KR101317379B1 - Method for producing microparticle of natural dye with increased stability - Google Patents

Abstract

PURPOSE: A production method of a natural pigment microcapsule is provided to improve the availability of a natural pigment, and to offer natural color and harmlessness to human body. CONSTITUTION: Whey protein is mixed with distilled water and stirred. Antocyanine obtained from mulberries is mixed with distilled water and stirred. The dissolved antocyanine is mixed with the whey protein in a ratio of 1:15-25 (w/w). The mixture is homogenized for 10-20 minutes using a homogenizer, and dried. An antocyanine microcapsule is in a sphere form having the diameter of 100-300 um. A personal care goods contains the antocyanine microcapsule as an active ingredient. The personal care goods is shampoo, soap, cosmetics, dying materials, or hair products.

Description

Method for producing microparticles of natural dye with increased stability

The present invention relates to a method for producing natural pigment microcapsules having enhanced stability, and more particularly, to improve the stability of natural pigments encapsulated by coating a natural pigment with whey protein, prepared by the above method. The present invention relates to a natural pigment microcapsule having enhanced stability, and a food and personal care product containing the natural pigment microcapsule having enhanced stability as an active ingredient.

Mulberry (mulberry) is a fruit of the mulberry (Morus alba L.), a deciduous tree belonging to the genus Mulacea (Moracea) is known as a heartache as a herbal medicine. Audi contains a large amount of glucose and fructose, and contains organic acids such as oxalic acid and citric acid, and anthocyanins and GABA (gamma amino butyric), which have strong antioxidant effects, which are functional ingredients. acid), flavonoids, moracin (moracin) derivatives, DNJ (1-deoxy-nojirimycin), containing resveratrol having an antimicrobial effect has recently increased its consumption as a health food. Audi has an anthocyanin pigment content of 170.47mg / 100g, which is much higher than apple 7.07mg / 100g and grape 48.57mg / 100g. Cyanidin 3-glucoside (C3G) and cyanidin 3-lutinoside (C3R) account for 90% of the anthocyanins of Audi, of which higher cyanidin 3-glucoside (C3G) content is reported. . Audi has been reported to have several effects on physiological functions such as neurostable, vision improvement, memory improvement, anti-aging, anti-hyperlipidemia, antioxidant activity, anti-hepatic toxicity. In general, vegetables and fruits contain natural pigments containing various kinds of anthocyanins, but in order to develop and prove a method for enhancing stability through the manufacture of natural pigment microcapsules, a relatively low anthocyanin type that can accurately represent individual anthocyanin content It was used as a raw material for the production of microcapsules.

On the other hand, the pigments used in various processed foods currently have a great influence on the 'appearance' related to the increase in appetite and decay of humans, and are an important part directly related to the consumer's desire to buy, and increase the degree of perfection in products, It also plays an important role in determining the first impression. Pigments include artificial coloring and natural coloring. Artificial colors are synthetic colors made from coal tar, a petroleum by-product, and natural colors are pigments extracted from nature and contain functional ingredients. It is a pigment, which is classified into three types according to the raw materials, and largely classified into animal, vegetable, and microorganisms. It is reported that pigments used in foods are harmful to humans because they are artificial colors and are made from coal tar, a by-product of petroleum.

With the increase of income and aging population, Hyundai changed to consumption pattern for health and preferred eco-friendly foods rather than general foods, and as modern people gradually pursue eco-friendly life that promotes naturalistic life, they use natural pigments rather than artificial colors. Products have become more preferred, and cosmetics have become more functional. Natural color used in this way is obviously harmless to the human body, and its color is also natural and functional. However, there is a disadvantage in that it is more expensive than tar and does not produce vivid colors when compared with synthetic colors. Above all, the stability of the pigment component is low, there are many restrictions in use. Gardenia pigment, a natural pigment used as a food pigment, has a very stable property and its use is increasing. However, natural pigments such as anthocyanins are easily destroyed by light, oxygen, temperature, pH, sugars, organic acids, and metal ions. .

In order to solve this problem, the present invention intends to apply a technique for encapsulating a pigment component using a protein which is a by-product of a cheese process called whey protein. Encapsulation is the coating of an internal material with a specific external material to protect the internal material from the external environment. The advantages of encapsulation are protection from light (ultraviolet radiation), protection from heat, odor blocking of aromatic ingredients, protection from reaction with other ingredients and above all powders of ingredients which are made only in liquid form.

Meanwhile, Korean Patent No. 0782984 discloses a method for preparing lactic acid bacteria multiple microcapsules, a microcapsule prepared by the method, and a product including the same. In Korean Patent No. 1145018, a water-in-oil type lip containing natural pigments is disclosed. Although a makeup composition has been disclosed, it is not known how to prepare natural pigment microcapsules having enhanced stability as in the present invention.

The present invention was derived by the above requirements, in the present invention, the anthocyanin derived from the natural pigments of the coat was prepared with a whey protein (whey protein) to prepare a microcapsule, the anthocyanin microcapsules (AWM) and the anthocyanin As a result of pH-specific stability assay and optical characteristic assay, the present invention was completed by confirming that it is a method for efficiently producing natural pigment microcapsules having enhanced stability.

In order to solve the above problems, the present invention provides a method for improving the stability of natural pigments, characterized in that the natural pigments are coated with a whey protein to microencapsulate the natural pigments.

In addition, the present invention provides a natural pigment microcapsules with enhanced stability prepared by the above production method.

In addition, the present invention provides a food containing the natural pigment microcapsules with enhanced stability as an active ingredient.

In addition, the present invention provides a personal care product containing the natural pigment microcapsules with enhanced stability as an active ingredient.

The microencapsulation of natural pigments of the present invention can increase the usability of natural pigments, which are often restricted in use due to low stability against light (ultraviolet), temperature, sugar, organic acid or metal ions, and are harmless to human body, and color is also natural. It can be used in the food and cosmetics industry using natural pigments because it can be usefully applied to the consumption pattern of modern society that prefers products using natural pigments.

Figure 1 shows the instrument used for the production of microencapsulation of anthocyanin derived from Audi using Whey protein and the result.
Figure 2 is a result of observing the appearance of anthocyanin-WPI microparticles (AWM) and C3G (Cyanidin-3-glucoside) using a scanning microscope. Cyanidin-3-glucoside (C3G) (A) 500X, (B) 1000X,; Anthocyanin-WPI Microparticles (AWM) (C) 500X, (D) 1000X.
3 shows UV-VIS spectra of anthocyanins, whey proteins and anthocyanins-WPI microparticles (AWM).
Figure 4 shows an analytical chromatogram of anthocyanin-WPI microparticles (AWM) and anthocyanin individual components included in anthocyanins, C3G and C3R. (A) Anthocyanin-WPI microparticle (AWM), (B) Anthocyanin, (C) C3G standard curve, R = 0.9397608, (D) C3R standard curve, R = 0.9029410
5 shows absorbance measurements over pH and time of anthocyanin-WPI microparticles (AWM) and anthocyanins. (A) pH 2, (B) pH 7, (C) pH 10
Figure 6 is a result of comparing the optical properties of 0h, 96h of anthocyanin-WPI microparticles (AWM) and anthocyanin at pH 10. (A) absorbance graph for each wavelength at AWM 0h, 96h; (B) Absorbance graph by wavelength when anthocyanin 0h, 96h
Figure 7 is a time zone anthocyanin release control result of AWM. (A) 0h, (B) 0.5h, (C) 1h, (D) 5h, (E) 10h, (F) 24h, (G) 48h, (H) 72h, (I) 96h, (J) standard C3G (cyanidin-3-glucoside, 100ppm), (K) standard C3R (cyanidin-3-rutinoside, 100ppm)
8 is a time zone anthocyanin release control result of AWM. (A) Contents of C3G (cyanidin-3-glucoside) and C3R (cyanidin-3-rutinoside) over time, (B) Graph enlarged between 0h and 10h

In order to achieve the object of the present invention, the present invention provides a method for enhancing the stability of natural pigments, characterized in that the natural pigments are coated with a whey protein to microencapsulate the natural pigments.

In the method according to an embodiment of the present invention, the natural pigment may preferably be a plant-derived pigment, and most preferably may be an anthocyanin derived from audi, but is not limited thereto.

Whey protein of the present invention is a spherical protein isolated from by-products of cheese making process, contains high levels of bioactive substances, promotes protein synthesis, shows anti-inflammatory and anti-cancer effects It is used as a supplement for health functional food materials or various diseases.

The microencapsulation technique of the present invention is a technique for packaging a desired substance or aroma component in solid, liquid and gas into microcapsules under specific conditions, and can be applied to pharmaceuticals, industrial materials, recording materials, agricultural materials and food fields. Through microencapsulation, it is possible to reduce the loss by protecting unstable substances such as incense burners and nutrients from external factors such as light, oxygen, moisture, and temperature. In addition, by encapsulating additives or useful substances in the food and cosmetic industries, it is possible to prevent oxidation and preservation of materials, to stabilize materials that are susceptible to deterioration, to block unnecessary odors, to control the release rate of materials, to improve manufacturing processes, and to improve physical properties. The effect can be obtained. Although a lot of research has been conducted on the technology of encapsulating vitamins and various trace elements in some food industries, the process of microencapsulating specific active ingredients is carried out only in some pharmaceutical industries, and is generally used to manufacture macrocapsules having a diameter of 5 mm or more. Only relevant studies were activated, which was inappropriate to use as food additives, and was difficult to apply and consume. Capsule coating material should have nontoxic, in vivo compatibility, biodegradable properties, alginate, chitosan, amylose and starch have been mainly used. Whey protein of the present invention is a kind of protein, it is characterized in that the core material is well protected when the capsule is made using it and dispersed in an aqueous solution.

In the method according to an embodiment of the present invention, the natural pigment microcapsules may be preferably a sphere having a diameter of 100 ~ 300㎛, but is not limited thereto.

In the method according to an embodiment of the present invention, the ratio of adding whey protein to natural pigment may be preferably 1:15 to 1:25 (w / w), and most preferably 1:20 (w / w), but is not limited thereto.

In the method according to an embodiment of the present invention, the pH conditions for maintaining the stability of the natural pigments may be preferably pH 2 to 7, and most preferably may be pH 5 to 6, but is not limited thereto. .

In the method according to an embodiment of the present invention, the natural pigment microcapsules may exhibit stability to light (ultraviolet), temperature, sugar, organic acids or metal ions, but is not limited thereto.

In addition, the present invention provides a natural pigment microcapsules with enhanced stability prepared by the above production method.

In addition, the present invention provides a food containing the natural pigment microcapsules with enhanced stability as an active ingredient.

When using the natural pigment microcapsules with enhanced stability of the present invention as a food additive, the natural pigment microcapsules may be added as it is, or may be used with other foods or food ingredients, and may be appropriately used according to conventional methods. . The amount of the active ingredient to be mixed can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, the natural pigment microcapsules of the present invention are added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on the raw materials in the manufacture of food or beverage. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Preferably, the food may be in the form of microcapsules containing natural pigments, and may also be used as an additive to other foods. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

In addition to the above, the natural pigment microcapsules as an active ingredient of the present invention are various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH regulators, stabilizers, Preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the natural pigment microcapsules of the active ingredient of the present invention may contain a flesh for preparing fruit drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the natural pigment microcapsules which is the active ingredient of the present invention.

In addition, the present invention provides a personal care product containing the natural pigment microcapsules with enhanced stability as an active ingredient.

In the personal care product according to an embodiment of the present invention, the personal care product may be a shampoo, rinse, conditioner, soap, cosmetic, dye or hair product, but is not limited thereto.

The personal care product of the present invention refers to a product used directly on the surface of the skin or hair for the purpose of maintaining clean, healthy and beautiful care for all parts of the human body. Personal care products are different from medicines and aim to beautify the health and beauty of a healthy person and to be beautiful. They are safe to use and have no side effects as they are used daily or over long periods in everyday life. . Personal care products include hair care products, shampoos, rinses, soaps, cosmetics, conditioners, mousses, styling gels, hair sprays, hair dyes, hair colorants, hair bleaches, hair waving agents, hair straighteners, Frizz control, hair volumizing, and the like, and include all personal care products in the conventional sense.

Personal care products in the form of tablets or pellets of the present invention also further comprise conventional additives, such as excipients, binders, glidants, and the like, for preparing tablets or pellets without departing from the object of the present invention. Such tablets or pellets may be prepared by conventional manufacturing methods well known in the art. For example, in the case of tablets, the prepared powder or granules are prepared by tableting in a uniform form using a conventional tableting machine. Solvents used in personal care products include water, acid or base ingredients for pH adjustment, thickening ingredients for adjusting the viscosity of the solvent, conditioning ingredients effective for hair and / or skin, catalysts to increase reactivity, chelate ingredients, preservatives, fragrances Or components included in conventional water-soluble preparations such as surfactants for solubilizing the oily conditioning components. More specifically, acetic acid, ammonium bicarbonate, ammonium carbonate, ammonium phosphate, ascorbic acid, benzoic acid, citric acid, formic acid, fumaric acid, maleic acid, potassium bicarbonate, potassium borate, potassium carbonate, potassium citrate, potassium phosphate , Sodium bicarbonate, sodium carbonate, sodium citrate, sodium fumarate, succinic acid, sodium succinate, sodium benzoate, sodium acetate, boric acid, sodium borate, phosphoric acid, sodium phosphate, lactic acid, sodium lactate, Conjugate or counterbase components, such as sodium phosphate (monobasic), sodium phosphate (dibasic), sodium hydroxide, alginate for the viscosity control of solvents, cellulose derivatives, queen seed gum, pectin, flulan, Xanthan gum, non-gum, carboxyvinyl polymer, acrylic acid polymer, lapona It may contain such components as.

In addition, hair personal care products include snopizilia extract, vitamin E additives, ylang-ylang, glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, and PEG (poly ethylene glycol) as an effective conditioning ingredient for hair or skin. Polyhydric alcohols such as (300, 400, 1500, 4000), saccharides such as hyaluronic acid salts, maltitol, and moisturizing components such as pyrrolidone carboxylic acid, and the like, and ester oils or vegetable oil components such as olive oil, jojoba oil and silicone. And various hair or skin conditioning ingredients such as oil components and vitamins, amino acid derivatives, animal and vegetable extracts, rosemary, hyaluronic acid, ceramides, collagen, elastin, cationic polymers, cationic surfactants, proteins, peptides, cedarwood, and the like. . On the other hand, in addition to the main components, solubilizers, fragrances and fragrances for solubilizing the oil components, fragrance and aroma effects, genariol, linariol or vegetable essential oil fragrance, methyl paraben for the microbial stability, ethyl paraben, propyl paraben, methylchloroiso Preservatives such as chiazolinone, methylisothiazolinone, and antifading agents for preventing the discoloration or discoloration of the contents may be contained together, and the composition intended to be applied to the hair or the skin may be any selected from additives conventionally used. A component can be included in the range which does not inhibit the objective of this invention.

In addition,

(a) adding distilled water to whey protein and stirring;

(b) adding distilled water to anthocyanin extracted from Audi and stirring to dissolve it;

(c) homogenizing the mixture prepared by adding the dissolved anthocyanin of step (b) to the stirred whey protein of step (a) for 10-20 minutes using a homogenizer; And

(d) it provides a method for producing an anthocyanin microcapsules with enhanced stability comprising the step of drying the homogenate of step (c).

In the method according to an embodiment of the present invention, the stability of the anthocyanin microcapsules characterized in that the whey protein is added to the anthocyanin of step (c) in a ratio of 1:15 to 1:25 (w / w) Method of preparation.

In the method according to an embodiment of the present invention, the ratio of adding whey protein to the anthocyanin of step (c) may be 1:15 to 1:25 (w / w), and most preferably 1 It may be: 20 (w / w), but is not limited thereto.

In addition, the present invention provides an anthocyanin microcapsules having improved stability prepared by the above production method. The anthocyanin microcapsules may be preferably a sphere having a diameter of 100 ~ 300㎛, but is not limited thereto.

Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Experimental Method

1. Extraction and Purification of Audi Anthocyanin

1) experimental material

After lyophilizing and lyophilizing, 80% EtOH containing 0.1% HCl corresponding to twice the volume of the Audi powder was added to 100 g of the pulverized mulberry powder, sonicated at 38 to 40 ° C for 30 minutes, and only the supernatant was taken to filter paper (Hyundai micro, No. 20, 300 mm), and the filtrate was concentrated under reduced pressure with a 38 ° C. rotary evaporator and then diluted with distilled water containing 0.1% HCl.

2) From Audi  Anthocyanin pigments

Two methods were used for the purification of anthocyanins.

① C18 Sep - Pak  Anthocyanin Pigmentation Using Cartridge

Two C18 Sep-Pak cartridges (Sep-Pak Vac 6 cc, 1 g, C18 Cartridges, Waters) were used for the anthocyanin pigment separation. 10 ml of ethyl acetate, 10 ml of anhydrous methanol, and 10 ml of 0.01 N aqueous HCl were eluted in a C18 Sep-Pak cartridge connected in two layers. 1 ml of Audi extract was added thereto, and 6 ml of 0.01 N HCl was added thereto to remove sugars, organic acids and water-soluble substances, and dried for 15 minutes with nitrogen gas. In order to remove non-anthocyanin, 20 ml of ethyl acetate was added and the eluted solution was obtained in a 20 ml vial, and then sealed and refrigerated. Methanol containing 0.1% HCl was added to extract pure anthocyanin, and the eluted solution was received in a 20 ml vial, and then concentrated under reduced pressure with a 35 ° C. rotary evaporator to dissolve in distilled water.

② Anthocyanin pigment purification using adsorption resin

The adsorption resin used was Pure Resin PB600. Resin (250 ml) activated overnight at 95% EtOH was charged into a glass column (length diameter 625 ml) and washed with 500 ml of 95% EtOH, 100 ml of 70% EtOH, 100 ml of 50% EtOH, 100 ml of 25% EtOH and 800 ml of distilled water. . After the concentration of the Audi extract was adjusted by dilution in the state of removing only EtOH was centrifuged for 15 minutes at 5000g, 4 ℃ to remove internal impurities. 300 ml of the supernatant was taken and loaded on the resin, and then 800 ml of distilled water was used to remove sugars, organic acids, and water-soluble substances. Add more distilled water, so washed with distilled water until tasteless. In order to elute the pigment, the pigment was eluted with EtOH (500 ml) containing 0.5% HCl until the color was not released. In order to reuse the used resin was washed several times with EtOH containing 4% HCl and then reactivated in 95% EtOH was used 3-4 times more. Purified anthocyanin was concentrated on a 35 ° C. rotary evaporator to completely remove the organic solvent and dilute with a small amount of distilled water to encapsulate it.

2. Whey  protein( Whey protein Encapsulation of anthocyanin Derived from Audi encapsulation Formulation development

1) Experimental material

For encapsulation of anthocyanin using whey protein, anthocyanin obtained from Audi was used as a core material, and WPI (Whey protein isolate, HILMAR 9410 INSTANTIZED, USA) was used as the encapsulating material.

2) encapsulation of anthocyanins ( encapsulation )

Each weight was measured using chemical balance to make anthocyanin and WPI (Whey Protein) at a ratio of 1:20. In other words, when anthocyanin is concentrated, the weight of anthocyanin is set to 1, and the weight of WPI is measured 20 times the weight of anthocyanin. Distilled water equivalent to 10 times the weight was added to the WPI, and the mixture was stirred. Anthocyanin was dissolved in a small amount of distilled water. The anthocyanin dissolved in the stirred WPI was added and the mixture was stirred at 300 rpm for 15 minutes using a homogenizer (WiseTis® HG-15A Homogenizer), followed by spray dryer (Flow rate 5ml / min, drying chamber 180 and 95 ° C). Dried.

3) Whey  Encapsulation of anthocyanin derived from audi using protein Encapsulation Investigation of the properties of the preparation

Scanning Electron Microscopy (SEM, S-4300, Hitachi, Tokyo, Japan) was used to observe the appearance of anthocyanin-WPI microparticles (AWM) prepared with anthocyanins and WPI 1:20. In order to observe the stability of encapsulation, the release of anthocyanin in the formulation encapsulated with whey protein, and the optical characteristics, absorbance by wavelength using UV-1650PC UV / VIS spectrophotometer (Shimadzu, Kyoto, Japan, 220-700nm) Was observed.

4) Anthocyanin WPI Microparticle  Encapsulation Efficiency ( Encapsulation  efficiency test

Encapsulation efficiency assay was performed to observe whether anthocyanin and WPI were present in a ratio of 1:20, and anthocyanins and actual encapsulation preparations that were considered to be included in the encapsulation preparation were assayed to test the efficiency. Samples were prepared using 1:20 encapsulated 1000ppm, anthocyanin 47.618ppm, WPI (Whey protein) 952.38ppm (Anthocyanin: WPI = 1: 20 = 47.618 ppm: 952.38 ppm).

① Absorbance Measurement

The absorbance was measured at 526 nm using a UV / VIS spectrophotometer to prepare a sample of the concentration to test the encapsulation efficiency by comparing the absorbance of AWM and anthocyanin. Encapsulation efficiency (% EE) was used in the following formula.

② freedom Radical Scatters  ( DPPH  analysis)

Encapsulation efficiency was assayed by comparing the radical scavenging activity of AWM with that of anthocyanin through DPPH radical scavenging. 400 μl of the sample, 1600 μl of Tris-HCl buffer (pH 7.0), and 2000 μl of 500 mM DPPH solution were reacted in the dark for 20 minutes, and the absorbance was measured at 517 nm using a UV / VIS spectrophotometer. Encapsulation efficiency (% EE) was used in the following formula.

③ HPLC  Encapsulated anthocyanins by analysis WPI Microparticle  Encapsulation Efficiency

In order to assay the encapsulation efficiency of anthocyanin-WPI microparticles, anthocyanins and actual encapsulation preparations that were believed to be included in the encapsulation preparation were analyzed by HPLC. 0.1N HCl was added to the anthocyanin-WPI microparticles, followed by centrifugation at 4 ° C., 15000 rpm for 15 minutes, and all the supernatants were recovered and filtered through a syringe filter (0.45 μm syringe-driven filter unit, Minisart RC 15). The machines used in the analysis included two LC-10AD pumps, SPD10A detector, CTO-10AS column oven, DGV-12A degasser, SIL-10AD autoinjector and YMC-Pack ODS-AM at 35 ° C (AM 12S05-2546WT, AM HPLC system consisting of SCL-10A system regulator (Shimadzu, Japan) with a column of -303, S-5 μm, 12 mm, 250 × 4.6 mm ID, No.0425004334CW) was used. A (1.5% phosphoric acid in water) and B (1.5% phosphoric acid, 20% acetic acid, 25% acetonitrile in water) were used for the mobile phase, and the flow rate was 1.0 ml / min. Mobile phase composition was analyzed for 100 minutes with 25% -5% mobile phase B of the concentration gradient. The chromatogram was measured at 530 nm and the standards were C3G (cyanidin-3-glucoside) and C3R (cyanidin-3-rutinoside) (Sigma). The total amount of anthocyanins (total 6) was obtained by adding all of the anthocyanin individual contents (C3G and C3R) obtained from the samples, respectively, to express the encapsulation efficiency (% EE).

5) Anthocyanins WPI Microparticles  Stability test

In order to investigate the effect of pH on the stability of anthocyanin-WPI microparticles, anthocyanin-WPI microparticles were adjusted to pH 2, 7, 10 using respective buffers. pH 2 is Clark-Lubs buffer (0.2 M KCl + 0.2 M HCl), pH 7 is Macllvaine buffer (0.1 M citrate + 0.2 M NaHPO), pH 10 is Clark-Lubs buffer (0.1 M KCl and HBO mixture + 0.1 M NaOH) was used. After diluting the sample with each pH solution, put 50ml into the vial, seal it, take 4ml of each sample at 0, 0.5, 1, 5, 10, 24, 48, 72, 96h, store in -78 ℃ freezer, and UV Absorbance change was measured at 526 nm using a / VIS spectrophotometer. In addition, the absorbance of each wavelength was scanned using a UV / VIS spectrophotometer to verify the optical characteristics of the difference between 0h and 96h at pH 10.

6) Anthocyanin WPI Microparticles  Emission control

To investigate the release of anthocyanin-WPI microparticles over time, anthocyanin-WPI microparticles were used at pH 7 and at pH 7 Macllvaine buffer (0.1 M citrate + 0.2 M NaHPO). After diluting the sample with a pH solution, 50 ml each was put in a vial, sealed, and 4 ml of each sample was taken at 0, 0.5, 1, 5, 10, 24, 48, 72, and 96 h, stored in a freezer at -78 ° C, and a syringe filter ( 0.45 ㎛ syringe-driven filter unit, Minisart RC 15) was filtered after analysis. The machines used for the analysis included two LC-10AD pumps, SPD10A detector, CTO-10AS column oven, DGV-12A degasser, SIL-10AD autoinjector and YMC-Pack ODS-AM at 35 ° C (AM 12S05-2546WT, AM HPLC system consisting of SCL-10A system regulator (Shimadzu, Japan) equipped with -303, S-5 μm, 12 mm, 250 × 4.6 mm ID, No. 0425004334CW) column was used. The mobile phase was A (1.5% phosphoric acid in water) and B (1.5% phosphoric acid, 20% acetic acid, 25% acetonitrile in water) and the flow rate was 1.0 ml / min. Mobile phase composition was analyzed for 100 minutes with 25% -5% mobile phase B of the concentration gradient. The chromatogram was measured at 530 nm.

Example  One. Whey  Characterization of Encapsulated Formulation of Anthocyanin Derived from Audi Using Protein

One) Whey  Encapsulation Formulation of Anthocyanins Derived from Audi Using Protein

① Anthocyanin using a scanning microscope WPI Microparticle  Appearance observation

The results of anthocyanin-WPI microparticles and C3G (cyanidin-3-glucoside) were examined using a scanning microscope. 3-glucoside) was irregular and entangled with each other (FIG. 2). This confirms that anthocyanins are well surrounded by whey protein. In addition, the particles become finer and more uniform than before they are encapsulated and can be easily absorbed by the human body.

② UV Of VIS  Anthocyanin Using Spectrophotometer WPI Microparticle  Optical property test

Wavelength absorbance was scanned using a UV / VIS spectrophotometer to verify with optical properties whether anthocyanins can be encapsulated in whey protein and preserved stably. Scanning the samples with absorbance 700-220 nm respectively, the anthocyanin showed a specific wavelength at the 260 nm point (Fig. 3). On the other hand, scanning the absorbance of the anthocyanin-WPI microparticles by wavelength showed similar wavelengths to the whey protein. Anthocyanin was encapsulated in the whey protein because no specific wavelength of anthocyanin was detected near 260 nm.

2) anthocyanin WPI Microparticle  Encapsulation Efficiency Assay

① Encapsulation efficiency test through absorbance measurement

Samples of concentrations prepared for assaying the encapsulation efficiency by comparing the absorbance of AWM and anthocyanin were measured at 526 nm using a UV / VIS spectrophotometer. As a result, the encapsulation efficiency was 84.336 ± 2.3216% (Table 1 (a)).

② freedom Radical Scatters  ( DPPH  Analysis) Encapsulation efficiency test through analysis

Efficiency can be assayed by comparing the radical scavenging activity of AWM with the radical scavenging activity of anthocyanin via DPPH radical scavenging ability. As a result of assaying the encapsulation efficiency using the DPPH analysis, the absorbance of the sample was 84.199 ± 5.5609%, similar to that of the measurement of the sample (Table 1 (b)).

③ HPLC  Encapsulated anthocyanins by analysis WPI Microparticle  Encapsulation Efficiency

To analyze the encapsulation efficiency of anthocyanin-WPI microparticles, anthocyanins and actual encapsulation preparations that were considered to be included in the encapsulation preparation were analyzed by HPLC. It was measured somewhat higher (Table 2, Figure 4). It is believed that the reason is that it is more accurate to measure the content of individual anthocyanins by HPLC and to compare with the content of total anthocyanins than that measured by absorbance.

3) anthocyanin WPI Microparticles  Stability test

① UV - VIS  Anthocyanin Using Spectrophotometer WPI Microparticle pH Star stability black

AWM was sampled hourly at pH 2, 7, and 10 for pH stability assays, and the absorbance was measured to analyze trends in UV-VIS spectrophotometry. There was no significant change in absorbance up to 96h, and at pH 7 and 10 it was seen that the absorbance decreased over time (Fig. 5). Therefore, it can be concluded that anthocyanin and AWM can be maintained at the most stable state for a long time at pH 2 than at pH 7 or 10, and that performing at low pH in sample storage or experimentation will prevent the destruction of anthocyanins.

In addition, as shown in FIG. 5, although there was a change in absorbance with time, AWM encapsulated with whey protein showed higher absorbance than anthocyanin which is a control group, and it can be seen that even at high pH, the rate dropped later than anthocyanin. This means that whey protein protects anthocyanins well, so if AWM is used to make cosmetics, it can be preserved for a long time even at pH 5 ~ 6, which is the acidity of cosmetics.

② Anthocyanin WPI Microparticle pH Optical properties according to

We measured the wavelength-specific absorbances of anthocyanin and AWM at 0 and 96 h at pH 10, the largest change in pH-specific stability assay. As a result, in the case of encapsulation, there was no big difference, but in the case of anthocyanin, a change in the specific wavelength of anthocyanin appeared around 260 nm (FIG. 6). In the first 0h, a sharp curve was formed from 260 to 300nm, but in the 96h, a gentle curve was observed, indicating that anthocyanin changed over time. As a result, unlike anthocyanins that are destroyed with pH and time, AWM is well conserved by whey protein because no specific wavelength of anthocyanin is detected.

Example  2. Anthocyanins WPI Microparticle  Emission control

To determine the release of encapsulated anthocyanin over time, AWM release control experiments at pH 7 were carried out in the two anthocyanins present in Audi, C3G (cyanidin-3-glucoside) and C3R (cyanidin-3-rutinoside). ) Was analyzed by HPLC (FIG. 7). As a result, in case of C3G, the content of anthocyanin increased from 0h to 0.5h, and then decreased sharply from 0.5h to 5h and then gradually decreased. In case of C3R, it increased slightly from 1h to 5h and then gradually decreased (FIG. 8). Therefore, at pH 7, both C3G and C3R were released early and anthocyanin was gradually broken.

Claims (12)

(a) adding distilled water to whey protein and stirring;
(b) adding distilled water to anthocyanin extracted from Audi and stirring to dissolve it;
(c) adding the stirred whey protein of step (a) to the dissolved anthocyanin of step (b) in a ratio of 1:15 to 1:25 (w / w), using a homogenizer, Homogenizing for 20 minutes; And
(d) a method for producing anthocyanin microcapsules having a spherical stability, including the step of drying the homogenate of step (c) and having a diameter of 100 to 300 µm. delete delete delete delete Stability is improved by the manufacturing method of claim 1, the anthocyanin microcapsules of the spherical diameter of 100 ~ 300㎛. Food containing the anthocyanin microcapsules of claim 6 as an active ingredient. A personal care product containing the anthocyanin microcapsules of claim 6 as an active ingredient. The personal care product of claim 8 wherein the personal care product is a shampoo, soap, cosmetic, colorant, or hair product. delete delete delete

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