Berry composition for scalp care as well as preparation method and application of berry composition

Abstract

The invention relates to the field of cosmetics, in particular to a berry composition for scalp care and a preparation method and application thereof, and the preparation method comprises the following steps: A, pretreating fruits to obtain berry freeze-dried powder; b, preparing a deep eutectic solvent; c, mixing the berry freeze-dried powder with the deep-eutectic solvent, and performing oscillation extraction to obtain a berry procyanidine crude extract; d, purifying the berry procyanidine crude extract with macroporous resin to obtain a first purified berry procyanidine product; e, removing the solvent from the first purified berry proanthocyanidins to obtain a second purified berry proanthocyanidins; and F, dissolving epicatechin, mixing the dissolved epicatechin with the second purified product of the berry procyanidine, uniformly heating in a vacuum environment, cooling after relieving the vacuum environment, mixing with amino acid hydrochloride, and preserving heat to obtain the berry extract.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, and in particular to a berry composition that can be used for scalp care, and a preparation method and application thereof. Background Art

[0002] Berries refer to juicy fleshy single fruits such as blueberries, blue indigo fruits, black fruited mountain ash, black wolfberries, and black currants. Berries are rich in nutrients such as amino acids, vitamins, and mineral elements, as well as rich in biologically active ingredients such as flavonoids, anthocyanins, and phenolic acids. They have antioxidant, anti-inflammatory, antibacterial, anti-radiation, and anti-aging effects.

[0003] Naturally occurring anthocyanins are generally formed by glycosidic bonds between anthocyanins and glucose, galactose, arabinose, xylose and rhamnose. Free anthocyanins have poor stability and are rarely found in plants. There are multiple phenolic hydroxyl groups in the structure of anthocyanins, which lead to high polarity, low bioavailability, and unstable anthocyanin structure. Other substrates in berries will also react with anthocyanins in a series of ways. During the pulping process, the berries will be mechanically damaged and undergo enzymatic browning reactions. Anthocyanin modification can be divided into three pathways: polymerization, cleavage and derivatization. The cleavage of anthocyanins leads to the production of colorless compounds, polymerization leads to browning, and derivatization reactions may produce other pigment molecules. This application uses amino acids to prepare a new type of low eutectic solvent, and uses cheap and green ethylene glycol as a raw material, which provides a new idea for promoting overcapacity. The low eutectic solvent is mainly composed of a hydrogen bond donor and a hydrogen bond acceptor. Like ionic liquids, it has a strong solubility, low toxicity, simple synthesis, good economy and high atomic utilization.

[0004] A large number of research reports show that type A proanthocyanidins can play a key role in preventing and treating recurrent urinary tract infections by inhibiting Escherichia coli and even adhering to the mucosal wall of urethral tissue. Type B proanthocyanidins, which lack an ether bond compared to type A proanthocyanidins, do not show any anti-adhesion activity. Proanthocyanidins in nature mainly exist in the form of type B. Type A proanthocyanidins in food sources are only found in cranberries and their products, while cranberries are only produced in North America, Northern Europe and parts of Daxinganling in my country, with limited production. Type A proanthocyanidins have few sources and their unique and safe health functions without side effects, so the price is relatively high. This patent realizes the conversion of type A proanthocyanidins in existing berry compositions, increases the content of type A proanthocyanidins in extracts, and successfully prepares amino acid derivatives of type A proanthocyanidins, changes their polarity and biological activity, improves their utilization, and applies them to scalp care products. It is found that they have good anti-inflammatory, antibacterial and oil control effects.

[0005] Scalp Ecology A healthy scalp ecological environment is maintained by three major balances: oil, flora, and metabolic balance. Oily scalp, itchy scalp, dandruff, dry hair, hair loss, etc. are all scalp problems caused by an imbalance in the scalp ecological environment. The scalp microecology is composed of host cells and secretions on the surface of the scalp, scalp microorganisms, and the environment in which they are located. From a macroscopic perspective, the physiological environment of the scalp, such as sebum content, water content, and pH, will reflect the health of the scalp and directly affect the growth of hair. The scalp secretes a certain amount of oil to ensure the moisture of the hair, but if the oil secretion is too much, it will cause greasy hair, easily induce dermatitis and even hair loss. The appropriate pH of the scalp is conducive to the healthy growth of hair, and the water content of the scalp skin also directly affects the health of the scalp and hair. A healthy scalp has a diverse microbiome and contains specific beneficial strains. The diversity of the microbiome of an oily scalp is reduced. When the flora is stimulated and unbalanced, it will cause skin diseases. In the process of scalp conditioning, if antibacterial agents are added to inhibit the excessive growth of harmful bacteria such as Malassezia, the ecological balance of the scalp cannot be maintained, because the antibacterial ingredients also inhibit the growth of beneficial bacteria on the scalp, destroying the ecological environment of the scalp. At present, the solutions for itching on the market mainly focus on the use of antifungal agents such as piroctone olamine, pyrithione zinc, climbazole and salicylic acid, selenium disulfide, sulfur, etc. to inhibit the massive growth of Malassezia. These raw materials are also slightly irritating, and for users with non-fungal itching, it is likely to increase the probability of itching. Summary of the invention

[0006] The purpose of the present invention is that, based on the fact that the current research on berries is mainly concentrated in the field of food processing, there are not many studies and applications in the field of cosmetics, there are few types of berry extracts in the catalog of used cosmetic raw materials, the stability of anthocyanins is poor, and due to the few sources of type A proanthocyanidins and their unique characteristics, their prices are high. The present invention inhibits the enzymatic browning reaction of berry compositions to a certain extent, improves the stability of berry compositions, and realizes the preparation of amino acid derivatives of type A proanthocyanidins in existing berry compositions, provides a reference scheme for improving the bioavailability of type A proanthocyanidins and anthocyanidins, and applies them to scalp care products, which have the effects of soothing and anti-inflammatory, controlling oil and reducing itchy dandruff, can regulate the scalp ecological environment, increase the diversity of microorganisms of oily scalp, and realize all-round care of the scalp.

[0007] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0008] A method for preparing a berry extract comprises the following steps:

[0009] A. Fruit pretreatment to obtain berry freeze-dried powder;

[0010] B. preparing a deep eutectic solvent;

[0011] C. mixing the berry freeze-dried powder with the low eutectic solvent, and extracting by shaking to obtain a crude extract of berry proanthocyanidins;

[0012] D. purifying the crude extract of berry proanthocyanidins using a macroporous resin to obtain a first purified product of berry proanthocyanidins;

[0013] E. removing the solvent from the first purified product of berry proanthocyanidins to obtain a second purified product of berry proanthocyanidins;

[0014] F. Dissolving epicatechin, mixing it with the second purified product of berry proanthocyanidins, uniformly heating it under vacuum, releasing the vacuum and cooling it, and then mixing it with amino acid hydrochloride and keeping it warm to obtain the berry extract.

[0015] Furthermore, the method also includes analyzing and detecting type A and type B proanthocyanidin derivatives: analyzing the reaction mixture by UPLC-DAD-EIS-MS, then subjecting the reaction mixture to ODS chromatography and eluting with a 0-100% methanol-water gradient, further chromatographing the eluted portion on Sephadex LH-20 and eluting with a 10%-100% methanol-water gradient to obtain proanthocyanidins and type A and type B proanthocyanidin-amino acid derivatives.

[0016] Preferably, the operation of step F comprises: dissolving the epicatechin in deionized water, determining the content of anthocyanin in the second purified product of berry proanthocyanidins, mixing the anthocyanin and the epicatechin in a molar ratio of 1:1-2, adjusting the pH value to 2-3.5 with an acid-base regulator and then evacuating the mixture, wherein the vacuum degree comprises -0.03-0.10 MPa, heating uniformly at 85-95°C for 80-120 min, releasing the vacuum environment, cooling to 60-65°C with cooling water, dissolving the reaction product and the amino acid hydrochloride in water in a mass ratio of 1:2-4, and continuing to keep warm and stir for 24-36 hours to obtain the berry extract; in step F, the acid-base regulator comprises one or more of citric acid, lactic acid, malic acid, sodium citrate, sodium bicarbonate, sodium hydroxide, and potassium hydroxide; and the amino acid hydrochloride comprises one or more of L-cysteine ​​hydrochloride, arginine hydrochloride, histidine hydrochloride, and lysine hydrochloride.

[0017] Preferably, the operation of step B includes: heating and mixing the amino acid and ethylene glycol to obtain the low eutectic solvent; the molar ratio of the amino acid to the ethylene glycol includes 1:5 to 10; and the amino acid includes one or more of arginine, alanine, and lysine.

[0018] Preferably, the operation of step A comprises: washing the fruit with running water, draining the water, freeze-drying and grinding, passing through a 60-80 mesh sieve to obtain the berry freeze-dried powder, and storing it in the dark for later use; the operation of step C comprises: adding the berry freeze-dried powder and the low eutectic solvent in a reaction container at a solid-liquid ratio of 1:2-10 g / mL, and starting ultrasonic-assisted oscillation extraction after adding a color protection agent, the ultrasonic power comprises 100-300 W, the extraction temperature comprises 45-55° C., extracting for 30-70 minutes, and evaporating at 60 00~10000r / min centrifugation for 15~30min, obtaining supernatant, and obtaining the crude extract of berry proanthocyanidins; the operation of step D comprises: eluting the crude extract of berry proanthocyanidins in the macroporous resin, the eluent comprises 60%~70% by volume ethanol, and obtaining the first purified product of berry proanthocyanidins; the operation of step E comprises: taking the first purified product of berry proanthocyanidins and performing vacuum rotary evaporation concentration, the temperature control comprises 58~75℃, recovering the eluent, and obtaining the second purified product of berry proanthocyanidins.

[0019] Preferably, in step A, the fruit includes one or more of blue indigo fruit, black fruit glandular ribbed aronia, blueberry, black currant, black wolfberry, and blackberry; in step C, the color protection agent includes one or more of ascorbic acid, L-cysteine, citric acid, and malic acid; in step D, the macroporous resin includes one or more of resin S-8, resin AB-8, resin D101, and resin X-5.

[0020] Preferably, in step D, the macroporous resin is pretreated before use; the pretreatment comprises: soaking the macroporous resin in anhydrous ethanol for 24 hours, rinsing it repeatedly with deionized water after sufficient swelling until the effluent is free of white turbidity, rinsing it with a 0.1 mol / L hydrochloric acid solution to acidity, and soaking it for 3 hours, then rinsing it with deionized water to neutrality, then washing the macroporous resin with a 0.1 mol / L NaOH solution to alkalinity, and soaking it for 3 hours, then rinsing it with deionized water to neutrality, and setting it aside.

[0021] Pretreatment of macroporous resin: Use a certain amount of macroporous resin and soak it in anhydrous ethanol for 24 hours to make the resin fully swell. After removing the floating resin and impurities, repeatedly rinse the swollen macroporous resin with deionized water until the effluent is free of white turbidity. Rinse the macroporous resin with 0.1mol / L hydrochloric acid solution to acidity and soak it in hydrochloric acid solution for 3 hours, then rinse it with deionized water to neutrality, then wash the macroporous resin with an equal volume of 0.1mol / L NaOH solution to alkalinity and soak it in NaOH solution for 3 hours, rinse it with deionized water to neutrality, and set aside.

[0022] A berry extract obtained by the preparation method of the berry extract.

[0023] The berry extract of the present invention is rich in B-type proanthocyanidins, A-type proanthocyanidins, catechin-amino acids, epicatechin-amino acids, A-type proanthocyanidin-amino acid derivatives and B-type proanthocyanidin-amino acid derivatives. A-type and B-type proanthocyanidins have broad application prospects as antioxidants. The antioxidant mechanism includes free radical scavenging, metal chelation and polyphenol oxidase inhibition. Proanthocyanidins can downregulate the expression of inflammatory cytokines (IL-6, IL-1β and TNF-α) and phosphorylation signal transduction and transcription activator (STAT3), achieving anti-inflammatory effects. The preparation of proanthocyanidin-amino acid derivatives changes its polarity, effectively improves its bioavailability, and applies it to scalp care products. It is found that it can completely improve the scalp problems of people with itchy dandruff by increasing the diversity of scalp microbiota and the proliferation of scalp beneficial bacteria, and under the synergistic effect of antibacterial agents, it will not recur.

[0024] An application of the berry extract is used in one or more of cosmetics and cosmetic additives.

[0025] A cosmetic additive obtained by the above application, comprising the following components in weight percentage:

[0026]

[0027] The sugar and its analogs include one or more of glucosamine polysaccharides, saccharide isomers, and β-glucan.

[0028] The healthy stratum corneum contains a certain amount of natural moisturizing factor, and its water content is generally 15% to 20%. When the water content is less than 10%, the skin appears dry, rough, and chapped. The maintenance of the moisture content in the stratum corneum of the skin depends on the hydration and barrier function of the skin. Hydration mainly depends on the natural moisturizing factor in the keratinocytes and between the matrix. The latter is a small molecule complex with water absorption properties, which can effectively absorb and lock moisture. The glucosamine polysaccharide / saccharide isomer / β-glucan selected by the present invention can synergize with the berry extract of the present invention at low pH, stimulate the synthesis of natural moisturizing factors, hyaluronic acid and ceramide in the skin, and effectively increase the water content of the scalp skin, reduce the aging of the scalp, so as to relieve itching and quickly repair the scalp barrier, and can effectively alleviate the scalp dryness, itching and tightness caused by the residual surfactant. The moisturizer selected by the present invention has good moisturizing effect, water retention capacity and skin surface permeability, resists scalp inflammation related to dandruff, and thus improves and strengthens the scalp.

[0029] Preferably, the moisturizer includes one or more of glycerol, propylene glycol, butylene glycol, sorbitol, glyceraldehyde, erythrose, maltitol, mannose, thulose, and arabinose; the glucosamine polysaccharide includes one or more of hyaluronic acid, sodium hyaluronate, and sodium chondroitin sulfate; the carbohydrate isomers include water-locking magnetite; the β-glucan includes one or more of yeast glucan and oat glucan; the zinc salt includes PCA zinc, zinc gluconate, zinc chloride, glycinate zinc, glycyrrhetinic acid The invention relates to one or more of zinc, zinc pyrithione, yeast / zinc fermentation product, zinc sulfide, zinc sulfate, zinc myristate, lactobacillus / milk / magnesium / zinc fermentation lysate, zinc lactate, zinc undecylenate, hydrolyzed collagen zinc, zinc salicylate, zinc carbonate, zinc carbonate hydroxide, zinc aspartate, zinc oxide, and Porphyridium / zinc fermentation product; the antibacterial agent comprises one or more of hexamidine di(hydroxyethylsulfonate) salt, salicylic acid and salicylate, piroctone olamine salt, climbazole, and zinc pyrithione.

[0030] The zinc salt of the present invention can cooperate with the berry extract to improve the composite stability of the composition at low pH, enhance the shelf life of the product, and also enhance the effect of the zinc component in reducing oil secretion and anti-inflammatory.

[0031] A hair cosmetic comprising a berry composition which can be used for scalp care.

[0032] Furthermore, the dosage form of the hair cosmetic is one of a solution, a suspension, and a solid.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] 1. There are few types of berry extracts in the catalog of used cosmetic raw materials. The invention provides ideas for the application of berry extracts in the field of cosmetics. This patent realizes the preparation of amino acid derivatives of type A proanthocyanidins in existing berry compositions, provides a reference scheme for improving the bioavailability of type A proanthocyanidins and anthocyanidins, and inhibits the enzymatic browning reaction of berry compositions to a certain extent, improves the stability of berry compositions, and applies them to scalp care products, which have the effects of soothing and anti-inflammatory, reducing itchy dandruff, regulating the scalp ecological environment, and increasing the diversity of microbiota of oily scalp, thus realizing all-round care of the scalp.

[0035] 2. The glucosamine polysaccharide / saccharide isomer / β-glucan selected by the present invention, combined with the berry extract of the present invention, can effectively increase the skin water content of the scalp, reduce scalp aging, quickly repair the scalp barrier, and effectively alleviate the problems of dry scalp, itchy scalp and tight scalp caused by surfactant residues.

[0036] 3. The zinc salt selected by the present invention can cooperate with the berry extract to improve the composite stability of the composition at low pH, thereby enhancing the shelf life of the product and also enhancing the effect of the zinc component in reducing oil secretion and anti-inflammatory. The composition of the present invention and the antibacterial agent can synergistically increase the diversity of the scalp microbiome and the proliferation of the beneficial scalp bacteria while inhibiting the proliferation of Malassezia, thus completely improving the scalp problems of people with itchy scalp and dandruff and preventing them from recurring. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a comparison chart of the retention rate of proanthocyanidins at 5°C.

[0038] Figure 2 This is a comparison chart of the retention rate of proanthocyanidins at 25°C.

[0039] Figure 3 This is a comparison chart of the retention rate of proanthocyanidins at 48°C.

[0040] Figure 4 This is a comparison chart of the retention rate of proanthocyanidins under light conditions.

[0041] Figure 5 This is a comparison chart of the retention rate of proanthocyanidins under light-proof conditions.

[0042] Figure 6 The scavenging rate of hydroxyl radicals, DPPH radicals, and ABTS in the total reducing capacity determination + Free radical scavenging rate comparison chart.

[0043] Figure 7 This is a comparison chart of absorbance values ​​in the total reducing capacity determination. DETAILED DESCRIPTION

[0044] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings.

[0045] Example 1

[0046] The berry extract was prepared as follows:

[0047] 1. Fruit pretreatment: Select plump, non-damaged and rotten blue indigo fruit and black fruit glandular rib aronia fruit in a weight ratio of 1:1, wash with running water, drain and freeze-dry, grind, pass through a 70-mesh sieve and store in dark for later use;

[0048] 2. Preparation of low eutectic solvent: weigh alanine and ethylene glycol in a molar ratio of 1:8, heat and stir until a uniformly mixed liquid is formed.

[0049] 3. Extraction: Add berry freeze-dried powder and low eutectic solvent in a reaction vessel at a solid-liquid ratio of 1:8 g / mL, and start ultrasonic-assisted extraction after adding 0.15% L-cysteine ​​and 0.15% citric acid. The ultrasonic power is 200 W, the extraction temperature is 50°C, and after extraction for 50 minutes, centrifuge at 8000 r / min for 15 minutes to obtain the supernatant, which is the crude extract of berry proanthocyanidins.

[0050] 4. Pretreatment of macroporous resin: Use a certain amount of X-5 macroporous resin and soak it in anhydrous ethanol for 24 hours to fully swell the resin. After removing the floating resin and impurities, repeatedly rinse the swollen macroporous resin with deionized water until the effluent is free of white turbidity. Rinse the macroporous resin with 0.1mol / L hydrochloric acid solution to acidity, soak it in hydrochloric acid solution for 3 hours, then rinse it with deionized water to neutrality, then wash the macroporous resin with an equal volume of 0.1mol / L NaOH solution to alkalinity, soak it in NaOH solution for 3 hours, rinse it with deionized water to neutrality, and set aside.

[0051] 5. Adsorption purification: Add the obtained crude extract to the container containing macroporous resin for macroporous resin purification. The eluent is 60% ethanol by volume, the sample solution pH is 2.5, the desorption solution pH is 1.5, the loading solution concentration is 0.2 mg / mL, the loading flow rate is 1.0 mL / min, and the desorption flow rate is 1.0 mL / min.

[0052] 6. Concentration: The filtrate is concentrated by vacuum rotary evaporation with the temperature controlled at 65°C to obtain berry extract 1 and recover ethanol.

[0053] 7. After determining the total anthocyanin content of the extract obtained in step 6, the extract and epicatechin are dissolved in deionized water respectively so that the molar ratio of anthocyanin to epicatechin is 1:1. The pH value of the anthocyanin and epicatechin solution is adjusted to 2.8 using an acid-base regulator, and then the solution is evacuated to a vacuum degree of -0.08 MPa. After uniform heating at 90°C for 100 minutes, the vacuum pump is turned off, the valve is opened, and then the cooling water is cooled to 60°C, and part of the extract is extracted to obtain berry extract 2. The remaining part is dissolved in water at a mass ratio of 1:3 with L-cysteine ​​hydrochloride, and the mixture is kept warm and stirred for 30 hours to obtain berry extract 3.

[0054] Example 2

[0055] The berry extract was prepared as follows:

[0056] 1. Fruit pretreatment: Select plump, non-damaged and rotten blue indigo fruit and black fruit glandular rib aronia fruit in a weight ratio of 1:1, wash with running water, drain and freeze-dry, grind, pass through an 80-mesh sieve and store in dark for later use;

[0057] 2. Preparation of low eutectic solvent: weigh lysine and ethylene glycol in a molar ratio of 1:10, heat and stir until a uniformly mixed liquid is formed.

[0058] 3. Extraction: Add berry freeze-dried powder and low eutectic solvent in a reaction vessel at a solid-liquid ratio of 1:10 g / mL, and start ultrasonic-assisted extraction after adding 0.15% ascorbic acid and 0.15% citric acid. The ultrasonic power is 300 W, the extraction temperature is 55 ° C, and after extraction for 30 minutes, centrifuge at 10000 r / min for 15 minutes to obtain the supernatant, which is the crude extract of berry proanthocyanidins.

[0059] 4. Pretreatment of macroporous resin: Use a certain amount of X-5 macroporous resin and soak it in anhydrous ethanol for 24 hours to fully swell the resin. After removing the floating resin and impurities, repeatedly rinse the swollen macroporous resin with deionized water until the effluent is free of white turbidity. Rinse the macroporous resin with 0.1mol / L hydrochloric acid solution to acidity, soak it in hydrochloric acid solution for 3 hours, then rinse it with deionized water to neutrality, then wash the macroporous resin with an equal volume of 0.1mol / L NaOH solution to alkalinity, soak it in NaOH solution for 3 hours, rinse it with deionized water to neutrality, and set aside.

[0060] 5. Adsorption purification: Add the obtained crude extract to the container containing macroporous resin for macroporous resin purification. The eluent is 70% ethanol by volume, the sample solution pH is 2.5, the desorption solution pH is 1.5, the loading solution concentration is 0.2 mg / mL, the loading flow rate is 1.0 mL / min, and the desorption flow rate is 1.0 mL / min.

[0061] 6. Concentration: The filtrate is concentrated by vacuum rotary evaporation with the temperature controlled at 75°C, and ethanol is recovered.

[0062] 7. After determining the total anthocyanin content of the extract obtained in step 6, the extract and epicatechin are dissolved in deionized water respectively so that the molar ratio of anthocyanin to epicatechin is 1:1. The pH value of the anthocyanin and epicatechin solution is adjusted to 3.0 using an acid-base regulator, and then vacuumed to a vacuum degree of -0.10 MPa. After uniformly heating at 95°C for 100 minutes, the vacuum pump is turned off, the valve is opened, and then the cooling water is cooled to 65°C. The reaction product and lysine hydrochloride are dissolved in water in a mass ratio of 1:4, and the mixture is kept warm and stirred for 36 hours to obtain berry extract 4.

[0063] Example 3

[0064] The berry extract was prepared as follows:

[0065] 1. Fruit pretreatment: Select plump, non-damaged and rotten blue indigo fruit and black fruit glandular rib aronia fruit in a weight ratio of 1:1, wash with running water, drain and freeze-dry, grind, pass through a 60-mesh sieve and store in dark for later use;

[0066] 2. Preparation of low eutectic solvent: Weigh arginine and ethylene glycol in a molar ratio of 1:5, heat and stir until a uniformly mixed liquid is formed.

[0067] 3. Extraction: Add berry freeze-dried powder and low eutectic solvent in a reaction vessel at a solid-liquid ratio of 1:8 g / mL, and add 0.1% L-cysteine, 0.1% citric acid and 0.1% ascorbic acid, then start ultrasonic-assisted extraction. The ultrasonic power is 100 W, the extraction temperature is 45°C, and after extraction for 70 minutes, centrifuge at 6000 r / min for 30 minutes to obtain the supernatant, which is the crude extract of berry proanthocyanidins.

[0068] 4. Pretreatment of macroporous resin: Use a certain amount of X-5 macroporous resin and soak it in anhydrous ethanol for 24 hours to fully swell the resin. After removing the floating resin and impurities, repeatedly rinse the swollen macroporous resin with deionized water until the effluent is free of white turbidity. Rinse the macroporous resin with 0.1mol / L hydrochloric acid solution to acidity, soak it in hydrochloric acid solution for 3 hours, then rinse it with deionized water to neutrality, then wash the macroporous resin with an equal volume of 0.1mol / L NaOH solution to alkalinity, soak it in NaOH solution for 3 hours, rinse it with deionized water to neutrality, and set aside.

[0069] 5. Adsorption purification: Add the obtained crude extract to the container containing macroporous resin for macroporous resin purification. The eluent is 60% ethanol by volume, the sample solution pH is 2.5, the desorption solution pH is 1.5, the loading solution concentration is 0.2 mg / mL, the loading flow rate is 1.0 mL / min, and the desorption flow rate is 1.0 mL / min.

[0070] 6. Concentration: The filtrate is concentrated by vacuum rotary evaporation, the temperature of which is controlled at 58°C, and ethanol is recovered.

[0071] 7. After determining the total anthocyanin content of the extract obtained in step 6, the extract and epicatechin are dissolved in deionized water respectively so that the molar ratio of anthocyanin to epicatechin is 1:1. The pH value of the anthocyanin and epicatechin solution is adjusted to 2.5 with an acid-base regulator, and then vacuumed to a vacuum degree of -0.07 MPa. After uniformly heating at 85°C for 100 minutes, the vacuum pump is turned off, the valve is opened, and then the cooling water is cooled to 60°C. The reaction product and arginine hydrochloride are dissolved in water in a mass ratio of 1:2, and the mixture is kept warm and stirred for 24 hours to obtain berry extract 5.

[0072] Effect Example 1

[0073] Study on the stability of berry extracts

[0074] The vanillin-hydrochloric acid method was used to determine the proanthocyanidin content, with the retention rate as an indicator. The proanthocyanidin retention rate was calculated as follows:

[0075]

[0076] Wherein: R is the retention rate of proanthocyanidins, %; C is the concentration of proanthocyanidins in the solution after treatment in different ways, g / mL; C0 is the initial proanthocyanidin concentration in the solution before treatment, mg / mL.

[0077] 1. Effect of temperature on the stability of berry extracts

[0078] A certain concentration of berry extract solution was prepared and placed at 5°C, 25°C, and 48°C for a 30-day thermal stability test. The proanthocyanidin content was measured every 5 days and its retention rate was calculated. The test results are as follows: Figure 1-3 shown.

[0079] Depend on Figure 1-3 It can be seen that the retention rate of proanthocyanidins in berry extracts gradually decreases with the increase of storage days at 5℃, 25℃, and 48℃, and the rate of decrease is faster under high temperature conditions. Figure 1 , Figure 2 , Figure 3 It can be seen that the thermal stability of the berry extracts at each experimental temperature is ranked as berry extract 3>berry extract 2>berry extract 1, indicating that the berry composition prepared according to the present invention can effectively improve the thermal stability of proanthocyanidins.

[0080] 2. Effect of light on the stability of berry extracts

[0081] A certain concentration of berry extract solution was prepared and placed under an incandescent lamp (lx>200) and dark conditions for a 30-day light stability test. The proanthocyanidin content was measured every 5 days and its retention rate was calculated. The test results are shown in Figure 4 and Figure 5 shown.

[0082] Depend on Figure 4 and Figure 5 It can be seen that the retention rate of proanthocyanidins in berry extracts gradually decreases with the increase of storage days under light and dark conditions, and the rate of decrease is faster under light conditions. Figure 4 and Figure 5 It can be seen that the stability ranking of the berry extracts under light and light-protected conditions is berry extract 3> berry extract 2> berry extract 1, indicating that the berry composition prepared according to the present invention can effectively improve the photostability of proanthocyanidins.

[0083] Effect Example 2

[0084] Antioxidant Properties of Berry Extracts

[0085] 1. Determination of hydroxyl radical scavenging rate

[0086] Take 50 μL of sample in a 96-well microplate, add 0.07 mL H2O2 (6.0 mmol / L), 0.1 mL FeSO4 solution (1.5 mmol / L), and 0.03 mL salicylic acid solution (20 mmol / L), mix well, and treat at a constant temperature for 1 h (37 ° C), measure the absorbance at 510 nm, and calculate the hydroxyl radical scavenging rate according to the following formula, V C As positive control.

[0087]

[0088] Where: Z is the hydroxyl radical scavenging rate, %; A S is the absorbance value of the sample; A0 is the absorbance value of the blank group (deionized water replaces the sample).

[0089] 2. Determination of DPPH free radical scavenging rate

[0090] Take 2 mL of sample with a concentration of 0.01 g / L, add 2 mL of DPPH solution (0.008 g of DPPH solid is dissolved in anhydrous ethanol and diluted to a 100 mL volumetric flask), shake and mix, and measure the absorbance at 517 nm after 30 minutes. C The DPPH free radical scavenging rate was calculated as follows:

[0091]

[0092] Wherein: P is the DPPH free radical scavenging rate, %; Ai is the absorbance value after the sample is mixed with the DPPH solution; Aj is the absorbance value after the sample is mixed with the solvent; Ac is the absorbance value after the DPPH solution is mixed with the solvent.

[0093] 3. Determination of ABTS+ free radical scavenging rate

[0094] Prepare a mixed solution of 7mmol / L ABTS and 2.45mmol / L potassium persulfate with deionized water, store it at 23℃ in the dark for 12-16h to obtain ABTS+ free radical basic liquid. Take 3.9mL ABTS working solution and add 5mL sample, let it stand for 6min, and measure the absorbance at 734nm. V C As positive control.

[0095] Deionized water was used as blank control.

[0096]

[0097] Where: H is the ABTS+ free radical scavenging rate, %; A1 is the absorbance value of the test group; A2 is the absorbance value of the blank group.

[0098] 4. Determination of total reducing capacity

[0099] Take 2.5 mL of sample, potassium ferrocyanide solution (mass fraction 1%), and phosphate buffer (0.2 mol / L, pH 6.6), mix well, place in a 50°C water bath for 20 min, add 2.5 mL of trichloroacetic acid solution (mass fraction 10%) after cooling, shake and mix, centrifuge at 3000 r / min for 10 min, take 2.5 mL of supernatant, add 2.5 mL of deionized water and 0.5 mL of ferric chloride solution (mass fraction 0.1%), mix well, and measure the absorbance at 700 nm.

[0100] Test results such as Figure 6 and Figure 7 shown.

[0101] Depend on Figure 6 and Figure 7 It can be seen that Vc has the best free radical scavenging effect. The hydroxyl free radical scavenging rate, DPPH free radical scavenging rate, and ABTS + The free radical scavenging rate and the total reducing capacity were ranked as follows: berry extract 3 > berry extract 2 > berry extract 1, indicating that the berry composition prepared according to the present invention can effectively improve the antioxidant properties of proanthocyanidins.

[0102] Effect Example 3

[0103] Comparison of the bioavailability of berry extracts

[0104] 1. Draw the standard curve of anthocyanins in berry extract

[0105] 1. Preparation of plasma freeze-dried powder: 30 Kunming mice were taken and divided into 3 groups, half male and half female in each group, 8 experimental groups, 1 blank blood group, and 1 normal saline group. They were fasted but not watered for 12 hours before the experiment. According to the weight of the mice, 0.2 ml / 10 g, 20 mg / ml berry extract was prepared with normal saline. After gavage, an appropriate amount of anticoagulant was added to the EP tube in advance. About 0.5 ml of blood was taken from the fundus at 0.5 h, 1 h, 2 h, 4 h, and 6 h after the time points were designed, and placed in a centrifuge tube. Immediately centrifuged at 4 ° C at 10000 rpm / min for 10 min, 200 μl of supernatant was taken in a 1.5 ml centrifuge tube, 200 μl of acetonitrile was added, and high-speed refrigerated centrifugation was performed at 10000 rpm / 10 min, and 200 μl of supernatant was taken, and 200 μl of acetonitrile was added, and high-speed refrigerated centrifugation was performed at 10000 rpm / 10 min, and 200 μl of supernatant was taken to prepare plasma samples. The plasma samples were frozen at -20°C and then placed in a vacuum freeze dryer to prepare freeze-dried plasma powder.

[0106] 2. HPLC detection of plasma samples: During the experiment, the lyophilized powder was dissolved in the mobile phase D with the same volume as the blood collection volume to mix evenly, and then mixed with a vortex mixer for 1 minute to fully dissolve it. After standing for 1 minute, it was centrifuged at 4°C for 16000rpm / 10min, and the supernatant was taken and filtered with a 0.22μm disposable filter. Then 200μl was taken and put into the injection bottle sleeve, and then put into the injection bottle and put into the HPLC injection plate for detection. The obtained data was calculated using Graphpad Prism 5 software, and its bioavailability values ​​are shown in Table 1.

[0107] Table 1

[0108] Sample / test time (h) 0.5 1 2 4 6 Berry Extract 1 9.21％ 5.12％ 1.02％ 0 0 Berry Extract 2 16.62％ 8.62％ 4.24％ 0.69％ 0 Berry Extract 3 45.53％ 32.69％ 23.12％ 8.31％ 2.11％

[0109] As can be seen from Table 1, at each test time point, the bioavailability of the three berry extracts in the blood of mice after oral gavage was ranked as berry extract 3>berry extract 2>berry extract 1, indicating that the berry composition prepared according to the present invention can effectively improve the bioavailability of proanthocyanidins.

[0110] Effect Example 4

[0111] In vitro skin permeation test

[0112] The vertical Franz diffusion cell method was used to conduct the in vitro mouse skin permeation experiment: the abdominal skin of SD male rats was fixed between the receiving chamber and the supply chamber, 1g of each berry extract was taken in the supply chamber, and 15% propylene glycol, 5% Tween 80 and 80% physiological saline were used as the receiving solution, and the mixture was stirred and diffused at 37°C. 0.5mL of the receiving solution was taken at 1, 2, 4, 6, 8, 10, and 12h, and an equal amount of constant temperature fresh receiving solution was immediately added. HPLC analysis was performed to calculate the cumulative permeation per unit area of ​​specific drugs at different times. After 12h, the skin was removed, washed, cut into pieces, ground into a homogenate, and an appropriate amount of receiving solution was added for centrifugation. The supernatant was taken for HPLC analysis to calculate the skin retention per unit area of ​​specific drugs. The drugs measured in this experiment were B-type proanthocyanidins, A-type proanthocyanidins, A-type proanthocyanidins-amino acid derivatives, and B-type proanthocyanidins-amino acid derivatives. The experimental results are shown in Table 2.

[0113] Table 2

[0114] Group Berry Extract 1 Berry Extract 2 Berry Extract 3 <![CDATA[Skin cumulative permeation amount (μg / cm 2 )]]> 91.2 132.1 246.2 <![CDATA[Skin retention amount (μg / cm 2 )]]> 38.2 46.2 76.8

[0115] From the experimental results in Table 2, it can be seen that the cumulative skin penetration of berry extract 1 after 12 hours is only 91.2 μg / cm2, and the cumulative skin penetration of berry extract 2 after 12 hours is only 132.1 μg / cm 2 The cumulative skin penetration of berry extract 3 after 12 hours was 246.2 μg / cm 2 , indicating that the cumulative penetration of the berry composition prepared according to the present invention in the skin is significantly improved.

[0116] Effect Example 5

[0117] Crowd testing and research

[0118] A berry composition that can be used for scalp care, the components are shown in Table 3, which is a formula table of 3 examples and 5 comparative examples of a berry composition that can be used for scalp care (unless otherwise specified, the unit is weight parts)

[0119] Table 3

[0120]

[0121] A certain mass of Examples 1-3 and Comparative Examples 1-6 were added to ordinary commercial shampoo to prepare samples 1-9 containing 10% of the composition, and ordinary commercial shampoo without the composition was used as sample 10. 300 subjects with similar scalp health conditions were selected and divided into 10 groups, with 30 subjects in each group. The performance of samples 1-10 was tested after using equal amounts of samples 1-10. The test results are shown in Table 4 below.

[0122] Table 4

[0123]

[0124]

[0125] All indicators in the test results of Sample 10 did not show scalp care efficacy.

[0126] By comparing the test results of sample 1 and samples 4 to 5, it can be seen that after adding the berry extract not prepared according to the scheme described in the present invention, the reduction rate of the immediate scalp TEWL value after use and the increase rate of the immediate scalp moisture content are significantly reduced, and the scalp tightness rate, the proportion of people with itchy scalp, the proportion of people with dandruff and the proportion of people with inflammatory reaction are significantly increased, indicating that the berry extract of the present invention can quickly repair the scalp barrier function on the basis of increasing the scalp moisture content and maintaining moisture, and also improves the utilization of the berry extract and the moisturizer.

[0127] Comparing the test results of sample 1 and samples 6 to 8, it can be seen that without adding the composition of the present invention, the reduction rate of the immediate scalp TEWL value after use and the increase rate of the immediate scalp moisture content are significantly reduced, and the scalp tightness ratio, the proportion of people with itchy heads, the proportion of people with dandruff, and the proportion of people with inflammatory reactions are significantly increased, indicating that the berry extract and other components can synergize to make the composition have a good moisturizing effect, can effectively increase the water content of the scalp, repair the scalp barrier, and can reduce the scalp dryness, itching, and scalp tightness caused by surfactant residues. Comparing the test results of sample 1 and sample 9, it can be seen that the berry extract can cooperate with the antibacterial agent to make the composition have a suitable antibacterial effect, which is beneficial to maintaining the balance of the scalp microecology, avoiding the excessive antibacterial effect of the antibacterial agent, inhibiting or killing the beneficial bacteria in the scalp, and destroying the scalp microecology.

[0128] It can be seen that the berry composition of the present invention has a mild formula, is nourishing and moisturizing, has a good antibacterial effect, soothes and relieves inflammation, has a good scalp barrier repair function, can help the scalp resist external stimuli, oil peroxidation and the effects of surfactant residues, and effectively relieves scalp problems such as itchy scalp, scalp inflammation, and tight scalp.

[0129] The above disclosure is only the preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. A method for preparing a berry extract, characterized in that: The following steps are involved: A. Fruit pretreatment to obtain berry freeze-dried powder; B. preparing a deep eutectic solvent; C. mixing the berry freeze-dried powder with the low eutectic solvent, and extracting by shaking to obtain a crude extract of berry proanthocyanidins; D. purifying the crude extract of berry proanthocyanidins using a macroporous resin to obtain a first purified product of berry proanthocyanidins; E. removing the solvent from the first purified product of berry proanthocyanidins to obtain a second purified product of berry proanthocyanidins; F. Dissolving epicatechin, mixing it with the second purified product of berry proanthocyanidins, uniformly heating it under vacuum, releasing the vacuum and cooling it, and then mixing it with amino acid hydrochloride and keeping it warm to obtain the berry extract.

2. The method for preparing the berry extract according to claim 1, characterized in that: The operation of step F includes: dissolving the epicatechin in deionized water, determining the content of anthocyanin in the second purified product of berry proanthocyanidins, mixing the anthocyanin and the epicatechin in a molar ratio of 1:1-2, adjusting the pH value to 2-3.5 with an acid-base regulator, and then evacuating the mixture, wherein the vacuum degree is -0.03-0.10 MPa, and heating uniformly at 85-95° C. for 80-120 min, releasing the vacuum environment, and then cooling the mixture to 60-65° C. with cooling water, dissolving the reaction product and the amino acid hydrochloride in water in a mass ratio of 1:2-4, and continuing to keep warm and stir for 24-36 hours to obtain the berry extract; in step F, the acid-base regulator includes one or more of citric acid, lactic acid, malic acid, sodium citrate, sodium bicarbonate, sodium hydroxide, and potassium hydroxide; and the amino acid hydrochloride includes one or more of L-cysteine ​​hydrochloride, arginine hydrochloride, histidine hydrochloride, and lysine hydrochloride.

3. The method for preparing the berry extract according to claim 1, characterized in that: The operation of step B includes: heating and mixing the amino acid and ethylene glycol to obtain the low eutectic solvent; the molar ratio of the amino acid to the ethylene glycol is 1:5-10; the amino acid includes one or more of arginine, alanine, and lysine.

4. The method for preparing the berry extract according to claim 1, characterized in that: The operation of step A comprises: washing the fruit with running water, draining the water, freeze-drying and grinding, passing through a 60-80 mesh sieve to obtain the berry freeze-dried powder, and storing it in the dark for later use; the operation of step C comprises: adding the berry freeze-dried powder and the low eutectic solvent in a reaction container at a solid-liquid ratio of 1:2-10 g / mL, and starting ultrasonic-assisted oscillation extraction after adding a color protection agent, the ultrasonic power comprises 100-300 W, the extraction temperature comprises 45-55° C., extracting for 30-70 minutes, and then evaporating at 6000 ~10000r / min centrifugation for 15~30min, obtaining a supernatant, and obtaining the crude extract of berry proanthocyanidins; the operation of step D comprises: eluting the crude extract of berry proanthocyanidins in the macroporous resin, the eluent comprises 60-70% by volume ethanol, and obtaining the first purified product of berry proanthocyanidins; the operation of step E comprises: taking the first purified product of berry proanthocyanidins and performing vacuum rotary evaporation and concentration, the temperature control comprises 58~75℃, recovering the eluent, and obtaining the second purified product of berry proanthocyanidins.

5. The method for preparing the berry extract according to claim 4, characterized in that: In step A, the fruit includes one or more of blue indigo fruit, black fruit glandular ribbed aronia, blueberry, black currant, black wolfberry, and blackberry; in step C, the color protection agent includes one or more of ascorbic acid, L-cysteine, citric acid, and malic acid; in step D, the macroporous resin includes one or more of resin S-8, resin AB-8, resin D101, and resin X-5.

6. The method for preparing the berry extract according to claim 1, characterized in that: In step D, the macroporous resin is pretreated before use; the pretreatment includes: soaking the macroporous resin in anhydrous ethanol for 24 hours, rinsing it repeatedly with deionized water after sufficient swelling until the effluent is free of white turbidity, rinsing it with a 0.1 mol / L hydrochloric acid solution to acidity, and soaking it for 3 hours, then rinsing it with deionized water to neutrality, then washing the macroporous resin with a 0.1 mol / L NaOH solution to alkalinity, and soaking it for 3 hours, and then rinsing it with deionized water to neutrality for standby use.

7. A berry extract obtained according to the method for preparing the berry extract according to claim 1.

8. A use of the berry extract according to claim 7, characterized in that: Used to include one or more of cosmetics and cosmetic additives.

9. A cosmetic additive obtained by the application according to claim 8, characterized in that: The composition comprises the following weight percentages: The sugar and its analogs include one or more of glucosamine polysaccharides, saccharide isomers, and β-glucan.

10. The cosmetic additive according to claim 9, characterized in that: The moisturizer includes one or more of glycerol, propylene glycol, butylene glycol, sorbitol, glyceraldehyde, erythrose, maltitol, mannose, thulose, and arabinose; the glucosamine polysaccharide includes one or more of hyaluronic acid, sodium hyaluronate, and sodium chondroitin sulfate; the carbohydrate isomers include water-locking magnetite; the β-glucan includes one or more of yeast glucan and oat glucan; the zinc salt includes PCA zinc, zinc gluconate, zinc chloride, zinc glycinate, zinc glycyrrhetinate, One or more of zinc pyrithione, yeast / zinc fermentation product, zinc sulfide, zinc sulfate, zinc myristate, lactobacillus / milk / magnesium / zinc fermentation lysate, zinc lactate, zinc undecylenate, hydrolyzed collagen zinc, zinc salicylate, zinc carbonate, zinc carbonate hydroxide, zinc aspartate, zinc oxide, and Porphyridium / zinc fermentation product; the antibacterial agent includes one or more of hexamidine di(hydroxyethylsulfonate) salt, salicylic acid and salicylate, piroctone olamine salt, climbazole, and zinc pyrithione.

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