Leontopodium alpinum callus extract as well as preparation method and application thereof

Abstract

The invention discloses a leontopodium alpinum callus extract as well as a preparation method and application thereof, and belongs to the technical field of plant active ingredient extraction. The leontopodium alpinum callus extract is prepared by the following steps: mixing leontopodium alpinum callus freeze-dried powder with a buffer solution, adding cellulase, pectinase and beta-glucanase, uniformly mixing, reacting to obtain primary enzymolysis slurry, adjusting the primary enzymolysis slurry to be neutral, adding phospholipase A2, stirring, reacting to obtain deep enzymatic hydrolysate, filtering, washing, and drying to obtain the leontopodium alpinum callus extract. Transferring into an extraction kettle, adding an entrainer, and extracting to obtain a leontopodium alpinum extract; dissolving soybean phospholipid and cholesterol in ethanol to obtain an organic phase; dispersing the leontopodium alpinum extract in a buffer solution to obtain an active matter water phase; and injecting the organic phase into the active matter water phase, stirring, carrying out reduced pressure distillation, and homogenizing. The preparation method provided by the invention can efficiently and completely obtain the active component, and the nano-liposome formed by phospholipid molecules can effectively improve the stability of the active component.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant active ingredient extraction, and particularly relates to a Leontopodium alpinum callus extract, a preparation method thereof and application thereof. BACKGROUND

[0002] Leontopodium alpinum, also known as Edelweiss, is a rare chrysanthemum herbaceous plant, which is famous for its unique pharmacological properties. The extract thereof is rich in flavonoids (such as luteolin, apigenin) and phenolic acids, and has excellent antioxidant, anti-inflammatory and anti-photoaging abilities. It shows strong antioxidant and anti-inflammatory activities in vitro and in vivo, and can reduce the formation of inflammation and wrinkles under UVB treatment.

[0003] The existing traditional extraction methods have some defects, such as high-temperature treatment which may lead to degradation of active ingredients, and use of organic solvents which brings safety and environmental problems. The traditional solvent extraction method (such as hot water extraction, organic solvent extraction) has limited ability to destroy cell structure. Although the callus cells have no cuticle, they still have tough cell walls, which makes it difficult for intracellular active ingredients to be fully released, resulting in low extraction rate. Traditional hot extraction or long-time concentration process may lead to oxidation and destruction of heat-sensitive components (such as flavonoids). In addition, a large amount of organic solvents such as petroleum ether, acetone and chloroform are used for extraction or defatting, which is effective but has the risk of toxic solvent residues.

[0004] Therefore, it is necessary to propose a new preparation method of Leontopodium alpinum callus extract. SUMMARY

[0005] Based on the deficiencies of the prior art, the purpose of the present application is to provide a Leontopodium alpinum callus extract, a preparation method thereof and application thereof.

[0006] The first aspect of the present application is to provide a preparation method of Leontopodium alpinum callus extract, comprising the following preparation steps: S1: mixing the freeze-dried powder form of Leontopodium alpinum callus with buffer solution I, and sequentially adding cellulase, pectinase and beta-glucanase to mix uniformly, to obtain primary enzymolysis slurry after reaction; S2: adjusting the pH value of the primary enzymolysis slurry to neutral, and then adding phospholipase A2 to stir and react to obtain a deep enzymolysis liquid; S3: transferring the deep enzymolysis liquid to an extraction kettle, adding an entraining agent, and introducing a supercritical fluid into the extraction kettle to extract to obtain a Leontopodium alpinum extract; S4: dissolving soybean phospholipid and cholesterol in an ethanol solvent to obtain an organic phase; S5: dispersing the Leontopodium alpinum extract in buffer solution II to obtain an active water phase; S6: injecting the organic phase into the active water phase, stirring, and then reducing pressure distillation and homogenization to obtain the Potentilla fruticosa L. callus extract.

[0007] In some embodiments, the buffer solution I is a citric acid-phosphate buffer solution with pH=4.5-5.5, and the mass amount of the buffer solution I is 13-15 times the mass amount of the Potentilla fruticosa L. callus lyophilized powder.

[0008] In some embodiments, the mass amount of the cellulase is 1-2% of the mass amount of the Potentilla fruticosa L. callus lyophilized powder, and the mass ratio of the cellulase, pectinase, β-glucanase and phospholipase A2 is 1-2:0.5-1.5:0.3-0.7:0.05-0.15.

[0009] In some embodiments, the entraining agent is selected from at least one of food-grade anhydrous ethanol and glycerol, and the mass amount of the entraining agent is 70-80% of the mass amount of the Potentilla fruticosa L. callus lyophilized powder.

[0010] In some embodiments, the mass ratio of the soybean phospholipid and cholesterol is 3-5:0.5-1.5, and the mass amount of the ethanol solvent is 10-12 times the mass amount of the soybean phospholipid and cholesterol.

[0011] In some embodiments, the buffer solution II is selected from at least one of a phosphate buffered saline solution and a 5% glycerol aqueous solution, and the mass amount of the buffer solution II is 18-22 times the mass amount of the Potentilla fruticosa L. extract.

[0012] In some embodiments, the mass ratio of the active water phase and the organic phase is 1:1.1-1.3.

[0013] In some embodiments, in S1, the reaction is specifically carried out at 85-95℃ for 8-12 min, and then the temperature is lowered to 35-45℃; in S2, the stirring reaction time is 1-2 h; in S3, the extraction pressure is 25-35 MPa, the extraction temperature is 40-50℃, and the extraction time is 2.5-3.5 h; in S4, the temperature of the organic solvent is 45-55℃; in S5, the temperature of the buffer solution II is 55-65℃; and in S6, the stirring temperature is 55-65℃, the stirring time is 1.5-2.5 h, and the homogenization pressure is 750-850 bar.

[0014] The second aspect of the present application is to provide a Potentilla fruticosa L. callus extract.

[0015] The third aspect of the present application is to provide an application of the Potentilla fruticosa L. callus extract in the field of cosmetics and / or skin care products.

[0016] It should be noted that the high mountain firebrush gynostemma pentaphyllum callus extract provided by the present application can be compounded with other antioxidant ingredients to prepare an essence, emulsion or gel, and the present application does not limit the compounded ingredients.

[0017] Compared with the prior art, the following beneficial effects are achieved: 1. The present application realizes efficient and complete extraction of active ingredients by combining complex enzymolysis and supercritical extraction technology. The complex enzymolysis first decomposes the cell wall skeleton structure of high mountain firebrush gynostemma pentaphyllum callus cells under acidic pH and mild heating conditions through the synergistic action of a complex enzyme system composed of cellulase, pectinase and beta-glucanase, and then specifically decomposes the phospholipid membrane structure of organelles through phospholipase, so as to realize the release of cell contents and efficient elution of active ingredients. The supercritical extraction technology uses a fluid in a supercritical state as a solvent to dissolve and extract the active ingredients released by enzymolysis, and is supplemented with an entrainer to enhance the solubility of polar ingredients.

[0018] 2. The present application uses liposome coating technology to dissolve soybean phospholipids and cholesterol in ethanol to form an organic phase, and then adds an aqueous phase containing active extract into the organic phase to form a water-in-oil state. The phospholipid molecules spontaneously assemble to form a nanoliposome with a bilayer structure, efficiently encapsulating the active ingredients of high mountain firebrush gynostemma pentaphyllum in the hydrophilic core and lipid bilayer. Finally, a stable system with uniform particle size is obtained through homogenization treatment, significantly improving the stability, skin permeability and bioavailability of active substances. DETAILED DESCRIPTION

[0019] The present application will be further described in detail below in combination with specific examples.

[0020] Example 1 A preparation method of high mountain firebrush gynostemma pentaphyllum callus extract includes the following preparation steps: S1: Mix high mountain firebrush gynostemma pentaphyllum callus freeze-dried powder with citric acid-phosphate buffer solution with pH=5, and then add cellulase, pectinase and beta-glucanase in sequence and mix uniformly, react at 90℃ for 10 min, and then cool to 40℃ to obtain primary enzymolysis slurry; wherein the mass amount of citric acid-phosphate buffer solution is 14 times the mass of high mountain firebrush gynostemma pentaphyllum callus freeze-dried powder, the mass amount of cellulase is 1.5% of the mass of high mountain firebrush gynostemma pentaphyllum callus freeze-dried powder, and the mass ratio of cellulase, pectinase, beta-glucanase and phospholipase A2 is 1.5:1:0.5:0.1; S2: Adjust the pH value of the primary enzymolysis slurry to neutral, and then add phospholipase A2 and stir for 1.5 h to obtain a deep enzymolysis solution; S3: transferring the deep enzymatic hydrolysate into an extraction kettle, adding food-grade anhydrous ethanol, introducing supercritical fluid into the extraction kettle, and extracting at 30 MPa and 45℃ for 3 h to obtain the extract of the Ledum palustre; wherein the mass amount of the food-grade anhydrous ethanol is 75% of the mass of the freeze-dried powder of the Ledum palustre callus; S4: dissolving soybean phospholipid and cholesterol in a mass ratio of 4:1 in anhydrous ethanol at 50℃ to obtain an organic phase; wherein the mass amount of the anhydrous ethanol is 11 times the mass of the soybean phospholipid and cholesterol; S5: dispersing the extract of the Ledum palustre in a phosphate buffer salt solution at 60℃ to obtain an active water phase; wherein the mass amount of the phosphate buffer salt solution is 20 times the mass of the extract of the Ledum palustre; S6: injecting the organic phase into the active water phase, stirring at 60℃ for 2 h, and homogenizing at 800 bar to obtain the extract of the Ledum palustre callus; wherein the mass ratio of the active water phase to the organic phase is 1:1.2.

[0021] The extract of the Ledum palustre callus prepared by the preparation method.

[0022] Application of the extract of the Ledum palustre callus in the field of cosmetics / skin care products.

[0023] Example 2 A preparation method of an extract of Ledum palustre callus, comprising the following preparation steps: S1: mixing the freeze-dried powder of the Ledum palustre callus with a citric acid-phosphate buffer solution with a pH of 5.5, sequentially adding cellulase, pectinase and β-glucanase, and uniformly mixing, then reacting at 95℃ for 12 min, and then cooling to 45℃ to obtain a primary enzymatic hydrolysis slurry; wherein the mass amount of the citric acid-phosphate buffer solution is 15 times the mass of the freeze-dried powder of the Ledum palustre callus, the mass amount of the cellulase is 2% of the mass of the freeze-dried powder of the Ledum palustre callus, and the mass ratio of the cellulase, the pectinase, the β-glucanase and the phospholipase A2 is 2:1.5:0.7:0.15; S2: adjusting the pH value of the primary enzymatic hydrolysis slurry to neutral, then adding phospholipase A2, and stirring for 2 h to obtain a deep enzymatic hydrolysate; S3: transferring the deep enzymatic hydrolysate into an extraction kettle, adding glycerol, introducing supercritical fluid into the extraction kettle, and extracting at 35 MPa and 50℃ for 3.5 h to obtain the extract of the Ledum palustre; wherein the mass amount of the glycerol is 80% of the mass of the freeze-dried powder of the Ledum palustre callus; S4: dissolving soybean phospholipid and cholesterol in a mass ratio of 5:1.5 in anhydrous ethanol at 55℃ to obtain an organic phase; wherein the mass amount of the anhydrous ethanol is 12 times the mass of the soybean phospholipid and cholesterol; S5: dispersing the high mountain fire extraction in 5% glycerol water solution at 65 ℃ to obtain active water phase; wherein, the mass of 5% glycerol water solution is 22 times of the mass of high mountain fire extraction; S6: injecting organic phase into the active water phase, stirring at 65 ℃ for 2.5 h, reducing pressure distillation, homogenizing at 850 bar to obtain high mountain fire callus extract; wherein, the mass ratio of active water phase and organic phase is 1:1.3.

[0024] The high mountain fire callus extract prepared by the preparation method.

[0025] The application of the high mountain fire callus extract in the field of cosmetics / skin care.

[0026] Example 3 A preparation method of high mountain fire callus extract, comprising the following preparation steps: S1: mixing high mountain fire callus freeze-dried powder with citric acid-phosphate buffer solution with pH value of 4.5, adding cellulase, pectinase and β-glucanase in sequence and mixing uniformly, reacting at 85 ℃ for 8 min, then cooling to 35 ℃ to obtain primary enzymolysis slurry; wherein, the mass of citric acid-phosphate buffer solution is 13 times of the mass of high mountain fire callus freeze-dried powder, the mass of cellulase is 1% of the mass of high mountain fire callus freeze-dried powder, and the mass ratio of cellulase, pectinase, β-glucanase and phospholipase A2 is 1:0.5:0.3:0.05; S2: adjusting the pH value of the primary enzymolysis slurry to neutral, then adding phospholipase A2 and stirring for 1 h to obtain deep enzymolysis liquid; S3: transferring the deep enzymolysis liquid to an extraction kettle, adding food-grade anhydrous ethanol, introducing supercritical fluid into the extraction kettle, extracting at 25 MPa and 40 ℃ for 2.5 h to obtain high mountain fire extraction; wherein, the mass of food-grade anhydrous ethanol is 70% of the mass of high mountain fire callus freeze-dried powder; S4: dissolving soybean phospholipid and cholesterol with a mass ratio of 3:0.5 in anhydrous ethanol at 45 ℃ to obtain organic phase; wherein, the mass of anhydrous ethanol is 10 times of the mass of soybean phospholipid and cholesterol; S5: dispersing the high mountain fire extraction in 5% glycerol water solution at 65 ℃ to obtain active water phase; wherein, the mass of 5% glycerol water solution is 22 times of the mass of high mountain fire extraction; S6: injecting organic phase into the active water phase, stirring at 65 ℃ for 2.5 h, reducing pressure distillation, homogenizing at 850 bar to obtain high mountain fire callus extract; wherein, the mass ratio of active water phase and organic phase is 1:1.3.

[0027] The high mountain firebrush callus extract prepared by the preparation method.

[0028] Application of the high mountain firebrush callus extract in the field of cosmetics / skin care products.

[0029] Example 4 A preparation method of a high mountain firebrush callus extract, comprising the following preparation steps: S1: freeze-dried powder of high mountain firebrush callus is mixed with citric acid-phosphate buffer solution with pH=5, cellulase, pectinase and β-glucanase are sequentially added and uniformly mixed, and then the mixture is reacted at 85℃ for 9min, and then the temperature is reduced to 35℃ to obtain primary enzymolysis slurry; wherein the mass amount of citric acid-phosphate buffer solution is 13 times the mass of freeze-dried powder of high mountain firebrush callus, the mass amount of cellulase is 1% of the mass of freeze-dried powder of high mountain firebrush callus, and the mass ratio of cellulase, pectinase, β-glucanase and phospholipase A2 is 2:1:0.3:0.1; S2: the pH value of the primary enzymolysis slurry is adjusted to neutral, and then phospholipase A2 is added and stirred for 1h to obtain a deep enzymolysis liquid; S3: the deep enzymolysis liquid is transferred to an extraction kettle, glycerol is added, and supercritical fluid is introduced into the extraction kettle, and the extraction is carried out at 30MPa and 50℃ for 2.5h to obtain a high mountain firebrush extract; wherein the mass amount of glycerol is 75% of the mass of freeze-dried powder of high mountain firebrush callus; S4: soybean phospholipid and cholesterol with a mass ratio of 3:1 are dissolved in anhydrous ethanol at 55℃ to obtain an organic phase; wherein the mass amount of anhydrous ethanol is 11 times the mass of soybean phospholipid and cholesterol; S5: the high mountain firebrush extract is dispersed in a 5% glycerol aqueous solution at 55-65℃ to obtain an active substance water phase; wherein the mass amount of 5% glycerol aqueous solution is 21 times the mass of the high mountain firebrush extract; S6: the active substance water phase is injected into the organic phase, stirred at 55℃ for 2.5h, and then subjected to reduced pressure distillation and homogenization at 850bar to obtain the high mountain firebrush callus extract; wherein the mass ratio of the active substance water phase and the organic phase is 1:1.2.

[0030] The high mountain firebrush callus extract prepared by the preparation method.

[0031] Application of the high mountain firebrush callus extract in the field of cosmetics / skin care products.

[0032] Example 5 A preparation method of a high mountain firebrush callus extract, comprising the following preparation steps: S1: The high mountain fireweed callus freeze-dried powder is mixed with a citric acid-phosphate buffer solution with pH=4.5, and cellulase, pectinase and β-glucanase are sequentially added and uniformly mixed, and then reacted at 95°C for 8 min, and then cooled to 45°C to obtain a primary enzymolysis slurry; wherein the mass amount of the citric acid-phosphate buffer solution is 13 times the mass of the high mountain fireweed callus freeze-dried powder, the mass amount of the cellulase is 1.5% of the mass of the high mountain fireweed callus freeze-dried powder, and the mass ratio of the cellulase, the pectinase, the β-glucanase and the phospholipase A2 is 1:1:0.7:0.05; S2: The pH value of the primary enzymolysis slurry is adjusted to neutral, and then phospholipase A2 is added and stirred for 2 h to obtain a deep enzymolysis liquid; S3: The deep enzymolysis liquid is transferred to an extraction kettle, food-grade anhydrous ethanol is added, and a supercritical fluid is introduced into the extraction kettle, and the extraction is carried out at 30 MPa and 50°C for 2.5 h to obtain a high mountain fireweed extract; wherein the mass amount of the food-grade anhydrous ethanol is 78% of the mass of the high mountain fireweed callus freeze-dried powder; S4: Soybean phospholipids and cholesterol with a mass ratio of 5:1 are dissolved in anhydrous ethanol at 50°C to obtain an organic phase; wherein the mass amount of the anhydrous ethanol is 10 times the mass of the soybean phospholipids and the cholesterol; S5: The high mountain fireweed extract is dispersed in a phosphate buffer salt solution at 65°C to obtain an active water phase; wherein the mass amount of the phosphate buffer salt solution is 22 times the mass of the high mountain fireweed extract; S6: The active water phase is injected into the organic phase, stirred at 60°C for 2 h, and then subjected to reduced pressure distillation and homogenization at 800 bar to obtain a high mountain fireweed callus extract; wherein the mass ratio of the active water phase to the organic phase is 1:1.3.

[0033] The high mountain fireweed callus extract prepared by the preparation method.

[0034] Application of the high mountain fireweed callus extract in the field of cosmetics / skin care products.

[0035] Comparative Example 1: Traditional extraction method The application is basically the same as Example 1, except that: The traditional extraction method is used, i.e., the high mountain fireweed callus freeze-dried powder is mixed with 70% ethanol, and then hot reflux extraction is carried out at 80°C for 2 h. After the reflux is completed, a Buchner funnel is used for vacuum filtration to collect the filtrate, and then the filtrate is transferred to a rotary evaporator, and concentrated under reduced pressure at a water bath temperature of 60°C to recover ethanol and remove residual water to obtain the high mountain fireweed callus extract.

[0036] Comparative Example 2: Single enzymolysis method The application is basically the same as Example 1, except that: S3 is replaced by the following steps: the deep enzymatic hydrolysate prepared in S2 is subjected to suction filtration, the solid residue is separated from the liquid part containing active ingredients, the filtrate is collected, and the filtrate is transferred to a rotary evaporator, concentrated under reduced pressure at a water bath temperature of 60°C, and the concentrated liquid is obtained as an active water phase.

[0037] Comparative Example 3 Single extraction method The preparation method of the high mountain firebrush callus extract according to the present application is basically the same as that of Example 1, except that: S1-S2 is replaced by the following steps: the high mountain firebrush callus freeze-dried powder is physically crushed and sieved through an 80-mesh sieve to obtain dry fine powder. The fine powder is replaced by the deep enzymatic hydrolysate in subsequent S3.

[0038] Comparative Example 4 Without using liposome coating technology The preparation method of the high mountain firebrush callus extract according to the present application is basically the same as that of Example 1, except that:

[0039] In order to prove that the preparation method of the high mountain firebrush callus extract according to the present application can extract more active ingredients and better ensure the stability of the active ingredients, the active ingredient content and stability tests of Examples 1-5 and Comparative Examples 1-4 are carried out, and the test data are shown in Table 1.

[0040] The high mountain firebrush callus extracts prepared in Examples 1-5 and Comparative Examples 1-4 are dissolved with methanol to prepare test solutions of the same concentration.

[0041] Active ingredient content test: the total flavonoid content is determined by aluminum nitrate-sodium nitrite colorimetry, rutin is used as the standard, a standard curve is drawn, the absorbance is measured at 510 nm after the sample is colored, and the total flavonoid content is calculated, which is expressed in milligrams of rutin equivalent per gram of extract (mg RE / g). The content of representative active ingredient, luteolin, is quantitatively analyzed by high performance liquid chromatography (HPLC), and the result is expressed in milligrams per gram of extract (mg / g).

[0042] Stability test: each group of test solutions is stored at 40°C under accelerated conditions for 30 days. Samples are taken at different time points (0, 15, and 30 days), and the chemical stability is investigated by determining the residual rate of luteolin.

[0043] Table 1 As can be seen from Table 1, the high mountain firebrush callus extract provided by the embodiment of the present application has high active ingredient content and good stability. It can be seen from the comparative examples that, due to insufficient release of cell contents and degradation of active ingredients caused by high-temperature extraction process, the total flavonoids and luteolin content of the traditional extraction method adopted in Comparative Example 1 is the lowest, and the stability is also poor; due to the absence of supercritical extraction, the subsequent hot solvent extraction and concentration steps destroy part of the activity, resulting in a decrease in the content of effective ingredients in Comparative Example 2; due to the absence of enzymatic pretreatment, the supercritical fluid is difficult to effectively penetrate the intact cell wall, resulting in a significant reduction in the extraction efficiency of Comparative Example 3; due to the absence of protection of the liposome coating technology, in the accelerated experiment, the active ingredients are directly exposed to a high-temperature environment, and the degradation speed is fast, resulting in a decrease in stability.

[0044] The above only describes some embodiments of the present application. For those skilled in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are within the scope of protection of the present application.

Claims

1. A method for preparing an extract from Edelweiss callus tissue, characterized in that, The preparation steps include the following: S1: The freeze-dried powder form of Edelweiss callus was mixed with buffer solution one, and cellulase, pectinase and β-glucanase were added in sequence and mixed evenly. After the reaction, the primary enzymatic hydrolysate was obtained. S2: Adjust the pH of the primary enzymatic hydrolysate to neutral, then add phospholipase A2 and stir to obtain a deep enzymatic hydrolysate; S3: Transfer the deep enzymatic hydrolysate to the extraction vessel, add an entrainer, and introduce supercritical fluid into the extraction vessel to extract the extract of Edelweiss extract. S4: Soybean lecithin and cholesterol are dissolved in ethanol solvent to obtain an organic phase; S5: Disperse the alpine edelweiss extract in buffer solution II to obtain the active ingredient aqueous phase; S6: Inject the organic phase into the aqueous phase of the active ingredient, stir, distill under reduced pressure, and homogenize to obtain the Edelweiss callus extract.

2. The method for preparing the *Edelweiss* callus extract according to claim 1, characterized in that, The first buffer solution is a citrate-phosphate buffer solution with a pH of 4.5-5.5, and the mass of the first buffer solution is 13-15 times the mass of the lyophilized powder of Edelweiss callus.

3. The method for preparing the *Edelweiss* callus extract according to claim 1, characterized in that, The amount of cellulase used is 1-2% of the mass of the freeze-dried Edelweiss callus powder; the mass ratio of cellulase, pectinase, β-glucanase and phospholipase A2 is 1-2:0.5-1.5:0.3-0.7:0.05-0.

15.

4. The method for preparing the Edelweiss callus extract according to claim 1, characterized in that, The entrainer is selected from at least one of food-grade anhydrous ethanol and glycerin, and the mass amount of the entrainer is 70-80% of the mass of the freeze-dried powder of Edelweiss callus.

5. The method for preparing the *Edelweiss* callus extract according to claim 1, characterized in that, The mass ratio of soybean lecithin to cholesterol is 3-5:0.5-1.5; the mass of ethanol solvent used is 10-12 times the mass of soybean lecithin and cholesterol.

6. The method for preparing the *Edelweiss* callus extract according to claim 1, characterized in that, The second buffer solution is selected from at least one of phosphate buffer solution and 5% glycerol aqueous solution, and the mass of the second buffer solution is 18-22 times the mass of the edelweiss extract.

7. The method for preparing the *Edelweiss* callus extract according to claim 1, characterized in that, The mass ratio of the active ingredient aqueous phase to the organic phase is 1:1.1-1.

3.

8. The method for preparing the Edelweiss callus extract according to claim 1, characterized in that, In step S1, the reaction is carried out at 85-95℃ for 8-12 minutes, and then cooled to 35-45℃; in step S2, the stirring reaction time is 1-2 hours; in step S3, the extraction pressure is 25-35 MPa, the extraction temperature is 40-50℃, and the extraction time is 2.5-3.5 hours; in step S4, the organic solvent temperature is 45-55℃; in step S5, the temperature of buffer solution II is 55-65℃; in step S6, the stirring temperature is 55-65℃, the stirring time is 1.5-2.5 hours, and the homogenization pressure is 750-850 bar.

9. An extract of Edelweiss callus tissue according to any one of claims 1-8.

10. The application of the Edelweiss callus extract according to claim 9 in the field of cosmetics and / or skin care products.

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