KR20150020513A - Method for Microencapsulation of Fat-Soluble Materials and Method for Functional Beverage by Using Microencapsulated Fat-Soluble Materials - Google Patents

Abstract

In the present invention, provided are a method for microencapsulating fat-soluble materials and a method for manufacturing a functional beverage by using microencapsulated fat-soluble materials. Provided are a microcapsule and a functional beverage manufactured by the method. The method for microencapsulating fat-soluble materials is allowed to mix water-soluble materials and fat-soluble materials with biological activity in a stable manner and prevent properties from being changed and functions from being damaged, caused by which fat-soluble materials directly contact the water-soluble materials. The method for microencapsulating fat-soluble materials minimizes bad smells of fat-soluble materials, thereby improving quality and reducing an amount of additives for removing the bad smell. The beverage manufactured by the method has an increasing effect on biological activity due to a stable connection of water-soluble materials and fat-soluble materials.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method of microencapsulating a lipid-soluble material and a method of manufacturing a functional beverage using the microencapsulated lipid-

The present invention relates to a method for microencapsulation of a lipid-soluble material and a method for producing a functional beverage using the microencapsulated lipid-soluble material.

The health of modern people is threatened by various adult diseases due to changes in eating habits and life patterns. One of the representative adult diseases is arteriosclerosis, a cardiovascular disease, and its incidence is increasing. Therefore, there are many studies on foods and medicines that can effectively control the blood cholesterol and triglyceride concentration, which are important for the treatment and prevention of cardiovascular diseases. The development of natural medicinal materials has great significance in the economic and industrial fields. Especially, development of natural medicinal materials which are effective for improving blood circulation and lowering cholesterol while having no adverse effects on the human body is desperately required.

Cholesterol levels are a major precursor of cardiovascular diseases, such as arterial occlusive disease, varicose veins, venous thrombosis, Burger's disease, lymphadenopathy, peripheral vascular disease, hypertension, hyperlipidemia, angina pectoris or stroke, The cholesterol found in the lipids of the bloodstream is used to produce cell membranes, some hormones and tissues. High levels of cholesterol in the blood (hypercholesterolemia) are the major risk factors for coronary heart disease, resulting in heart failure.

Omega-3 fatty acid oils are useful for the treatment of autoimmune and inflammatory diseases such as inflammatory bowel diseases such as rheumatoid arthritis, psoriasis, Crohn's disease and ulcerative colitis; Immunosuppressive therapy; Prevention of hypertension in normal human and heart transplant patients; Coronary heart disease; Hyperlipidemia; And triglycerides; Have the property that they can be used for numerous therapeutic advantages including improved kidney function and improved nephrotoxicity reduction. The use of omega-3 fatty acid oil is disclosed in U.S. Patent No. 5,034,415 (diabetes), U.S. Patent No. 4,843,095 (rheumatoid arthritis), Japanese Patent No. 2253629 (anticancer), U.S. Patent No. 4,879,312 1290625 (Enhancement of cerebral function), U.S. Patent No. 5,457,130 (Malignant tumor, inhibition of lipolytic activity) and U.S. Patent No. 5,436,269 (Hepatitis).

The omega-3 can be classified into animal omega-3 extracted from fish and vegetable omega-3 extracted from algae, perilla, flaxseed, and the like.

Onion is a lily of the 2nd century, belonging to the lily family, the diameter of which is 10 cm and is round or spherical. In September, a large flower grows from the tip of the flower, and many flowers with a sack run in a mountain shape. On the other hand, according to the statistics of the Ministry of Food, Agriculture, Forestry and Fisheries in 2008 (2008 vegetable greenhouse status and vegetable production) of onion, the domestic production of seasoning vegetables is the first place (about 1 million tons) It is a very important agricultural product to reach. Quercetin 3-O-glucoside, quercetin 3,4'-glucoside, quercetin 4'-glucoside and the like are known as components of onion, and propyl allyldisulfide It was found that a large number of compounds of the aryl sulfide group were contained. Quercetin is almost completely absent in edible parts, but it is known to be contained in brown crusts (about 8.3 mg per g of dry matter) and its content is several tens to several hundred times as high as that of edible parts. Quercetin has anti- , Brain cell protective activity, anticancer activity, antidiabetic activity, anti-obesity, antihypertensive and the like.

Blueberries are a deciduous or evergreen, low-calorie fruit tree of azaleas and (Ericaceae) Vaccinium, and are known as one of the world's top 10 longevity foods by the New York Times. Anthocyanins, which are abundant in blueberries, contribute to the health of the eyes by promoting the resynthesis of the pigment called rhodopsin in the human eye's retina. Rhodopsin is a key substance that transmits visual information to the brain through decomposition and recombination by light stimuli. When lack of rhodopsin is present, eye fatigue, decreased vision, cataracts, and cancer can be induced. Recently, interest in blueberries has been increasing in Korea recently. It has been recently revealed that blueberry fruit contains high quality physiologically active materials having various excellent bio-controlling functions, and also has excellent functionalities for preventing and healing various adult diseases This is a trend.

Asaiberry is known to contain 33 times more anthocyanins than wine, and antioxidant activity is 7.7 times higher than that of blueberries.

Vitamins are nutrients that perform a unique function to promote health maintenance and growth, and are essential substances to support the growth and recovery of normal tissues and normal physiological functions. Vitamins do not constitute energy sources or body tissues, but they are inevitably synthesized in the animal body and consumed from outside. Vitamin A, D, E, and K are fat-soluble vitamins, and all processes such as digestion, absorption, transport, and dressing depend on the fat.

Microcapsules are made of micrometer-sized capsules using a coating material for the purpose of protecting a specific substance from the external environment or releasing it at a desired or desired point in time. In recent years, it has been studied for the purpose of encapsulating functional materials, additives or microorganisms to enhance the efficiency thereof, and to improve the materials of products by encapsulating materials having undesirable taste or flavor. In the conventional microencapsulation process, Materials. Although various natural materials are used as microcapsule materials, there are few products that have reached commercialization because they are mainly related to microencapsulation process.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have intensively studied to make a lipid-soluble substance having physiological activity into a food material using a microencapsulation technique, and a method of effectively microencapsulating a lipid-soluble substance and using the microencapsulated lipid-soluble substance as an extract The inventors of the present invention have completed the present invention by confirming that a functional beverage having a remarkably increased physiological activity can be produced while reducing the deformation of the property and the loss of function caused by the direct contact of the liposoluble substance with the water-soluble substance.

Accordingly, it is an object of the present invention to provide a method of microencapsulation of a lipophilic substance.

It is another object of the present invention to provide a method for producing a functional beverage using a microencapsulated lipid-soluble material.

Another object of the present invention is to provide microcapsules prepared by the above method.

Another object of the present invention is to provide a functional beverage produced by the above method.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention and claims.

According to one aspect of the present invention, the present invention provides a method of microencapsulation of a lipophilic material comprising the steps of:

(a) preparing a capsule solution comprising a lipid-soluble substance having a physiological activity; And

(b) preparing a microcapsule by adding a coating material to the capsule solution and homogenizing the capsule, wherein the coating material is gelatin, agar, water or a combination thereof.

According to another aspect of the present invention, the present invention provides a method of making a functional beverage using a microencapsulated lipophilic material comprising the steps of:

(a) preparing a capsule solution comprising a lipid-soluble substance having a physiological activity;

(b) preparing a microcapsule by adding a coating material to the capsule solution and homogenizing the microcapsule, wherein the coating material is gelatin, agar, water or a combination thereof; And

(c) spraying the microcapsules onto the fruit extract.

The present inventors have intensively studied to make a lipid-soluble substance having physiological activity into a food material using a microencapsulation technique, and a method of effectively microencapsulating a lipid-soluble substance and using the microencapsulated lipid-soluble substance as an extract It has been found that a functional beverage having a remarkably increased physiological activity can be produced while reducing the deformation of the property and the loss of function caused by the direct contact of the liposoluble substance with the water-soluble substance.

The microencapsulation method and the functional beverage manufacturing method of the liposoluble material of the present invention will be described in detail in the following respective steps:

Step (a): emulsification of liposoluble material

According to the present invention, first, a capsule solution containing a lipid-soluble substance having physiological activity is prepared.

According to one embodiment of the present invention, the capsule solution is prepared by mixing an emulsifier with a lipophilic substance having physiological activity.

The liposoluble material used in the microencapsulation of the present invention may be various liposoluble materials having physiological activity known in the art.

The lipophilic material used in the microencapsulation of the present invention may be an omega fatty acid, a lutein, a fat soluble vitamin or a combination thereof.

According to one embodiment of the present invention, the omega fatty acid used in the microencapsulation of the present invention is omega-3, omega-6 or omega-9.

According to another embodiment of the present invention, the fat-soluble vitamin used in the microencapsulation of the present invention is one or more vitamins selected from the group consisting of vitamin A, vitamin D, vitamin E, vitamin F, vitamin K and vitamin U.

The emulsifier used in the oil-soluble substance emulsification of the present invention is an edible-grade emulsifier known in the art. According to one embodiment of the present invention, the emulsifier used in the present invention is a fatty acid ester, more preferably a sorbitan fatty acid ester.

According to another embodiment of the present invention, sorbitan fatty acid ester is added to the fat-soluble substance of the present invention and emulsified in a water bath at 60 ° C.

According to one embodiment of the present invention, the fat-soluble substance and the emulsifier are mixed in a weight ratio of 50: 1 to 30: 1.

According to certain embodiments of the present invention, the lipophilic substance and the emulsifier are mixed in a weight ratio of 40: 1.

Step (b): The coating solution is added to homogenize

The coating solution obtained through the above step (a) is added to the capsule solution and homogenized.

According to the present invention, the coating solution is a mixture of gelatin, agar, water or a material selected therefrom.

The present invention uses dull, odorless agar and gelatin as a coating agent and minimizes the use of deodorizing additives by minimizing the odor of lipophilic substances and improving the functionality.

According to an embodiment of the present invention, a mixture of the capsule solution obtained in step (a) and agar and gelatin in a volume ratio of 3: 1 is mixed at a volume ratio of 6: 4 to 8: 2 and homogenized.

According to another embodiment of the present invention, the capsule solution obtained through step (a) and agar are mixed at a volume ratio of 6: 4 to 8: 2 and homogenized.

The homogenization method used in the production of the microcapsule of the present invention can use various homogenization methods known in the art.

According to an embodiment of the present invention, homogenization of the resultant product and the coating solution is carried out using a homomixer. The homogenized product is sprayed onto purified water, and the solution is dried or filtered to separate only the microcapsules.

Step (c): The microcapsules are sprayed onto the seed extract

The microcapsules obtained in step (b) are used to prepare functional beverages.

The microcapsules obtained as a result of the step (b) are sprayed onto the fruit extract.

According to one embodiment of the present invention, the microcapsules are sprayed onto the fruit and vegetable extracts by a spray chilling method using an airless spryer.

According to the present invention, the aqueous solution for spraying the microcapsules of the present invention is an iced tea extract containing a physiologically active substance.

The fruit and vegetable extracts that can be used in the present invention include various fruit and vegetable extracts known in the art, and are not particularly limited.

For example, the fruit and vegetable extracts which can be used in the present invention include blueberry extract, Asaiberi extract, blackberry extract, raspberry extract, cranberry extract, Audi extract, bokbunja extract and onion extract.

According to one embodiment of the present invention, the fruit and vegetable extract used in the production of the functional beverage of the present invention is an onion extract fermentation broth fermented with lactic acid bacteria.

The onion extract fermentation liquid used in the composition of the present invention is obtained by fermenting an onion extract. When the onion extract is obtained by treating the onion with an extraction solvent, various extraction solvents can be used. According to one embodiment of the present invention, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o -xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

According to another embodiment of the present invention, the extraction solvent used in the present invention may be selected from the group consisting of (a) water, (b) an anhydrous or hydric alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, (E) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane, and (j) Diethyl ether.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the onion extract includes not only those obtained by using the above-mentioned extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained by the purification method is also included in the onion extract of the present invention.

The fermentation of the onion extract in the present invention can be carried out according to various fermentation methods known in the art.

The fermentation may be carried out by (i) directly inoculating the fermenting microorganism into the extract, (ii) using a culture medium (for example, crude enzyme solution) in which the fermenting microorganism has been cultured, or (iii) can do.

The fermented product used in the present invention can be obtained by fermenting an onion first before extraction and then obtaining an extract from it, or by obtaining an extract from an onion extract and fermenting it. Thus, the term " fermentation product " as used herein has the meaning that it encompasses both of the above two types of fermentation product.

The microorganism used for fermentation is not particularly limited, and preferably includes lactic acid bacteria.

Lactic acid bacteria suitable for the present invention include various lactic acid bacteria known in the art. As used herein, the term " lactic acid bacteria " refers to a group of bacteria that produce lactic acid as a major product of carbohydrate metabolism. Lactic acid bacteria, Lactococcus (Lactococcus), Lactobacillus bacteria (Lactobacillus), flow Pocono stock (Leuconostoc), Pro are pleased tumefaciens (Propionibacterium), Enterococcus (Enterococcus), Bifidobacterium (Bifidobacterium) used in the present invention, Streptococcus (Streptococcus) and Pediococcus . ≪ / RTI >

According to one embodiment of the present invention, the lactic acid bacteria used for the fermentation of the onion extract in the present invention may be selected from the group consisting of Lactobacillus plantarum , Lactobacillus alimentarius , Lactobacillus sakei , Lactobacillus acidophilus , Lactobacillus casei , Lactobacillus gasseri , Lactobacillus delbrueckii , Lactobacillus fermentum , Lactobacillus bulgaricus , Lactobacillus spp ., Lactobacillus spp ., Lactobacillus spp. , Lactobacillus bulgaricus , Lactobacillus helveticus , Leuconostoc mensenteroides , Streptococcus thermophilus , Streptococcus lactis , Enterococcus faecium, (Enterococcus faecium), Enterococcus wave Consisting faecalis (Enterococcus faecalis), Bifidobacterium William BP bonus (Bifidobacterium bifidum), Bifidobacterium William Infante Tees (Bifidobacterium infantis), Bifidobacterium William Bra Entebbe (Bifidobacterium brave) and Bifidobacterium William ronggeom (Bifidobacterium longum) Lt; / RTI > According to the present invention, one or more mixed lactic acid bacteria of the above-mentioned lactic acid bacteria can be used for the onion extract fermentation of the present invention.

According to another embodiment of the present invention, the lactic acid bacterium used for the fermentation of the onion extract in the present invention is Lactobacillus plantarum or Lactobacillus casei .

According to a specific embodiment of the present invention, the onion extract is sterilized after the pH is adjusted, then inoculated with Lactobacillus frutarium or Lactobacillus casei, fermented at 40 ° C for 6 to 24 hours, and stored at 4 ° C.

According to one embodiment of the present invention, an emulsifier is added to omega-3 and mixed (coating material A), and gelatin, agar, xanthan gum and purified water are mixed (coating material B) In a ratio of 8: 2, and then spray-dried on a cooled fermented onion extract to spray-dried or cooled purified water. Onion extract fermentation broth containing microencapsulated omega-3 prepared by the above method has high emulsion stability.

According to one embodiment of the present invention, the homogenized coating materials A and B are sprayed with 0.01-5.0 wt% of the onion extract fermentation liquid or purified water.

As demonstrated in the following examples, the compositions of the present invention comprising omega-3 microcapsules and an onion extract fermentation solution significantly reduced the specific discomfort (e.g., attack) of omega-3. Therefore, the functional beverage of the present invention is excellent in the sensibility.

According to the present invention, the functional beverage of the present invention prepared using the onion extract fermentation liquid has an antioxidative activity because it has a scavenging ability to free radicals.

According to a preferred embodiment of the present invention, the functional beverage of the present invention may further comprise a blood circulation improving agent in addition to the microencapsulated omega-3 and onion extract fermentation broth.

Preferably, the blood circulation improving agent is omega-6 fatty acid, omega-7 fatty acid, omega-9 fatty acid or lecithin, and the omega-6 fatty acid is, for example, octadecadienoic acid, octadecatrienoic acid, Eicosatetraenoic acid, docosadienoic acid, docosatetraenoic acid, docosapentaenoic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, and octadecatrienoic acid are preferable. 7 fatty acids include, for example, dodecenoic acid, tetradecenoic acid, hexadecenoic acid, decenoic acid, eicosanoic acid, docosanoic acid and tetracosenoic acid, and the omega-9 fatty acids include, for example, , Octadecenoic acid, eicosanoic acid, eicosatrienoic acid, docosanoic acid and tetracosenoic acid.

According to another embodiment of the present invention, the fruit and vegetable extract used in the production of the functional beverage of the present invention is a blueberry extract.

As demonstrated in the following examples, the functional beverage prepared using vitamin A as a lipid-soluble material using the blueberry extract as the fruit extract showed higher antioxidative activity than the blueberry extract, and the synergistic And showed a remarkable effect for lowering the incidence of inflammation in the body.

 According to another embodiment of the present invention, the fruit and vegetable extract used in the production of the functional beverage of the present invention is an Asaiberi extract.

As demonstrated in the following examples, the functional beverage prepared using vitamin E as a lipid soluble material using Asaiberi extract as an apple and vegetable extract showed a synergistic effect on the antioxidative effect as compared with a single substance.

The fruit and vegetable extracts used in the present invention include not only the extracts extracted with the extraction solvent but also their fermented products.

According to one embodiment of the present invention, the fruit and vegetable extract containing the physiologically active substance of the present invention may further contain guar gum or xanthan gum as a stabilizer.

When the microcapsule solution is mixed with the cotyledon extract obtained by adding guar gum or xanthan gum as a stabilizer, the dispersibility and stability of the microcapsule are maintained.

According to another aspect of the present invention, there is provided a microcapsule prepared by the above method of the present invention.

According to one embodiment of the present invention, the microcapsules of the present invention have a diameter of 1-50 mu m.

According to another embodiment of the present invention, the microcapsules of the present invention have a diameter of 7-30 탆.

According to certain embodiments of the present invention, the microcapsules of the present invention have a diameter of 10-20 占 퐉.

According to another aspect of the present invention, the present invention provides a functional beverage produced by the above method of the present invention.

According to the present invention, the functional beverage of the present invention is a microcapsule in which the microcapsules are dispersed in a fruit and vegetable extract containing a physiologically active substance.

In the functional beverage of the present invention, a mineral-containing compound (Ca, Mg, Zn, etc.), phospholipid (lecithin, etc.), maltol, oligosaccharide and amino acid may be used in addition to the main components. Among them, It is more preferable because it can reinforce the biological activity effect.

In addition to the above components, natural additives such as natural flavors such as plum, lemon flavor, pineapple flavor or herb flavor, natural fruit juice, natural pigments such as Chlorophyllin and Flovonoid in order to enhance the taste as a known additive, , Honey, sugar alcohol, sugar, citric acid, and sodium citrate.

The functional beverage according to the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, And carbonating agents used in carbonated drinks.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a method for microencapsulation of a lipid-soluble material and a method for producing a functional beverage using the microencapsulated lipid-soluble material.

(b) The present invention provides microcapsules and functional beverages by the method of the present invention.

(c) The microencapsulation method of the present invention can stably mix water-soluble and lipophilic substances having physiological activity, and it is possible to minimize the loss of properties and loss of properties that may be caused by the direct contact of the lipophilic substance with the water- .

(d) The present invention minimizes the use of deodorant additives by minimizing odor of lipophilic materials used in microencapsulation to improve functionality.

(e) The functional beverage produced by the method of the present invention exhibits a synergistic effect on physiological activity through stable binding of a water-soluble substance and a lipid-soluble substance.

1 is a schematic diagram of a spray chiller method.
Figure 2 is a photograph of the spray-drying process.
Figure 3 is a micrograph of particle size according to three experimental conditions. (A) agar + gelatin; (B) carrageenan + gum arabic; And (C) xanthan gum + gelatin + gum arabic. Each sample is magnified 1,000 times.
FIG. 4 shows the result of measuring the degree of emulsion stability of the emulsion according to the coating material. (A) agar: gelatin = 3: 1; (B) xanthan gum: gelatin: gum arabic = 5: 2: 50; (C) Carrageenan: Gum arabic = 1: 10.
Figure 5 shows the results of microencapsulation yields for the coating materials. (A) agar: gelatin = 3: 1, (B) xanthan gum: gelatin: gum arabic = 5: 2: 50.
6 shows the result of measurement of DPPH free radical scavenging effect of onion extract fermentation broth. Each value is expressed as ± standard deviation (n = 2).
FIG. 7 shows the result of measuring the antioxidative activity of onion extract fermentation broth according to fermentation strains in ABTS.
Each value is expressed as ± standard deviation (n = 2).
Fig. 8 shows the result of measuring the antioxidative activity of onion extract fermentation broth according to fermentation strains in FRAP.
Each value is expressed as ± standard deviation (n = 2).
FIG. 9 shows the result of measuring the blood circulation improving activity of the omega-3-added onion extract fermentation broth.
B: omega-3, C: fermented onion extract (DKL 128) + omega-3
Fig. 10 shows the results of comparative sensory evaluation of onion extract / fermented onion extract (a) and fermented onion extract (b) containing omega-3 fermented onion extract / microencapsulated omega-3.
Fig. 11 is a photograph showing the dispersibility and stability observed after the microcapsules were directly injected into the onion extract fermentation broth.
Fig. 12 is a photograph showing the stability and dispersibility of the onion extract fermentation liquid and the microcapsule solution mixed with the stabilizer after the preparation of the product.
FIG. 13 is a photograph of a blueberry extract obtained by extracting purified water and 80% alcohol. FIG.
14 is a graph showing the anthocyanin content of the blueberry extract by solvent.
Fig. 15 shows the result of measuring the antioxidative activity of blueberry extract by solvent.
Fig. 16 is a photograph of the sphere formation of vitamin A + sorbitan fatty acid ester.
Fig. 17 is a photograph showing the capsule formation of agar and vitamin A. Fig.
18 is a photograph of a vitamin A capsule observed using an optical microscope.
19 is a graph showing the encapsulation yield of vitamin A measured.
20 is a graph showing the antioxidative activity of the blueberry mixture.
FIG. 21 shows the results of measurement of blood total cholesterol by high fat diet and blueberry extract intake group.
FIG. 22 is a graph showing blood lipid levels in high fat diet and blueberry extract intake groups.
23 is a graph showing the HDL-c content by high fat diet and blueberry extract intake group.
24 is a graph showing the LDL-c content by high fat diet and blueberry extract intake group.
FIG. 25 is a graph showing blood coagulation time associated with extracellular pathway by high fat diet and blueberry extract intake group. FIG.
FIG. 26 is a graph showing blood coagulation time associated with the phosphorylation mechanism in the high fat diet and blueberry extract intake groups. FIG.
FIG. 27 is a graph showing the content of glutathione in the high fat diet and blueberry extract intake groups. FIG.
FIG. 28 is a graph showing the content of myeloperoxidase by high fat diet and blueberry extract intake group. FIG.
29 is a graph showing the content of prostaglandin E2 by high fat diet and blueberry extract intake group.
30 is a photograph showing a mixture of a blueberry extract and a vitamin A capsule.
31 is a photograph showing the capsule state of a mixture composed of a blueberry extract and a vitamin A capsule.
32 shows the result of measuring the antioxidative effect of a mixture composed of an Asai mixed-berry extract and a vitamin E capsule.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example 1: Microencapsulation of onion extracts

I. Materials and Methods

1. Fermentation of Onion Extract

Onion extract

Ethanol, methanol and water were added to the onion extracts, respectively, and extracted three times for 4 hours with an extraction device equipped with a reflux condenser on a water bath at 60 ° C. The filtrate was collected and concentrated with a rotary vacuum concentrator (Rotavapor R-3, buchi, Swiss), and the weight of the completely dried was measured.

Used strain

Onion extract fermented from Lactobacillus plantarum ssp. DKL 119 (hereinafter referred to as DKL 119) and Lactobacillus casei DKL 128 (hereinafter referred to as DKL 128) were subcultured three times.

Measure the number of lactic acid bacteria

The number of lactic acid bacteria in the prepared samples was measured by APHA method using BCP medium (Eiken Co., Japan) at 37 ° C for 24 hours. For this fermentation, strains DKL 109, 119, 121 and 128 were used.

Measurement of pH and titratable acidity

The pH was measured using a pH meter (Orion Co., USA). The titratable acidity was determined by adding APA to 9 g of sample and adding 3-4 drops of 0.1% phenolphtalein to 0.1 N NaOH and the consumption was calculated.

2. Measurement of quercetin content of fermented onion extract

Preparation of Test Solution

For analysis, the samples were weighed and dissolved by adding 40 mL of 60% ethanol and 5 mL of 6 N HCl and refluxing at 95 ° C for 2 hours. The solution was concentrated with a rotary evaporator (buchi-korea), and then adjusted to 50 mL with 60% ethanol and filtered through a 0.45 μm filter.

Preparation of standard solutions

Standard crude quercetin (Sigma Co., USA) was prepared with ethanol to a concentration of 1 mg / mL, and used as a standard stock solution. This was diluted stepwise with 0.001, 0.002, 0.005, 0.01, 0.05 and 0.1 mg / mL in water: 5% acetic acid: acetonitrile (40:30:30, v / v) and filtered through a 0.45 μm membrane filter As a standard solution, quercetin content was determined from a standard calibration curve prepared using a standard solution.

HPLC analysis

Crozier, Wang and Helliwell, and analyzed using the HPLC system (Waters, USA) under the same conditions as in Table 1.

HPLC conditions for the analysis of quercetin in onion extract - Condition column YMC Hydrosphere C18 Mobile phase Water: 5% Acetic acid: Acetonitrile (40%: 30%: 30%) detector UV detector, 370 nm Flow rate 1.0 mL / min Injection volume 20 μL

3. Antioxidant activity measurement of onion extract fermentation broth

DPPH radical scavenging activity

DPPH (2,2-Diphenyl-1-picrylhydrazyl) is able to measure antioxidative activity by utilizing the property that the nitrogen atom of hydrazide is in an unstable state and easily loses its color due to acceptance of hydrogen atom. The DPPH radical scavenging activity method is a method of measuring electron donating ability, and it is possible to measure the reducing power and the antioxidative activity of the measuring substance based on the degree of reduction of DPPH. DPPH radical scavenging activity was measured as follows. To 0.8 mL of the pretreated test solution, add 0.8 mL of 0.2 mM DPPH and 0.8 mL of ethanol, react at room temperature for 20 minutes, and centrifuge at 3,000 rpm for 10 minutes. The supernatant was measured for absorbance at 517 nm on a spectrophotometer (SP-2000 UV Spectrum, Korea). The results of the radical scavenging ability were calculated by the following equation.

Scavenging activity (%) = 100 × (A control - A sample) / (A control)

Antioxidant activity measurement by ABTS method

After adding 88 μL of 140 mM potassium sulfide (K ₂ S ₂ O ₈ ) to 5 mL of 7 mM ABTS, the ABTS stock solution was prepared in a dark room for 12 hours and mixed with 0.1 M potassium phosphate buffer (pH 7.4) Solution. The concentration of the ABTS radical working solution was adjusted so that the initial absorbance value at 734 nm was 0.70 ± 0.02. The ABTS radical scavenging activity was determined by measuring the absorbance at 734 nm and reacting with 1 mL of a certain concentration of the sample and 1 mL of ABTS radical walking solution. The ABTS radical scavenging activity was determined by the following equation.

ABTS radical scavenging activity (%)

FRAP assay

Ferrous Reducing Antioxidant Power (FRAP) is one of the methods to measure antioxidant activity. It is a method to measure the antioxidant ability by reducing power of iron ion, unlike the method of measuring radical scavenging ability using DPPH. The FRAP assay and the DPPH assay were reported to show a high correlation. It is possible to measure the antioxidant activity of the test substance by the reducing power of iron ion by substituting the absorbance value measured at 593 nm into the FeSO ₄ slope and measuring the electron donating ability.

Ferric-reducing antioxidant potential ability (FRAP) was measured according to the method of Benzie and Strain (16). The FRAP reagent was prepared by heating 25 mL of acetate buffer (300 mM, pH 3.6) at 37 ° C and adding 10 mM 2,4,6-tris (2-pyridyl) -s-tri-azine (TPTZ, 2.5 mL of Sigma Chemical Co.) and 2.5 mL of 20 mM FeSO ₄ . The prepared FRAP reagent and the onion extract fermentation broth were mixed in a ratio of 19: 1 and reacted in a cow for 30 minutes. Absorbance was measured at 593 nm using a spectrophotometer.

Blank was measured by adding distilled water instead of the sample. The Fe ²⁺ concentration was converted to the standard curve of FeSO ₄ prepared at concentrations of 25, 50, 100, 200, 400, 800 and 1,600 mM.

4. Measurement of vitality improvement activity of onion extract fermentation broth

And was measured using a fibrin plate method. 10 mL of a solution of 0.5% fibrinogen dissolved in 67 mM sodium phosphate buffer (pH 7.4) was added and mixed. Add 0.2 mL of thrombin (50 U / ml) and place the paper disc on a hardened fibrin plate. 20 μL of the test solution is dispensed on a paper disk and reacted at 37 ° C for 17 hours. The diameter of the transparent ring produced after the reaction was measured and the control group was plasmin (2.0 U / ml). The blood circulation improving vitality measurement was converted at a rate relative to the plasmin using the following equation.

Fibrinolytic activity (%)

= Transparent diameter of the sample / Transparent diameter of the plasmin x 100

5. Microencapsulation of omega-3

Omega-3 and Emulsifiers

(DHA 40% vegetable omega-3, DHA + EPA 30% animal omega-3) supplied from Korea (Korea) were used. The HLB (Hydrophile Lipophile Balance) values were 4.3, 4.7, 14.9 and 15.0 , Sorbitan monooleate, Ilshin emulsifier co., LTD., Korea), span 80 (sorbitan monooleate, Ilshin emulsifier co., LTD., Korea), Tween 60 (polyoxyethylene sorbitan monostearate, Ilshin emulsifier Ltd.), Tween 80 (polyoxyethylene sorbitan monostearate, Ilshin emulsifier co., LTD., Korea), Tween 20 (polyoxyethylene sorbitan monostearate, Ilshin emulsifier co.

Microencapsulation of omega-3

The capsule solution (0.5% gelatin, 1.5% agar, 77.4% purified water, 0.1% xanthan gum) was added to the omega-3 (20%) after adding the edible grade emulsifier (span60: 0.5% , Homogenized at 10,000 rpm for 3 minutes using a homomixer, sprayed on a cooled fermented onion extract by spray chilling using an airless spryer, or sprayed on cooled purified water It is mixed with liquid of different material. The microcapsules were sprayed on the purified water, and the solution was dried or filtered to separate only the microcapsules.

6. Observation of microcapsule size using optical microscope

The size of the microcapsules was observed using an optical microscope (BX 51TF, Olympus Co., Japan) and a calibrator (Konvison microscope co., LTD., Korea) and an alternative lens scale (Konvison microscope co., LTD., Korea).

7. Measurement of emulsion stability

The volume of the aqueous solution separated from the emulsion after measuring the volume of the emulsion was measured in an oven at 100 ° C. after the emulsion was placed in a 50-mL measuring cylinder. The measured value was substituted with the emulsion stability index (ESI ).

Emulsion stability index (ESI) = (1-) × 100

8. Measurement of microencapsulation yield

Pretreatment for yield measurement

10 g of the sample was placed in a sealed container, shaken vigorously with boiling water, and vigorously shaken. Add 60 mL of a solution prepared by mixing chloroform (Sigma-Aldrich) and methanol (Sigma-Aldrich) at a ratio of 2: (Sigma-Aldrich) was added to each well, and 10 g of each sample was taken into a separatory funnel, and 20 mL of petroleum ether (Sigma-Aldrich) was added thereto, followed by shaking for 5 minutes And the mixture was allowed to stand for 10 minutes. The lower layer was taken in another aliquot, and the resultant solution was combined with the second-extracted solution in the same manner as above to obtain a fatty acid extract. The resulting extract was evaporated under reduced pressure (80 ° C, 120 rpm) to remove the solvent. 15 ml of 1 N KOH ethanol was added to the remaining lipid. The reaction was allowed to proceed for 1 hour in a 90 ° C water bath connected with a cooling tube, transferred to a separatory funnel, And the mixture was allowed to stand for 5 minutes. Then, the upper layer from which wax, sterole, etc., which are non-adherent substances, was removed and repeatedly extracted by the same method. 1.6 mL of 1N HCl was added to the filtrate After adjusting to pH 2.0, 20 mL of petroleum ether was added again, and the mixture of fatty acids was extracted three times to remove moisture. The methyl ether of the fatty acid was prepared by methylation with 14% BF ₃ - methanol solution as follows. After adding 1 mL of 14% BF ₃ -methanol solution to the round-neck flask containing the fatty acid obtained above, add 3 mL of saturated NaCl solution, add hexane, 3 mL was added thereto to separate the hexane layer, and then anhydrous sodium sulfate was added to remove water to prepare a sample for GC analysis.

Measurement of fatty acids by GC

For the identification of omega - 3 fatty acids, Sigma 's standard fatty acids were purchased and used. The retention times of standard fatty acids and extracted fatty acids were compared to determine the concentration of omega - 3 fatty acids at the same time. The analytical conditions of the GC used in the experiment are shown in Table 2 and the yield was measured by the following equation.

TF: Fatty acid concentration of total capsule solution

NF: Uncapsulated fatty acid concentration in the capsule solution

detector FID column FUSED slica capillary column (60 m × 0.25 mm × 0.2 μm film thickness) Flow rate Separation rate 10: 1 Carrier gas (He) 1 mL / min air 450 mL / min H ₂ 45 mL / min Temperature Injection port 260 ℃ Detector 260 ℃ column 5 minutes at 180 ° C → 10 ° C per minute, 260 ° C for 20 minutes Injection volume 1.0 μL

9. Measurement of blood circulation improving activity of onion extract fermentation broth and microcapsule omega-3 mixture

The fibrin plate method was used to measure the blood circulation improving activity of a mixture of onion extract fermentation broth and microcapsule omega-3. The fermented onion extract was dissolved in DMSO (Sigma-Aldrich), and 10 mL of a solution in which 0.5% fibrinogen was dissolved in 67 mM sodium phosphate buffer (pH 7.4) was added and mixed. 0.2 mL of thrombin (50 U / ml) was added and the paper disk was placed on a hardened fibrin plate. A 5 mm hole was made in the fibrin plate, and 20 μL of the test solution was injected, followed by reaction at 37 ° C. for 17 hours. After the reaction, the diameter of the generated transparent ring was measured, and the control group was plasmin (2.0 U / ml) (Sigma-Aldrich). The blood circulation improving vitality measurement was converted at a rate relative to the plasmin using the following equation.

Fibrinolytic activity (%) = clear zone size of sample / clear zone size of plasmin × 100

10. Comparative sensory evaluation of onion extract and fermented onion extract

The sensory evaluation of onion and fermented onion extracts were conducted in 20 college students and workers.

II. Experiment result

1. Fermentation of Onion Extract

Establishment of onion fermentation condition by measuring the number of lactic acid bacteria

As shown in Table 1 below, the number of lactic acid bacteria in the onion extract fermented with DKL 109, 119, 121, and 128 strains was 10 ⁹ cfu / ml in the fermentation broth after fermentation for 12 hours, mL or more. Since the viable cell count is maintained at about 10 ⁹ cfu / mL even when fermentation at 12 or higher, the optimum fermentation time of the onion extract based on the viable cell count is determined to be 12 hours.

Measurement of viable cell counts of onion extracts according to fermentation time using DKL strain system Number of living cells (cfu / mL) 0 hours 12 hours 24 hours DKL 109 2.0 × 10 ⁷ 1.2 × 10 ⁹ 1.1 × 10 ⁹ DKL 119 2.0 × 10 ⁷ 1.1 × 10 ⁹ 1.0 × 10 ⁹ DKL 121 2.0 × 10 ⁷ 1.0 × 10 ⁹ 1.0 × 10 ⁹ DKL 128 2.0 × 10 ⁷ 1.0 × 10 ⁹ 1.0 × 10 ⁹

Measurement of pH and titratable acidity

As a result of pH measurement, the pH of DKL 109, 119, 121 and 128 showed a value of 4.92-4.97 at 0 hour of fermentation. The value of pH was lowered at 3.5-2.57 at 24 hours of fermentation. No significant change was observed.

Titratable acidity also showed a slight increase in the titratable acidity of each strain but increased until 24 hours after fermentation, but not significantly after 24 hours.

Establishment of onion fermentation conditions to increase effective substance (quercetin)

Onion extract was obtained and concentrated under reduced pressure using a rotary evaporator. The onion concentrate was diluted to 1/6 (purified water), adjusted to pH 7.5, sterilized at 85 ° C for 15 minutes, cooled at 40 ° C, and inoculated with 2% of DKL 109, DKL 119, DKL 121 and DKL 128 For 12 hours.

The content of quercetin increased after fermentation regardless of fermentation strain. Among them, quercetin content of onion extract fermented with DKL128 strain showed the greatest change from 48.133 to 75.336.

Changes in the content of quercetin in onion extract fermentation broth according to fermentation conditions - 0 hours (ppm) 12 hours (ppm) DKL 109 48.597 49.843 DKL 119 49.061 65.135 DKL 121 48.590 53.676 DKL 128 48.133 75.336

2. Establishment of fermentation onion analysis conditions

To analyze the content of quercetin in the fermented onion, 50 mL of EtOH and 21.4 mL of 3 M HCl were mixed with 1 g of the fermented onion concentrate. The mixture was concentrated under reduced pressure at 95 ° C for 105 minutes. To 0.3 mL of the concentrated sample, 2.7 mL of distilled water was mixed, After filtration with a filter, HPLC was performed.

HPLC analysis conditions for determination of quercetin content in fermented onion - Condition column YMC Pack C18 Eluent Acetonitrile _{/ 10 mM H 3 PO 4 (} 30/70) Flow rate 1 mL / min Temperature 40 ℃ Measure 260 nm UV Injection 5 μL

3. Antioxidant activity measurement of onion extract fermentation broth

DPPH radical scavenging activity

Antioxidant activity of onion extract fermentation broth according to fermentation strain was measured by DPPH assay. As shown in FIG. 6, the DPPH radical scavenging activity of DKL 109 was highest at 68.8% at 0 hour and 73.9% at 12 hours after fermentation, and DKL128 showed 68.8% at 0 hour and 73.3% at 12 hours of fermentation. However, DKL119 and DKL121 did not increase DPPH radical scavenging ability after fermentation. The DPPH radical scavenging activity of onion extract fermented by DKL 109 and DKL 128 was 73.9 and 73.3%, respectively.

ABTS method

The ABTS ^{+ ㆍ} scavenging activity of the onion extract fermentation broth according to the fermentation strain was as shown in FIG. In the ABTS assay antioxidant activity it showed more ABTS ^{+ and} scavenging effect of onion extract after fermentation 12 hours compared to the fermentation 0 hours, regardless of the fermentation strain higher, using DKL121 a 12-hour antioxidant activity of fermented onion extract broth 69.2 % (Fig. 7).

FRAP assay

The results of measuring the antioxidative activity of the onion extract fermentation broth according to the fermentation strain using the FRAP assay were as shown in FIG. DKL109, DKL119 and DKL121, except DKL128, showed a tendency to decrease antioxidative activity at 12 hours of fermentation compared to 0 hours of fermentation.

Antioxidant activity of onion extract fermented by strain was measured by DPPH, ABTS and FRAP. In the DPPH assay, DKL109 and DKL128 showed an antioxidant activity at 12 hours of fermentation. In the ABTS assay, antioxidant activity was increased by fermentation in four strains. However, in the FRAP assay, the antioxidant activity of onion extract fermentation broth tended to decrease with fermentation in the three strains except DKL128. Fermentation of lactic acid bacteria with onion extract increased the free radical scavenging ability such as DPPH radical, ABTS ^{+ ㆍ} radical, but decreased the ability of reducing Fe ³⁺ to Fe ²⁺ .

4. Microencapsulation of omega-3 fatty acids

Measurement of emulsion stability

The emulsion stability of omega-3 emulsions with agar: gelatin = 3: 1, xanthan gum: gelatin: gum arabic = 5: 2: 50 and carrageenan: gum arabic = 1: The emulsion stability index of agar: gelatin = 3: 1 as a coating material was 82%, which was significantly higher than other samples. On the other hand, microcapsules with carrageenan and gum arabic coating materials showed the lowest emulsion stability and were excluded from the following experiments.

Observation of microcapsule size using optical microscope

Measurement of microcapsule size number Size (㎛) Micro Capsule 1 One Micro Capsule 2 10 Micro Capsule 3 20 Micro Capsule 4 50 Micro Capsule 5 100

Yield measurement

A prepared by mixing omega-3 fatty acids and span 60 (sorbitan fatty acid ester) in a water bath at 60 ° C and a coating material B prepared by mixing 1.5% agar and 0.5% gelatin were each mixed at 8: 2, , And the microencapsulation yield was calculated using the following equation by spray-drying the A and B homogeneous mixture in a fermented onion solution (stored at 4 ° C, 12.9 brix) for 3 minutes.

Microencapsulation yield (%) = (total omega-3 oil-non-encapsulated omega-3 oil) / total omega-3 oil x 100

C _{17: 0} (heptadecaenoic acid) was identified by gas chromatography to determine the yield of omega-3 microcapsules. As shown in Fig. 5, C _{17: 0} (heptadecaenoic) was able to confirm the peak in the minute.

As shown in FIG. 4A, the yield of omega-3 microcapsules (A) containing agar and gelatin as the coating material, which showed the highest emulsification stability index in terms of emulsion stability, was 99%, and the coating materials of xanthan gum, gelatin and gum arabic B), respectively. The size of capsules, emulsion stability and yield were finally determined to be 3: 1 mixture of agar and gelatin.

5. Effect of onion extract fermentation and microcapsule omega-3 mixture on blood circulation

Improvement of blood circulation activity of omega-3 added onion extract fermentation broth

In previous experiments, it was confirmed that DKL 128 increased the content of quercetin in the fermentation process of onion extract and increased the blood circulation improving vitality. When omega - 3 was added to fermented onion extract fermented with DKL 128 strain, the change of blood circulation improving activity was measured. As shown in Table 7 and FIG. 9, when omega-3 alone was treated, the blood circulation improving vitality was slightly increased, and when omega-3 was added to the fermented onion extract, blood circulation improving vitality was further increased I could confirm.

Transparent diameter according to the mixture of onion extract fermentation broth and microcapsule omega-3 Sample Relative activity (%) Clear zone (mm) Plasmin 100 11 Fermented onion extract (DKL 128) 181 20 Omega-3 113 12 Fermented onion extract (DKL 128) + Omega-3 200 22

6. Comparative sensory evaluation of onion extract and fermented onion extract

The sensory evaluation of 20 students of university students and workers showed that the fermented onion extract had a higher total flavor than the onion extract. Onion extract showed higher onion extract than fermented onion extract, (Fig. 10A). Microencapsulated omega-3 fermented onion extracts showed higher overall taste than omega-3 fermented onion extracts, and seaweed encapsulated omega-3 showed reduced catch (Fig. 10b).

7. Stability of omega-3 microcapsule added onion extract fermentation broth

 When the omega-3 microcapsules were sprayed directly onto the onion extract fermentation broth, it was observed that the microcapsules could not be uniformly dispersed in the onion fermentation broth. To improve this, a method of preparing a product by adding a certain amount of pectin, xanthan gum and guar gum as an edible stabilizer to onion extract fermentation broth, spraying microcapsules, and a method of mixing microcapsule solution with onion extract fermentation solution And stability and dispersibility were observed. As shown in FIG. 11, when the microcapsules were directly sprayed on the fermentation broth of the onion extract, it was confirmed that stability was maintained when 0.5% of pectin was added. 11 shows the microcapsules directly sprayed on the onion extract fermentation liquid as in the method of Fig. In the case of xanthan gum and guar gum, it was observed that they tangled due to the reaction with the coating material of the capsules, indicating that they were not suitable as stabilizers.

However, when the product was prepared by mixing the microcapsule solution with the fermentation broth of the onion extract supplemented with the stabilizer, the dispersibility and stability were maintained when xanthan gum or guar gum was used as stabilizer rather than pectin (Fig. 12) . FIG. 12 shows microcapsules prepared by injecting microcapsule solution into purified water and mixing the microcapsules in an onion extract fermentation solution added with a stabilizer. In manufacturing the product, guar gum or xanthan gum was added to the onion extract fermentation broth, and the microcapsule solution was mixed in consideration of the convenience and stability of the preparation.

GC analysis (Clarus 500; PerkinElmer, USA) - Condition detector FID column FUSED slica capillary column (60 m × 0.25 mm × 0.2 μm film thickness) Flow rate Separation rate 10: 1 Carrier gas (He) 1 mL / min air 450 mL / min H ₂ 45 mL / min Temperature Injection port 260 ℃ Detector 260 ℃ column 5 minutes at 180 ° C → 10 ° C per minute, 260 ° C for 20 minutes Injection volume 1.0 μL

Composition ratio of microcapsule composition Kinds Composition ratio Omega-3 Oil 0.01-30% Distilled water 50-80% Baby 0.1-5.0% gelatin 0.1-3.0% Emulsifier 0.1-3.0% Xanthan gum 0.1-3.0%

Example 2 Microencapsulation of Blueberry Extract

I. Materials and Methods

1. Blueberry Extract

The blueberry used as a sample in this experiment was dried and used as the first blueberry purchased from Nutra Phanax. Purified water and 80% alcohol were used as the blueberry extracting solvent. Blueberry extraction was carried out using a magnetic stirrer. The blueberry samples were extracted with 5 times of purified water and 80% of alcohol, respectively, and then filtered through a filter paper (Whatman No. 2) to obtain a first blueberry extract. Each of the extracts obtained was concentrated by a vacuum rotary condenser, Respectively.

2. Analysis of active ingredient of blueberry extract

For analysis of anthocyanin content of blueberry extract, anthocyanin was extracted from the sample using an extraction solvent (MeOH: 0.1M HCl = 85: 15). The extraction method is as follows: The sample treated with the extraction solvent is allowed to stand at room temperature (25 ° C) for 30 minutes, then subjected to primary filtration with Whatman No 2 filter paper and subjected to secondary filtration with a syringe filter (0.45 μm) Respectively.

Content of anthocyanin (mg / g) in analytical sample = Y 'x V / S

Y ': Concentration of the analytical sample obtained by substituting in the calibration equation

V: Total volume (ml) of the extraction solution

S: weight of sample used for extraction (g)

3. Measurement of physiological activity of blueberry extract

Antioxidant activity measurement

To determine the scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical, the blueberry extract was mixed with the same amount of distilled water and suspended for 1 hour. The filtrate was concentrated under reduced pressure, centrifuged (1200 rpm, 10 minutes) to 100 ml, and used as a sample for electron donating ability test. Electron Donating Ability (EDA) To 0.2 ml of sample extract, 0.8 ml of 0.04 ml DPPH reagent was added, vortexed for 10 seconds, and left at room temperature for 10 minutes. Absorbance was measured at 525 nm.

EDA (%) = [1- (OD value of sample / OD value of blkn)] × 100

4. Establish optimal extraction conditions of blueberry

The optimal extraction conditions were established based on the results of extraction solvent, extraction method, component analysis, and physiological activity measurement.

5. Microencapsulation of fat-soluble vitamin A

5-1. Selection of optimal buffer for fat-soluble vitamin A

Vitamin A was first supplied with vitamin A palmitate 1.0 MIU / g (VIT A PALMITATE 55.5%, Diel alpha tocopherol 1%, Pinus oil 43.5%; Switzerland) and sorbitan fatty acid ester was edible grade, .

Sorbitan fatty acid ester was applied as a buffer to stabilize the capsule shape after pulling the fat-soluble vitamin A into the capsule (minimizing the standard value used in the existing food).

The optimum mixing ratio of fat - soluble vitamin A and sorbitan fatty acid ester was selected and the shape of capsule composed of fat soluble vitamin A and sorbitan fatty acid ester was observed with an optical microscope.

5-2. Selection of optimal coating for fat-soluble vitamin A

Agar was applied as a coating material to form a fat soluble vitamin A capsule. Soluble vitamin A and sorbitan fatty acid ester were dissolved in a water bath at 60 ° C. The coating material, agar, was added and microencapsulated using a homomixer at 13,000 rpm for 3 minutes using homogenized microcapsule equipment (GRACO, USA).

5-3. Size of fat-soluble vitamin A capsule

The morphology of capsules composed of fat soluble vitamin A and agar, a coating material, was observed with an optical microscope. Capsule size was measured using an optical microscope (BX 51TP, Olympus Co., Japan) and a calibrator (Konvison microscope co., LTD., Korea) and an alternative lens scale (Konvison microscopr co., LTE, Korea).

5-4. Measurement of encapsulation yield of vitamin A

C for measuring the encapsulation yield of vitamin A _{17: 0} microencapsulation of heptadecaenoic acid

The capsule material was sufficiently dissolved in distilled water, and then maintained at 60 DEG C. 200 mg of heptadecanoic acid (C17 _{: 0} , Sigma, USA) was added to 50 g of this solution. And homogenized in a 60 ° C water bath using a homomixer at 13,000 rpm for 3 minutes. 240 g of distilled water was sprayed with 10 g each using microcapsule equipment.

Pretreatment for yield measurement

10 g of the sample was placed in a sealed container, shaken vigorously by boiling in a hot water bath, and 60 mL of a solution prepared by mixing chloroform and methanol at a ratio of 2: 1 was added. After cooling tube was connected in 80 ℃ water bath and reaction was completed for 1 hour, the capsules were completely decomposed to extract the fat. 10 g each of the thus treated and untreated samples were taken in a separatory funnel, 20 mL of petroleum ether was added, and the mixture was shaken for 5 minutes and allowed to stand for 10 minutes. The lower layer was taken in another aliquot and the upper layer was subjected to secondary extraction in the same manner as described above, and the resultant solution was combined with the primary extract to obtain a fatty acid extract. The obtained extract was evaporated under reduced pressure (80 DEG C, 120 rpm) using a rotary evaporator to remove the solvent, and 15 mL of 1N-KOH ethanol was added to the remaining lipid. The lipid added with 15 mL of 1N-KOH ethanol was reacted in a 90 ° C water bath with a cooling tube for 1 hour and cooled. The cooled sample was transferred to a separatory funnel, 20 mL of ethyl ether was added, and the mixture was shaken for 10 minutes and allowed to stand for 5 minutes. The upper layer from which the non-adherent substances such as wax, sterol and the like were extracted was removed. The non-immiscible material was extracted by repeating the same procedure twice, and 1.6 mL of 1N HCl was added to the filtrate to adjust the pH to 2.0. 20 mL of petroleum ether was again added to the sample adjusted to pH 2.0, and this was repeated three times to extract the fatty acid mixture to remove moisture and obtain a fatty acid.

The methyl ester of the obtained fatty acid was prepared by methylation with a 14% BF ₃ -methanol solution as follows. To the ground-necked flask containing the fatty acid obtained above, 1 mL of 14% BF ₃ -methanol solution was added. A cooling tube was connected and reacted in a water bath at 90 ° C. for 3 minutes. 3 mL of a saturated NaCl solution was added, and 3 mL of hexane was added to separate the hexane layer. Anhydrous sodium sulfate was added to remove water and used as a sample for GC analysis.

Measurement of fatty acids by GC

For the identification of vitamin A, Sigma's standard retinyl palmitate was purchased and used. The retention time of the extracted vitamin A was compared with that of the standard material, and the concentration at the same time was measured. The analytical conditions of the GC used in the experiment are shown in Table 4 and the yield is estimated by the following equation.

    TF: Fatty acid concentration of total capsule solution

NF: Uncapsulated fatty acid concentration in the capsule solution

Gas chromatographic conditions of fatty acids Detector Flame ionization detector (FID) Column FUSED slica capillary column
(60 m x 0.25 mm x 0.2 m film thickness) Flow rate Split Ratio 10: 1 Carrier gas (He) 1 mL / min Air 450 mL / min H ₂ 45 mL / min Temperature Injection port 260 ℃ Detector 260 ℃ Column 180 ° C for 5 min
→ incresing by 10 ° C / min
260 ° C for 20 min Injection volume 1.0 μL

6. Mix of blueberry extract and microcapsule vitamin A

The optimal blending ratio of the blueberry extract and the vitamin A microcapsule and the content of the vitamin A microcapsule in the blueberry extract were set.

7. Measurement of physiological activity of the mixture

7-1. Antioxidant activity measurement

To determine the scavenging activity of the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical, the blueberry extract was mixed with an equal volume of distilled water and suspended. After filtration for 1 hour, the filtrate was concentrated under reduced pressure and centrifuged (1,200 rpm, 10 minutes) to 100 ml, and used as a sample for electron donating ability test. Electron Donating Ability (EDA) 0.2 ml of the sample extract, 0.8 ml of 0.04 ml DPPH reagent, was vortexed for 10 seconds, allowed to stand at room temperature for 10 minutes, and absorbance was measured at 525 nm.

EDA (%) = [1- (OD value of sample / OD value of blkn)] × 100

7-2. Evaluation of efficacy using SD rats

Duration 7 weeks (1 week of purification period, 6 weeks of experiment) Experimental animal Male SD Rat process HFD 45%, blueberry extract, vitamin A, lutein Inspection items HDL-c, LDL-c, Tc, TG, glutathione, myeloperoxidase, protaglandin E2 Experimental group Group 6 (5 animals / group) Sample Processing Oral administration One N + W 1000 μl / rat / day The standards for sample processing are set based on the 'Health Functional Food Standards and Specifications' (KFDA).

N: general formula
H: Highland choking
W: water
B: blueberry extract
A: vitamin A
L: lutein 2 H + W 1000 μl / rat / day 3 H + B 1000 μl / rat / day 4 H + B + A 1,000 + 1 μl / rat / day 5 H + B + L 1,000 + 10 μl / rat / day 6 H + B + A + L 1,000 + 1+ 10 μl / rat / day

8. Verification of the stability of the mixture of blueberry and vitamin A capsules

To verify the stability of the mixture consisting of blueberry and vitamin A capsules, the mixture was left at room temperature and observed to be stable.

9. Establishment of production process of blueberry mixture containing vitamin A capsules

The blueberry extract + vitamin A capsule raw material was mixed at 65 캜 using an agitator. The mixed solution was sterilized at 85 ± 3 ° C for 30 minutes. The prototype was produced through the product automatic packaging process.

II. Experiment result

1. Blueberry Extract

Both the purified water extract of the blueberry and the 80% alcohol extract had no significant difference in visual or sensory characteristics (Fig. 13).

2. Analysis of active ingredient of blueberry extract

The anthocyanin content of the blueberry purified water extract was 75.30 per mg / 100g, and that of the blueberry 80% alcohol extract was 78.80, which was higher than that of the purified water extract (FIG. 14).

Determination of anthocyanin content by solvent Sample mg / 100g Purified water 75.30 ± 1.06 80% alcohol 78.80 + - 10.03

Measurements were expressed as ± SD, p <0.05

3. Measurement of physiological activity of blueberry extract

Antioxidant activity measurement

The EDA value of the blueberry purified water extract was 80.42 and that of the blueberry 80% alcohol extract was 82.80, which was superior to that of the purified water extract (FIG. 15).

Determination of Antioxidative Activity by Solvent Sample EDA Purified water 80.42 + - 0.93 80% alcohol 82.80 ± 1.03

(0.1% ascorbic acid - 91.03%)

4. Establish optimal extraction conditions of blueberry

The blueberry purifying water extract showed lower anthocyanin content and antioxidant activity than the blueberry 80% alcohol extract.

Anthocyanin Content and Antioxidative Effect of Extracts with Different Extraction Solvents Sample mg / 100g EDA Purified water 75.30 ± 1.06 80.42 + - 0.93 80% alcohol 78.80 + - 10.03 82.80 ± 1.03

Based on the above results, the 80% alcohol extraction method was given priority, but there was no significant difference in the active ingredient and antioxidant effect, and the purified water extraction method was selected as the basic extraction method in consideration of the actual product production method and production cost. When the product is used in the production of the product, the purchase cost of the alcohol causes the cost increase, and the second step of removing the solvent after stirring extraction should be added.

5. Microencapsulation of fat-soluble vitamin A

5-1. Selection of Optimal Buffer of Vitamin A

As a result of the experiment to select the optimal ratio of vitamin A and buffer, the ratio of the sorbitan fatty acid ester (buffer), which allows the vitamin A to remain in a stable form after entering the capsule, is the ratio of vitamin A: buffer = 40: 1 It is possible to obtain an optimal shape (Fig. 16).

The ratio of vitamin A to sorbitan fatty acid ester Material ratio Vitamin A 40 Sorbitan fatty acid ester One

5-2. Selection of optimal coating for fat-soluble vitamin A

The ratio of microcapsule coating agent was selected.

The ratio of agar to vitamin A division ratio Agar: Vitamin A 1: 13

Agar was applied as a coating material for encapsulating vitamin A, and a stable capsule form was obtained when the ratio of agar to vitamin A was 1: 13 (Fig. 17).

5-3. Size of fat-soluble vitamin A capsule

Capsule size of vitamin A was measured using an alternate lens scale (Konvison microscopr co., LTE, Korea).

Vitamin A capsule size division size Vitamin A capsule 20 μm or less

5-4. Measurement of encapsulation yield of vitamin A

The yield of vitamin A in the capsules was about 98% (Fig. 19).

6. Mix of blueberry extract and microcapsule vitamin A

The optimal blending ratio of the blueberry extract and the vitamin A microcapsule and the content of the vitamin A microcapsule in the blueberry extract were set.

Blueberry extract and vitamin A capsule ratio division ratio Blueberry extract: Vitamin A capsule 110: 1

7. Measurement of physiological activity of the mixture

7-1. The DPPH radical scavenging ability of the mixture

The blueberry extract and the vitamin A capsule had a high antioxidative activity when mixed with the blueberry extract and vitamin A (FIG. 20).

Antioxidant activity measurement of mixture Sample EDA Blueberry extract 79.22 ± 1.08 Blueberry Extract + Vitamin A Capsule 88.37 + - 4.61

7-2. Evaluation of efficacy using SD rats

Blood total cholesterol

The results of total cholesterol analysis were as follows: (1) 61.81, (2) high - fat diet + 89.85, (3) high cholesterol + blueberry extract intake 87.28, (4) high cholesterol + blueberry + Vitamin A capsule supplementation group was 87.59, (5) high fat diet + Lutein capsule supplementation group was 88.36, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsule supplementation group was 88.81 (FIG. The lutein capsules were prepared by mixing vitamin A and lutein in a ratio of 1: 3, mixing the mixture with a sorbitan fatty acid ester in a ratio of 40: 1, and mixing the mixture with agar in a ratio of 13: 1.

Total cholesterol (Tc) Experimental group Tc (mg / dL) Sample Processing One N + W 66.81 + - 3.44 1000 μl / rat / day 2 H + W 89.85 8.10 1000 μl / rat / day 3 H + B 87.28 ± 15.73 1000 μl / rat / day 4 H + B + A 87.59 + - 8.97 1,000 + 1 μl / rat / day 5 H + B + L 88.36 +/- 11.93 1,000 + 10 μl / rat / day 6 H + B + A + L 88.81 + 9.07 1,000 + 1+ 10 μl / rat / day

(N: a general formula, H: a hyperphylla, W: water, B: blueberry extract, A: vitamin A

L: lutein)

The total cholesterol level was lower in all other groups than the group fed only high fat diet, but there was no significant difference. The total cholesterol level (3) was lowest in the high cholesterol + blueberry extract group. The blueberry extract is believed to have the effect of reducing total cholesterol.

Blood neutral lipid

The results of the neutral lipid analysis showed that (1) the general diet diet was 124.03, (2) the high fat diet was 176.47, (3) the high fat diet + the blueberry extract was 125.54, (4) + Vitamin A capsule ingestion group 125.24, (5) high fat diet + lutein capsule ingest group 149.84, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsule ingest group 110.59 (FIG.

Triglyceride (TG) analysis Experimental group TG (mg / dL) Sample Processing One N + W 124.03 + - 9.37 1000 μl / rat / day 2 H + W 176.47 ± 10.31 1000 μl / rat / day 3 H + B 125.54 + - 11.02 1000 μl / rat / day 4 H + B + A 125.24 + - 8.24 1,000 + 1 μl / rat / day 5 H + B + L 149.34 ± 5.34 1,000 + 10 μl / rat / day 6 H + B + A + L 110.59 ± 12.72 1,000 + 1+ 10 μl / rat / day

(N: general formula, H: high lipid cholesterol, W: water, B: blueberry extract, A: vitamin AL: lutein)

The triglyceride levels in all other groups were significantly lower than those in the high fat diet group. In the case of neutral lipid levels (5), the highest lipid level was found in the high fat diet + blueberry extract + vitamin A capsules + lutein capsules. It is considered that the combination of blueberry extract, vitamin A capsule and lutein capsule acts to increase the effect in lowering neutral lipids.

High-density lipoprotein cholesterol (HDL-c)

High density lipoprotein cholesterol analysis showed that (1) general diet diet 34.09, (2) high fat diet + water ingestion 39.09, (3) high fat diet + blueberry extract intake 38.28, (4) The extracts of vitamin A and vitamin A capsules were 39.13, 39.13, 40.19, and 6, respectively. The high frequency lipoprotein + blueberry extract + vitamin A capsules + lutein capsules intake group was 39.55 (FIG. 23).

High-density lipoprotein cholesterol (HDL-c) analysis Experimental group HDL-c (mg / dL) Sample Processing One N + W 34.09 ± 1.38 1000 μl / rat / day 2 H + W 39.09 + - 3.42 1000 μl / rat / day 3 H + B 38.28 + - 4.58 1000 μl / rat / day 4 H + B + A 39.13 + 1.91 1,000 + 1 μl / rat / day 5 H + B + L 40.19 + - 3.64 1,000 + 10 μl / rat / day 6 H + B + A + L 39.55 + 2.87 1,000 + 1+ 10 μl / rat / day

(N: general formula, H: high lipid cholesterol, W: water, B: blueberry extract, A: vitamin AL: lutein)

(5) High fat diet + Blueberry extract + Lutein capsules intake, (6) High fat diet + Blueberry extract + Vitamin A capsules + Lutein capsules But there was no significant difference. In the case of HDL-c, synergy of blueberry extract, vitamin A capsule, and lutein capsule does not appear.

Low density lipoprotein cholesterol (LDL-c)

The results of the low density lipoprotein cholesterol analysis showed that (1) general diet diet was 9.91, (2) high fat diet was 23.30, (3) high fat diet + high fat diet + blueberry extract diet was 21.47, (4) high fat diet + (20%), vitamin A (20%), vitamin A (20%), vitamin A (20%) and vitamin A (20%), respectively.

Low-density lipoprotein cholesterol (LDL-c) analysis Experimental group LDL-c (mg / dL) Sample Processing One N + W 9.91 + 1.78 1000 μl / rat / day 2 H + W 23.30 ± 5.33 1000 μl / rat / day 3 H + B 21.47 ± 5.15 1000 μl / rat / day 4 H + B + A 20.89 ± 5.96 1,000 + 1 μl / rat / day 5 H + B + L 19.10 ± 2.48 1,000 + 10 μl / rat / day 6 H + B + A + L 20.43 + - 3.58 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

LDL-c levels were lower in all groups than in the group fed only high-fat diets, and (5) LDL-c levels were lowest in high fat diet + blueberry extract + lutein capsules. In the case of LDL-c, the inhibitory effect is maximized through the binding of the blueberry extract and the lutein capsule.

5. PT (prothrombin Time)

The results of (1) general feeding diet group, (9), (2) high water diet, (+) water ingestion group, 8.55, (3) high fat diet + blueberry extract group, 8.88 (4) High fat diet + blueberry extract + vitamin A capsules intake 8.88, (5) high fat diet + lutein capsule intake group 8.64, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsules Group 8.04 (FIG. 25).

Analysis of foreign coagulation mechanism (PT) Experimental group PT (sec) Sample Processing One N + W 9.12 ± 0.54 1000 μl / rat / day 2 H + W 8.55 0.63 1000 μl / rat / day 3 H + B 8.88 + - 0.67 1000 μl / rat / day 4 H + B + A 8.88 ± 0.80 1,000 + 1 μl / rat / day 5 H + B + L 8.64 ± 0.23 1,000 + 10 μl / rat / day 6 H + B + A + L 8.04 0.36 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

There was no significant difference in the time taken for blood clotting according to the externalization mechanism in all groups compared to the group receiving high fat diet alone. However, (3) blueberry extract group and (4) blueberry extract + vitamin A capsule ingestion group were found to have little possibility of slowing the coagulation time according to the foreign body coagulation mechanism.

6. Activated Partial Thromboplastin Time (aPTT)

(1) general diet diet group, (2) high fat diet group, (+) water ingestion group, (20.66), (3) high fat diet group, + blueberry extract group, 18.88 , (4) high fat diet + blueberry extract + vitamin A capsule ingestion group 20.42, (5) high fat diet + lutein capsule ingestion group 18.64, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsule ingestion Group 18.24 (FIG. 26).

My handover coagulation assay (aPTT) Experimental group aPTT (sec) Sample Processing One N + W 19.10 ± 0.47 1000 μl / rat / day 2 H + W 20.66 ± 2.65 1000 μl / rat / day 3 H + B 18.88 +/- 0.72 1000 μl / rat / day 4 H + B + A 20.42 ± 2.26 1,000 + 1 μl / rat / day 5 H + B + L 18.64 ± 0.53 1,000 + 10 μl / rat / day 6 H + B + A + L 18.24 + - 0.72 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

In all groups, there was no significant difference in the time taken for blood clotting according to extrinsic mechanism, or rather the prolongation of clotting time was found to be ineffective for the transplantation - related mechanism.

7. Glutathione

(1) dietary intake of 143.96, (2) high fat diet + 106.59, (3) high fat diet + blueberry extract intake 121.44 (4) high fat diet + high fat diet + vitamin A capsule ingestion group 123.66, (5) high lipid diet + lutein capsule intake group 114.49, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsules Group 125.87 (FIG. 27).

Results of glutathione analysis Experimental group Glutathione
(/ Ml) Sample Processing One N + W 143.96 ± 22.78 1000 μl / rat / day 2 H + W 106.59 ± 19.74 1000 μl / rat / day 3 H + B 121.44 + - 11.30 1000 μl / rat / day 4 H + B + A 123.66 ± 16.73 1,000 + 1 μl / rat / day 5 H + B + L 114.49 ± 9.36 1,000 + 10 μl / rat / day 6 H + B + A + L 125.87 ± 26.06 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

Compared with the group fed only with high fat diet, all groups showed a high level of reactive oxygen scavenging activity. (3) blueberry extract, (4) blueberry extract + vitamin A capsule, and (6) blueberry extract + vitamin A capsule + lutein capsule. I was able to find something very good. In particular, it was shown that the mixture of blueberry extract, vitamin A capsule and lutein capsule showed the most effective oxygen scavenging effect, whereas in the group treated with only lutein capsule, the oxygen scavenging effect was very low. Blueberry extract, Vitamin A capsule and Lutein capsule complex can increase the active oxygen removal efficiency through interaction.

8. Myeloperoxidase (Myeloperoxidase)

(1) general diet diet group 7.53, (2) high fat diet + water ingestion group 10.04, (3) high fat diet + blueberry extract (6.4), (6.) High cholesterol diet + blueberry extract + vitamin A capsules + 6.77, (4) high cholesterol diet + Lutein capsule ingestion group was 6.65 (FIG. 28).

Myeloperoxidase assay results Experimental group Myeloperoxidase (ng / ml) Sample Processing One N + W 7.53 + - 0.82 1000 μl / rat / day 2 H + W 10.04 + - 1.15 1000 μl / rat / day 3 H + B 6.47 1.87 1000 μl / rat / day 4 H + B + A 6.77 3.80 1,000 + 1 μl / rat / day 5 H + B + L 7.42 ± 0.64 1,000 + 10 μl / rat / day 6 H + B + A + L 6.65 ± 0.65 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

In all groups, the inflammatory response was lower than in the group receiving only high fat diet. (3) blueberry extract, (4) blueberry extract + vitamin A capsule, and (6) blueberry extract + vitamin A capsule + lutein capsule. I can see that this is very good. Analysis of the degree of inflammation, which is an indicator of antioxidant efficacy, showed that blueberry, vitamin A capsules and lutein capsules are highly effective in reducing inflammation in the body through interaction, while lutein It can be seen that when the capsule alone is used, the effect is halved.

9. Prostaglandin E2 (Prostaglandin E2)

(2) high fat diet + 238.86, (3) high fat diet + blueberry extract intake group 237.02, (2) high fat diet + high fat intake group, (4) high-fat diet + blueberry extract + vitamin A capsules 218.12, (5) high fat diet + lutein capsules intake 232.60, (6) high fat diet + blueberry extract + vitamin A capsules + lutein capsules 215.53 (FIG. 29).

Results of prostaglandin E2 analysis Experimental group Prostaglandin E2
(pg / ml) Sample Processing One N + W 216.07 ± 15.12 1000 μl / rat / day 2 H + W 238.86 + - 7.35 1000 μl / rat / day 3 H + B 237.02 ± 9.99 1000 μl / rat / day 4 H + B + A 218.12 + - 21.36 1,000 + 1 μl / rat / day 5 H + B + L 232.60 ± 12.76 1,000 + 10 μl / rat / day 6 H + B + A + L 215.53 + - 12.45 1,000 + 1+ 10 μl / rat / day

B: blueberry extract, A: vitamin A, L: lutein)

In all groups, the inflammatory response was low compared to the group receiving only high fat diet. In particular, (4) blueberry extract + vitamin A capsule ingestion group, (6) blueberry extract + vitamin A capsule + lutein capsule ingestion group showed significant difference and the inflammation suppressing effect was remarkable. As a result of analyzing the degree of inflammation, which is an index of antioxidant efficacy, Blueberry, Vitamin A capsule and Lutein capsule are considered to be very effective in reducing inflammation in the body through interaction, When using only berry + lutein capsules or blueberries, the effect is halved.

Comprehensive analysis

Efficacy evaluation index Interaction Blueberry extract Blueberry Extract Vitamin A Capsule Blueberry Extract Lutein Capsule Blueberry Extract Vitamin A Capsule
Lutein Capsule Tc + + - - TG +++ +++ ++ ++++ HDL-c - - + - LCL-c ++ ++++ ++++ +++ PT - + + - aPTT - - - - Active oxygen scavenging ability ++ +++ + ++++ Inflammation inhibition Ⅰ ++++ +++ ++ +++ Inflammation inhibition Ⅱ + ++++ + ++++ Total (total score 36) 13 19 12 18

10. Verification of stability of mixture composed of blueberry and vitamin A capsules

The mixture (blueberry extract and vitamin A capsule) was stored at room temperature for 180 days to confirm the stability of the mixture composed of blueberry extract and vitamin A capsule. As a result, it was confirmed that the mixture was maintained stably (FIGS.

Example 3 Microencapsulation of Acai Berry Extract

1. Raw material preparation

Prepare water-soluble Asaiberi (Experimental), DSM Nutri- tional Products (Switzerland), Edible agar (Meishin agar), Gelatin (Geltech), and Fatty acid ester (ILSHINWELLS).

2. Preparation of microcapsule solution

Fat-soluble vitamin E, edible agar, gelatin and fatty acid esters were uniformly mixed in accordance with the ratios shown in Table 30. The ratio of vitamin A to fatty acid ester (buffer) can be optimized for vitamin E: buffer = 40: 1.

Selection of Vitamin E Microencapsulation Experiment division Furtherance Vitamin E oil 30g Fatty acid ester 0.5% (from oil + coating) (0.75 g) Coating material Total (120 g)
Agar 1.5% (1.8g) Gelatin 0.5% (0.6g)
Distilled water (117.6 g)

3. Microencapsulation

The homogenized capsule solution was sprayed under the conditions shown in Table 31. Experiments were conducted with purified water to select a solution suitable for microcapsule preparation, and the purified water was finally selected as a solution suitable for microcapsule production.

Injection of homogenized capsule solution division Furtherance Injection pressure 10 psi Injection nozzle Radial Injection time 1 min Injection capacity 200 mL

4. Asai berry and microcapsules

Acai Berry and Vitamin E + coating materials were microencapsulated at 24: 1.

5. Stability test

In order to test the thermal stability of the Asai berry and vitamin E microcapsules, they were allowed to stand at 85 ° C for 30 minutes. As a result, the Aceberry and vitamin E microcapsules showed high stability.

6. Preparation of prototype material

(Mass production) Asaieri mixed concentrate for the prototype was prepared.

7. Raw material mixing

Asaiibery mixed concentrate and vitamin E microcapsules were mixed at 65 캜 using a quiescent period.

8. Sterilization and product packaging

The mixture solution was sterilized at 85 ± 3 ° C. for 30 minutes and the product was packed in an automatic packing machine.

9. Characteristics and effects of products

Asai Berry is particularly well-known in the antioxidant product line. In the past, when using antioxidant-related products, it was inconvenient to separately ingest water-soluble fruit juice and fat-soluble vitamins. However, this product included both of the above, thus satisfying the consumers' most important ingestion ease. In addition, the modification of the property and the loss of the function that could be caused by the direct contact of the lipid soluble substance with the water soluble substance was fundamentally compensated, and the chemical substance was replaced with the natural agar as the microcapsule material.

On the other hand, through the microencapsulation technique, a substance which does not dissolve in a water-soluble substance is stably mixed by the combination of a water-soluble substance and a lipid-soluble substance. The microencapsulation process minimizes the emulsifier (by 80-97% or more), so that the fat soluble vitamin E is mixed with the water soluble berry extract, minimizing the odor of the lipophilic material and improving the functionality, thus minimizing the use of the additive removing additive . In addition, the micro-sized capsules were applied to the product to minimize the foreign body sensation to the capsules to facilitate the ingestion (100 μm → 10 μm or less), and the synergy effect through the combination of the water-soluble substance and the lipophilic substance was obtained Antioxidant synergistic effect: water soluble antioxidant + fat-soluble vitamin E) (FIG. 32).

The product manufactured by the above method can exhibit synergistic effects of various functions through binding of Asaiberi and Vitamin E. This product can meet consumers' purchase desire. It is a product that can appeal to the younger generation who considers the spread of consumer trend to maintain health through fruit juice drink and skin health. Most of the products used for skin health at home and abroad are limited to external preparations, but at a point when they are gradually shifting toward food intake, this product can be positioned as a functional food that has no side effects and can be easily ingested can do.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

A method of microencapsulation of a lipophilic material comprising the steps of:
(a) preparing a capsule solution comprising a lipid-soluble substance having a physiological activity; And
(b) preparing a microcapsule by adding a coating solution to the capsule solution and homogenizing the capsule solution, wherein the coating solution is gelatin, agar, water or a combination thereof.
A method for preparing a functional beverage using a microencapsulated liposoluble material comprising the steps of:
(a) preparing a capsule solution comprising a lipid-soluble substance having a physiological activity;
(b) preparing a microcapsule by adding a coating solution to the capsule solution and homogenizing the capsule solution, wherein the coating solution is gelatin, agar, water or a combination thereof; And
(c) spraying the microcapsules onto the fruit extract.
3. The method according to claim 1 or 2, wherein the fat-soluble substance is selected from the group consisting of omega fatty acids, lutein and fat soluble vitamins.
4. The method according to claim 3, wherein the omega fatty acid is omega-3, omega-6 or omega-9.
4. The method according to claim 3, wherein the fat-soluble vitamin is one or more vitamins selected from the group consisting of vitamin A, vitamin D, vitamin E, vitamin F, vitamin K and vitamin U.
3. The method according to claim 1 or 2, wherein the microcapsules have a diameter of 1-50 [mu] m.
A microcapsule prepared by the method of claim 1.
A functional beverage made by the method of claim 2.

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