientific researchers.
Researches show that ginseng has relatively remarkable effects of resisting radiation and oxidation aging, and has high research value in the direction of developing novel anti-radiation damage medicaments and skin care products.
The invention compares the skin aging resistance of the upper and lower parts of the ginseng by antioxidant experiments (DPPH method) and cell anti-radiation screening including cck-8 method for detecting survival rate, ELISA method for detecting related indexes and flow cytometry for detecting apoptosis and period, and the result shows that the upper part of the ginseng has better anti-radiation effect, thus providing theoretical basis for further developing anti-radiation damage medicines and skin care products.
The invention provides a preparation method of an extract of the overground part of ginseng.
The invention provides an extract of the overground part of ginseng, which is suitable for preparing cosmetics.
The above-ground part has a more significant effect on anti-skin aging than the underground part.
Disclosure of Invention
The invention provides preparation of an active ingredient of a ground part of ginseng with an anti-aging effect and application research of the ground part of the ginseng in cosmetics.
The ginseng preparation disclosed by the invention is characterized in that the ground upper part of ginseng is prepared by an ultrahigh pressure extraction technology, and the activity of the ground part of ginseng is researched; the invention screens the effective concentration through optimization by establishing a cell radiation model. The ground part of the ginseng prepared by detection and extraction has the functions of resisting radiation, oxidation and aging.
The technical scheme of the invention is realized as follows: is prepared by extracting aerial parts (stem, leaf, and flower) of Ginseng radix. The overground part of ginseng is, for example, stem, leaf, flower, etc.
In the present application, the overground part of ginseng refers to one of stems, leaves and flowers of ginseng, or a mixture of the two or three.
Further, different solvents were used for extraction without active ingredient.
Further, the antioxidant activity of active ingredients extracted from different solvents was investigated using the DPPH method.
Further, the screened active ingredients are utilized to carry out the efficacy research of radiation resistance and aging resistance.
The invention provides a preparation method of an anti-aging ginseng overground part active ingredient extract, which comprises the steps of treating the ginseng overground part with an organic solvent, and extracting the anti-aging ginseng overground part active ingredient extract under the condition of ultrahigh pressure.
Preferably, the overground part of the ginseng is root, stem and leaf of the ginseng.
Preferably, the overground part of ginseng is the overground part of ginseng in 6 months of flowering season.
Preferably, the overground part of ginseng is dried at a low temperature of 45 ℃, extracted with ethanol under ultrahigh pressure, centrifuged, filtered, and extracted with different solvents to obtain active ingredients.
The extraction process parameters are that the ethanol concentration is 72 percent, the extraction pressure is 423Mpa, the material-liquid ratio is 1 (g: ml), and the retention time is 4 minutes.
The extraction process parameters may also be in the following ranges: the ethanol concentration is 40-90%, the extraction pressure is 100-600Mpa, the material-liquid ratio is 1.
The organic solvent for extracting aerial parts of Ginseng radix is ethyl acetate.
The ethyl acetate extract has antioxidant effect.
An application of ground active component extract of Ginseng radix with antiaging effect in cosmetics is provided. Is applied in radiation resistance, oxidation resistance and aging resistance.
The anti-aging ginseng overground part active ingredient extract is prepared by a method which comprises the steps of treating the ginseng overground part with an organic solvent, and extracting the anti-aging ginseng overground part active ingredient extract under the condition of ultrahigh pressure.
The invention discloses an anti-aging active ingredient of overground part of ginseng, which has the following beneficial effects:
1. the overground part of the ginseng with the anti-aging effect is prepared by an ultrahigh pressure extraction technology. The extraction process was optimized by orthogonal. Extracting with different extractants to obtain extracts with different polarities, and drying and storing.
2. And (3) evaluating the activity of the extract by a DPPH method on the ground part extracts of the ginseng with different polarities, and screening out the extract with good activity.
3. The ground ginseng part has good radiation resistance and anti-aging effects by measuring the radiation of the screened ground ginseng part extract.
Drawings
FIG. 1 shows the effect of different UVB radiation doses on HaCaT cell survival. The abscissa is the irradiation dose. The ordinate is the cell survival rate.
FIG. 2 shows the effect of different concentrations of ethyl acetate on the survival rate of HaCaT cells in the above-ground and underground parts of ginseng. The abscissa shows the concentration of ethyl acetate phase in the overground and underground parts of ginseng, and vitamin A is a positive control drug. The ordinate is the cell survival rate.
FIG. 3 shows the effect of different concentrations of ethyl acetate on HaCaT cell survival in above-ground and underground ginseng. In the figure, each group 1,2,3 represents a drug concentration of 12.5, 50, 100 μ g/mL, which caused a very significant decrease in cell viability (P < 0.01) after UVB irradiation treatment of the cells.
FIG. 4 shows the effect of different concentrations of ethyl acetate on the above-ground and underground parts of ginseng on the induction of activity of SOD, MDA, COX-2, and MMP-9 in apoptotic cells by UVB irradiation. In the figure, each group 1,2 and 3 represents the drug concentration of 12.5, 25 and 50 mug/mL, compared with a blank control group, the SOD activity of the UVB model group is obviously reduced (P < 0.01), the MDA level is obviously increased (P < 0.01), and the COX-2 and MMP-9 levels are obviously reduced (P < 0.01); compared with the UVB model group, the SOD activity of the administration treatment group (ethyl acetate phase of the overground and underground parts of the ginseng and vitamin A as the positive drug) is obviously increased (P is less than 0.01), the MDA level is obviously reduced (P is less than 0.01), the COX-2 and MMP-9 levels are obviously increased (P is less than 0.01), and the contents of SOD, COX-2 and MMP-9 are obviously increased and the SOD content is obviously reduced after the three drugs are irradiated on HaCaT cells, wherein the administration group with 50mg/L is most obvious and has the same result as the vitamin A as the positive drug.
FIG. 5 shows the result of detecting apoptosis by flow cytometry. HaCaT cells were 0.72J/cm compared to blank 2 After UVB radiation, the number of apoptotic cells is remarkably increased (P is less than 0.01), and the number of apoptotic cells can be remarkably reduced (P is less than 0.01) after drug treatment, wherein the overground part of ginseng has better effect of inhibiting apoptosis compared with the underground part.
FIG. 6 shows the results of flow cytometry measurements of the cell cycle. HaCaT compared to blank groupCells were passed through 0.72J/cm 2 After UVB irradiation, a significant S-phase block occurs. After the treatment of adding ethyl acetate phase of underground part of ginseng, the number of cells in S phase is increased, and no statistical difference exists (P is more than 0.05); after the treatment of adding ethyl acetate phase and vitamin A as a positive drug to the overground part of the ginseng, the number of cells in the S phase is remarkably reduced (P is less than 0.01), which shows that the overground part of the ginseng can obviously reduce the block of HaCaT cell cycle after UVB radiation.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Examples
1. Extraction process parameters
The optimal extraction process parameters are that the ethanol concentration is 72%, the extraction pressure is 423MPa, and the feed-liquid ratio is 1:50 (g: mL) and a dwell time of 4min.
2. The effectiveness of each example and comparative example was analyzed by measuring antioxidant activity using DPPH.
Accurately weighing samples of ground and underground polar parts (petroleum ether phase, chloroform phase, ethyl acetate phase, and n-butanol phase) of Ginseng radix, respectively, dissolving with anhydrous ethanol, and preparing into a series of sample solutions with gradient concentration for testing. DPPH free radical scavenging ability and IC of different polar substances in different parts of ginseng 50 The values are expressed by evaluating the antioxidant activity in vitro using 2,6-di-tert-butyl-p-methylphenol (BHT) as a positive control.
The difference of DPPH free radical scavenging ability of different polar parts is obvious, wherein ethyl acetate on the ground part of ginseng has the best antioxidant ability compared with other parts (Table 1).
Table 1 the DPPH radical scavenging ability of the different polar sites of ginseng (n =3,
g/L)
3. effect of different UVB irradiation doses on HaCaT cell survival Rate
The results are shown in FIG. 1, the cell survival rate is in a descending trend along with the increase of the irradiation dose, and the irradiation dose of 0.72J/cm2 is selected for the experiment when the cell viability is inhibited by 50%, and the subsequent experiment is carried out.
4. Influence of ethyl acetate with different concentrations on overground and underground parts of ginseng on survival rate of HaCaT cells
The results are shown in FIG. 2, and the survival rate of the ethyl acetate phase of the overground and underground parts of the ginseng on HaCaT cells is more than or equal to 90 percent in the concentration range of 0 mg/L-100 mg/L. Therefore, the drug was safe for cells in this concentration range (fig. 1). Vitamin A is a positive control drug.
5. Influence of ethyl acetate with different concentrations on overground and underground parts of ginseng on survival rate of HaCaT cells
The results are shown in FIG. 3 (each group 1,2,3 in the figure represents drug concentrations of 12.5, 50, 100 μ g/mL), which caused a very significant decrease in cell viability (P < 0.01) following UVB irradiation of the treated cells. Compared with a UVB irradiation group, the survival rate of HaCaT cells (P is less than 0.05) can be improved by 50mg/L ethyl acetate phase of ginseng roots and seeds, and the survival rate of HaCaT cells (P is less than 0.01) can be remarkably improved by 50mg/L ethyl acetate phase of ginseng overground and underground parts and vitamin A serving as a positive medicine.
FIG. 3 is a graph of the effect of varying concentrations of ethyl acetate on the survival of apoptotic HaCaT cells induced by UVB irradiation.
Note: Δ P <0.05, Δ Δ P <0.01; p <0.05, P <0.01 compared to model group.
6. Influence of ethyl acetate with different concentrations on overground and underground parts of ginseng on activities of SOD, MDA, COX-2 and MMP-9 of apoptosis cells induced by UVB irradiation
The results are shown in FIG. 4 (each group 1,2,3 in the figure represents drug concentrations of 12.5, 25, 50 μ g/mL), compared with the blank control group, the SOD activity of the UVB model group is obviously reduced (P < 0.01), the MDA level is obviously increased (P < 0.01), and the COX-2 and MMP-9 levels are obviously reduced (P < 0.01); compared with the UVB model group, the SOD activity of the administration treatment group (ethyl acetate phase of the overground and underground parts of the ginseng and vitamin A as the positive drug) is obviously increased (P < 0.01), the MDA level is obviously reduced (P < 0.01), the COX-2 and MMP-9 levels are obviously increased (P < 0.01), and the contents of SOD, COX-2 and MMP-9 are obviously increased and the SOD content is obviously reduced after the three drugs are irradiated on HaCaT cells, wherein the administration group with 50mg/L is most obvious and has the same result as the vitamin A as the positive drug, so the concentration is used as the subsequent experimental concentration.
FIG. 4 is a graph of the effect of varying concentrations of ethyl acetate on the indication associated with UVB irradiation induced apoptotic cells.
(a.SOD b.MDA c.COX-2d.MMP-9)
Note: Δ P <0.05, Δ Δ P <0.01; compared to the model group, P <0.05, P <0.01.
7. Flow cytometry for detecting apoptosis results
As can be seen from FIG. 5 and Table 2, compared with the blank group, haCaT cells have significantly increased numbers of apoptotic cells (P < 0.01) after being irradiated by UVB at 0.72J/cm2, and have significantly decreased numbers of apoptotic cells (P < 0.01) after being treated with drugs, wherein the above-ground parts of ginseng have better apoptosis-inhibiting effects than the underground parts.
Fig. 5 is an apoptosis analysis after UVB irradiated HaCaT cell drug treatment (a. Control group b. UVB model group c. Ginseng above ground group d. Ginseng below ground group e. Vitamin a group).
Table 2 Annexin v-FITC/PI method for detecting apoptosis (n =3,
％)
note: the ratio to placebo # P <0.05, # # P <0.01; compared to the model group, P <0.05, P <0.01.
8. Flow cytometry detection of cell cycle results
As can be seen from FIG. 6 and Table 3, haCaT cells were significantly blocked in S phase after 0.72J/cm2 of UVB irradiation compared to the blank. After the treatment of adding ethyl acetate phase of underground part of ginseng, the number of cells in S phase is increased, and no statistical difference exists (P is more than 0.05); after the treatment of adding ethyl acetate phase and vitamin A as a positive drug to the overground part of the ginseng, the number of cells in the S phase is remarkably reduced (P is less than 0.01), which shows that the overground part of the ginseng can obviously reduce the block of HaCaT cell cycle after UVB radiation.
FIG. 6 is a cell cycle analysis of UVB irradiated HaCaT cells after drug treatment (a. Control group b. UVB model group c. Group d. Group of ginseng above ground part e. Group of vitamin A)
Table 3 Annexin v-FITC/PI method for detecting apoptosis (n =3,
％)
note: the ratio # P <0.05, # P <0.01 to blank; p <0.05, P <0.01 compared to model group.
The present invention has been described in detail and exemplified by the embodiments with reference to the accompanying drawings, and it is within the scope of the invention to make modifications and changes without departing from the spirit and substance of the invention. The scope of the invention is indicated only by the claims. Various technical schemes of the invention can be combined with each other, and the described technical features can be transplanted and combined among different technical schemes, so that more technical schemes can be generated, and the technical schemes are within the protection scope of the invention.
All other embodiments obtained by a person skilled in the art based on the manufacturing process and embodiments of the present invention without any creative effort belong to the protection scope of the present invention.
Claims (5)
1. A preparation method of an anti-aging ginseng overground part active ingredient extract comprises the steps of drying ginseng overground parts at a low temperature of 45 ℃, extracting active ingredients by using ultrahigh pressure and ethanol in combination, carrying out centrifugal filtration, and extracting the active ingredients by ethyl acetate, wherein the overground parts of ginseng are flowers, stems and leaves of ginseng, and the extraction process parameters are ethanol concentration of 72%, extraction pressure of 423MPa, material-liquid ratio of 1.
2. The method according to claim 1, wherein the ginseng aerial parts are ginseng aerial parts at 6 months of flowering.
3. The method according to claim 1, wherein the ethyl acetate extract has antioxidant activity.
4. Use of the above-ground ginseng extract having anti-aging effect prepared according to the preparation method of claim 1 for preparing cosmetics.
5. The use according to claim 4, in the fields of radiation protection, antioxidation and anti-aging.
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