Preparation method and application of high-water-solubility dihydroquercetin for cosmetics

Abstract

The invention discloses a preparation method and application of high-water-solubility dihydroquercetin for cosmetics, and belongs to the technical field of compound extraction.The preparation method comprises the steps that larch roots are dried and smashed, soaked in cold water and filtered, filter residues are extracted through plate-frame pressure filtration, an extracting solution is concentrated, alcohol precipitation is conducted, an alcohol solution is concentrated and refrigerated till crystals are separated out, and the high-water-solubility dihydroquercetin is obtained. Adding the crystal into an inclusion solution, filtering after reaction, adding a composite decolorizing agent into the filtrate, filtering again, adjusting the pH value to be neutral, diluting until the solid content of the filtrate is 3 + / -0.5%, passing through a column, concentrating, and spray-drying to obtain the high-water-solubility dihydroquercetin for cosmetics, which is in a powder state and light in color. The composition has a good anti-wrinkle effect, a tightening effect and a soothing effect, and can be used for preparing cosmetics.

Description

Technical Field

[0001] The invention belongs to the technical field of compound extraction, and in particular relates to a preparation method and application of highly water-soluble dihydroquercetin for cosmetics. Background Art

[0002] The roots of larch are rich in natural antioxidants (dihydroquercetin). The existing traditional extraction processes mainly include organic solvent extraction and boiling water decoction, etc.; the purification processes mainly include column chromatography and crystallization, etc., but few people pay attention to its water solubility, conductivity value, stability and product color in room temperature water, which greatly limits its application in cosmetics, biomedicine and other industries. As in patent application CN 103951645 A, larch extract is prepared by raw material selection, enzymatic treatment, extraction, concentration, hydrolysis, extraction, recrystallization, spray drying and other processes, and the prepared product is almost difficult to dissolve in room temperature water. Patent application CN 109305953 A mentions: Step 1: Put high-purity dihydroquercetin into boiling water to fully dissolve, then stir and mix with a certain proportion of saponin and arabinogalactan, add boiling water after mixing, stir, make saponin and arabinogalactan fully dissolved, mix the aqueous solution of dihydroquercetin with the aqueous solution of saponin and arabinogalactan, stir into a suspension, and set aside. Step 2: Pass the suspension in step 1 through an ultra-high pressure homogenizer, homogenize 3-6 times, and the homogenization pressure is 50-600MP to obtain a homogenous solution. Step 3: Spray dry the homogenized solution in step 2 to obtain highly water-soluble dihydroquercetin. According to the full text, it is understood that the technology only increases the room temperature water solubility of dihydroquercetin by 5-40 times; the highest solubility in the embodiment is only 15.23g / L, and it is also impossible to understand the stability of the product aqueous solution according to the full text, and whether long-term placement will cause effective components to precipitate. Patent application CN 109574976 A discloses a method for industrially extracting high-purity dihydroquercetin from larch roots. The method uses the abandoned larch roots from artificial forests selected or clear felled as raw materials, and obtains pure dihydroquercetin through twelve steps: raw material selection, raw material cleaning, washing, raw material treatment before extraction, extraction, concentration, sedimentation, secondary concentration, primary crystallization, primary centrifugal separation, recrystallization, secondary centrifugal separation, freeze drying, and crushing. The method utilizes the abundant resources of abandoned larch roots (main roots and lateral roots) after felling, and the dihydroquercetin content is relatively rich (1.5-1.9%). The patent uses n-hexane as the solvent, and only turpentine is produced during the extraction process, and the stability of the extract is not paid attention to. Summary of the invention

[0003] Based on the problems that dihydroquercetin extracted by traditional methods is poorly soluble in water, has a dark powder color, and a high conductivity value, the present invention proposes a preparation method and application of highly water-soluble dihydroquercetin for cosmetics.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] One of the technical solutions of the present invention:

[0006] A method for preparing highly water-soluble dihydroquercetin for cosmetics comprises the following steps:

[0007] The larch root is dried and crushed, soaked in cold water, filtered, the filter residue is extracted by plate and frame filter pressing, the extract is concentrated, alcohol precipitation is performed, the alcohol solution is concentrated and refrigerated until crystals are precipitated, the crystals are added to the inclusion solution, filtered after reaction, a composite decolorizing agent is added to the filtrate, filtered again, the pH is adjusted to neutral, diluted to a solid content of 3±0.5% in the filtrate, passed through a column, concentrated, and spray-dried to obtain the highly water-soluble dihydroquercetin for cosmetics.

[0008] Preferably, the larch roots are crushed and then passed through a 60-80 mesh sieve.

[0009] Preferably, the cold water soaking is soaking in water at 10-20° C. for 12-24 hours. Since dihydroquercetin is almost insoluble in cold water, cold water soaking can leach out water-soluble impurities in the larch roots.

[0010] Preferably, the method of extracting the filter residue by plate and frame filter pressing is: using the plate and frame filter pressing extraction method, using 40-80 times boiling water of the raw material (here the raw material refers to the powder obtained by drying and crushing the larch root) to extract the filter residue 2-3 times. Since dihydroquercetin can be dissolved in boiling water, using boiling water extraction can more effectively extract dihydroquercetin, and can further remove water-insoluble components.

[0011] Preferably, the alcohol used in the alcohol precipitation is 80% (mass percentage, the same below) ethanol.

[0012] Preferably, the inclusion solution is a mixed solution of cyclodextrin derivatives, alginate, sodium bisulfite, hydroxyethyl cellulose and water.

[0013] Preferably, the mass ratio of the cyclodextrin derivative, alginate, sodium bisulfite, hydroxyethyl cellulose and water is (40-50):(0.3-0.7):(0.5-1.5):(0.3-0.7):(160-200), more preferably 40:0.5:1.0:0.4:180.

[0014] Preferably, the cyclodextrin derivative is hydroxypropyl-β-cyclodextrin or hydroxypropyl-γ-cyclodextrin.

[0015] Preferably, the alginate is sodium alginate.

[0016] The invention adds sodium bisulfite into the inclusion solution, which can play a stabilizing and protective role, so that dihydroquercetin is not destroyed by high temperature. The cyclodextrin derivative has a hydrophilic exterior and a hydrophobic internal cavity, and can form an inclusion compound with dihydroquercetin to increase its solubility and stability, and can also cover up bad smells. Moreover, the cyclodextrin derivative can form a complex with alginate and hydroxyethyl cellulose to increase the inclusion amount and control the release rate of the inclusion compound. The invention uses the mixture, which not only has a better inclusion effect than using a single cyclodextrin, but also can greatly increase the product content and increase the water solubility of the product. In addition, the water solubility of dihydroquercetin can also be greatly increased.

[0017] Preferably, the composite decolorizing agent is a mixture of activated carbon, basic alumina and perlite.

[0018] Preferably, the mass ratio of the activated carbon, alkaline alumina and perlite is (2-4): (0.5-1): (1-3), more preferably 3: 0.8: 2. Activated carbon is a highly porous material with strong adsorption capacity, alkaline alumina has adsorption, decolorization and impurity removal effects, and perlite is a porous volcanic rock. The porous structure of perlite increases the surface area in contact with other components, which helps to improve the decolorization effect and efficiency of the composite decolorizer. The combination of these components can provide a more comprehensive decolorization effect, in which the activated carbon is mainly responsible for adsorption and decolorization, the alkaline alumina is responsible for adsorption, decolorization and pH adjustment, and the perlite acts as a filter medium and increases the contact area. This composite effect can more effectively remove impurities and colors in the larch extract and improve the purity of dihydroquercetin in the larch extract and the clarity of the aqueous solution.

[0019] Preferably, the column is firstly passed through a cationic resin, and then the column liquid is passed through an anionic resin, preferably firstly passed through a LSD001 macroporous strong acid cationic resin, and then the column liquid is passed through a LSD396 decolorizing anionic resin. The two-step column can significantly remove inorganic salt impurities in larch, reduce the conductivity of the product aqueous solution, and improve the stability of the product aqueous solution.

[0020] The second technical solution of the present invention:

[0021] The present invention also provides a highly water-soluble dihydroquercetin for cosmetics prepared according to the above preparation method. The highly water-soluble dihydroquercetin for cosmetics obtained by the present invention is actually a microencapsulated dihydroquercetin, which includes pure dihydroquercetin and external embedding capsule materials and other impurities. The highly water-soluble dihydroquercetin for cosmetics prepared by the present invention has a solubility in water of up to 200-300 g / L (25±2°C), and can increase the solubility of dihydroquercetin in room temperature water to 48-80 g / L (as shown in Example 1: the finished product (i.e., highly water-soluble dihydroquercetin for cosmetics) has a dihydroquercetin content of 32.69%; the maximum solubility in water is 244.7 g / L. Then relative to pure dihydroquercetin, its solubility in water is calculated as follows: 244.7×32.69%=79.99 g / L); the room temperature aqueous solution is placed for a long time without precipitation. The product has the advantages of high water solubility, light color of finished powder, high content of active ingredients, good stability of aqueous solution and low conductivity value, and can be used in a wide range of applications.

[0022] The highly water-soluble dihydroquercetin for cosmetics prepared by the present invention has good anti-wrinkle effect, firming effect and soothing effect, and can be used for preparing cosmetics.

[0023] The third technical solution of the present invention:

[0024] The present invention also provides application of the dihydroquercetin in preparing cosmetics.

[0025] Compared with the prior art, the present invention has the following advantages and technical effects:

[0026] The invention can effectively extract the effective ingredient dihydroquercetin from the crushed larch root raw material through cold water immersion for impurity removal and boiling water plate-frame circulation extraction; at the same time, alcohol precipitation and cold storage crystallization can be used to crudely crystallize the effective ingredient dihydroquercetin; and because of the defect of low water solubility of dihydroquercetin, cyclodextrin+sodium alginate+sodium bisulfite+hydroxyethyl cellulose+deionized water solution are used to carry out inclusion treatment on the crude crystals, and then a composite decolorizing agent is used for decolorization, and the mixture is passed through a column and spray-dried to obtain the highly water-soluble dihydroquercetin for cosmetics. During the water-soluble treatment, cyclodextrin, sodium alginate, sodium bisulfite and hydroxyethyl cellulose cooperate with each other to make it have a higher embedding rate and make the product have higher water solubility. Sodium alginate can effectively encapsulate cyclodextrin and dihydroquercetin to improve the embedding rate of cyclodextrin, and sodium bisulfite can effectively act as a stabilizer, mainly to prevent oxidation and discoloration of dihydroquercetin; hydroxyethyl cellulose acts as a suspending agent to promote the embedding of cyclodextrin to dihydroquercetin. Cyclodextrin, sodium alginate and hydroxyethyl cellulose can also play a synergistic role to form a composite system to further optimize the inclusion efficiency, solubility, stability and release characteristics of the product. The present invention also uses a composite decolorizing agent, in which activated carbon is mainly used as a decolorizing agent, and alkaline alumina can provide a weakly alkaline environment for the overall solution, which can appropriately increase the solubility of dihydroquercetin in water. At the same time, alkaline alumina can appropriately decolorize and remove impurities, and its combination with activated carbon can achieve a good decolorizing effect; perlite, as a traditional filter aid, can effectively filter out activated carbon and alkaline alumina. By comparison, perlite has almost no adsorption on dihydroquercetin and will not cause loss, while diatomaceous earth, which is also a filter aid, shows an adsorption effect on dihydroquercetin, resulting in the loss of effective ingredients; anionic and cationic resins The fat string is placed in the last step for treatment, which can effectively remove inorganic salt impurities in the product without causing loss of effective ingredients of the product (taking Example 1 as an example, after liquid phase detection, the content of highly water-soluble dihydroquercetin for cosmetics before passing through the resin is 32.74% (the highly water-soluble dihydroquercetin for cosmetics of the present invention contains other ingredients such as cyclodextrin, so this content refers to the content of dihydroquercetin therein), and after passing through the column, it is 32.69%, and almost no obvious loss is observed; and the conductivity value of 1wt% aqueous solution drops from 471.7μS / cm to 83.58μS / cm, proving that most of the free impurities such as inorganic salts can be removed by passing through the column). BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The drawings constituting a part of the present application are used to provide a further understanding of the present application. The illustrative embodiments and descriptions of the present application are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0028] Figure 1 This is the liquid phase spectrum of the highly water-soluble dihydroquercetin for cosmetics in Example 1;

[0029] Figure 2 The effect of highly water-soluble dihydroquercetin for cosmetics on the inhibition rate of AGEs in Example 1, wherein "**" indicates a significant difference compared with the negative control (p<0.01);

[0030] Figure 3 This is a physical picture of the highly water-soluble dihydroquercetin for cosmetics in Example 1;

[0031] Figure 4 The actual pictures are 1wt% (left) and 5wt% (right) aqueous solutions of highly water-soluble dihydroquercetin for cosmetics in Example 1;

[0032] Figure 5 This is a physical picture of a 1 wt % aqueous solution of the low-content extract in Example 1. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0038] Unless otherwise specified, the normal temperature in the present invention is 25±2°C.

[0039] All raw materials used in the examples of the present invention are commercially available.

[0040] In the embodiment of the present invention, the origin of larch roots is Russia.

[0041] In an embodiment of the present invention, the method for detecting the content of dihydroquercetin is: testing according to the method for determining the content of taxifolin under the item of Polygonum multiflorum in Part I of the Pharmacopoeia of the People's Republic of China (2020 edition).

[0042] The recovery rate of dihydroquercetin is calculated as follows: (content of dihydroquercetin in the obtained highly water-soluble dihydroquercetin for cosmetics × mass of the obtained highly water-soluble dihydroquercetin for cosmetics) / (content of dihydroquercetin in the larch root raw material × mass of the larch root raw material used) × 100%.

[0043] The embedding rate is calculated as follows: (content of dihydroquercetin in the obtained highly water-soluble dihydroquercetin for cosmetics × mass of the obtained highly water-soluble dihydroquercetin for cosmetics) / (content of dihydroquercetin in the obtained yellow crystals × mass of the obtained yellow crystals) × 100%.

[0044] The technical solution of the present invention is further illustrated by the following embodiments.

[0045] Example 1

[0046] Dry the larch root, crush it through a 60-mesh sieve, weigh 1 kg of the larch root powder, add it to 20 kg of 10°C cold water and soak it for 14 hours, filter it with a plate and frame filter, and recover the filtrate as a low-content extract (tests show that the total flavonoid content in the low-content extract can reach more than 40%, with good water solubility, and can be used in health food or other fields). Weigh 60 times the weight of the raw material in deionized water, boil the water in a reactor, connect the pipe to the plate and frame filter press equipment, circulate the extraction for 1 hour, and set the filtrate aside. Repeat the above operation twice, combine the filtrates and concentrate them under reduced pressure at 70°C to 3 times the weight of the raw material (i.e., larch root powder). Anhydrous ethanol was added to the concentrated solution to make its alcohol concentration reach 80%, and it was allowed to stand at room temperature for 12 hours. The supernatant was extracted and concentrated to a solid content of more than 70%, and refrigerated for 12 hours to precipitate yellow crystals; 45g of hydroxypropyl-β-cyclodextrin, 0.5g of sodium alginate, 1g of sodium bisulfite, 0.5g of hydroxyethyl cellulose and 180g of deionized water were weighed and mixed and dissolved completely, and the above yellow crystals were added to the above mixed solution and stirred at room temperature for 2 hours, and the mixture was filtered through medium-speed filter paper under reduced pressure. 20g of composite decolorizing agent (activated carbon: alkaline alumina: perlite = 3:0.8:2, mass ratio, the same below) was added to the filtrate, and the mixture was stirred and decolorized at 70°C for 4 hours; the decolorized solution was filtered through medium-speed filter paper, and the pH of the filtrate was adjusted to neutral using hydrochloric acid. , diluted to a solution with a solid content of 3% (i.e., 3g of solid in 100g of solution), first passed through LSD001 type macroporous strong acid cation resin, passed through the column liquid and then passed through LSD396 type decolorizing anion resin, the column liquid was concentrated, and spray-dried to obtain highly water-soluble dihydroquercetin for cosmetics. The finished product was tested to have a dihydroquercetin content of 32.69%, a dihydroquercetin recovery rate of 82.73%, and an embedding rate of 98.89%. The obtained product was a white powder, tested by a whiteness tester, with a whiteness of 45.6, a maximum solubility in water of 244.7g / L (25±2°C, the same below), and a 1% deionized water solution conductivity (25°C, the same below) of 83.58μS / cm.

[0047] The actual picture of 1 wt% aqueous solution of low-content extract in this example is shown in Figure 5 The physical picture of the highly water-soluble dihydroquercetin for cosmetics in this embodiment is shown in Figure 3 The actual pictures of the 1wt% (left) and 5wt% (right) aqueous solutions of highly water-soluble dihydroquercetin for cosmetics in this embodiment are shown in Figure 4 It can be seen that the finished powder of highly water-soluble dihydroquercetin for cosmetics obtained by the method of the present invention has the characteristics of light color, high water solubility and good stability of aqueous solution.

[0048] Example 2

[0049] The larch root was dried and crushed through an 80-mesh sieve. 1 kg of the larch root powder was weighed and added to 20 kg of 15°C cold water and soaked for 24 hours. The filtrate was recovered as a low-content extract. Deionized water 40 times the mass of the raw material was weighed and boiled in a reactor. The pipe was connected to the plate and frame filter press equipment for 1 hour of cyclic extraction. The filtrate was set aside. The above operation was repeated 3 times. The filtrates were combined and concentrated at 70°C to 3 times the weight of the raw material. Anhydrous ethanol was added to the concentrate to make its alcohol concentration reach 80%. The concentrate was allowed to stand at room temperature for 12 hours. The supernatant was extracted and concentrated to a solid content of more than 70%. It was refrigerated for 12 hours to precipitate yellow crystals. 40 g of hydroxypropyl-γ-cyclodextrin, 0.7 g of sodium alginate, 1.5 g of sodium bisulfite, 0.3 g of hydroxyethyl cellulose and 160 g of deionized water were weighed and mixed and dissolved completely. The yellow crystals were added to the mixed solution and stirred at room temperature for 2 hours. , pass through medium-speed filter paper under reduced pressure, add 20g of composite decolorizing agent (activated carbon: alkaline alumina: perlite = 2: 0.5: 3) to the filtrate, and stir at 70°C for 4h for decolorization; pass the decolorized solution through medium-speed filter paper, adjust the pH of the filtrate to neutral with hydrochloric acid, dilute it into a solution with a solid content of 3.5%, first pass through LSD001 type macroporous strong acid cation resin, pass through the column liquid and then pass through LSD396 type decolorizing anion resin, concentrate the column liquid, and spray dry to obtain highly water-soluble dihydroquercetin for cosmetics, with a content of 30.57%, a recovery rate of dihydroquercetin of 80.34%, and an embedding rate of 98.79%. The obtained product is a white powder, tested by a whiteness tester, with a whiteness of 42.7, a solubility of 241.6g / L, and a conductivity of 103.49μS / cm.

[0050] Example 3

[0051] The larch root was dried and crushed through an 80-mesh sieve. 1 kg of the larch root powder was weighed and added to 20 kg of 20°C cold water and soaked for 12 hours. The filtrate was recovered as a low-content extract. Deionized water 80 times the mass of the raw material was weighed and boiled in a reactor. The pipe was connected to the plate and frame filter press equipment for cyclic extraction for 1 hour. The filtrate was set aside. The above operation was repeated 3 times. The filtrates were combined and concentrated at 70°C to 3 times the weight of the raw material. Anhydrous ethanol was added to the concentrated solution to make its alcohol concentration reach 80%. The concentrated solution was allowed to stand at room temperature for 12 hours. The supernatant was extracted and concentrated to a solid content of more than 70%. The solution was refrigerated for 12 hours to precipitate yellow crystals. 50 g of hydroxypropyl-γ-cyclodextrin, 0.3 g of sodium alginate, 0.5 g of sodium bisulfite, 0.7 g of hydroxyethyl cellulose and 200 g of deionized water were weighed and mixed and dissolved completely. The yellow crystals were added to the mixed solution and stirred at room temperature for 2 hours. The mixture was filtered through medium-speed filter paper under reduced pressure, 20 g of a composite decolorizing agent (activated carbon: alkaline alumina: perlite = 4:1:1) was added to the filtrate, and the mixture was stirred at 70°C for 4 h for decolorization; the decolorized solution was filtered through medium-speed filter paper, the pH of the filtrate was adjusted to neutral using hydrochloric acid, and the solution was diluted to a solution with a solid content of 2.5%, and the solution was first filtered through a LSD001 macroporous strong acid cation resin, and then filtered through a column liquid and then filtered through a LSD396 decolorizing anion resin. The column liquid was concentrated and spray-dried to obtain highly water-soluble dihydroquercetin for cosmetics, with a content of 28.3%, a recovery rate of dihydroquercetin of 81.46%, an embedding rate of 99.81%, and a white powder. The whiteness of the product was 45.8 as tested by a whiteness tester, the solubility in water was 237.81 g / L, and the conductivity was 94.68 μS / cm.

[0052] In addition, it has been tested that the highly water-soluble dihydroquercetin for cosmetics obtained by the present invention can be stored for three years at room temperature.

[0053] Comparative Example 1

[0054] The same as Example 1, except that only hydroxypropyl-β-cyclodextrin is added during inclusion, that is, the mass of sodium alginate, sodium bisulfite and hydroxyethyl cellulose is replaced by hydroxypropyl-β-cyclodextrin. The embedding efficiency of dihydroquercetin obtained in this comparative example is 47.69%, and the solubility in water is 189.68 g / L.

[0055] Comparative Example 2

[0056] The same as Example 1, the only difference is that the mass of sodium bisulfite is replaced by hydroxypropyl-β-cyclodextrin during inclusion. The dihydroquercetin obtained in this comparative example has an embedding rate of 98.89% and a solubility in water of 241.5 g / L; however, because there is no sodium bisulfite, the color of the product after final drying is light yellow, and the final product has poor stability and is very likely to discolor after being placed.

[0057] Comparative Example 3

[0058] The same as Example 1, except that only hydroxypropyl-β-cyclodextrin and sodium alginate are added during inclusion, that is, the mass of sodium bisulfite and hydroxyethyl cellulose is replaced by sodium alginate. The dihydroquercetin obtained in this comparative example has an embedding rate of 78.36%, and a solubility in water of 173.6 g / L, but because there is no sodium bisulfite, the color of the product after final drying is light yellow, and the final product has poor stability, and is very likely to discolor after being placed. At the same time, without hydroxyethyl cellulose, the solution cannot be fully suspended, resulting in a significant reduction in the embedding rate.

[0059] Comparative Example 4

[0060] The same as Example 1, except that no basic alumina was added during decolorization. Without adding basic alumina, the content of dihydroquercetin would be reduced, because the weakly alkaline environment can promote the solubility of dihydroquercetin in water, and without adding basic alumina, some dihydroquercetin would be precipitated and lost. At the same time, because the amount of activated carbon added needs to be increased to achieve an off-white powder, the activated carbon will adsorb a part of the dihydroquercetin, resulting in a significant reduction in the content of dihydroquercetin in the final product.

[0061] Comparative Example 5

[0062] The same as Example 1, except that hydroxyethyl cellulose was not added during inclusion, the dihydroquercetin encapsulation efficiency obtained in this comparative example was 73.21%, and the solubility in water was 201.36 g / L. Because hydroxyethyl cellulose was not added, the solution could not be fully suspended, resulting in a significant reduction in the encapsulation efficiency.

[0063] Comparative Example 6

[0064] The same as Example 1, except that the mass of the composite decolorizing agent is replaced by activated carbon. The dihydroquercetin obtained in this comparative example is a white powder, the recovery rate of dihydroquercetin is 51.21%, and the whiteness is 61.5. Although increasing the amount of activated carbon added can increase the whiteness of the powder, there is a problem of excessive adsorption of dihydroquercetin, resulting in excessive loss of dihydroquercetin.

[0065] Comparative Example 7

[0066] The same as Example 1, except that the mass of perlite in the composite decolorizing agent is replaced by diatomaceous earth. The content of dihydroquercetin obtained in this comparative example is 18.74%.

[0067] Comparative Example 8

[0068] The same as Example 1, except that the larch root powder is directly subjected to plate and frame filter pressing extraction without cold water soaking.

[0069] The content of dihydroquercetin obtained in this comparative example is 17.67%, and the conductivity of dihydroquercetin is 271.68μS / cm. If cold soaking is not performed, a large amount of water-soluble pigments and impurities will be extracted, resulting in a darker color of the finished product. At the same time, too much water-soluble impurities will inevitably increase ionicity.

[0070] Comparative Example 9

[0071] The same as Example 1, except that the solvent is replaced by anhydrous ethanol from boiling water during the plate and frame filter pressing extraction.

[0072] The content of dihydroquercetin obtained in this comparative example is 16.89%. Since most of the extracted substances are fat-soluble substances, although an inclusion process is added later, many fat-soluble impurities will be included, which reduces the content of the product. In addition, due to the influence of the alcohol extract, the stability of the final product aqueous solution is greatly reduced.

[0073] The dihydroquercetin prepared in each embodiment of the present invention can be used to prepare cosmetics. Taking the highly water-soluble dihydroquercetin for cosmetics in Example 1 as an example, its performance is tested:

[0074] 1. In vitro anti-wrinkle efficacy test:

[0075] This test was conducted in accordance with the "In vitro efficacy evaluation method for cosmetics / cosmetic raw materials-Advanced glycation end product inhibition test" (CTC-SOP-012-2024) of China Inspection and Quarantine (Beijing) Cosmetics Technology Co., Ltd.

[0076] (I) Main instruments:

[0077] Microplate reader (2020SW0107): Thermo Scientific, Varioskan fisher LUX N / M, USA.

[0078] Constant temperature incubator (2015SW0015): Shanghai Yiheng, DHP-9272B, China.

[0079] Electronic balance (2014LH0016): Mettler, XSE204, Switzerland.

[0080] (II) Main reagents:

[0081] BSA: Source Leaf Biotechnology, S07HS193768.

[0082] Glucose:Aladdin, F2209126.

[0083] PBS: ViVaCell, 2327212.

[0084] Aminoguanidine hydrochloride: Aladdin, G1909194.

[0085] (III) Test samples and control samples:

[0086] Test samples: The highly water-soluble dihydroquercetin for cosmetics in Example 1 was diluted with PBS to a concentration of 0.10% (mass percentage, the same below), 0.50%, 1.00%, 1.50%, and 2.00%, and then filtered and sterilized for later use.

[0087] Negative control: PBS solution.

[0088] Positive control: 10% (mass percentage) aminoguanidine hydrochloride solution.

[0089] (IV) Test methods

[0090] Preparation of reaction solution: 160 mg / mL bovine serum albumin solution and 480 mg / mL glucose solution were prepared using PBS solution, the two were mixed at a volume ratio of 1:1, and filtered through a 0.22 μm filter membrane to serve as the 2× glycosylation reaction solution.

[0091] Experimental grouping: The experiment sets up sample group and sample blank, negative control and negative blank, positive control and positive blank.

[0092] Preparation of reaction system: 2× glycosylation reaction solution and corresponding samples were added to sample group, negative control and positive control, and PBS solution and corresponding samples were added to sample blank, negative blank and positive blank.

[0093] ELISA detection: After the reaction system is mixed evenly, it is incubated at 55°C for 7 days. After the reaction is completed, a fluorescence ELISA reader is used to detect the changes in AGEs content under the conditions of excitation wavelength of 370nm and emission wavelength of 440nm.

[0094] Test result judgment criteria: SPSS software was used for analysis. Compared with the negative control, the AGEs fluorescence intensity value of the positive control decreased, and the AGEs inhibition rate increased and had a statistically significant difference (p<0.05), indicating that the test conditions were effective. Compared with the negative control, the AGEs inhibition rate of the sample test group increased and had a statistically significant difference (p<0.05), indicating that the sample had the ability to inhibit AGEs at this concentration, indicating that the sample had anti-wrinkle effect.

[0095] Test results: The original data fluorescence intensity values ​​are shown in Table 1, and the result analysis is shown in Table 2 and Figure 2 .

[0096] Table 1. Raw data of AGEs inhibition test

[0097]

[0098] Table 2 Analysis of AGEs inhibition test results

[0099]

[0100] This experiment evaluates the anti-wrinkle effect of the sample by testing the inhibitory effect of the sample on AGEs in the glycation reaction solution. The causes of wrinkles and loss of elasticity of the skin are divided into internal and external factors. The internal factors are mainly the natural aging of the organism, bad eating habits and living habits, such as staying up late and lack of exercise; the external factors are mainly ultraviolet rays and environmental pollution. The combined action of endogenous and exogenous factors will accelerate the glycation reaction in the body. Glycation reaction, also known as Maillard reaction, refers to the non-enzymatic condensation reaction between the carbonyl group on the reducing sugar (such as glucose) and the free amino group on the macromolecules such as protein, lipid or nucleic acid. The covalent adducts generated by the glycation reaction are collectively referred to as advanced glycation end products (AGEs). Excessive advanced glycation end products in the body will undergo glycation cross-linking reactions with the elastin and collagen of the skin, causing the skin to lose elasticity and generate wrinkles. AGEs have fluorescent properties, and obvious fluorescence peaks can be observed at an excitation wavelength of 370nm and an emission wavelength of 440nm.

[0101] 2. Firming efficacy test

[0102] Experimental organisms: AB wild zebrafish larvae (4 dpf).

[0103] Test water: standard dilution water (temperature: 28.5°C, pH value: 6.91).

[0104] Blank control: standard dilution water.

[0105] Positive control: 0.02 g / mL carnosine.

[0106] Experimental method: The zebrafish were divided into three groups, including a blank control group, a positive control group (carnosine) and a test group. The blank control group was not treated with anything, the positive control group was added with 0.02g / mL carnosine, and the test group was added with different concentrations of the test products. After 24 hours of adding the test products, samples were collected from each group of zebrafish, RNA was extracted using a kit, and the transcription levels of the target genes eln1 and eln2 were detected using q-PCR. The firming effect of the test product was evaluated by the relative expression of the zebrafish elastin genes eln1 and eln2.

[0107] Experiment 2: Classification of zebrafish test results:

[0108] 1. “Ineffective” or “No significant effect”: The test result is lower than 115% of the negative control group value.

[0109] 2. “Mild effect”: The test result is higher than 115% of the blank control group value, but lower than 150%.

[0110] 3. “Moderate effect”: The test result is higher than 150% but lower than 200% of the blank control group value.

[0111] 4. “Significant effect”: The test result is 200% higher than the blank control group value.

[0112] The test results are shown in Table 3.

[0113] Table 3 Relative expression of elastin genes eln1 and eln2

[0114]

[0115]

[0116] Note: "Test concentration" refers to the concentration of the highly water-soluble dihydroquercetin for cosmetics added in Example 1 (the same below).

[0117] From the above results, it can be seen that the relative expression levels of zebrafish elastin genes eln1 and eln2 in the treatment with different concentrations of highly water-soluble dihydroquercetin for cosmetics in Example 1 are higher than those in the blank control group. Therefore, based on the above test results, it can be shown that the highly water-soluble dihydroquercetin for cosmetics of the present invention has a firming effect.

[0118] 3. Soothing efficacy test

[0119] Experimental organisms: AB wild zebrafish larvae (24 hpf).

[0120] Test water: standard dilution water (temperature: 28.5°C, pH value: 6.91).

[0121] Blank control: standard dilution water.

[0122] Model control: 0.01% (mass percentage) sodium dodecylbenzene sulfonate solution.

[0123] Positive control: 0.09% (mass percentage) dipotassium glycyrrhizinate solution.

[0124] Test method: The zebrafish were divided into four groups, including a blank control group, a model control group, a positive control group and a test substance group. The blank control group was not treated with anything, the model control group was added with 0.01% sodium dodecyl sulfate solution, the positive control group was added with 0.09% dipotassium glycyrrhizinate solution and 0.01% sodium dodecyl sulfate solution, and the test substance group was added with different concentrations of test substance solution and 0.01% sodium dodecyl sulfate solution. After 15 minutes, the zebrafish embryos were observed and recorded under an optical microscope, and the number of spin movements within 30 seconds was calculated.

[0125] The grading of zebrafish test results in Experiment 3:

[0126] 1. “Ineffective” or “No significant effect”: There is no significant difference in the number of spinning exercises between the group A and the model control group.

[0127] 2. “Mild effect”: The number of spinning movements is less than 20% of the value in the model control group.

[0128] 3. “Moderate effect”: The number of spinning movements is less than 50% of the value in the model control group.

[0129] 4. “Significant effect”: The number of spinning movements was less than 70% of the value in the model control group.

[0130] The number of spin movements of zebrafish embryos at different test concentrations is shown in Table 4.

[0131] Table 4 Spin movement times of zebrafish embryos at different test concentrations

[0132]

[0133] The average number of spinning movements per tail of zebrafish in the control group is shown in Table 5.

[0134] Table 5 Average number of spin movements per tail of zebrafish in the control group

[0135]

[0136] Example 1 The number of embryonic spinning movements (tail) in 30s for highly water-soluble dihydroquercetin for cosmetics, positive control, model control and blank control is shown in Table 6.

[0137] Table 6 Example 1 Highly water-soluble dihydroquercetin for cosmetics, positive control, model control and blank control 30s embryo spin movement times (tail)

[0138]

[0139] Note: A1-F7 in Table 6 represent the groups of zebrafish.

[0140] According to the test results, the number of spin movements within 30 seconds in zebrafish embryos treated with different concentrations of highly water-soluble dihydroquercetin for cosmetics in Example 1 was lower than that in the model control group. Therefore, based on the above test results, it can be shown that the highly water-soluble dihydroquercetin for cosmetics of the present invention has a significant soothing effect.

[0141] From the above content, it can be seen that the highly water-soluble dihydroquercetin for cosmetics prepared by the present invention has good anti-wrinkle effect, firming effect and soothing effect, and can be used for preparing cosmetics.

[0142] The above are only preferred specific implementations of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present application should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. A method for preparing highly water-soluble dihydroquercetin for cosmetics, characterized in that: The following steps are involved: The larch root is dried and crushed, soaked in cold water, filtered, the filter residue is extracted by plate and frame filter pressing, the extract is concentrated, alcohol precipitation is performed, the alcohol solution is concentrated and refrigerated until crystals are precipitated, the crystals are added to the inclusion solution, filtered after reaction, a composite decolorizing agent is added to the filtrate, filtered again, the pH is adjusted to neutral, diluted to a solid content of 3±0.5% in the filtrate, passed through a column, concentrated, and spray-dried to obtain the highly water-soluble dihydroquercetin for cosmetics.

2. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 1, characterized in that: The cold water soaking is soaking in water at 10-20° C. for 12-24 hours.

3. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 1, characterized in that: The method for extracting the filter residue by plate and frame filter pressing is: using the plate and frame filter pressing extraction method, using boiling water 40-80 times the raw material to extract the filter residue 2-3 times.

4. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 1, characterized in that: The inclusion solution is a mixed solution of cyclodextrin derivatives, alginate, sodium bisulfite, hydroxyethyl cellulose and water.

5. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 4, characterized in that: The mass ratio of the cyclodextrin derivative, alginate, sodium bisulfite, hydroxyethyl cellulose and water is (40-50): (0.3-0.7): (0.5-1.5): (0.3-0.7): (160-200).

6. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 5, characterized in that: The cyclodextrin derivative is hydroxypropyl-β-cyclodextrin or hydroxypropyl-γ-cyclodextrin.

7. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 1, characterized in that: The composite decolorizing agent is a mixture of activated carbon, alkaline alumina and perlite.

8. The method for preparing highly water-soluble dihydroquercetin for cosmetics according to claim 1, characterized in that: The column is passed through the cationic resin first, and the column liquid is passed through the anionic resin.

9. A highly water-soluble dihydroquercetin for cosmetics, characterized in that: Prepared according to the preparation method according to any one of claims 1 to 8.

10. Use of the dihydroquercetin according to claim 9 in the preparation of cosmetics.

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