Glycyrrhiza glabra root extract with anti-inflammatory and skin-brightening effects as well as preparation method and application of glycyrrhiza glabra root extract

Abstract

The invention relates to a glycyrrhiza glabra root extract with anti-inflammatory and skin-brightening effects as well as a preparation method and application of the glycyrrhiza glabra root extract. The preparation method comprises the following steps: (1) mixing glycyrrhiza glabra root powder with a solution containing a deep eutectic solvent, adjusting the pH value of the mixed solution to 8-10, carrying out hot extraction and filtration, and taking the filtrate to obtain a glycyrrhiza glabra root crude extract; and (2) adjusting the pH value of the glycyrrhiza glabra root crude extract to 3-4, and carrying out secondary hot extraction to obtain the glycyrrhiza glabra root extract. According to the preparation method disclosed by the invention, the content of functional components in the glycyrrhiza glabra root is greatly increased by matching a specific hot extraction solvent with multiple pH regulation, so that the anti-inflammatory and skin-brightening effects of the prepared glycyrrhiza glabra root extract are improved.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to a Glycyrrhiza glabra root extract with anti-inflammatory and skin-lightening effects, a preparation method and an application thereof. Background Art

[0002] The root and rhizome tissue cells of Glycyrrhiza glabra contain functional ingredients with strong antioxidant activity that can depigment the skin and lighten spots. These ingredients are also the main components, including glycyrrhizin and glycyrrhizin. Compared with artificially synthesized chemical ingredients, they are milder and have soothing and anti-inflammatory effects, making them suitable for sensitive skin and acne-prone skin. In addition, glycyrrhizic acid in the root of Glycyrrhiza glabra can significantly inhibit the expression of skin inflammatory factors and has soothing and anti-inflammatory effects. These characteristics make its application in skin care products and cosmetics particularly prominent.

[0003] However, due to the different preparation methods of Glycyrrhiza glabra root extract, the types and contents of its effective ingredients are also different, and the anti-inflammatory and skin-lightening effects are uneven. The effects of Glycyrrhiza glabra root extract are affected to a certain extent, and the product application is also limited.

[0004] CN117362308A discloses a method for preparing ultra-high-purity glabridin, comprising soaking Glycyrrhiza glabra in pure water and hydrochloric acid, adding the acid-washed Glycyrrhiza glabra to a sodium hydroxide solution, and filtering to obtain a first filtrate; adjusting the pH value of the first filtrate to precipitate solids, filtering to obtain a second filtrate, washing the filter residue, and drying to obtain crude glabridin; heating and concentrating the second filtrate, cooling it to room temperature to precipitate solids, and filtering to obtain a third filtrate; adjusting the pH value of the third filtrate with hydrochloric acid to obtain crude glycyrrhizic acid; purifying the crude glycyrrhizic acid to obtain pure glycyrrhizic acid; recrystallizing the crude glabridin to obtain pure glabridin; and purifying the pure glabridin to obtain ultra-high-purity glabridin. However, although this method obtains high-purity glabridin, it can only exert the single efficacy of glabridin during application, and cannot improve the anti-inflammatory and skin-lightening effects of Glycyrrhiza glabra extract from multiple angles.

[0005] Therefore, developing a method for preparing a Glycyrrhiza glabra root extract so that the prepared Glycyrrhiza glabra root extract has excellent anti-inflammatory and skin-lightening effects has become one of the technical problems that need to be solved urgently. Summary of the Invention

[0006] To address the shortcomings of the existing technology, the present invention aims to provide a Glycyrrhiza glabra root extract with anti-inflammatory and skin-lightening effects, as well as its preparation method and application. The Glycyrrhiza glabra root extract prepared by this preparation method can reduce localized pigmentation and lighten skin spots, while also inhibiting the release of inflammatory factors and alleviating skin inflammatory reactions, demonstrating soothing and anti-inflammatory effects.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing a Glycyrrhiza glabra root extract having anti-inflammatory and skin-lightening effects, the preparation method comprising the following steps:

[0009] (1) mixing the powder of Glycyrrhiza glabra root with a solution containing a deep eutectic solvent, adjusting the pH value of the mixture to 8-10, performing hot extraction, filtering, and taking the filtrate to obtain a crude extract of Glycyrrhiza glabra root;

[0010] (2) The pH value of the crude extract of Glycyrrhiza glabra root was adjusted to 3-4, and a secondary hot extraction was performed to obtain the Glycyrrhiza glabra root extract.

[0011] In the present invention, by adjusting the pH value to 8-10, glycyrrhizic acid, glycyrrhetinic acid and other components in the root of Glycyrrhiza glabra are fully dissolved, and the root of Glycyrrhiza glabra is subjected to hot extraction using a solution containing a low eutectic solvent, which can better increase the content of the active ingredients. At the same time, in step (2), the pH value of the crude extract of the root of Glycyrrhiza glabra is adjusted to 3-4 for secondary hot extraction, which can increase the dissolution of active ingredients such as flavonoids and phenolic substances. The present invention greatly increases the content of the active ingredients in the root of Glycyrrhiza glabra by combining a specific hot extraction solvent and multiple pH adjustments, thereby improving the anti-inflammatory and skin-lightening effects of the root extract of Glycyrrhiza glabra.

[0012] Preferably, the raw materials for preparing the solution containing a low eutectic solvent in step (1) include a low eutectic solvent and water.

[0013] Preferably, the deep eutectic solvent comprises choline chloride and glycerol.

[0014] The present invention creatively discovers that using choline chloride as a hydrogen bond acceptor and glycerol as a hydrogen bond donor to form a deep eutectic solvent can better achieve the extraction effect of the effective components in the roots of Glycyrrhiza glabra.

[0015] Preferably, the molar ratio of choline chloride to glycerol is 1:(2-7), for example, it can be 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, etc.

[0016] Preferably, the mass ratio of the deep eutectic solvent to water is (1-5):1, for example, it can be 1:1, 2:1, 3:1, 4:1, 5:1, etc.

[0017] Preferably, the material-liquid ratio of the Glycyrrhiza glabra root powder to the solution containing the deep eutectic solvent in step (1) is 1:(15-20) g / ml, for example, it can be 1:15 g / ml, 1:16 g / ml, 1:17 g / ml, 1:18 g / ml, 1:19 g / ml, 1:20 g / ml, etc.

[0018] Preferably, the solution containing a deep eutectic solvent is prepared by a preparation method comprising the following steps:

[0019] Choline chloride and glycerol are mixed, reacted at 70-90° C. for 1-5 hours, cooled to 20-40° C., and then mixed with water to obtain a solution containing a deep eutectic solvent.

[0020] Among them, the specific point values ​​in 70-90℃ can be selected from 70℃, 75℃, 80℃, 85℃, 90℃, etc., the specific point values ​​in 20-40℃ can be selected from 20℃, 25℃, 30℃, 35℃, 40℃, etc., and the specific point values ​​in 1-5h can be selected from 1h, 2h, 3h, 4h, 5h, etc.

[0021] Preferably, the temperature of the hot extraction in step (1) is 55-65° C., and the time of the hot extraction is 5-8 hours.

[0022] Among them, the specific point values ​​in 55-65℃ can be selected from 55℃, 57℃, 59℃, 61℃, 63℃, 65℃, etc., and the specific point values ​​in 5-8h can be selected from 5h, 5.5h, 6h, 6.5h, 7h, 7.5h, 8h, etc.

[0023] Preferably, the temperature of the secondary hot extraction in step (2) is 70-80° C., and the time of the secondary hot extraction is 0.5-2 h.

[0024] Among them, the specific point values ​​in 70-80℃ can be selected from 70℃, 72℃, 74℃, 76℃, 78℃, 80℃, etc., and the specific point values ​​in 0.5-2h can be selected from 0.5h, 1h, 1.5h, 2h, etc.

[0025] Preferably, after the secondary hot extraction in step (2), the process further comprises filtering the product of the secondary hot extraction and collecting the filtrate.

[0026] Preferably, the filtering method includes microfiltration.

[0027] Preferably, the filter membrane of the microfiltration is 0.01-0.03 μm, for example, it can be 0.01 μm, 0.015 μm, 0.02 μm, 0.025 μm, 0.03 μm, etc.

[0028] Preferably, the filtrate is concentrated after the filtration.

[0029] In a second aspect, the present invention provides a Glycyrrhiza glabra root extract prepared by the method for preparing the Glycyrrhiza glabra root extract with anti-inflammatory and skin-lightening effects as described in the first aspect.

[0030] In a third aspect, the present invention provides a composition with anti-inflammatory and skin-lightening effects, comprising the Glycyrrhiza glabra root extract as described in the second aspect, acetyl glucosamine, panthenol and resveratrol.

[0031] Acetyl glucosamine is an N-acetyl derivative of glucosamine. It has good permeability and can penetrate into the skin, stimulating the synthesis of collagen in the skin. It has anti-wrinkle, anti-aging, and spot-lightening effects. Panthenol, as a common moisturizer, has a deep moisturizing effect. It can also stimulate cell division and proliferation, and can help repair skin tissue. Resveratrol is a non-flavonoid polyphenol compound that can delay skin aging, improve uneven skin tone, and has whitening, spot removal, and anti-aging effects.

[0032] The present invention creatively discovered that Glycyrrhiza glabra root extract, acetyl glucosamine, panthenol and resveratrol have significant synergistic effects in skin lightening and anti-inflammation, and can synergistically reduce local pigmentation and lighten skin spots, while inhibiting the release of inflammatory mediators and alleviating the skin's inflammatory response.

[0033] In addition, the Glycyrrhiza glabra root extract prepared by the specific method of the first aspect of the present invention can better exert a synergistic effect with acetyl glucosamine, panthenol, and resveratrol, further enhancing the skin-lightening and anti-inflammatory effects of the composition.

[0034] Preferably, the mass ratio of the Glycyrrhiza glabra root extract, acetyl glucosamine, panthenol and resveratrol is (3-8):(1-3):(2-4):(0.5-2).

[0035] Among them, the specific point values ​​in 3-8 can be selected from 3, 4, 5, 6, 7, 8, etc., the specific point values ​​in 1-3 can be selected from 1, 1.5, 2, 2.5, 3, etc., the specific point values ​​in 2-4 can be selected from 2, 2.5, 3, 3.5, 4, etc., and the specific point values ​​in 0.5-2 can be selected from 0.5, 0.7, 0.9, 1.1, 1.3, 1.5, 1.7, 2, etc.

[0036] Based on the synergistic effects of Glycyrrhiza glabra root extract, acetyl glucosamine, panthenol and resveratrol in skin lightening and anti-inflammatory, when the components in the composition are compounded according to the above proportions, the skin lightening and anti-inflammatory effects of the composition can be better exerted.

[0037] In a fourth aspect, the present invention provides a use of the Glycyrrhiza glabra root extract as described in the second aspect or the composition with anti-inflammatory and skin-lightening effects as described in the third aspect in cosmetics.

[0038] The numerical range described in the present invention includes not only the point values ​​listed above, but also any point values ​​between the above numerical ranges that are not listed. Due to space limitations and for the sake of simplicity, the present invention no longer exhaustively lists the specific point values ​​included in the range.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] The invention greatly increases the content of effective components in the root of Glycyrrhiza glabra by combining a specific hot extraction solvent and multiple pH adjustments, thereby improving the anti-inflammatory and skin-lightening effects of the Glycyrrhiza glabra root extract.

[0041] Furthermore, using choline chloride as a hydrogen bond acceptor and glycerol as a hydrogen bond donor to form a low eutectic solvent can better achieve the extraction effect of the effective components in the roots of Glycyrrhiza glabra.

[0042] Furthermore, the Glycyrrhiza glabra root extract prepared by the present invention has a significant synergistic effect with acetyl glucosamine, panthenol and resveratrol in terms of skin lightening and anti-inflammation, and can synergistically reduce local pigmentation and lighten skin spots, while inhibiting the release of inflammatory mediators and reducing the inflammatory response of the skin. DETAILED DESCRIPTION

[0043] In order to further illustrate the technical means and effects adopted by the present invention, the technical solutions of the present invention are further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0044] The sources of the raw materials in the following specific implementation manner are shown in Table 1:

[0045] Table 1

[0046]

[0047] Preparation Example 1

[0048] This preparation example provides a Glycyrrhiza glabra root extract, and the preparation method thereof is as follows:

[0049] (1) Preparing a solution containing a deep eutectic solvent: choline chloride and glycerol were mixed in a molar ratio of 1:5, reacted at 80°C for 3 hours, cooled to 30°C, and then mixed with water in a mass ratio of 3:1 to obtain a solution containing a deep eutectic solvent.

[0050] (2) Selecting Glycyrrhiza glabra root that is free of mold and impurities, clean it, dry it, and grind it into a Glycyrrhiza glabra root powder with a particle size of 1-2 mm; mixing the Glycyrrhiza glabra root powder with a solution containing a deep eutectic solvent at a solid-liquid ratio of 1:18 g / ml, adjusting the pH to 9, and hot-extracting at 60°C for 7 h. Filtering the mixture through a 200-mesh nylon filter cloth, collecting the filtrate, and obtaining a crude Glycyrrhiza glabra root extract.

[0051] (3) The pH of the crude extract of Glycyrrhiza glabra root was slowly adjusted to 5.5, and then further adjusted to 3.5. A second hot extraction was performed at 75°C for 1 hour. The secondary extraction product was microfiltered using a 0.02 μm microfiltration membrane, and the filtrate was evaporated and concentrated in a vacuum concentrator at 70°C until the volume was reduced to 10% of the original volume. The concentrate was sterilized by heating at 85°C for 15 minutes and cooled to obtain the Glycyrrhiza glabra root extract.

[0052] Preparation Example 2

[0053] This preparation example provides a Glycyrrhiza glabra root extract, and the preparation method thereof is as follows:

[0054] (1) Preparing a solution containing a deep eutectic solvent: choline chloride and glycerol were mixed in a molar ratio of 1:2, reacted at 70°C for 5 h, cooled to 20°C, and then mixed with water in a mass ratio of 2:1 to obtain a solution containing a deep eutectic solvent.

[0055] (2) Selecting Glycyrrhiza glabra root that is free of mold and impurities, clean it, dry it, and grind it into a Glycyrrhiza glabra root powder with a particle size of 1-2 mm; mixing the Glycyrrhiza glabra root powder with a solution containing a deep eutectic solvent at a solid-liquid ratio of 1:15 g / ml, adjusting the pH to 8, and heat-extracting at 55°C for 8 h. Filtering through a 100-mesh nylon filter cloth, collecting the filtrate, and obtaining a crude Glycyrrhiza glabra root extract.

[0056] (3) The pH of the crude extract of Glycyrrhiza glabra root was slowly adjusted to 6, and then further adjusted to 4. A second hot extraction was performed at 80°C for 0.5 h. The secondary extraction product was microfiltered using a 0.01 μm microfiltration membrane, and the filtrate was evaporated and concentrated in a vacuum concentrator at 60°C until the volume was reduced to 15% of the original volume. The concentrate was sterilized by heating at 90°C for 10 min and cooled to obtain the Glycyrrhiza glabra root extract.

[0057] Preparation Example 3

[0058] This preparation example provides a Glycyrrhiza glabra root extract, and the preparation method thereof is as follows:

[0059] (1) Preparing a solution containing a deep eutectic solvent: choline chloride and glycerol were mixed in a molar ratio of 1:7, reacted at 90°C for 1 hour, cooled to 40°C, and then mixed with water in a mass ratio of 5:1 to obtain a solution containing a deep eutectic solvent.

[0060] (2) Selecting Glycyrrhiza glabra root that is free of mold and impurities, clean it, dry it, and grind it into a Glycyrrhiza glabra root powder with a particle size of 1-2 mm; mixing the Glycyrrhiza glabra root powder with a solution containing a deep eutectic solvent at a solid-liquid ratio of 1:20 g / ml, adjusting the pH to 10, and hot-extracting at 65°C for 5 h. Filtering the mixture through a 200-mesh nylon filter cloth, collecting the filtrate, and obtaining a crude Glycyrrhiza glabra root extract.

[0061] (3) The pH of the crude extract of Glycyrrhiza glabra root was slowly adjusted to 5, and then further adjusted to 3. A second hot extraction was performed at 70°C for 2 h. The secondary extraction product was microfiltered using a 0.01 μm microfiltration membrane, and the filtrate was evaporated and concentrated in a vacuum concentrator at 65°C until the volume was reduced to 20% of the original volume. The concentrate was sterilized by heating at 90°C for 15 min and cooled to obtain the Glycyrrhiza glabra root extract.

[0062] Preparation Example 4

[0063] This preparation example provides a Glycyrrhiza glabra root extract, which differs from Preparation Example 1 only in that the glycerol in step (1) is replaced by an equimolar amount of propionic acid, that is, the low eutectic solvent is adjusted to a molar ratio of choline chloride and propionic acid of 1:5. The rest of the preparation method is consistent with Preparation Example 1.

[0064] Preparation Example 5

[0065] This preparation example provides a Glycyrrhiza glabra root extract, which differs from Preparation Example 1 only in that the choline chloride in step (1) is replaced with an equimolar amount of betaine, that is, the deep eutectic solvent is adjusted to a molar ratio of betaine and glycerol of 1:5. The rest of the preparation method is consistent with Preparation Example 1.

[0066] Comparative Preparation Example 1

[0067] This comparative preparation example provides a Glycyrrhiza glabra root extract, which differs from Preparation Example 1 only in that step (1) of preparing a solution containing a deep eutectic solvent is not performed, and step (2) is adjusted to mixing Glycyrrhiza glabra root powder and water at a material-liquid ratio of 1:18 g / ml. The remaining steps are consistent with Preparation Example 1.

[0068] Comparative Preparation Example 2

[0069] This comparative preparation example provides a Glycyrrhiza glabra root extract, which differs from Preparation Example 1 only in that step (2) adjusts the pH value to 10.5, and step (3) slowly adjusts the pH value of the crude Glycyrrhiza glabra root extract to 6.5 and then continues to adjust the pH value to 4.5. The remaining steps are consistent with Preparation Example 1.

[0070] Comparative Preparation Example 3

[0071] This comparative preparation example provides a Glycyrrhiza glabra root extract, which differs from Preparation Example 1 only in that step (2) adjusts the pH value to 7.5, and step (3) slowly adjusts the pH value of the crude Glycyrrhiza glabra root extract to 4.5 and then continues to adjust the pH value to 2.5. The remaining steps are consistent with Preparation Example 1.

[0072] Example 1

[0073] This embodiment provides a composition, the formula of which is: 5 parts of the Glycyrrhiza glabra root extract prepared in Preparation Example 1, 2 parts of acetyl glucosamine, 3 parts of panthenol and 1 part of resveratrol.

[0074] Example 2

[0075] This embodiment provides a composition, the formula of which is: 3 parts of the root extract of Glycyrrhiza glabra prepared in Preparation Example 2, 3 parts of acetyl glucosamine, 2 parts of panthenol and 0.5 parts of resveratrol.

[0076] Example 3

[0077] This embodiment provides a composition, the formula of which is: 8 parts of the extract of Glycyrrhiza glabra root prepared in Preparation Example 3, 1 part of acetyl glucosamine, 4 parts of panthenol and 2 parts of resveratrol.

[0078] Example 4

[0079] This embodiment provides a composition, which differs from Example 1 only in that the composition does not include resveratrol, that is, the formula is: 5 parts of the Glycyrrhiza glabra root extract prepared in Preparation Example 1, 2 parts of acetyl glucosamine and 3 parts of panthenol.

[0080] Example 5

[0081] This embodiment provides a composition, which differs from Example 1 only in that the composition does not include panthenol, that is, the formula is: 5 parts of the Glycyrrhiza glabra root extract prepared in Preparation Example 1, 2 parts of acetyl glucosamine and 1 part of resveratrol.

[0082] Example 6

[0083] This embodiment provides a composition, which differs from Example 1 only in that the composition does not include acetyl glucosamine, that is, the formula is: 5 parts of the Glycyrrhiza glabra root extract prepared in Preparation Example 1, 3 parts of panthenol and 1 part of resveratrol.

[0084] Comparative Example 1

[0085] This comparative example provides a composition, which differs from Example 1 only in that the composition does not include the Glycyrrhiza glabra root extract, that is, the formula is: 2 parts of acetyl glucosamine, 3 parts of panthenol and 1 part of resveratrol.

[0086] Application Example 1

[0087] This application example provides an essence water, which includes, by mass percentage, 3% of the root extract of Glycyrrhiza glabra prepared in Preparation Example 1, 5% of polydimethylsiloxane, 3% of squalane, 2% of isohexadecane, 3% of glycerin, 5% of butylene glycol, 3% of hyaluronic acid, 0.3% of carbomer, and the balance is water.

[0088] Application Examples 2-5

[0089] Application Examples 2-5 each provide an essence water, which differs from Application Example 1 only in that the glycyrrhiza glabra root extract prepared in Preparation Example 1 is replaced by the glycyrrhiza glabra root extract prepared in Preparation Examples 2-5 in equal mass, and the rest of the formula remains consistent with Application Example 1.

[0090] Application Examples 6-11

[0091] Application Examples 6-11 each provide an essence water, which differs from Application Example 1 only in that the Glycyrrhiza glabra root extract prepared in Preparation Example 1 is replaced with the compositions of Examples 1-6 in equal mass, and the rest of the formula remains consistent with Application Example 1.

[0092] Comparative Application Examples 1-3

[0093] Comparative Application Examples 1-3 each provide an essence water, which differs from Application Example 1 only in that the glycyrrhiza glabra root extract prepared in Preparation Example 1 is replaced by the glycyrrhiza glabra root extract prepared in Comparative Preparation Examples 1-3 in equal mass, and the rest of the formula remains consistent with Application Example 1.

[0094] Comparative Application Example 4

[0095] This comparative application example provides an essence water, which differs from Application Example 1 only in that the mass of the Glycyrrhiza glabra root extract prepared in Preparation Example 1 is replaced by the composition of Comparative Example 1, and the rest of the formula remains consistent with Application Example 1.

[0096] Test Example 1

[0097] This test example evaluates the brightening and whitening effects of the essence water in each application example:

[0098] (1) Test subjects: 150 healthy women (aged 30-45 years old), divided into 15 groups, each with 10 people; each group used the products of Application Examples 1-11 and Comparative Application Examples 1-4.

[0099] (2) Test environment: temperature 20-22°C, humidity 40-60%.

[0100] (3) Testing instrument: Delfin SkinColorCatch (Finland).

[0101] (4) Test method:

[0102] After cleansing and toning your face in the morning and evening, apply an appropriate amount of the essence water from the application example and the comparative application example to your face, evenly apply, and massage until absorbed. Continue using for 28 days. Test your skin's L* value (brightness) and ITA° value (color individual type angle, representing overall chromaticity changes; a larger ITA° value indicates a lighter skin tone) before and after use. Results are expressed as the rate of change in L* and ITA° values ​​before and after use.

[0103] Change rate of L* value = (L* value after 28 days of use - L* value before use) / L* value before use × 100%;

[0104] ITA° value change rate = (ITA° value after 28 days of use - ITA° value before use) / ITA° value before use × 100%.

[0105] The test results of the change rate of L* value and ITA° value are shown in Table 2.

[0106] Table 2

[0107]

[0108]

[0109] From the comparison of the data of Application Example 1 and Application Examples 4-5, it can be seen that the present invention uses choline chloride as a hydrogen bond acceptor and glycerol as a hydrogen bond donor to form a deep eutectic solvent, which can better achieve the extraction effect of the whitening functional components in the root of Glycyrrhiza glabra.

[0110] From the comparison of the data of Application Example 1 and Comparative Application Example 1, it can be seen that compared with pure water extraction, hot extraction of Glycyrrhiza glabra root using a solution containing a low eutectic solvent can better increase the content of active ingredients; from the comparison of the data of Application Example 1 and Comparative Application Examples 2-3, it can be seen that first adjusting the pH value of the mixed solution to 8-10 to obtain a crude extract of Glycyrrhiza glabra root, and then adjusting the pH value to 3-4, and performing a secondary hot extraction, can better increase the content of active ingredients, and the prepared Glycyrrhiza glabra root extract has better whitening and skin lightening effects.

[0111] From the data comparison of Application Example 6 with Application Examples 9-11 and Comparative Application Example 4, it can be seen that the Glycyrrhiza glabra root extract prepared by the present invention has a significant synergistic effect with acetyl glucosamine, panthenol and resveratrol in lightening spots and brightening skin, and can more effectively reduce local pigmentation and lighten skin spots.

[0112] Test Example 2

[0113] This test example evaluates the anti-inflammatory effect of the prepared Glycyrrhiza glabra root extract and composition:

[0114] (1) Test samples: the root extracts of Glycyrrhiza glabra prepared in Preparation Examples 1-5 and Comparative Preparation Examples 1-3, and the compositions of Examples 1-6 and Comparative Example 1.

[0115] (2) Test method:

[0116] IL1-α and IL-6 are cellular inflammatory factors. ELISA was used to detect the inflammatory factors to evaluate the anti-inflammatory effect of the test samples.

[0117] (2.1) Experimental groups and treatment methods:

[0118] Blank group: HaCat cells without any treatment were cultured in 12-well plates;

[0119] Model group: HaCat cells were stimulated with 1 μg / ml LPS and cultured in 12-well plates for 24 h;

[0120] Sample group: After stimulating HaCat cells with 1 μg / ml LPS for 24 h, 100 μl of the above test sample at 1% concentration was added to a 12-well plate;

[0121] Each experimental group was cultured in a 37°C, 5% CO2 constant temperature incubator for 72 hours, with three replicates per experimental group. The cell culture supernatant was collected and the expression levels of IL1-α and IL-6 were detected using ELISA kits, and the results were averaged.

[0122] Taking the content detected in the blank group as the reference amount (ie 100%), the relative content results of IL1-α and IL-6 in each experimental group are shown in Table 3.

[0123] Table 3

[0124]

[0125]

[0126] From the comparison of the data of Preparation Example 1 and Preparation Examples 4-5, it can be seen that the present invention uses choline chloride as a hydrogen bond acceptor and glycerol as a hydrogen bond donor to form a deep eutectic solvent, which can better achieve the extraction effect of the effective ingredient with anti-inflammatory effect in the root of Glycyrrhiza glabra.

[0127] From the comparison of the data of Preparation Example 1 and Comparative Preparation Example 1, it can be seen that compared with pure water extraction, hot extraction of Glycyrrhiza glabra root using a solution containing a low eutectic solvent can better increase the content of the active ingredient; from the comparison of the data of Preparation Example 1 and Comparative Preparation Examples 2-3, it can be seen that first adjusting the pH value of the mixed solution to 8-10 to obtain a crude extract of Glycyrrhiza glabra root, and then adjusting the pH value to 3-4, and performing a secondary hot extraction, can better increase the content of the active ingredient, and the prepared Glycyrrhiza glabra root extract has better soothing and anti-inflammatory effects.

[0128] From the comparison of the data of Example 1 with Examples 4-5 and Comparative Example 1, it can be seen that the Glycyrrhiza glabra root extract prepared by the present invention has a significant synergistic effect with acetyl glucosamine, panthenol and resveratrol in anti-inflammatory aspects, can significantly reduce the expression level of inflammatory factors, and has better soothing and anti-inflammatory effects.

[0129] The applicant declares that the present invention is illustrated by the above-described embodiments, but the present invention is not limited to the above-described embodiments. This does not mean that the present invention must rely on the above-described embodiments in order to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements for raw materials in the present invention, additions of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0130] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0131] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for preparing a Glycyrrhiza glabra root extract having anti-inflammatory and skin-lightening effects, characterized in that: The preparation method comprises the following steps: (1) mixing the powder of Glycyrrhiza glabra root with a solution containing a deep eutectic solvent, adjusting the pH value of the mixture to 8-10, performing hot extraction, filtering, and taking the filtrate to obtain a crude extract of Glycyrrhiza glabra root; (2) The pH value of the crude extract of Glycyrrhiza glabra root was adjusted to 3-4, and a secondary hot extraction was performed to obtain the Glycyrrhiza glabra root extract.

2. The preparation method according to claim 1, characterized in that The raw materials for preparing the solution containing the deep eutectic solvent in step (1) include the deep eutectic solvent and water; Preferably, the deep eutectic solvent comprises choline chloride and glycerol; Preferably, the molar ratio of choline chloride to glycerol is 1:(2-7); Preferably, the mass ratio of the deep eutectic solvent to water is (1-5):1; Preferably, the solid-liquid ratio of the Glycyrrhiza glabra root powder to the solution containing a deep eutectic solvent in step (1) is 1:(15-20) g / ml.

3. The preparation method according to claim 2, characterized in that The solution containing the deep eutectic solvent is prepared by a preparation method comprising the following steps: Choline chloride and glycerol are mixed, reacted at 70-90° C. for 1-5 hours, cooled to 20-40° C., and then mixed with water to obtain a solution containing a deep eutectic solvent.

4. The preparation method according to any one of claims 1 to 3, characterized in that The temperature of the hot extraction in step (1) is 55-65° C., and the time of the hot extraction is 5-8 hours.

5. The preparation method according to any one of claims 1 to 4, characterized in that The temperature of the secondary hot extraction in step (2) is 70-80° C., and the time of the secondary hot extraction is 0.5-2 h.

6. The preparation method according to any one of claims 1 to 5, characterized in that After the secondary hot extraction in step (2), the method further comprises filtering the product of the secondary hot extraction and collecting the filtrate; Preferably, the filtering method includes microfiltration; Preferably, the filter membrane of the microfiltration is 0.01-0.03 μm; Preferably, the filtrate is concentrated after the filtration.

7. A Glycyrrhiza glabra root extract having anti-inflammatory and skin-lightening effects obtained by the method for preparing the Glycyrrhiza glabra root extract according to any one of claims 1 to 6.

8. A composition with anti-inflammatory and skin-lightening effects, characterized in that: The composition comprises the Glycyrrhiza glabra root extract according to claim 7, acetyl glucosamine, panthenol and resveratrol.

9. The composition according to claim 8, characterized in that The mass ratio of the Glycyrrhiza glabra root extract, acetyl glucosamine, panthenol and resveratrol is (3-8):(1-3):(2-4):(0.5-2).

10. Use of the Glycyrrhiza glabra root extract according to claim 7 or the composition with anti-inflammatory and skin-lightening effects according to claim 8 or 9 in cosmetics.

Images