CN114224795B

CN114224795B - A Ganoderma extract for cosmetics and its preparation method

Info

Publication number: CN114224795B
Application number: CN202210082453.8A
Authority: CN
Inventors: 刘倩; 吴震生; 亓玉锋; 亓云吉
Original assignee: Jinan Zerun Biotechnology Co ltd
Current assignee: Jinan Zerun Biotechnology Co ltd
Priority date: 2022-01-24
Filing date: 2022-01-24
Publication date: 2023-04-25
Anticipated expiration: 2042-01-24
Also published as: CN114224795A

Abstract

The invention discloses a preparation method of ganoderma lucidum extract for cosmetics, which is characterized by comprising the following steps of: step one, adding lucid ganoderma, an extraction solvent and a compound enzyme solution into a flash extractor, vacuumizing the flash extractor, and extracting for 4-7 min at 1000-3500 r/min by using the flash extractor to obtain a mixed solution; reflux extracting the mixed solution with ethanol, filtering to obtain primary lucid ganoderma extract and lucid ganoderma residues, adding the lucid ganoderma residues into ethanol, and carrying out microwave extraction to obtain secondary lucid ganoderma extract; mixing the first and second ganoderma lucidum extract, cooling, vacuum filtering, rotary steaming to remove the extracting solvent, obtaining ganoderma lucidum extract and weighing; and step four, dissolving the lucid ganoderma extract to obtain lucid ganoderma dissolving liquid, adding diatomite into the lucid ganoderma dissolving liquid, stirring, standing overnight, vacuum filtering, supplementing 1,3 propylene glycol, and stirring until the lucid ganoderma extract is clear and transparent, thus obtaining the lucid ganoderma extract. The invention combines enzymolysis with flash extraction and microwave extraction, and the obtained product has good antiallergic effect and can be widely applied to cosmetics.

Description

A Ganoderma extract for cosmetics and its preparation method

Technical Field

The invention relates to the technical field of cosmetic production, in particular to a ganoderma lucidum extract for cosmetics and a preparation method thereof.

Background

Ganoderma (Ganoderma) is a fungus belonging to the genus Ganoderma (GanodermaKarst) of the family Polyporaceae, and also called Ganoderma lucidum (Ganoderma lucidum). Ganoderma lucidum is flat in nature, bitter in taste and nontoxic, and is used as a large-scale fungus for both medicine and food, so that the ganoderma lucidum is more and more valued for the remarkable effects of nourishing and strengthening body, prolonging life and the like. Ganoderma contains polysaccharide, triterpene, alkaloid, ergosterol, etc., and has effects of resisting tumor, enhancing immunity, scavenging free radicals, resisting aging, resisting thrombosis, and lowering blood sugar.

The ganoderma lucidum is used as a medicinal material with extremely high medicinal value, and the cost-saving efficient extraction of the effective components becomes a technical key. In recent years, microwave-assisted extraction of polysaccharides has become a research hotspot. The traditional Chinese medicine extraction method comprises a water decoction method, an immersion method, a percolation method, an alcohol extraction method and the like, and the microwave-assisted extraction has the characteristics of high extraction rate, solvent saving, rapidness, high efficiency, low pollution, simple process and the like, and is widely applied to the extraction of natural plant components. Ethanol is one of the most commonly used solvents, has excellent dissolving capacity, has strong penetrating power on Chinese herbal medicine cells, short extraction time, less dissolved water-soluble impurities, low toxicity and low cost, and can extract specific substances through ethanol solvents with different concentrations.

Inflammation and aging are two major problems of the skin, propionibacterium acnes are considered to be a key element in causing acne, and many enzymes and active substances generated in metabolic activities have damaging effects on skin cells and tissues, so that the barrier of the hair follicle funnel is destroyed, and an acne inflammatory reaction is caused. When human immune cells are subjected to the action of inflammatory factors, some small molecular weight soluble proteins or polypeptides which can transmit information among cells and have specific immunoregulatory functions, such as NO, TNF-alpha, IL-6, PGE-2, IL-1 and the like, are secreted by the body and can participate in or cause inflammatory reactions. The excessive expression of MMP-1 by skin cells is the most important enzyme for causing skin to have aging symptoms such as wrinkles, fine lines and the like, and can cause the damage of extracellular matrixes of dermis layers, especially normal structures of collagen fibers and elastic fibers, so that the anti-inflammatory and anti-aging work is performed, and the skin diseases such as acne, seborrheic dermatitis, aging and the like can be effectively assisted and prevented.

The prior art fails to provide an extraction method for improving the antiallergic property and the activity of ganoderma lucidum.

Disclosure of Invention

In order to solve the above problems, the present invention aims to provide a method for preparing a ganoderma lucidum extract for cosmetics, which can effectively increase the content of active ingredients and thus promote anti-inflammatory and anti-aging activities, comprising the steps of:

step one, adding lucid ganoderma, an extraction solvent and a compound enzyme liquid into a flash extractor, wherein the enzyme activity ratio of the compound enzyme liquid is (2-6): (0.5-2) xylanase and pectase, vacuumizing the flash extractor, and extracting for 4-7 min at 1000-3500 r/min by using the flash extractor to obtain a mixed solution;

reflux extracting the mixed solution with ethanol, filtering to obtain primary lucid ganoderma extract and lucid ganoderma residues, adding the lucid ganoderma residues into the ethanol, and extracting for 1-3 times with the aid of microwaves, wherein the power of the microwaves is 350-600 w, and the extracting time is 10-30 min each time to obtain secondary lucid ganoderma extract;

mixing the primary lucid ganoderma extract and the secondary lucid ganoderma extract, cooling, vacuum filtering, removing the extraction solvent by rotary evaporation to obtain lucid ganoderma extract, and weighing;

and step four, stirring and dissolving the lucid ganoderma extract, deionized water and 1,3 propylene glycol to obtain lucid ganoderma dissolution liquid, adding diatomite into the lucid ganoderma dissolution liquid, stirring, standing overnight, vacuum filtering, supplementing 1,3 propylene glycol, and stirring until the mixture is clear and transparent to obtain the lucid ganoderma extract.

Further, the extraction solvent is water.

In the first step, the compound enzyme solution is 3% of the ganoderma lucidum by mass.

Further, in the first step, the total enzyme activity in the complex enzyme solution is about 12 ten thousand U/g.

Further, in the first step, the enzyme activity ratio of the complex enzyme solution is 4:1, xylanase and pectase.

Further, in the first step, the flash extractor extracts for 6min at 2000 r/min.

In the second step, the reflux extraction is carried out for 2 times in a water bath at the water bath temperature of 60 ℃ for 2 hours each time.

Further, in the second step, the microwave extraction is performed for 2 times, the power of the microwave is 500w, and the extraction time is 20min each time.

On the other hand, the application also provides the ganoderma lucidum extract prepared by the preparation method.

On the other hand, the application also provides application of the lucid ganoderma extract prepared by the preparation method of the lucid ganoderma extract in preparing cosmetics, wherein the cosmetics comprise anti-inflammatory and anti-aging cosmetics.

The invention has the following beneficial effects:

according to the preparation method, the ganoderma lucidum is extracted by adopting a flash extraction method capable of being extracted at normal temperature, wherein the flash extraction is particularly suitable for directly extracting plants with more fibers, and the active ingredients in the ganoderma lucidum can be more effectively reserved by combining the normal temperature and enzymolysis auxiliary extraction modes;

according to the preparation method, vacuum is pumped to discharge air during flash extraction, so that the content of plants is largely exposed due to the fact that the flash extraction is highly broken on the plant materials, strong vortex is formed at the center of the inner edge of the flash extractor, and the crushed materials are driven to turn inside and outside, so that a violent stirring effect is generated, the extract of the plants is largely contacted with the outside air to oxidize and react, and even inactivate, and the vacuum pumping air discharging mode is adopted, so that the contact of active substances in ganoderma lucidum, such as ergosterol and the like, with the air can be effectively reduced, and deterioration caused by the reaction of the active substances with the air is avoided;

according to the preparation method, the microwave assistance is carried out when the ganoderma lucidum residues are extracted by ethanol, so that heat conduction in the ganoderma lucidum residues is quickened, the ethanol extraction time and the solvent consumption are relatively shortened, the ganoderma lucidum utilization rate is improved, the ganoderma lucidum extract with good activity and small irritation is obtained through diatomite adsorption and decolorization, and the cell degranulation experiment test shows that the number of degranulation cells at the concentration of 0.1563%,0.3125% and 0.625% is obviously reduced, so that the ganoderma lucidum extract has obvious antiallergic capability;

according to the preparation method provided by the invention, the prepared ganoderma lucidum extract adopts a traditional Chinese medicine plant source, has the advantages of good naturalness, no toxic or side effect and the like, remarkably improves the content of antibacterial and anti-aging active ingredients in the extract, and further improves the anti-inflammatory and anti-aging activity of the ganoderma lucidum extract.

According to the preparation method of the ganoderma lucidum extract, the obtained product can effectively inhibit the over-expressed IL 1 beta, TNF alpha and NO inflammatory factors caused by the breeding of anaerobic acne bacillus in skin hair follicles, can regulate and control the content of the most main MMP-1 and the content of the enzyme of the protocol tagen-1, which cause the appearance of aging symptoms such as wrinkles, fine lines and the like, has the anti-inflammatory and anti-aging effects, lays a foundation for the application of the ganoderma lucidum extract in the field of cosmetics, has the advantages of good naturalness, NO toxic or side effect and the like, has the advantages of high extraction efficiency, good purity, NO pollution in the extraction process, and can be recovered by using ethanol as an extraction solvent.

Drawings

The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiments of the application and together with the description serve to explain the application and do not constitute an undue limitation to the application. In the drawings:

fig. 1 is a microscopic view of the degranulation process of the ganoderma lucidum extract provided by the application.

Detailed Description

In the following description, numerous specific details are set forth in order to provide a more thorough understanding of the present invention. It will be apparent, however, to one skilled in the art that the invention may be practiced without one or more of these details. In other instances, well-known features have not been described in detail in order to avoid obscuring the invention.

The vacuum extractor is a product of Shanghai bright type electromechanical equipment limited company, the model is BDR16, and the vacuum degree is less than or equal to 0.05Pa; the flash extractor is a product of Shanghai vanadium precise equipment limited company, the model is SC 200, and the motor rotation speed is that: 0-6000 r/min; the rotary evaporation evaporator is specifically manufactured by Kexing instruments, the model is R1020, the temperature is controlled to be between room temperature and 99 ℃, and the vacuum degree is less than or equal to 399.9Pa.

The materials and instruments used in the examples below are commercially available, and unless otherwise specified, the raw material grades in the examples below are cosmetic grades and are commercially available.

In addition, the "water" in the present invention includes any feasible water that can be used in the cosmetic field such as deionized water, distilled water, ion-exchanged water, double distilled water, high purity water, purified water, and the like.

Example 1

The embodiment provides a preparation method of ganoderma lucidum extract for cosmetics, which comprises the following specific steps:

firstly, shearing 200kg of dried ganoderma lucidum to obtain ganoderma lucidum fragments with the diameter of 0.8cm, adding the ganoderma lucidum fragments into a flash extractor, mixing the ganoderma lucidum fragments by taking water as a solvent, adding 3% by mass of compound enzyme liquid of ganoderma lucidum into the solvent, wherein the total enzyme activity in the compound enzyme liquid is about 12 ten thousand U/g, and the enzyme activity ratio is 4:1, vacuumizing a flash extractor to 0.05Pa, and extracting the solution containing ganoderma lucidum fragments with the flash extractor at 2500r/min for 6min to obtain a mixed solution;

extracting the mixed solution obtained in the second step with 70% wt of ethanol for 2 times in a water bath at a water bath temperature of 60 ℃ for 2 hours each time, filtering with a 200-mesh screen to obtain a first lucid ganoderma extract and lucid ganoderma residues, and adding the lucid ganoderma residues into a solution with a feed liquid ratio of 1: extracting with 70% ethanol of 20% weight for 2 times under the assistance of microwave with microwave power of 500w for 20min to obtain secondary Ganoderma extractive solution;

thirdly, mixing the primary lucid ganoderma extract and the secondary lucid ganoderma extract, cooling to 40 ℃, carrying out vacuum suction filtration on diatomite cakes prepared from diatomite accounting for 5% of the total weight of the lucid ganoderma extract and EDTA-2Na accounting for 1%, and carrying out rotary evaporation in a water bath kettle at 70 ℃ under the vacuum degree of 150mbar to remove ethanol solvent until the mixed extract is in an extractum shape, thus obtaining lucid ganoderma extractum and weighing;

fourthly, taking lucid ganoderma extract, wherein the dosage ratio of the lucid ganoderma extract to deionized water to 1,3 propylene glycol is 3:4:12 stirring and dissolving to obtain ganoderma lucidum solution, adding diatomite accounting for 5% of the total weight of the ganoderma lucidum solution, stirring for 2 hours till standing overnight, vacuum filtering diatomite cakes prepared from diatomite accounting for 5% of the total weight of the ganoderma lucidum solution and EDTA-2Na accounting for 1,3 propanediol of the sample solution after the vacuum filtering till 90% of the total weight of the ganoderma lucidum solution, stirring till the sample solution is clear and transparent, heating and sterilizing for 2 hours at 70 ℃, cooling to 40 ℃, and filling to obtain the ganoderma lucidum extract with stable color.

Examples 2 to 7

Examples 2 to 7 are similar to the preparation method of example 1, except that the enzyme activity ratio in the complex enzyme solution is different, the rotation speed of the flash extractor is different, and the power of the microwave-assisted extraction is different.

Comparative example 1

The comparative example is substantially identical to the preparation method of example 1, except that the complex enzyme solution obtained in the first step is a plant refined enzyme purchased from the company of the scientific development of Jiaming, beijing.

Comparative example 2

This comparative example is substantially identical to the preparation of example 1, except that the microwave is used for the first step of flash extraction, the time and power of which are identical to those of example 1.

The ganoderma lucidum extract prepared in the embodiment is subjected to a cell degranulation experiment, mast cells are used as a cell model, and a detection means is a microscopic observation method. As shown in fig. 1 and table 1, the NC degranulation cell number significantly increased compared to the BC group, indicating that the stimulation conditions were effective in this experiment. Compared with the NC group, the number of degranulation cells of the PC group is obviously reduced, which proves that the positive control detection is effective, and the result shows that the degranulation cells of the ganoderma lucidum extract (0.1563%, 0.3125% and 0.625%) of the embodiment 1 are obviously reduced, thus the ganoderma lucidum extract of the embodiment 1 has obvious antiallergic capability. Further efficacy experiments were performed on the ganoderma lucidum extract obtained in example 1 as the most preferred example.

TABLE 1 test results of particle removal rate

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As can be seen from the data in table 1, when the enzyme activity ratio of the complex enzyme solution in the first step is 4: 1. the rotating speed of the flash extractor is 2500r/min, and when the microwave auxiliary preheating power in the second step is 500W, the obtained ganoderma lucidum extract has better antiallergic property. In addition, in the preparation method provided by the application, the enzyme type of the complex enzyme solution and the auxiliary extraction method and the cooperation of the extraction method have important influences. Wherein example 1 is the preferred example in the present application.

Inhibition of inflammatory factor experiments:

in this experiment, anti-inflammatory effect of the sample of example 1 to be tested was evaluated by detecting inflammatory factors (IL-6, TNF-. Alpha., NO) secreted in cell supernatants by ELISA kit using LPS-induced RAW264.7 cells as an in vitro inflammatory cell model.

TABLE 2 comparison of IL-6 differences between samples in different groups

# indicates that the difference was statistically significant (p < 0.05) compared to the negative control group (NT)

* The expression is compared with the model control group (M), and the difference has statistical significance (p < 0.05)

As can be seen from the data in Table 2, the IL-6 content of the negative control group (NT) was significantly reduced by 35.07% (P < 0.05) compared to the model group (M); compared with the positive control group (PC) and the model group, the relative content of IL-6 is obviously reduced by 37.03 percent (P is less than 0.05), which indicates that the modeling is successful; at concentrations of 0.60%, 0.20% and 0.06% of the ganoderma lucidum extract of sample group (TA) of example 1, the relative content of IL-6 was significantly reduced by 29.54%,22.18% and 30.86% compared to the model group (P < 0.05).

TABLE 3 comparison of TNF-alpha differences for samples in different groups

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As can be seen from the data in Table 3, the negative control (NT) had a significant reduction in TNF-. Alpha.content of 75.89% (P < 0.05) compared to the model (M); compared with the positive control group (PC) and the model group, the relative content of TNF-alpha is obviously reduced by 77.77 percent (P is less than 0.05), which indicates that the modeling is successful; at concentrations of 0.60%, 0.20% and 0.06% for the ganoderma lucidum extract of sample group (TA) of example 1, the relative TNF- α content was significantly reduced by 78.56%,66.54% and 40.05% compared to the model group (P < 0.05).

Table 4 comparison of NO differences between samples in different groups

As can be seen from the data in table 4, the negative control group (NT) had significantly reduced NO content by 78.64% (P < 0.05) compared to the model group (M); compared with the model group, the positive control group (PC) has the advantages that the relative NO content is obviously reduced by 44.48 percent (P is less than 0.05), which indicates that the modeling is successful; at concentrations of 0.60% and 0.06% of the ganoderma lucidum extract of sample group (TA) of example 1, the relative NO content was significantly reduced by 32.10% and 44.94%, respectively, compared to the model group (P < 0.05).

The ganoderma lucidum extract samples showed anti-inflammatory activity in this test. By inhibiting the three pro-inflammatory factors, the problem of skin inflammation caused by the breeding of anaerobic bacteria acne bacillus can be prevented or improved.

Anti-aging experiment:

MMP-1 is the most important enzyme for degrading type I and type III collagens, and the excessive expression of MMP-1 by skin cells can cause the damage of extracellular matrixes of dermis layers, especially the normal structure of collagen fibers and elastic fibers, so that MMP-1 is the most important enzyme for causing aging symptoms such as wrinkles, fine lines and the like of skin, and the content of the Procolagen-1 is also an important index for reflecting the aging condition of cells. The anti-aging effect of the test substance is evaluated by detecting the contents of the two.

TABLE 5 Effect of samples on cellular MMP-1 levels

From the data in Table 5, the relative MMP-1 content of model group (M) was significantly increased (P < 0.05) compared to negative control group (NT); compared with the model group, the relative content of MMP-1 in the positive control group is obviously reduced (P is less than 0.05), which indicates that the modeling is successful; at a concentration of 0.63% of the ganoderma lucidum extract of the sample group (TA) of example 1, the MMP-1 relative content of the sample group of example 1 was significantly reduced (P < 0.05) compared to the model group.

TABLE 6 Effect of samples on cellular Procolagen-1 levels

As can be seen from the data in Table 6, the relative content of Procolagen-1 was significantly reduced (P < 0.05) in the model group (M) compared with the negative control group (NT); compared with the model group, the positive control group has obviously increased relative content of the Procolagen-1 (P is less than 0.05), which indicates that the modeling is successful; at a concentration of 0.63% of the ganoderma lucidum extract of the sample group (TA) of example 1, the relative content of the Procollagen-1 of the sample group of example 1 was significantly increased (P < 0.05) compared with the model group.

The ganoderma lucidum extract samples showed anti-aging activity in this test. By inhibiting MMP-1 and increasing the relative content of the Procolagen-1, the skin can be provided with aging problems such as wrinkles, fine lines and the like.

Cell degranulation assay:

TABLE 7 test results of particle removal rate

And (3) performing a cell degranulation experiment on the prepared ganoderma lucidum extract, and adopting mast cells as a cell model, wherein a detection means is a microscopic observation method. As shown in fig. 1 and table 7, the NC degranulation cell number significantly increased compared to the BC group, indicating that the stimulation conditions were effective in this experiment. Compared with NC group, the number of degranulation cells in PC group is obviously reduced, which proves that the positive control detection is effective, and the result shows that the prepared ganoderma lucidum extract has obvious antiallergic capability.

In conclusion, the ganoderma lucidum extract provided in embodiment 1 of the application adopts Chinese herbal medicine sources, and has the advantages of good naturalness, no toxic or side effect, mildness and no stimulation. According to laboratory tests, the effects of mildly and efficiently extracting the ganoderma lucidum extract with high bioactivity are achieved by combining microwaves and alcohol extraction, the IL-1 beta, TNF-alpha and NO inflammatory factors which are over expressed due to the breeding of anaerobic acne bacillus in skin hair follicles can be inhibited, the content of MMP-1 enzyme which is the most important in the aging symptoms such as wrinkles and fine lines of skin is inhibited, the relative content of the Procolagen-1 is improved, both the symptoms and root causes can be treated, and the effects of anti-inflammatory, antibacterial and antiallergic are achieved, so that a foundation is laid for the application of the ganoderma lucidum extract in the field of anti-inflammatory, anti-aging skin care products.

The foregoing is merely exemplary of the present application and is not intended to limit the present application. Various modifications and changes may be made to the present application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc. which are within the spirit and principles of the present application are intended to be included within the scope of the claims of the present application.

Claims

1. A method for preparing a ganoderma lucidum extract for cosmetics, which is characterized by comprising the following steps:

step one, adding lucid ganoderma, an extraction solvent and a compound enzyme liquid into a flash extractor, wherein the enzyme activity ratio of the enzyme in the compound enzyme liquid is (2-6): (0.5-2) xylanase and pectase, vacuumizing the flash extractor, and extracting for 4-7 min at the speed of 1000-3500 r/min by using the flash extractor to obtain a mixed solution; the extraction solvent is water;

reflux-extracting the mixed solution with ethanol, filtering to obtain primary lucid ganoderma extract and lucid ganoderma residues, adding the lucid ganoderma residues into the ethanol, and extracting for 1-3 times with the aid of microwaves, wherein the power of the microwaves is 350-600 w, and the extracting time is 10-30 min each time to obtain secondary lucid ganoderma extract;

and step four, stirring and dissolving the lucid ganoderma extract, deionized water and 1, 3-propylene glycol to obtain lucid ganoderma dissolution liquid, adding diatomite into the lucid ganoderma dissolution liquid, stirring, standing overnight, vacuum filtering, supplementing 1, 3-propylene glycol, and stirring until the lucid ganoderma dissolution liquid is clear and transparent to obtain the lucid ganoderma extract.

2. The method for preparing a ganoderma lucidum extract for cosmetics according to claim 1, wherein in the first step, the complex enzyme solution is 3% of the mass of ganoderma lucidum.

3. The method according to claim 1, wherein in the first step, the total enzyme activity in the complex enzyme solution is about 12U/g.

4. The method for preparing a ganoderma lucidum extract for cosmetics according to claim 1, wherein in the first step, the enzyme activity ratio of the enzyme in the complex enzyme solution is 4:1, xylanase and pectase.

5. The method for preparing a ganoderma lucidum extract for cosmetics according to claim 1, wherein in the first step, the flash extractor extracts for 6min at 2000 r/min.

6. The method of claim 1, wherein in the second step, the reflux extraction is performed 2 times in a water bath at 60 ℃ for 2h times.

7. The method for preparing a ganoderma lucidum extract for cosmetics according to claim 1, wherein in the second step, the microwave extraction is performed 2 times, the power of the microwave is 500w, and the extraction time is 20min each time.

8. The ganoderma lucidum extract prepared by the preparation method of any one of claims 1 to 7.

9. Use of the ganoderma lucidum extract according to claim 8 for preparing cosmetics, wherein the cosmetics comprise anti-inflammatory and anti-aging cosmetics.