阅读说明：本技术 大黄类药材中白皮杉醇的提取方法、微胶囊分子包埋方法及其在化妆品中的应用 (Method for extracting piceatannol from rheum officinale, microcapsule molecule embedding method and application of piceatannol in cosmetics ) 是由 尹开云 胡琳 吴明一 任杨帆 胡群 方震东 于 2020-07-07 设计创作，主要内容包括：本申请公开了一种大黄类药材中白皮杉醇的提取方法、微胶囊分子包埋方法及其在化妆品中的应用,包括以下步骤：以大黄类药材粉体为原料,向所述原料喷洒复合酶溶液后酶解,得到酶解物；酶解物中加入10wt％的碳酸钠溶液调节pH至10后加入乙醇,形成醇水体系溶剂后在常温下进行微波萃取,过滤得到滤液,在滤液中加入浓度5wt％的乳酸水,调节pH值至中性；减压浓缩得到浸膏,回溶浸膏得到不溶于水的白色沉淀,对所述白色沉淀进行1～3次水洗重结晶,得到结晶体物质,所述结晶体物质中白皮杉醇含量≥98％(w/w)；该方法以大黄类药材为原料,采用对皮肤无刺激、性能温和的pH值调节试剂,无需使用强碱和盐酸,提取结晶体中白皮杉醇含量大于98％,所得物质尤其适用于化妆品内物质中使用。(The application discloses a method for extracting piceatannol from rhubarb medicinal materials, a method for embedding microcapsule molecules and application of the piceatannol in cosmetics, which comprises the following steps: spraying a complex enzyme solution on rhubarb medicinal material powder serving as a raw material, and performing enzymolysis to obtain an zymolyte; adding 10 wt% of sodium carbonate solution into the zymolyte to adjust the pH value to 10, adding ethanol, forming an alcohol-water system solvent, performing microwave extraction at normal temperature, filtering to obtain a filtrate, adding lactic acid water with the concentration of 5 wt% into the filtrate, and adjusting the pH value to be neutral; concentrating under reduced pressure to obtain extract, dissolving the extract back to obtain water-insoluble white precipitate, and washing and recrystallizing the white precipitate for 1-3 times to obtain a crystalline substance, wherein the content of piceatannol in the crystalline substance is more than or equal to 98% (w/w); the method uses rhubarb as raw materials, adopts a pH value adjusting reagent which has no stimulation to skin and mild performance, does not need strong alkali and hydrochloric acid, extracts the piceatannol content in crystals to be more than 98 percent, and the obtained substance is particularly suitable for being used in substances in cosmetics.)

1. A method for extracting piceatannol from rhubarb medicinal materials is characterized by comprising the following steps:

spraying a complex enzyme solution on rhubarb medicinal material powder serving as a raw material, and performing enzymolysis to obtain an zymolyte;

adding 10 wt% of sodium carbonate solution into the zymolyte to adjust the pH value to 10, adding ethanol, forming an alcohol-water system solvent, performing microwave extraction at normal temperature, filtering to obtain a filtrate, adding lactic acid water with the concentration of 5 wt% into the filtrate, and adjusting the pH value to be neutral;

concentrating under reduced pressure to obtain extract, dissolving the extract back to obtain water-insoluble white precipitate, and washing and recrystallizing the white precipitate for 1-3 times to obtain a crystalline substance, wherein the content of piceatannol in the crystalline substance is more than or equal to 98% (w/w);

the complex enzyme solution is prepared by dissolving cellulase, amylase and pectinase in warm water; according to the cellulose: amylase: and (3) pectinase: the mass ratio of the raw materials is 0.015: 0.01: 0.005:1

The radix et rhizoma Rhei is at least one of radix et rhizoma Rhei Himalayan, radix et rhizoma Rhei Lhasa or radix et rhizoma Rhei Shangri.

2. The method for extracting piceatannol from rhubarb as claimed in claim 1, wherein the rhubarb is Rheum palmatum L.

3. The method for extracting piceatannol from rhubarb as claimed in claim 1, wherein the step of enzymolysis is carried out by keeping the temperature at 42 ℃ for 36 hours.

4. The method for extracting piceatannol from rhubarb as claimed in claim 1, wherein the concentration of ethanol is 73% (v/v); the adding amount of the ethanol is that the mass ratio of the raw materials to the ethanol is 1: 12 are added.

5. The method for extracting piceatannol from rhubarb as claimed in claim 1, wherein the microwave extraction is performed under 650W microwave power for 180 s.

6. A method for microencapsulation molecular encapsulation of a crystalline substance as defined in any one of claims 1 to 5, comprising the steps of:

dissolving the crystal substance in ethanol to prepare a piceatannol alcohol solution, and preparing a glucosyl-beta-cyclodextrin water solution;

dropwise adding the piceatannol alcohol solution into the glucose-beta-cyclodextrin aqueous solution within 30 minutes while stirring at 25 ℃, and continuously stirring for 30 minutes after the addition to obtain a mixed solution;

and freeze-drying the mixed solution for 24 hours to obtain the powdery piceatannol microcapsule molecule embedding body.

7. The method for microencapsulating a crystalline substance as in claim 6, wherein the mass concentration of said alcohol solution of piceatannol to the aqueous solution of glucosyl- β -cyclodextrin is 1: 2.

8. The method for microencapsulating a crystalline substance as in claim 6, wherein said glucose- β -cyclodextrin is prepared by the following steps:

step 1, mixing 20% maltose syrup and beta-cyclodextrin powder according to a mass ratio of 4: 1 mixing to prepare slurry, adding pullulanase (pullulanase) with the total weight of 1-3% (w/w), stirring and terminating enzyme conversion under the conditions of constant temperature and constant humidity to obtain an intermediate product: a glucose-based- β -cyclodextrin slurry;

and 2, adding 1-3% (w/w) of saccharifying amylase into the intermediate product to obtain a glucose-beta-cyclodextrin slurry material, uniformly stirring and mixing, keeping the temperature and stirring in a constant-temperature and constant-humidity box with the relative humidity of 60% at 40 ℃ for 48 hours, and hydrolyzing the glucose-beta-cyclodextrin under the action of the saccharifying amylase to generate the glucose-beta-cyclodextrin.

9. The method for embedding microcapsule molecules of crystalline substances according to claim 8, further comprising step 3, purifying and separating the glucose and glucose- β -cyclodextrin mixture by using a UF4050 ultrafiltration membrane, concentrating, and freeze-drying to obtain glucose- β -cyclodextrin with content of 98% or more.

10. The application of the paclitaxel microcapsule molecular inclusion complex obtained by the method according to any one of claims 6 to 9 in cosmetics, wherein the cosmetics are any one of face cream, hand cream, eye cream, body lotion, moisturizing lotion, essence, hydrolat, bath lotion, shampoo, hair conditioner, hair oil, massage oil, essence oil, body oil, facial mask, eye mask, sunscreen cream and sunscreen spray.

Technical Field

The application relates to a method for extracting piceatannol from rhubarb medicinal materials, a microcapsule molecule embedding method and application of the microcapsule molecule embedding method in cosmetics, and belongs to the field of traditional Chinese medicinal materials and cosmetics.

Background

Quzhu or Da Huang (Rhei Radix et Rhizoma) is a folk Tibetan medicine used for thousands of years in Tibetan region, and the most important two classical medical books in Tibetan, four medical classics and Jingzhu materia Medica, make detailed records on the use of the medicine. Meanwhile, the medicine is also a traditional Chinese medicine, the medical records of the medicine are originally seen in Shen nong's herbal Jing, about 45 plants in rhubarb are available, in the traditional Chinese medicine application, except three ' genuine ' Rheum palmatum L, Tang Gu Da Huang (R.. tonggutium Maxim. ex Balf.) and medicinal Rheum officinale (R.. offisinale Baill.) carried in the Chinese pharmacopoeia are mainly used, and other more than 40 ' non-genuine ' Rheum officinale in the same genus hardly contains or only contains a small amount of anthraquinone compounds, so that the effect of the medicine under the purgation is weak or none, and most of the Rheum palmatum L and rhubarb are not well developed and utilized.

Piceatannol (Piceatannol) has a chemical formula of C₁₄H₁₂O₄Chinese alias: 3' -hydroxy resveratrol; a bisabolol; 4- [2- (3, 5-dihydroxyphenyl) ethenyl]-1, 2-benzenediol; (E) -3,3',4,5' -tetrahydroxystilbene; (E)4- [2- (3, 5-dihydroxyphenyl) ethenyl]-1, 2-benzenediol; (E) -3,3',4,5' -stilbenetetraol. Pure piceatannol is generally white crystal, has amorphous powder, is sensitive to light and is easy to be oxidized and discolored. Piceatannol is a compound with a chemical structure similar to that of resveratrol, widely exists in vegetables and fruits such as grapes, blueberries, passion fruits and the like, has the main function basically consistent with that of resveratrol, but has the effects of resisting oxidation and removing free radicals which are 4-5 times that of resveratrol due to the fact that the piceatannol has one more hydroxyl group than the resveratrol.

Disclosure of Invention

The application provides an extraction method of piceatannol in rhubarb medicinal materials, a microcapsule molecule embedding method and application of the piceatannol in cosmetics, and aims to solve the technical problems that the existing natural piceatannol molecule has poor water solubility, is easy to oxidize and discolor when being used in food and cosmetic products, is separated out in a layering manner, and influences the absorption effect of a human body on the piceatannol.

One aspect of the application provides a method for extracting piceatannol from rhubarb, which comprises the following steps:

spraying a complex enzyme solution on rhubarb medicinal material powder serving as a raw material, and performing enzymolysis to obtain an zymolyte;

adding 10 wt% of sodium carbonate solution into the zymolyte to adjust the pH value to 10, adding ethanol, forming an alcohol-water system solvent, performing microwave extraction at normal temperature, filtering to obtain a filtrate, adding lactic acid water with the concentration of 5 wt% into the filtrate, and adjusting the pH value to be neutral;

concentrating under reduced pressure to obtain extract, dissolving the extract back to obtain water-insoluble white precipitate, and washing and recrystallizing the white precipitate for 1-3 times to obtain a crystalline substance, wherein the content of piceatannol in the crystalline substance is more than or equal to 98% (w/w);

the complex enzyme solution is prepared by dissolving cellulase, amylase and pectinase in warm water; according to the cellulose: amylase: and (3) pectinase: the mass ratio of the raw materials is 0.015: 0.01: 0.005:1

The radix et rhizoma Rhei is at least one of radix et rhizoma Rhei Himalayan, radix et rhizoma Rhei Lhasa or radix et rhizoma Rhei Shangri.

The method takes three non-genuine rhubarb (Rheum weibbianum Royle) and Rheum lhasaense (Rheum lhasaense A.J.Li et P.K.Hsiao), Rheum officinale in Tibet or Rheum Grangladesh (Rheum bethicum Maxim.ex HK.f.) as raw materials, and adopts microwave technology and alcohol-water system solvent to realize extraction and separation without strong base and hydrochloric acid under specific pH value and normal temperature conditions, so that the obtained product has high purity of the piceatannol crystal compound, does not contain strong base, hydrochloric acid and other substances, can be better absorbed and utilized by the skin, and does not cause allergic reaction of the skin.

The method only needs sodium carbonate and lactic acid to adjust the pH value, the substances are easy to decompose and have small skin sensitization, and the obtained compound is particularly suitable for being used in cosmetic products. According to the method, crystals with high purity can be obtained only by purifying the obtained precipitate for 1-3 times, and compared with the existing method for extracting piceatannol from rhubarb traditional Chinese medicines, the method needs to carry out purification for multiple times, so that the purification efficiency is effectively improved.

The crystalline substance obtained by the above method contains piceatannol (formula I) or its derivatives

The natural piceatannol or its derivatives described herein include cis, trans or a mixture of cis and trans isomers of the compound, as well as crystalline or amorphous structures of the compound.

Preferably, the rhubarb is Lhasa rhubarb. The extracted compound has the highest piceatannol content.

Preferably, the enzymolysis step is carried out at 42 ℃ for 36 hours. The extract obtained by enzymolysis under the condition has high piceatannol derivative content, which is beneficial to the release of the piceatannol derivative from plant cells. The temperature of warm water used for dissolving the enzyme preparation is also 42 ℃ to ensure no activity.

Preferably, the ethanol concentration is 73% (v/v); the adding amount of the ethanol is that the mass ratio of the raw materials to the ethanol is 1: 12 are added. The ethanol solution used contains water as solvent. The material-liquid ratio here means that 12kg of ethanol is added for 1000g of rhubarb internal medicine.

Preferably, the microwave extraction conditions are extraction time 180s at microwave power of 650W. The extraction is carried out according to the condition, the content of impurities in the crystal after extraction is low, the adjustment of the pH value can be realized only by lactic acid, and simultaneously the water-soluble impurities obtained after extraction can be combined with the lactic acid to form precipitate and removed, thereby improving the purity of the final crystal.

Another aspect of the present application further provides a method for embedding the above-mentioned crystalline substance into a microcapsule molecule, comprising the following steps:

dissolving the crystal substance in ethanol to prepare a piceatannol alcohol solution, and preparing a glucosyl-beta-cyclodextrin water solution;

dropwise adding the piceatannol alcohol solution into the glucose-beta-cyclodextrin aqueous solution within 30 minutes while stirring at 25 ℃, and continuously stirring for 30 minutes after the addition to obtain a mixed solution;

and freeze-drying the mixed solution for 24 hours to obtain the powdery piceatannol microcapsule molecule embedding body.

After the piceatannol is embedded by the method, the content of the piceatannol in the obtained embedding body is 25 wt%, and the embedding rate and the solubility of the microcapsule molecule embedding body are measured by an infrared spectrum method and an ultraviolet spectrum method. The piceatannol extracted by the method is extracted by adopting glucosyl-beta-cyclodextrin, and the water solubility of the obtained embedding body is higher than that of other piceatannol embedding bodies. The glucosyl-beta-cyclodextrin has high hydroxyl content, and can be combined with lactic acid in the crystal substance obtained by the method, so that the content of piceatannol in the embedding body can be increased.

The glucosyl-beta-cyclodextrin (Glu-beta-CD) used in the present application was prepared according to the existing method.

The obtained compound is subjected to microcapsule molecule preparation by adopting a biological high molecular material glucose-beta-cyclodextrin obtained by an enzymatic preparation process so as to improve the water solubility of the piceatannol and prevent oxidative discoloration, and the natural piceatannol microcapsule molecule embedding body is obtained.

The method provided by the application carries out molecular embedding on the piceatannol by using a glucosyl-beta-cyclodextrin parent cavity to prepare piceatannol microcapsule molecules, thereby improving the water solubility of the piceatannol, simultaneously protecting and preventing the compound obtained by extraction from oxidative discoloration, and enabling the compound to be applied to products in cosmetics in a large amount.

Glucosyl-beta-cyclodextrin (Glu-beta-CD) is a derivative of beta-cyclodextrin, and is prepared from the beta-cyclodextrin by a biological enzyme method. The parent compound, beta-Cyclodextrin (CD), is a complex of 6, 7, 8 or more D-glucopyranose units linked by a-1, 4 glycosidic bonds (referred to in turn as alpha, beta, gamma-CD), and has a structure that resembles a tapered cylinder with a "tapered cylindrical cavity". Through modification treatment of the molecular structure of the beta-cyclodextrin, a glucose molecule is introduced to the outer side to replace a hydrogen atom, so that an intramolecular hydrogen bond is broken, the water solubility of the compound can be greatly improved, and the lipid solubility of the inner side is improved.

Preferably, the mass concentration of the piceatannol alcohol solution and the mass concentration of the glucosyl-beta-cyclodextrin aqueous solution are 1: 2. For example, the mass concentration of the piceatannol alcohol solution is 10%; the mass concentration of the aqueous solution of the glucosyl-beta-cyclodextrin is 20 percent. The encapsulation is carried out according to the proportion, so that the effective encapsulation of the piceatannol by the glucosyl-beta-cyclodextrin can be realized, and partial materials can be prevented from being not embedded.

Preferably, the glucosyl-beta-cyclodextrin is prepared by the following steps:

step 1, mixing 20% maltose syrup and beta-cyclodextrin powder according to a mass ratio of 4: 1 mixing to prepare slurry, adding pullulanase (pullulanase) with the total weight of 1-3% (w/w), stirring and terminating enzyme conversion under the conditions of constant temperature and constant humidity to obtain an intermediate product: a glucose-based- β -cyclodextrin slurry;

and 2, adding 1-3% (w/w) of saccharifying amylase into the intermediate product to obtain a glucose-beta-cyclodextrin slurry material, uniformly stirring and mixing, keeping the temperature and stirring in a constant-temperature and constant-humidity box with the relative humidity of 60% at 40 ℃ for 48 hours, and hydrolyzing the glucose-beta-cyclodextrin under the action of the saccharifying amylase to generate the glucose-beta-cyclodextrin.

Preferably, the method also comprises a step 3 of purifying and separating the mixed material of the glucose and the glucosyl-beta-cyclodextrin by using a UF4050 ultrafiltration membrane, concentrating, and freeze-drying to obtain the glucosyl-beta-cyclodextrin with the content of more than or equal to 98%. By adopting the step, the glucose contained in the substance obtained in the step 2 can be removed, and the product purity is improved.

Preferably, the conditions of constant temperature and humidity stirring and enzyme conversion termination are as follows: the enzyme conversion was terminated by stirring the samples while maintaining the temperature in a constant temperature and humidity chamber at 38 ℃ and a relative humidity of 60% for 72 hours, and then maintaining the temperature in a constant temperature chamber at 50 ℃ for 2 hours.

The application also provides an application of the paclitaxel microcapsule molecular inclusion body obtained by the method in cosmetics, wherein the cosmetics are any one of face cream, hand cream, eye cream, body lotion, moisturizing water, essence, hydrolat, body wash, shampoo, hair conditioner, hair oil, massage oil, essence oil, body oil, facial mask, eye mask, sun cream and sun spray.

The beneficial effects that this application can produce include:

1) according to the method for extracting piceatannol from rhubarb medicinal materials, rhubarb medicinal materials are used as raw materials, a pH value adjusting reagent which is nonirritating to skin and mild in performance is adopted, strong alkali and hydrochloric acid are not needed, the piceatannol content in extracted crystals is greater than 98%, and the obtained substances are particularly suitable for substances in cosmetics.

2) According to the method for embedding the microcapsule molecules of the piceatannol crystal, the piceatannol crystal and the glucose-beta-cyclodextrin are subjected to microcapsule molecule embedding, compared with the existing method for embedding the piceatannol by using the beta-cyclodextrin, a large number of hydroxyl groups contained in the glucose-beta-cyclodextrin can interact with neutralized lactic acid, the embedding rate and the water solubility of the piceatannol are greatly improved, after embedding and embedding, the content of the piceatannol in the obtained material can reach 25%, the embedding rate is 80%, and the solubility is 25%. The embedding effect is better.

3) According to the microcapsule molecule embedding method of the piceatannol crystal, the microcapsule molecule obtained by the method can effectively protect piceatannol, prevent piceatannol from being oxidized and discolored and influence the efficacy of piceatannol in cosmetics, and can stably exist in face cream.

Detailed Description

The present application will be described in detail with reference to examples, but the present application is not limited to these examples.