Lycium barbarum extract based on eutectic solvent as well as preparation method and application of Lycium barbarum extract

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a fructus lycii extract based on a deep-eutectic solvent and a preparation method and application thereof.The preparation method comprises the following steps that a hydrogen bond donor and a hydrogen bond acceptor are mixed according to the molar ratio of (1-3): 1, and the deep-eutectic solvent is obtained; the method comprises the following steps: adding a deep eutectic solvent into Chinese wolfberry according to a solid-liquid ratio of 1g: 10mL-30mL, extracting at 40-80 DEG C for 30-120 minutes, and separating supernate, namely the Chinese wolfberry extract. The lycium barbarum extract prepared by the invention can remarkably inhibit the release of tryptase in a zebra fish allergy model induced by Compound 48 / 80, and the relieving effect is superior to that of a positive control drug sodium cromoglycate, so that the lycium barbarum extract has an excellent anti-allergy effect; meanwhile, the extract can remarkably promote repair of a zebra fish tail fin injury model and effectively promote tissue regeneration, and it is proved that the extract has the repair effect.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a wolfberry extract based on a eutectic solvent, its preparation method, and its application. Background Technology

[0002] Goji berries are a traditional Chinese medicine and health food, rich in active ingredients such as goji polysaccharides, carotenoids, and flavonoids, which are known to have antioxidant and immunomodulatory effects. In recent years, their application in cosmetics to achieve skincare benefits has become a research hotspot.

[0003] Currently, goji berry extracts are mostly extracted using traditional solvents such as water and ethanol. These methods have several drawbacks: water extraction is relatively efficient for polysaccharides, but has a low extraction rate for fat-soluble components such as flavonoids; while ethanol extraction is better for fat-soluble components, it uses toxic solvents such as methanol, posing a risk of residue, and the high-temperature extraction process can easily destroy heat-sensitive active ingredients. These factors result in goji berry extracts obtained using existing technologies not exhibiting sufficiently prominent and stable anti-allergic and repairing skincare effects. Meanwhile, cosmetic raw materials used for anti-allergy and repair purposes, such as dipotassium glycyrrhizate and bisabolol, while effective, are either expensive or have limited availability. Therefore, developing a highly efficient and specific extraction method for enriching anti-allergic and repairing active ingredients in goji berries has significant application value. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preparing wolfberry extract based on a eutectic solvent. This method achieves efficient and green extraction of anti-allergic and repairing active ingredients from wolfberry by optimizing the combination of hydrogen bond donors and hydrogen bond acceptors and the extraction process.

[0005] It solves the problems existing in the current technology.

[0006] The technical solution adopted in this invention is: This invention provides a method for preparing wolfberry extract based on a eutectic solvent, the steps of which are as follows: The eutectic solvent is obtained by mixing hydrogen bond donors and hydrogen bond acceptors in a molar ratio of 1 to 3:1. Add a eutectic solvent to wolfberries at a material-to-liquid ratio of 1g:10mL~30mL, extract at 40℃~80℃ for 30min~120min, separate the supernatant, and the obtained supernatant is wolfberry extract.

[0007] Preferably, the hydrogen bond donor is any one of lactic acid, gluconic acid, citric acid, urea, and glycerol; The hydrogen bond acceptor is either choline chloride or betaine.

[0008] Preferably, the molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 2:1.

[0009] Preferably, the material-to-liquid ratio is 1g:20mL.

[0010] Preferably, the stirring extraction is performed at 70°C for 1 hour.

[0011] Preferably, the supernatant is further freeze-dried after being obtained.

[0012] A second aspect of the present invention provides a wolfberry extract prepared using the aforementioned preparation method.

[0013] A third aspect of the present invention provides an application of the wolfberry extract, wherein the wolfberry extract is used to prepare cosmetics with anti-allergic and / or repairing effects.

[0014] A fourth aspect of the present invention provides a soothing and repairing essence, wherein the soothing and repairing essence comprises the aforementioned wolfberry extract; The soothing and repairing serum is formulated according to the following weight percentages: Glycerin 4%~6%, Sodium hyaluronate 0.08%~0.1%, Goji berry extract 0.8%~1%, 1,3-butanediol 3%~4%, Carbomer 0.15%~0.2%, Preservative 0.5%, the remainder is water to make up to 100%.

[0015] The preparation method of the soothing and repairing essence is as follows: Carbomer was dispersed in water and swollen. Glycerin and sodium hyaluronate were added, and the mixture was stirred until homogeneous. The mixture was then cooled to 40°C. Goji berry extract and butylene glycol were added. Finally, the pH was adjusted to 5.5-6.0, and preservatives were added to obtain the soothing and repairing essence.

[0016] Compared with the prior art, the beneficial effects of the present invention are: This invention provides a method for preparing wolfberry extract based on a eutectic solvent, the steps of which are as follows: A eutectic solvent is obtained by mixing hydrogen bond donors and hydrogen bond acceptors at a molar ratio of 1-3:1. The eutectic solvent is then added to wolfberries at a material-to-liquid ratio of 1g:10mL-30mL, and extraction is performed at 40℃-80℃ for 30-120 minutes. The supernatant is then separated, and the resulting supernatant is the wolfberry extract. The wolfberry extract described in this invention is extracted using a eutectic solvent composed of hydrogen bond donors and hydrogen bond acceptors. Experimental results show that the wolfberry extract prepared in this invention can significantly inhibit the release of trypsin-like proteins in a zebrafish allergy model induced by Compound48 / 80, exhibiting a better soothing effect than the positive control sodium cromoglycate, demonstrating its excellent anti-allergic efficacy. Simultaneously, the extract can significantly promote the repair of zebrafish tail fin damage, effectively promoting tissue regeneration, demonstrating its repair efficacy. The wolfberry extract provided by this invention is green, safe, and highly active, and can be widely used in cosmetics such as serums, lotions, and creams to soothe sensitive skin and repair the skin barrier. Attached Figure Description

[0017] Figure 1 The figure shows the effect of wolfberry extract on the relative expression level of trypsin in zebrafish.

[0018] Figure 2 To evaluate the anti-allergic efficacy of the wolfberry extracts prepared in Example 1 and Comparative Example 1 based on a zebrafish model.

[0019] Figure 3 The results show the effect of wolfberry extract on the growth rate of zebrafish tail fin repair. A is a representative image; B is the statistical result of image A.

[0020] Figure 4 To evaluate the repair efficacy of the wolfberry extracts prepared in Example 1 and Comparative Example 1 based on a zebrafish tail fin injury model. Detailed Implementation

[0021] The present invention will be further illustrated below with specific embodiments, but these embodiments do not limit the scope of the invention. Modifications or substitutions to the details and form of the technical solutions of the present invention may be made without departing from the spirit and scope of the invention, but all such modifications or substitutions fall within the protection scope of the present invention.

[0022] The inventive concept of this invention is as follows: Eutectic solvents (DES) are eutectic mixtures formed by hydrogen bond donors and acceptors through hydrogen bond complexation. They offer advantages such as simple preparation, low cost, high designability, and environmental friendliness. In recent years, DES has shown great potential in the field of natural product extraction, with its unique physicochemical properties potentially leading to higher selectivity and extraction efficiency for specific types of active ingredients. However, its application in wolfberry extraction, significantly enhancing its anti-allergic and restorative effects, has not yet been reported.

[0023] To enable those skilled in the art to better understand and implement the technical solutions of this invention, the invention will be further described below with reference to specific embodiments. Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0024] Example 1 A method for preparing wolfberry extract based on a eutectic solvent is as follows: S1. Preparation of eutectic solvent: The eutectic solvent is obtained by mixing hydrogen bond donors and hydrogen bond acceptors.

[0025] Weigh choline chloride and lactic acid in a molar ratio of 1:2, place them in a round-bottom flask, and stir at 60°C until a colorless and transparent liquid is formed. This yields the eutectic solvent, DES.

[0026] S2. Preparation of wolfberry extract.

[0027] Ningxia wolfberries were dried, pulverized, and passed through a 60-mesh sieve. 10g of wolfberry powder was weighed and added to 200mL of the prepared DES at a material-to-liquid ratio of 1g:20mL. The mixture was stirred and extracted in a 70℃ water bath for 1 hour. After extraction, the mixture was centrifuged at 8000r / min for 10 minutes, and the supernatant was collected. The supernatant was filtered through a 0.22μm microporous membrane to obtain the wolfberry DES extract, i.e., the wolfberry extract, which was named NXU-C. NXU-C was freeze-dried to obtain the wolfberry extract powder for easy storage and weighing.

[0028] Example 2 A method for preparing wolfberry extract based on a eutectic solvent is as follows: S1. Preparation of eutectic solvent: The eutectic solvent is obtained by mixing hydrogen bond donors and hydrogen bond acceptors.

[0029] Weigh choline chloride and lactic acid in a molar ratio of 1:1, place them in a round-bottom flask, and stir at 60°C until a colorless and transparent liquid is formed. This yields the eutectic solvent, DES.

[0030] S2. Preparation of wolfberry extract.

[0031] Ningxia wolfberries were dried, pulverized, and passed through a 60-mesh sieve. 10g of wolfberry powder was weighed and added to 100mL of the prepared DES at a material-to-liquid ratio of 1g:10mL. The mixture was stirred and extracted in a 40℃ water bath for 30min. After extraction, the mixture was centrifuged at 8000r / min for 10min, and the supernatant was collected. The supernatant was filtered through a 0.22μm microporous membrane to obtain the wolfberry DES extract, i.e., the wolfberry extract, which was named NXU-C. NXU-C was freeze-dried to obtain the wolfberry extract powder for easy storage and weighing.

[0032] Example 3 A method for preparing wolfberry extract based on a eutectic solvent is as follows: S1. Preparation of eutectic solvent: The eutectic solvent is obtained by mixing hydrogen bond donors and hydrogen bond acceptors.

[0033] Weigh choline chloride and lactic acid in a molar ratio of 1:3, place them in a round-bottom flask, and stir at 60°C until a colorless and transparent liquid is formed. This yields the eutectic solvent, DES.

[0034] S2. Preparation of wolfberry extract.

[0035] Ningxia wolfberries were dried, pulverized, and passed through a 60-mesh sieve. 10g of wolfberry powder was weighed and added to 300mL of the prepared DES at a material-to-liquid ratio of 1g:30mL. The mixture was then stirred and extracted in an 80℃ water bath for 2 hours. After extraction, the mixture was centrifuged at 8000r / min for 10min, and the supernatant was collected. The supernatant was filtered through a 0.22μm microporous membrane to obtain the wolfberry DES extract, i.e., the wolfberry extract, which was named NXU-C. NXU-C was freeze-dried to obtain the wolfberry extract powder for easy storage and weighing.

[0036] Comparative Example 1.

[0037] A method for preparing wolfberry extract is as follows: Following the method of Example 1, pure water was used instead of DES for extraction, and the rest of the process was exactly the same as in Example 1.

[0038] Example 4 An application of wolfberry extract based on eutectic solvent In this invention, the wolfberry extracts prepared in Examples 1 to 3 showed no significant differences in various properties; therefore, only the results of Example 1 and Comparative Example 1 are presented here. The following experiments were conducted to verify the wolfberry extracts prepared in Examples 1 and Comparative Example 1:

[0039] 1. Evaluation of anti-allergy efficacy based on zebrafish model.

[0040] 1.1 Experimental Objective: To investigate the soothing effects of wolfberry extracts by detecting their ability to inhibit trypsin expression in zebrafish.

[0041] 1.2. Test methods.

[0042] 1.2.1 Grouping.

[0043] A blank control group, a model control group (Compound48 / 80), a positive control group (sodium cromoglycate), and a test sample group (NXU-C) were set up. The concentration of NXU-C was 0.125%, as detailed in Table 1.

[0044] Table 1 Grouping In Table 1, "-" indicates that this item is not present.

[0045] The preparation method of 0.125% NXU-C is as follows: Weigh the lyophilized NXU-C and dilute it with ultrapure water to 0.125% (wt).

[0046] 1.2.2, Steps.

[0047] First, an allergy model was constructed: wild-type AB strain zebrafish juveniles developed to 96 hours old were exposed to 10 µg / mL Compound 48 / 80 solution and incubated for 30 min to establish the allergy model. Compound 48 / 80 is a known mast cell activator that induces mast cell degranulation, mimicking an allergic response in organisms. The success of the model was indicated by a significantly increased relative expression level of trypsin in the zebrafish juveniles compared to the blank control group, confirming the successful induction of the allergic response.

[0048] Grouping: Place 96hpf zebrafish juveniles in a 96-well plate and pat dry the culture water in the 96-well plate.

[0049] Blank control group: 100 μL of standard dilution water was added to each well of zebrafish that had not undergone Compound 48 / 80 treatment; Model control group: 100 μL of Compound 48 / 80 solution was added to each well to construct an allergy model; Positive control group: 100 μL of Compound 48 / 80 solution was added to each well. After the allergy model was successfully constructed, 100 μL of sodium cromoglycate solution was added to each well. In the sample group: 100 μL of Compound 48 / 80 solution was added to each well. After the allergy model was successfully constructed, NXU-C was added to each well.

[0050] Meanwhile, corresponding solutions were set up as zero-adjustment control groups, and each group was incubated in a 28℃ constant temperature incubator for 30 min. After 30 min of incubation, 50 μL of supernatant was taken into a 96-well plate, and 50 μL of 1 mg / mL Na-benzoyl-DL-arginine-p-nitrobenzamide hydrochloride (BAPNA) was added to each well. After incubation at 37℃ for 1 hour, the absorbance was measured at a wavelength of 405 nm, and the relative expression level of trypsin was calculated.

[0051] Statistical analysis: Data are expressed as mean ± SD, and t-tests were used for inter-group comparisons. p A value <0.05 is considered statistically significant.

[0052] 1.3. Test Results.

[0053] The results are as follows Figure 1 As shown in Table 2, the expression level of trypsin-like enzymes in the model control group was significantly higher than that in the blank control group. p A value <0.001 indicates that the allergy model was successfully established. Sodium cromoglycate in the positive control group significantly inhibited trypsin expression, with an inhibition rate of 89.15%. p <0.001. The NXU-C of this invention, at a concentration of 0.125%, exhibits an inhibition rate of up to 99.22% against trypsin expression, demonstrating extremely significant efficacy. p The concentration of the extract was less than 0.001, and its soothing effect was even better than that of the positive control drug. This indicates that the extract of the present invention has excellent anti-allergic potential.

[0054] Table 2. Effects of NXU-C on relative expression levels of trypsin in zebrafish, x±s, n=5 Note: ### indicates p < 0.001 compared to the model control group. "-" indicates this option is not available.

[0055] 2. The anti-allergic efficacy of the wolfberry extracts prepared in Example 1 and Comparative Example 1 was evaluated based on the zebrafish model.

[0056] The same method was used to construct the zebrafish allergy model as described above. The allergic zebrafish were divided into three groups:

[0057] One group served as the model control group: allergic zebrafish were cultured normally; another group was given NXU-C prepared in Example 1 at a concentration of 0.125% wt; and the third group was given Lycium barbarum extract prepared in Comparative Example 1 at a concentration of 0.125% wt. The preparation methods were the same as above.

[0058] The results are as follows Figure 2 As shown, compared with the wolfberry extract prepared in Comparative Example 1, the expression level of trypsin in zebrafish was significantly reduced after treatment with the same concentration of NXU-C prepared in Example 1, indicating that NXU-C has a stronger soothing effect.

[0059] 3. Evaluation of repair efficacy based on zebrafish tail fin injury model.

[0060] 3.1. Purpose of the experiment.

[0061] The effects of different wolfberry extracts on the growth of damaged tail fins in juvenile zebrafish were investigated to explore the restorative effects of wolfberry extracts.

[0062] 3.2. Test methods.

[0063] 3.2.1 Grouping.

[0064] A blank control group was set up consisting of standard dilution water and the test sample group containing 0.08% NXU-C.

[0065] The preparation method of 0.08% NXU-C is as follows: Weigh the lyophilized NXU-C and dilute it with ultrapure water to 0.08% wt.

[0066] Procedure: 72 hpf zebrafish juveniles were taken, anesthetized, and part of their caudal fin was removed. After a 2-hour recovery period, they were treated with standard diluted water and 0.08% NXU-C for 72 hours respectively. Microscopic images were taken, and the caudal fin growth rate was measured using Image-J software.

[0067] Statistical analysis: Data are expressed as mean ± SD, and t-tests were used for inter-group comparisons. p A value <0.05 is considered statistically significant.

[0068] 3.3. Test Results.

[0069] The results are as follows Figure 3 As shown in Table 3, the natural growth rate of the caudal fin in the blank control group was 100%. After treatment with 0.08% NXU-C for 72 hours, the caudal fin growth rate of zebrafish reached 135.96%, which was a highly significant increase of 35.96% compared with the blank control group. p <0.001. This fully demonstrates that the extract of the present invention has a significant ability to promote tissue repair and regeneration.

[0070] Table 3. Effects of NXU-C on zebrafish tail fin repair, x±s, n=20 Note: "-" in Table 3 indicates that this item is not present.

[0071] 4. The repair efficacy of the wolfberry extract prepared in Example 1 and Comparative Example 1 was evaluated based on the zebrafish tail fin injury model.

[0072] Procedure: 72 hpf zebrafish juveniles were anesthetized, and part of their caudal fin was removed. After a 2-hour recovery period, they were treated for 72 hours with 0.08% NXU-C prepared in Example 1 and 0.08% Lycium barbarum extract prepared in Comparative Example 1, respectively. Microscopic images were taken, and the caudal fin growth rate was measured using Image-J software. The preparation methods for 0.08% NXU-C and 0.08% Lycium barbarum extract prepared in Comparative Example 1 were the same as above.

[0073] The results are as follows Figure 4 As shown, in the repair test, compared with the wolfberry extract prepared in Comparative Example 1, the DES extract at the same concentration had a significantly enhanced repair ability.

[0074] Example 5 A soothing and repairing serum containing goji berry extract, as detailed below: Based on the wolfberry extract prepared in Example 1, this example provides a soothing and repairing serum, formulated according to the following weight percentages: Phase A: 5% glycerol, 0.1% sodium hyaluronate.

[0075] Phase B: 1% of the wolfberry extract prepared in Example 1, on a solid basis; 4% of 1,3-butanediol; and 89.2% of water.

[0076] C phase: Carbomer 0.2%.

[0077] Phase D: Adjust the pH to 5.5-6.0 using triethanolamine and add 0.5% benzyl alcohol as a preservative.

[0078] Preparation process: Disperse phase C in water (phase B), allow it to swell, then add phase A and heat to 75°C while stirring until homogenized. After homogenization, cool to 40°C, add the remaining components from phase B, and finally adjust the pH with phase D and add preservatives. Stir until well combined. This serum has excellent soothing and repairing effects and is suitable for sensitive skin.

[0079] The wolfberry extract based on a eutectic solvent provided by this invention, through a unique extraction process, exhibits unexpectedly significant enhancements in its anti-allergic and repairing effects. In vivo experimental data from zebrafish fully demonstrate its effectiveness, providing a high-quality raw material solution for the development of highly effective and safe soothing and repairing cosmetics.

[0080] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0081] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A method for preparing a low eutectic solvent-based wolfberry extract, characterized by, The steps are as follows: The hydrogen bond donor and the hydrogen bond acceptor are mixed at a molar ratio of 1-3:1 to obtain the eutectic solvent; According to the solid-liquid ratio of 1g:10mL-30mL, the eutectic solvent is added to the wolfberry, and the supernatant is separated after extraction at 40℃-80℃ for 30min-120min. The obtained supernatant is the wolfberry extract.

2. The production method according to claim 1, wherein The hydrogen bond donor is any one of lactic acid, gluconic acid, citric acid, urea and glycerol; The hydrogen bond acceptor is any one of choline chloride and betaine.

3. The production method according to claim 1, wherein The molar ratio of the hydrogen bond donor and the hydrogen bond acceptor is 2:

1.

4. The production method according to claim 1, wherein The solid-liquid ratio is 1g:20mL.

5. The production method according to claim 1, wherein The stirring extraction conditions are extraction at 70℃ for 1h.

6. The production method according to claim 1, wherein After obtaining the supernatant, it needs to be freeze-dried.

7. A wolfberry extract prepared by the preparation method of claim 1.

8. The wolfberry extract for use according to claim 7, wherein the wolfberry extract is administered in an amount of 0.1 to 1.5 g per day. The wolfberry extract is used for preparing a cosmetic with anti-allergic and / or repairing effect.

9. A soothing repair serum, characterized in that, The soothing repair serum contains the wolfberry extract of claim 7; The soothing repair serum is prepared according to the following weight percentage: Glycerol 4%-6%, sodium hyaluronate 0.08%-0.1%, wolfberry extract 0.8-1%, 1,3 butanediol 3%-4%, carbomer 0.15%-0.2%, preservative 0.5%, and the rest is water to 100%.

10. The method for preparing the soothing and repairing essence as described in claim 9, characterized in that, The steps are as follows: Carbomer is dispersed in water, and after swelling, glycerol and sodium hyaluronate are added. After stirring uniformly, the temperature is lowered to 40℃, wolfberry extract and butanediol are added, and finally the pH value is adjusted to 5.5-6.0, and preservative is added to obtain the soothing repair serum.

Images