WO2014051353A2

WO2014051353A2 - Microcapsule comprising glycoprotein derived from plants

Info

Publication number: WO2014051353A2
Application number: PCT/KR2013/008632
Authority: WO
Inventors: 임형준; 김한별; 김도훈; 신송석; 박영호
Original assignee: (주)아모레퍼시픽
Priority date: 2012-09-28
Filing date: 2013-09-26
Publication date: 2014-04-03
Also published as: WO2014051353A3

Abstract

The present invention relates to a microcapsule containing a glycoprotein derived from plants. Also, the present invention provides a microcapsule having oxidation ability and which is easy to prepare. Through said antioxidant capacity, oxidation and decomposition of an internally contained material can be prevented. According to the microcapsule of the present invention, the active components, in a form which was previously stabilized by a synthetic polymer derived from crude oil or by a surfactant, can be stabilized by naturally derived components. Also, since an organic solvent is not additionally used during the capsule preparation process, the preparation is simple and eco-friendly.

Description

【Specification】

[Name of invention]

Microcapsule containing plant-derived glycoprotein

Technical Field

The present invention relates to microcapsules comprising plant derived glycoproteins. [Background]

New stabilized capsules are being developed to develop colloidal stabilization systems consisting of edible biopolymers and protein complexes. If the glycoproteins with plant peptide origin have antioxidant power, they will be able to more reliably protect the substances carried in the capsule. In particular, efforts have been made in the art to attempt stabilization by introducing biopolymers derived from nature from the dimension replacing existing petroleum-derived synthetic polymers.

The present inventors have completed the present invention by confirming that capsules containing plant glycoproteins together with polysaccharides and proteins can be prepared simply and conveniently, and have an antioxidant capacity, which greatly increases the stability of the supported material.

[Detailed Description of the Invention]

[Technical problem]

An object of the present invention is to provide a microcapsule having an antioxidant capacity and easy to manufacture.

Technical Solution

In order to achieve the above object, the present invention provides a polysaccharide having a total charge ( _ne t charge) of a negative charge; Provided is a protein having an isoelectric point (PI) of 4 to 6; and a plant-derived glycoprotein having a net charge of negative charge.

Advantageous Effects

The microcapsules of the present invention have the advantage of stabilizing active ingredients of an emulsifier type stabilized by conventional petroleum-derived synthetic polymers or surfactants with naturally occurring ingredients. In addition, in the manufacturing process of the capsule, since the organic solvent is not used separately, the manufacturing is simple and more environmentally friendly. In addition, the microcapsule of the present invention can pursue the stabilization of the potency as a soft microcapsules such as hydrogel, rather than a hard microcapsule, it is useful in showing the effect of the potent component. In addition, the microcapsules of the present invention are reversible generation and breakdown of stable particles according to pH It has the advantage that it can be used as a pH sensitive carrier. The microcapsules of the present invention also have antioxidant properties, and can prevent oxidation and decay of the internally supported materials.

[Brief Description of Drawings]

1 is a result of analyzing the particle size.

2 shows the result of analyzing the surface potential.

3 is a photograph taken to compare the degree of turbidity and precipitate formation.

4 to 6 show results of checking particle size and uniformity. (Scale bar is 10 micrometers)

<π> Figure 7 shows the results of confirming the antioxidant capacity of the plant-derived glycoprotein.

8 shows the results of analyzing the particle size of the ISP nanoparticles (A) and photograph (B) of the ISP aqueous solution.

9 shows Examples 24 to 15 taken to compare the degree of turbidity and precipitate formation.

It is a photo of 26.

10 is a photograph of Examples 23 and 27 to 30 taken to compare the degree of turbidity and precipitate generation.

11 to 15 show results of confirming particle sizes and uniformities of Examples 24 to 26 and 35 to 50 (scale bars are 10 micrometers).

[Best form for implementation of the invention]

In one aspect, the present invention provides a polysaccharide having a net charge of negative charge; Proteins having an isoelectric point (PI) of 4 to 6; and microcapsules comprising plant derived glycoproteins whose net charge is negatively charged.

In the present specification, the 'polysaccharide' is a total charge in the presence of an aqueous solution or moisture.

(net charge) means a polysaccharide having a negative charge, specifically, the 'polysaccharide' in the present specification may have an anionic functional group, such as C00H, but is not limited to the functional group, the charges of the polysaccharide functional group If the total charge has a negative charge, it is applicable without limitation. Specifically, the polysaccharide is pectin, xanthan, bit pectin (beet pectin), carrageenan (carrageenan), ^chitosan, acacia (gum arabic), inulin (inulin), Oz, xanthan gum, methyl selreul, flaxseed gum (flaxseed gum ), Kappa-carrageenan (κ— carrageenan), iota-carrageenan (L—carrageenan), gellan gum, dextran sulfate, galactomannans, alginate, and the like. But it is not limited thereto. In the present specification, the 'protein' refers to a protein having a surface potential that is changed according to pH change, that is, having an isoelectric point. Specifically, the protein of the present specification means a protein having an isoelectric point between pH 4 and 6, and these proteins may have a positive charge at pH 7 and then change to an acidic condition. In the present specification, the 'protein' may have a positive total charge between pH 4 and 6 to form a complex with polysaccharides and plant-derived glycoproteins, thereby producing a microcapsule. In the present specification, the 'protein' may be used as a surfactant. For example, the protein herein may include, but is not limited to, soy protein, casein, ovalbumin, lactoglobulin, and the like.

In the present specification, the isoelectric point of the protein may be 4 or more, 4.2 or more, 4.6 or more, 4.8 or more or 4.9 or more and 6 or less, 5.8 or less, 5.6 or less, 5.4 or less, 5.2 or less, or 5.1 or less.

In this specification, soy protein is not limited to include soy protein, but may include all proteins that can be obtained from beans.

<2i> In the present specification, soybean is a soybean (67yc / / 2e max (L.) Merr.), White kidney beans (/ ¾as? / S multif lorus Wi 1 Id. For.a I bus Bailey), Haenyeo beans (CaAS s / s I i neat a (Thunberg) DC), Pigeon pea, Cajanus cajan (L.) Mi lisp), ^ r \ ^ r (Dipteryx odora t aAAubl.) Willd), Big Fox 0 y cA s / a acuminati folia Makino, Carob beans (Ceratonia siliqua (L.) Taub.), Swallow beans (aZ? / b purpureus (L.) Sweet), Silkworm beans (Vicia faba L.), Egyptian beans (C / ce arietinu L.), °-^-^-(Rhynchosai volubi lis Loureira), ¾ ^■ ° 1 (Rhynchos ia nulubilis), Glycine soja Sieb. et

Zucc), Sweet Pe. Oa Ajr s odoratus Linne), 1] (Amphicarpaea bracteata subsp.edgeworthii (Benth.) H.Ohashi), Rake (7c / a angustifol ia Linne var. Segetilis (Thui 11.) K. Koch.), ^ ^ Lathurus vaniotii Leveille.), Bijindo bean (Dumas iat rune at a Sieb. et Zucc.),-^-^^^^-(Phaseolus multif lous Wild.), Bee ^ ¾ "° 1 (Lotus corniculatus Linne var. japonicus Rege 1), Baekdu 0¾ // ¾c» s lab lab L. ), ^u l ^" ^ ^" Sfl ^^-(Amphicarpaea bracteata), fountain bean 0¾cAras 3 japonica Hook fi 1. ex Regel.), Lee] "bean 0¾as«? / i / s lunatus L.), lentil culinaris Medik ), ^^ ^■ (Arachis hypogaea L.), ^-^ \ (Glycine max Merrill),-(Glycine soja Sieb. et Zucc), Dried Canava a gladiata (Jacq.) DC), Honeysuckle Beans (/ ¾aseo / "s vulgaris), Beans (^ σ // 7ί / 5 luteus), Green Bean radiatus L. var. aurea), Black beans (67yc /? E max (L.) Merr.), And kidney beans 0¾aseo / s vulgaris L.).

The soy protein or soy protein in the present specification may be an isolated soy protein (ISP). In the present specification, the 'plant-derived glycoprotein' is also a net charge in the presence of an aqueous solution or water. Means negatively charged glycoprotein. Specifically, in the present specification, the 'plant-derived glycoprotein' may have an anionic functional group, for example, C00H, to the functional group of the sugar or protein, but is not limited thereto, and the total charge sum of the charges of the functional group has a negative charge. If so, it can be applied without limitation. Specifically, the plant-derived glycoprotein may include, but is not limited to, Virginia-derived glycoprotein, green tea-derived glycoprotein, ginseng-derived glycoprotein, pine needle-derived glycoprotein, Rhodiola-derived glycoprotein, and Abis-derived glycoprotein. It is not. The 'plant-derived glycoprotein' of this specification also has an antioxidant ability, thereby preventing the oxidation of the material carried in the microcapsules.

Microcapsules of one aspect of the present invention can be stably prepared in combination with a positively charged protein, a negatively charged glycoprotein and a polysaccharide under acidic conditions. Glycoprotein as a natural component plays a role of a surfactant, so there is little irritation, and since it does not require a separate organic solvent for its preparation, it is easy to manufacture and environmentally friendly. In addition, the microcapsules, which is an aspect of the present invention, have the advantage that particles which are stable according to pH are reversibly generated and decomposed, and thus can be used as a pH sensitive carrier.

In the microcapsules of one aspect of the present invention, the plant-derived glycoprotein is Virginia-derived glycoprotein, green tea-derived glycoprotein, ginseng-derived glycoprotein, pine needle-derived glycoprotein, Rhodiola-derived glycoprotein, and Avis-derived sugar. At least one selected from the group consisting of proteins. The glycoprotein has an antioxidant ability to prevent oxidation and decay of the water contained in the capsule, as well as less irritation as a natural derived material.

In the microcapsules of one aspect of the present invention, the plant-derived glycoprotein may have a carboxylic acid functional group. The plant-derived glycoprotein may have a carboxylic acid functional group on a sugar or protein.

In the microcapsules of one aspect of the present invention, the polysaccharide comprises pectin, xanthan, beet pectin, carrageenan, chitosan, gum arabic, inulin, and methyl cells. Rhodes, xanthan gum, flaxseed gum, kappa— ^ 1 "la ^' κ-carrageenan, ^{0 0} ] Ota-carrageenan (ι -carrageenan), gellan gum (gellan) gum, dextran sulfate, galactomannans, and alginate. Specifically, the pectin may be high methoxy pectin (HME). The high or low hydroxypectin is determined by how much methyl carboxyl groups are substituted by the carboxyl groups in the main chain. In the present specification, the high methoxy pectin (HME) refers to a pectin in which at least 50% of carboxyl groups are substituted with methyl ester groups. Specifically, the high methoxypectin (HME) may refer to pectin in which carboxyl groups of 69 to 74% are substituted with methyl ester groups.

In a microcapsule which is an aspect of the present invention, the protein is a soy protein.

(soy protein), casein, ovalbumin and lactoglobulin may be one or more selected from the group consisting of.

In the microcapsule which is an aspect of the present invention, the weight ratio of the polysaccharide to the protein may be 1 to 9: 1. When preparing the microcapsules in the weight ratio, it is possible to produce a larger number of microcapsules having a more uniform particle size. In view of the above, the weight ratio of the polysaccharide: protein is from 1 to 8: 1, from 1 to 7: 1, from 1 to 6: 1, from 1 to 5: 1, 1 to 4: 1, 1 to 1 to 1 or 1 ˜2: 1, specifically 1-2.3: 1.

In the microcapsules according to the aspect of the present invention, the weight ratio of the polysaccharide: protein: sugar protein may be 1 to 9: 1: 0.01-0.99. When the microcapsule is produced at the increase ratio, a larger number of microcapsules having a more uniform particle size may be prepared, and the antioxidant capacity of the microcapsule may be maximized. In view of the above, the weight ratio of the polysaccharide: protein: sugar protein may be 1-7: 1: 0.01 0.9, 1-5: 1: 0.01-0.8, or 1-3: 1: 0.01-0.7. _.

In the microcapsules of one aspect of the present invention, the microcapsules may include a fat-soluble substance in the coating. Microcapsules, which are aspects of the present invention, have an antioxidant capability, and have an advantage of preventing oxidation and decay of the fat-soluble substance loaded therein.

In the present specification, the 'lipophilic material' includes a material which is not mixed with the water-soluble material and is separated and exists as a layer, and may include an extract or a compound having a fat-soluble property. The fat-soluble substance may mean a substance capable of producing a desired effect with an active ingredient or an active ingredient. In the manufacture of cosmetics, medicines or health foods, the active ingredients having fat-soluble properties are supported, manufactured, distributed and sold. If you do, you will be able to prevent oxidation and will have fewer side effects, including plant-derived glycoproteins. Specifically, the 'fat-soluble substance' in the present specification is not limited as long as it is a soluble active ingredient that can be dissolved in ester-based oil, triglyceride-based oil, silicone-based oil, or hydrocarbon-based oil, the efficacy of terpenoid-based, flavonoid-based It also contains ingredients.

In microcapsule which is an aspect of the present invention, the fat-soluble substance is coenzyme

It may be one or more selected from the group consisting of Q10, Carnosic acid, omega-3 and beta carotene.

In another aspect, the present invention,

(A) mixing an oil soluble substance and water to prepare an oil drop;

(B) the oil drop and

Polysaccharides with a net charge of negative charge;

A protein having an isoelectric point (PI) of 4 to 6; and

The present invention relates to a method for preparing microcapsule, comprising reacting a plant-derived glycoprotein having a net charge with a negative charge at pH 4 to pH 6.

In one aspect of the invention, the step (a) and (b) has a pH of 6 to

It may be performed in the state of seven.

In the method which is one aspect of this invention, the said method is

(C) lowering the second emulsified emulsion to pH 4 to 5.5 to form a microcapsule.

In one aspect of the invention, the pH of step (c) is pH 4.2 to 5.5, pH 4.4 to 5.5, pH 4.5 to 5.5, pH 4.6 to 5.4, pH 4.7 to 5.3, pH 4.8 to 5.2 or pH 4.9 to 5.1. Preferably the pH of step (c) is around 5, specifically, the pH may be 4.8 to 5.2.

In the present specification, the 'water-soluble material' may include a material that is not completely integrated with the fat-soluble material and is present in different layers. Specifically, the water-soluble substance of the present specification may include water, but is not limited thereto.

In the present specification, the 'oil drop' means that the oil-soluble substance is present as a drop, that is, a drop between the water-soluble substances. Microcapsules, which is an aspect of the present invention, may surround the oil drop with a surfactant to make the oil drop's internal environment more completely anhydrous, and a polysaccharide such as high viscosity pectin is added thereto. Water-soluble by forming bonds Intrusion of powder can be prevented more strongly.

In the manufacturing method of a microcapsule which is an aspect of the present invention,

Step (b) may include reacting the polysaccharide and the plant-derived glycoprotein after reacting the oil drop with the protein. In the preparation method, the oil drop and the protein are first reacted to provide a more powerful anhydrous environment. In the manufacturing method of the microcapsules, which is an aspect of the present invention, the polysaccharide and plant-derived sugar The step of reacting the protein may first react the plant-derived glycoprotein and then react the polysaccharide. When the plant-derived glycoprotein is reacted first, the plant-derived glycoprotein is present closer to the supported material, thereby maximizing the antioxidant capacity of the supported material, and the polysaccharide with the negative charge is present externally. Because of the characteristics of the particles having a surface having a negative charge it is possible to prevent the coalescence between the particles, etc. has the advantage of increasing the stability of the particles.

In the method of manufacturing a microcapsule, which is an aspect of the present invention, the fat-soluble substance of step (a) may include a mixture of oil-soluble substance and oil. It may include carbon-based, triglyceride-based or ester-based oils, and any substance capable of dissolving the active ingredient fat-soluble substance to a high content can be used without limitation. In the method of producing a microcapsule, which is an aspect of the present invention, the plant-derived glycoprotein is Virginia-derived glycoprotein, green tea-derived glycoprotein, ginseng-derived glycoprotein, pine needle-derived glycoprotein, Rhodiola-derived glycoprotein, and Abis-derived. At least one selected from the group consisting of glycoproteins.

In the method of preparing microcapsule, which is an aspect of the present invention, the plant-derived sugar protein may include a carboxylic acid functional group.

In the method for preparing microcapsule, which is an aspect of the present invention, the polysaccharide is pectin, xanthan, beet pectin, carrageenan, chitosan, gum arabic, inulin ), Methyl salorose, xanthan gum, flaxseed gum, ^！ ^" ᅳ ^ 1" ^raginan (κ ᅳ carrageenan), ^ό 1 " ⁰ 1 ota_garrageenan (ι_ carrageenan), gellan gum (gel lan gum), dextran sulfate, galactomannans, and alginate.

In the method of manufacturing a microcapsule which is an aspect of the present invention, the protein is a group consisting of soy protein, casein, ovalbumin, and lactoglobulin. It may be one or more selected from.

In the manufacturing method of microcapsule which is one aspect of this invention, the said polysaccharide: Protein The increase in quality can range from 9 to 1: 1. When preparing the microcapsules in the weight ratio, it is possible to produce a larger number of microcapsules having a more uniform particle size. In view of the above, the weight ratio of the polysaccharide: protein is from 1 to 8: 1, 1-7: 1, 1 to 6: 1, 1 to 5: 1, 1-4: 1, 1-3: 1 or 1 ˜2: 1, specifically 1-2.3: 1.

In the method of preparing a microcapsule, which is an aspect of the present invention, the weight ratio of the polysaccharide: protein: sugar protein may be 1 to 9: 1: 0.01 to 0.99. When preparing the microcapsules by the weight ratio, it is possible to produce a larger number of microcapsules having a more uniform particle size, the anti-oxidation capacity of the microcapsules can be maximized. In view of the above, the weight ratio of the polysaccharide: protein: sugar protein may be 1 to 7: 1: 0.01-0.9, 1 to 5: 1: 0.01 to 0.8, or 1 to 3: 1: 0.01 to 0.7.

In one aspect, the present invention may be an emulsion including microcapsules prepared according to the method for preparing microcapsules or microcapsules according to the present specification.

In one aspect of the invention, the protein may be 0.1 to 1% by weight based on the total weight of the final composition comprising the microcapsule. Also in one aspect of the invention, the protein is 0.01 to 5% by weight, 0.05 to 4.5% by weight, 0.1 to 4% by weight, 0.2 to 3.5% by weight based on the total weight of the final composition comprising microcapsule , 0.3 to 3% by weight, 0.4 to 2.5% by weight, 0.5 to 2% by weight, 0.6 to 1.5% by weight or 0.7 to 1% by weight.

In one aspect of the invention, the polysaccharide may be from 1 to 3% by weight based on the total weight of the final composition containing the microcapsules. In addition, in one aspect of the present invention, the polysaccharide is 0.1 to 5% by weight 0.5 to 4% by weight, 1 to 3.5% by weight, 1.3 to 3% by weight, 1.6 based on the total weight of the final composition comprising microcapsule To 2.7 weight percent or 2 to 2.4 weight percent.

In one aspect of the invention, the fat-soluble substance comprises a final microcapsule

^' Can be from 0.01 to 30% by weight based on the total weight of the composition. In addition, according to one aspect of the present invention, fat-soluble ^substance, 0.001 to 30% by weight based on the total weight of the final composition, 1 to 25% by weight, 10 to 20 parts by weight ¾ containing microcapsules>, 12 to 18 increases It may be an amount% or 14 to 16% by weight.

Another aspect of the present invention is a method of manufacturing microcapsule, the method

I) Performing primary emulsification with a protein having an isoelectric point (PI) of 4 to 6 Preparing a primary oAv emulsion; And

Ii) adding a polysaccharide having a net charge of negative charge and a plant-derived glycoprotein having a net charge of negative charge in the prepared primary emulsion to secondary emulsification;

It relates to a method comprising a.

In one aspect of the invention, the method may be performed at room temperature.

In one aspect of the present invention, the steps i) and ii) may be performed at a pH of 6 to 7.

In a method of one aspect of the invention, the method

Iii) lowering the second emulsified emulsion to pH 4 to 5.5 to form microcapsules.

In one aspect of the invention, the pH of step iii) is pH 4.2 to 5.5, pH 4.4 to 5.5, pH 4.5 to 5.5, pH 4.6 to 5.4, pH 4.7 to 5.3, pH 4.8 to 5.2 or pH It may be 4.9 to 5.1, preferably the _P H of the step Hi) is around 5, specifically may be pH 4.8 to 5.2.

Hereinafter, the present invention will be described in more detail with reference to examples. These examples are only for illustrating the present invention, and the scope of the present invention is not interpreted to be limited by these examples. It will be obvious to those of ordinary skill in the industry.

[67] [Example]

Example I Preparation of HMP / SC Capsule

The experiment was conducted with the following composition. High-methoxy pectin used in

Genuee Pectin (CP Kelco, Denmark), which is a pectin with 69 to 74% carboxyl group substituted with methyl ester, and Sodium caseinate is obtained from American Casein Company.

<70> [Table 1]

<71> [Table 2]

<? 2> HMP powder was dissolved in water with stirring to prepare a 3% aqueous solution.

1 ~ 3¾> is suitable). Since pH is low, HOH 3% aqueous solution was prepared by adding NaOH and adjusting to pH 7. The SC powder was dissolved in water with stirring to prepare a 3> aqueous solution (the concentration of SC was adjusted below the HMP concentration). After agitation, the micelle water solution composed of SC is prepared and has an average particle size of about 100-200 nm. The pH is prepared in the vicinity of 7 and when the pH is high or low, the NaOH or citric acid was adjusted to pH 7 using the prepared SC 3% aqueous solution. The HMP 3% aqueous solution and SC 3% aqueous solution prepared above were mixed by ratio (9: 1/7: 3/5: 5) to make a mixed aqueous solution and stirred.

While stirring the mixture at a constant speed, the aqueous solution of citric acid is slowly added while the pH is lowered to around 5, and the negative charge on the surface of the SC micelle is changed to positive charge, forming a complex with the negative charge of HMP. A capsule is produced (HMP coated on the SC micelle surface).

[Example Π] HMP / SC / glycoprotein Capsule Preparation

Example 75 In the process of mixing the HMP aqueous solution and the SC aqueous solution of the production method of Example I, the green tea-derived glycoprotein powder (ISAI-016, Dure Co., Ltd.) was completely dissolved together and the pH was adjusted to adjust the pH of the green tea-derived glycoprotein. Preparation of microcapsules introduced as a capsule component Was

<76> [Table 3]

<77> [Table 4]

[Example m] Preparation of HMP / SC / glycoprotein capsule having oil in the particle core Instead of the aqueous SC solution used in the above example, an aqueous solution of o / w emulsion form using SC as a surfactant was used. For the preparation of this, the oil is added to the water first, and an aqueous solution having oil drops is made through a homer mixer, and a certain amount of SC powder is added to the homer for mixing with homer. Was prepared.

Table 5

<81> [Table 6]

Example IV Preparation of Isolated Soy Protein (ISP) Aqueous Solution

While stirring the water at a suitable speed with an _ag mixer at room temperature, Fuji Pro NKJILIN FUJI PROTEIN, China, an isolated soy protein, was added to the final composition of the aqueous solution to 3% by weight. It was left to stand for 6 hours while stirring at room temperature. The prepared suspension was separated using a centrifuge to dissolve in water (7000 rpm, 30 minutes). Then, the submerged portion was discarded without dissolving in water and only the supernatant was taken. The ISP aqueous solution thus obtained contained 1.8% by weight of soy protein.

Example V Preparation of HMP / ISP Capsule

The experiment was conducted with the following composition.

<86> [Table 7]

MP powder was dissolved in water with stirring to prepare a 3% aqueous solution (concentration of HMP

1 ~ 3% is appropriate), since the pH is low, NaOH was added to adjust the pH to near 7 to prepare a HMP 3% aqueous solution. The HMP 3% aqueous solution prepared above and the ISP 1.8% aqueous solution prepared according to Example IV were mixed (1: 1/2: 1/4: 1) by weight ratio to make a mixed aqueous solution and stirred.

While stirring the mixture at a constant speed, slowly adding a citric acid solution and lowering the pH to around 5, the negative charges on the surface of the ISP nanoparticles are reduced and the negative charges of HMP and HMP are relatively high. Microcapsules are produced by forming a complex (HMP is coated on the surface of ISP particles or ISP particle clusters).

Example VI Preparation of HMP / ISP / Glycoprotein Capsules

In the process of mixing the HMP aqueous solution and the ISP aqueous solution in the preparation method of Example V, the green tea-derived glycoprotein powder (ISAI-016, Dure Co., Ltd.) was dissolved with 0.01-0.1% by weight. As the pH was adjusted (using 1M citric acid), green tea-derived glycoprotein was prepared as a microcapsule. The sugar portion of the green tea-derived glycoprotein used here is neutral sugar (49.3 weight¾>) + uronic acid (50.7 Weight percent) (the negative charge of uronic acid is thought to be complex with the positive charges on the surface of the ISP nanoparticles).

Table 8

Examples 27 to 30 according to Table 9 correspond to samples in which 1M citric acid was slowly added while stirring the aqueous solution of ISP prepared in Example 23, and the pH was lowered to around 6, 5.5, 5, and 4.5, respectively.

Table 9

[Example VII] Preparation of HMP / ISP (/ glycoprotein) Capsule-Emulsion with Oil in Particle Core

Instead of the aqueous ISP solution used in the above example, an aqueous solution in the form of an o / w emulsion using ISP as a surfactant was used. A primary o / w emulsion was prepared using a Homer mixer while a certain amount of oil was added to an ISP aqueous solution. After the first emulsification, HMP aqueous solution (+ glycoprotein) is added to make a mixture, and the aqueous solution of citric acid is slowly added and the pH is lowered to around 5. As the negative charge of the glycoprotein forms a complex, an HMP-coated capsule or emulsion is prepared having an oil core primarily emulsified with the ISP. It is possible to reduce the size of the emulsion particles by using a high pressure emulsifier to reduce the particle size to increase the stability of the emulsion.

<96>

Examples 31 to 34 according to Table 10 below are CEH as an ester oil, CSA as a triglyceride oil, DC200 100cs as a silicone oil, and L L14E as a hydrocarbon oil. It was prepared by homomixing (5 min, 7500 rpm) while adding oil.

<98> [Table 10]

Examples 35 to 38 according to Table 11 of Example 1 to Examples 31 to 34

Add HMP aqueous solution and green tea-derived glycoprotein to make a mixture, homomix (5 min, 5000rpm) to lower _P H with citric acid and electrostatic bond on the surface of o / w emulsion particles firstly emulsified by ISP. It was prepared by fixing the glycoprotein and HMP using.

<ιοο> This process can increase the stability of emulsified particles using ISP.

Structural characteristics of the emulsion with multi-layer structure and negative charge by HMP on the outermost layer can greatly improve the stability of o / w emulsification using only ISP.

<ιοι> 【Table 11】

Examples 39 to 42 according to Table 12 below were fixed with csa, a triglyceride-based oil, and given a change in the oil / ISP / HMP content to observe the size change of the finally produced emulsion particles. ISP was used as a primary emulsifier and the HMP and green tea-derived glycoproteins were coated on the surface of the emulsion particles thus formed to maintain higher HMP content than ISP content. The method for preparing this was the same as in the 35 to 38 in the embodiment.

<103> [Table 12]

Example 43 according to Table 13 is the same as the preparation method of Examples 35 to 3S 15% of three kinds of triglyceride oils are first emulsified with ISP, and correspond to a sample coated with primary milk particles with HMP and glycoprotein. Example 44 is a sample using an additional high-pressure emulsifier to reduce the particle size of the final emulsified particles of Example 43, the high-pressure emulsifier used a model name APV 2000, as soon as the preparation of Example 43 to 1000 bar using APV 2000 High pressure emulsification was turned 4 cycles.

<105> [Table 13]

Examples 45 to 47 according to Table 14 change the CSA content to 5%, 10%, and 15%, and the ratio of ISP and HMP is fixed, and Examples 48 to 50 are only kinds of triglyceride oils. The oil content of 15% and ISP and HMP were fixed and the production conditions and methods were the same as those of Examples 35 to 38 and the composition was the same as below. In addition, all of Examples 45 to 50 additionally used a high pressure emulsifier.

<107> [Table 14]

Test Example 1 Particle Size Analysis

Particle size analysis of SC 1.5% aqueous solution prepared in Example 3 and ISP 1.8% aqueous solution prepared in Example IV (pH 6, 3) was carried out using a dynamic light scattering apparatus (device name: Malvern Corporation Nano-ZS). It was.

As a result, in case of 1.5% aqueous solution of SC (Fig. 1), average particle size was 262.6 nm, PE ⁾ I was obtained 0.289, and SC formed SC micelles with the following particle size distribution in aqueous solution. .

<ιιι> Also, in case of ISP 1.8% aqueous solution (A of FIG. 8), the average particle size was 287 nm and PDI was obtained at 0.428, and ISP confirmed that ISP micelles were formed in the below particle size distribution in aqueous solution.

Test Example 2 Surface Potential

<ιΐ3> The SC micelle surface potential of the SC 1.5% aqueous solution prepared in Example 3 and the ISP micelle surface potential of the ISP 1.8% aqueous solution prepared in Example IV (pH 6.3) were measured using a dynamic light scattering device (registered name: Malvern Corporation Nano). -ZS), using an automatic titrator, lowered the pH of the sample near the initial pH to pH 5 and confirmed the SC micelle surface potential at a specific pH.

As a result, in the case of the SC micelle surface potential (FIG. 2), it can be seen that the surface potential of the SC micelle having a negative surface potential at high pH is changed to a positive charge near the isoelectric point 4.89 as the pH is lowered.

<ιΐ5> Also, for ISP micelle surface potential, -12.9 mV (SD: 3.02) at pH 7, -9.33 mV (SD: 2.69) at pH 6, -6.31 mV (SD: 4.08) at pH 5.5, and pH 5 It was confirmed that-(). 86 mV (SD: 6.65) at -4.54 mV (SD: 5.64) and pH 4.5. In the case of ISP micelles, as with SC micelles, the negative charge tends to decrease gradually as the pH is lowered.

<116>

<Π7> [Test Example 3] Turbidity and precipitate formation

As a result of photographing with a digital camera in order to compare the turbidity and precipitate generation degree of the above embodiment, the same results as in FIGS. 3, 9 and 10 were obtained.

Examples 1 and 2 were prepared in a translucent aqueous solution. Example 3 confirmed that the precipitate occurred in Example 4, while the semi-transparent sample.

In Examples 5-7, the translucent aqueous solution was shown, but in Examples 8-10, the sample appearance changed in the form of an opaque emulsion. This is because the positive charge of HMP complexes with the negative charge of SC micelles and coats the surface as the pH is lowered. In addition, as the HMP: SC concentration ratio was changed from 9: 1 to 5: 5, the amount of complex formation increased, and thus samples with high turbidity were obtained. <i2i> Examples 12 and 13 had a slight color change according to the pH change, but the apparent difference could not be confirmed. On the other hand, in Examples 14 and 15 it was confirmed that the turbidity changes according to the pH change and a small amount of precipitate is formed. Therefore, it was confirmed that a specific complex was formed between green tea glycoprotein and SC micelle.

According to Examples 16 and 17, it was confirmed that the three components of HMP / SC micelles / green tea glycoproteins together form a microcapsule having a predetermined particle size.

In addition, as shown in the photographs of Examples 18 to 20, the microcapsules of the present invention showed a similar appearance to general emulsion dogs, and no phenomenon such as precipitation / separation / creaming was observed.

The photographs of Examples 21 and 22 showed that HMP / SC / tea green tea glycoprotein microparticles were well formed without sedimentation / separation / creaming.

In addition, as shown in the photographs of Examples 24 to 26 (Fig. 9), the amount of the composite is formed as the weight ratio of P: ISP increases from 1: 1 to 2: 1 ᅳ 4: 1. As the turbidity decreased, samples with low turbidity were obtained. According to Examples 24 to 26, it was confirmed that three components of Li P / ISP nanoparticles / green tea glycoproteins constitute microcapsules having a constant particle size.

As can be seen in the photographs of Examples 23 and 27 to 30 (FIG. 10), the ISP aqueous solution can be confirmed to precipitate and precipitate while losing hydrophilicity as the pH is lowered from 7 to 4.5. This is because the negative charges in the ISP's positive and negative charges are combined with the hydrogen cations by the pH drop and lose their hydrophilicity as they lose their charges. On the other hand, if the ISP and HMP aqueous solutions are mixed in a certain ratio and the pH is lowered, The ISP is not precipitated and the positive charges of the ISP and the negative charges of the HMP form a subsequent composite, thereby stably dispersing microparticles as shown in FIG. 9.

<127>

Test Example 4 Confirmation of Particle Size and Uniformity

The formation and the structure of the microparticles were confirmed by optical microscopic observation of the solution containing the microcapsule prepared by each of the above examples.

The optical microscopy images (FIG. 4) corresponding to Examples 8 to 10 showed that the frequency of the generated microcapsules was changed as the ratio of HMP: SC was changed. As the pH was adjusted, it was found that a stable and relatively uniform particle size microcapsule (in the form of HMP coated on SC micelles) was prepared. <i3i> As can be seen from the optical micrograph (FIG. 5) corresponding to Example 15

Particles found in the microcapsule optical images of the HMP and SC complexes were also identified in the green tea glycoproteins and the SC complexes. This indirectly indicates that some green tea glycoproteins can be coated on the surface of SC micelles through ionic bonds. However, it was difficult to observe uniform, spherical and uniform particle size distributions, such as microcapsule through HMP and SC complex. On the other hand, as shown in the optical micrograph (FIG. 5) corresponding to Example 17, the microcapsules prepared by the combination of HMP / SC / tea green tea glycoproteins were spherical microcapsules having a very uniform particle size. It can be seen that manufactured. Therefore, it was found that green tea glycoprotein and HMP were uniformly coated by ionic bond on SC micelles to form capsules.

In addition, looking at the optical microscope image (Fig. 6) of Example 18, it can be seen that the emulsified particles have a particle size of several micrometers ~ 10 micrometers by the SC and has an o / w emulsion dog form. there was. Looking at the optical microscope image (FIG. 6) of Example 20, it can be seen that the particle size of the particles compared with Example 18 is slightly increased, it was also confirmed the multi-layer structure seen as a coating layer of HMP.

In addition, the optical image (FIG. 6) corresponding to Example 22 was confirmed that the microcapsule of the P-SC / green tea glycoprotein containing CSA in the core was well prepared.

34) In particular, both Examples 20 and 22 from the core of the capsule to CSA (oil), SC micelles, 腿 P (+

Green tea glycoprotein) was confirmed to have a multi-layered structure.

In the optical microscope image (FIG. 11) of Examples 24 to 26, HMP aqueous solution and ISP

It can be seen that the particle size of the resulting ion complex microcapsules changes according to the mixing ratio of the aqueous solution. From Example 24 to 26, it can be seen that the amount of ISP that can be considered to form the core side of the capsule is smaller, and the particle size of the microcapsule observed by the microscopic image is also reduced according to this tendency. Therefore, it was confirmed that the larger the amount of ISP, the larger particles are formed, and vice versa, smaller particles are formed. In addition, when the negative charges of ISP particles are combined with hydrogen cations and lose their hydrophilicity, these micrographs show that when HMP is present, the negative charges of HMP and positive charges of ISP are ion-bonded to coat the ISP, resulting in spherical structure. Eggplant was again confirmed that the water is dispersed in capsule form.

Example 35 The optical microscope image (Fig. 12) of the magnetic domain 38 shows that the emulsified particles of Examples 35 and 36 using triglyceride-based or ester-based oils used Example 37 and silicone-based or hydrocarbon-based oils. Remarkably smaller than 3 emulsified particles Confirmed. Therefore, in general, the smaller the emulsified particles can be considered to increase the overall stability of the emulsification system, so using emulsified particles (microcapsules) and emulsions containing triglyceride-based or ester-based oils It was confirmed that it was advantageous in terms of emulsion stability.

The optical microscope images (FIG. 13) of Examples 39 to 42 showed that the emulsion particles or emulsions having the composition of Example 40 were the most advantageous in terms of the degree of emulsification. . The content of ISP covered by the experiments of Examples 39 to 42 was about 0.178% to 0.81% based on the final content, the HMP content was about 1.35 to 2.4%, and even lower than this to prepare a high content Emulsified particles that appear to be sufficiently stable are expected to be produced. However, the higher the content of ISP / HMP is expected to be able to produce a more stable emulsion particles, but considering the final product and the formulation used, it is not recommended to raise the content at random. Therefore, in the case of an emulsion or the like, the oil content is about 0 to 30% by weight, the ISP content is about 0.1 to 1% by weight, and the HMP content is about 1 to 3% by weight based on the total weight of the final product or the formulation. It is considered to be preferable.

Looking at the optical microscope images (FIG. 14) of Examples 43 and 44, it could be seen that in Example 44 using a high pressure emulsifier, the size of the emulsified particles is much smaller than that of Example 43 which is not. In the case of treatment, it was confirmed that the final emulsion stability increased. In addition, in the case of Example 44, it was possible to obtain a low viscosity (<100 cps) suspension emulsion content that is not easy to prepare a general emulsifier.

Referring to the optical microscope image (FIG. 15) of Examples 45 to 50, the size of the emulsified particles gradually increased as the content of the oil phase of the o / w emulsion was increased from 5% to 15% in Examples 45 to 47. The tendency to increase was confirmed. In addition, in Examples 48 to 50, even if the type of oil is changed in the case of the same series, when the content is maintained at about 15% it was confirmed that there is no significant effect on the size of the emulsion particles.

<140>

Test Example 5 Confirmation of Antioxidant Activity

In order to evaluate the antioxidant activity, a DPPH assay was used. Samples used in the experiment (Dure) are as follows.

Table 15.

<i44> DPPH assay is an experiment to measure the scavenging ability of radicals by the sample to be tested after initiation of oxidation with an oxidizing agent called DPPH. When DPPH encounters a substance with antioxidant activity, it releases electrons while giving off electrons. The color changes from purple to yellow so that the oxidation activity can be easily observed with the naked eye.

Each sample was placed in a 96 well plate by lOul and 190ul by lOOuM DPPH. The reaction was carried out at 37 ^° C. for 30 minutes and the absorbance at 517 nm was measured. Vitamin C was used as a positive control (10, 5, 2.5 ug / ml).

As a result (FIG. 7), Virginia glycoprotein (ISAI-021)> green tea glycoprotein (ISAI-016) = abis glycoprotein (ISAI-019)> pine needle glycoprotein (ISAI-018) in order of excellent antioxidant It was confirmed to have a force.

Since neither casein sodium (ISAI-022) nor pectin (ISAI-023), which are the main constituents of the capsule, have free radical scavenging ability, by introducing the above glycoproteins to enhance the antioxidant activity, Will be able to improve.

<148>

[Test Example 6] Checking the stability of the components inside the oil core

The HMP / SC / green tea glycoprotein multilayer microcapsule having the oil core prepared above was stabilized with coenzyme-Q10, which is one of cosmetic efficacy ingredients, and stabilized with coenzyme-Q10 stabilized in a general o / w formulation. We compared and evaluated.

<i5i> Example 52 was used to prepare a P / SC / green tea glycoprotein microcapsule having an oil core.

Example 52

<153> [Table 16]

> Coenzyme-QIO (QIO) 10% is dissolved in CSA, as in Example 18 previously prepared

Example 51 was prepared by emulsifying 20% Q10 10% containing oil to 3% SC. The Q10 / CSA / SC emulsion (Example 51) thus prepared was mixed 1: 1 with the HMP 3% aqueous solution of Example 1, and 0.1% of green tea glycoprotein was further dissolved therein. The HMP / SC / green tea glycoprotein microcapsules having C10 oil dissolved in Q10 in the core were prepared by slowly adding 1M citric acid and titrating the pH from about 7 to about .5 with gentle stirring.

In addition, as a comparative example, the microcapsules stabilized Q10 1% prepared before (Example

As a control of 52), Comparative Example 1 in which Q10 1% was stabilized in general o / w emulsification was prepared as follows. The composition of the comparative example 1 is as follows.

Table 17

<i57> As a result of measuring the Q10 stability of the sample corresponding to Example 52 and Comparative Example 1 prepared at room temperature and 40 degrees for 4 weeks, the stability results for 4 weeks compared to the initial Q10 content was as follows. The following results show that the Q10 stability of the microencapsulated raw material is superior. ·

<158> [Table 18]

Experimental Example 7 Evaluation of Skin Safety

18 adult women and men to confirm the skin safety of the embodiments of the present invention

A patch test of 12 subjects (average 32.5 years) was applied to evaluate the skin safety of the composition of the present invention.

In <i6i>, the patch was removed 28 hours after the patch was attached, the first reading was made 30 minutes later, and the second reading was made after 96 hours. In order to determine the intensity of skin irritation of the sample, the skin was given a weight according to the degree of positive reaction of the skin, and the skin average reaction intensity was obtained to visually examine the skin irritation of the sample. The results are shown in the table below.

<162> [Table 19]

> As shown in the above table, Examples 8, 9, 10, 17, 20, 22, 52, and Comparative Example 1 Examples 35 to 50 all confirmed that they do not irritate the skin.

Therefore, the cosmetic composition of the present invention can be determined to be superior to the safety for the skin.

> The specific parts of the present invention have been described in detail above, and for those skilled in the art, these specific descriptions are only preferred embodiments, and thus the scope of the present invention is not limited thereto. Will be obvious. Therefore, the substantial scope of the present invention will be determined by the appended claims and their equivalents.

Claims

[Range of request]

[Claim 1]

Polysaccharides with a net charge of negative charge;

Proteins having an isoelectric point (PI) of 4 to 6; And

A microcapsule comprising a plant derived glycoprotein with a net charge of negative charge.

[Claim 2]

The method of claim 1,

The plant-derived glycoprotein is one or more selected from the group consisting of Virginia-derived glycoprotein, green tea-derived glycoprotein, pine needle-derived glycoprotein, and Abis-derived glycoprotein, and microcapsules.

[Claim 3]

The microcapsule of claim 1, wherein the plant-derived glycoprotein has a carboxylic acid functional group.

[Claim 4]

The method of claim 1, wherein the polysaccharide is pectin, xanthan, beet pectin, carrageenan, chitosan, gum arabic, inulin, methyl salulose, xanthan gum, flaxseed Flaxseed gum, kappa-carrageenan, l-carrageenan, gellan gum, dextran sulfate, galactomannans, and alginate One or more microcapsules selected from the group consisting of:

[Claim 5]

The microcapsule of claim 1, wherein the protein is at least one selected from the group consisting of soy protein, casein, ovalbumin, and lactoglobulin.

[Claim 6]

The microcapsule of claim 5, wherein the soy protein is soy protein.

[Claim 7]

The microcapsule according to claim 1, wherein the weight ratio of polysaccharide to protein is 9 to 1: 1.

[Claim 8]

The microcapsule according to claim U, wherein the weight ratio of the polysaccharide: protein: sugar protein is 9-1: 1: 0.01-0.99.

[Claim 9]

The microcapsule of claim 1, wherein the microcapsule comprises a fat-soluble substance in the film.

[Claim 10]

The microcapsule of claim 9, wherein the fat-soluble substance is a fat-soluble substance that can be dissolved in an ester oil, a triglyceride oil, a silicone oil, or a hydrocarbon oil.

[Claim 11]

In the method for preparing microcapsules

i) preparing a primary o / w emulsion by first emulsifying the fat-soluble substance with a protein having an isoelectric point (PI) of 4 to 6; And

ii) secondary emulsification by adding a polysaccharide having a net charge of negative charge and a plant derived glycoprotein having a net charge of negative charge to the prepared primary emulsion;

How to Include

[Claim 12]

The method of claim 11,

steps i) and Π) are carried out at pH 6-7;

The method is

iii) lowering the secondary emulsified emulsion to pH 4.8 to 5.2 to form microcapsules.

[Claim 13]

Emulsion composition comprising the microcapsules according to any one of claims 1 to 10.

[Claim 14]

The emulsion composition of claim 13, wherein the protein is 0.1 to 1% by weight based on the total weight of the emulsion composition comprising microcapsules.

[Claim 15]

14. The emulsion composition of claim 13, wherein said polysaccharide is 1-3 weight percent based on the total weight of the emulsion composition comprising microcapsules.

[Claim 16]

The emulsion composition of claim 13, wherein the fat soluble material is from 0.01 to 30% by weight based on the total weight of the emulsion composition comprising microcapsule.