KR100666830B1 - Alginate microcapsule encapsulated extract of horseradish and preparation method thereof - Google Patents

Abstract

The present invention relates to alginate microcapsules encapsulated with wasabi extract and a method for producing the same. Alginate microcapsules collected from the horseradish extract of the present invention has an antimicrobial activity while being shielded from the strong irritation and high volatility of wasabi extract to improve the usability of wasabi extract can be usefully used in various fields.

Wasabi extract, alginate, microcapsules, AIT (Allylisothiocyanate)

Description

Alginate microcapsule encapsulated extract of horseradish extract and preparation method

1 is a schematic diagram showing a cross-sectional view of an air atomizer that can be used in the manufacturing process of the present invention.

* Explanation of symbols for main parts of the drawings

1: tank 2: spray nozzle

3: calcium chloride solution 4: air filter

Figure 2 is a photograph of the alginate microcapsules encapsulated wasabi extract according to the present invention with an electron microscope (SEM).

The present invention relates to alginate microcapsules encapsulated with wasabi extract and a method for producing the same. More specifically, the present invention relates to alginate microcapsules encapsulated with water extract of concentrated wasabi and a method for producing the same.

Wasabi is classified as a plant belonging to the Brassicaceae (十字花科) as Japanese horseradish (Wasabia japonica) and standing production wasabi (Cocholearia armoracia). Japanese horseradish is also called wasabi (Korean name, horseradish), and its root is mainly used as a spice. Wasabi has the effect of removing fishy fish smell and at the same time, enhancing the taste.It has the effect of improving appetite, enhancing vitamin B₁ synthesis, promoting beta amylase activity, antioxidant activity, and antibacterial activity, and therefore gaining attention as a health food. have. In particular, it is common in Korea and Japan to consume with sashimi or sushi.

The activity of horseradish is due to isothiocyanates produced by hydrolysis by the hydrolytic enzyme myrosinase when the isothiocyanate glycoside (Sinigrin) present in the cells is disrupted. do. In particular, the antibacterial activity of horseradish is exerted by isothiocyanate (RN = C = S), which is mainly composed of volatile allylisothiocyanate (AIT). Isothiocyanate (RN = C = S) in horseradish is a Helicobacter pylori known as a causative agent of rot, gastritis or gastric ulcers, such as cholera bacteria, food poisoning bacteria such as typhoid fever, and pathogenic yeasts associated with rancidity of other foods. ( Helicobacter pylori ) It is known to exhibit antimicrobial activity against and the like. In addition to the AIT, there are various isothiocyanates having different R groups of glycosides, and 6-methylsulfinylhexyl isothiocyanate (6-MeS-NCS), which is mainly contained in the roots and leaves of horseradish, is soy sauce. Has also been shown to increase the activity of glutathione S-transferase and quinone reductase, enzymes that inhibit chemical stress or inhibit their formation in the early stages of tumor formation. .

Due to these various physiological activities wasabi was mainly applied to food for the purpose of improving the freshness or preservation. For example, Korean Patent Laid-Open Publication No. 2002-26929 discloses a freshness-retaining agent of meat containing horseradish extract, and Korean Patent Laid-Open Publication No. 2003-94194 discloses kimchi in addition to the main ingredients and seasonings to improve the preservation of kimchi. Wasabi kimchi prepared by adding wasabi is disclosed.

In addition to horseradish, wasabi has been applied to soap, toothpaste, and the like. For example, Korean Patent Publication No. 2003-46545 discloses an antibacterial soap containing horseradish extract, and Korean Patent Publication No. 2003-64902 discloses an antibacterial toothpaste containing horseradish extract.

However, in the case of using it as an extract or in the form of extract without processing wasabi as described above, there is a limit in applying to the product due to the strong irritability and high volatility of wasabi. Therefore, there is a need for a method for shielding strong stimulation and high volatility while maintaining various physiological activities of wasabi.

Microencapsulation technology is a process of encapsulating or coating liquid, solid or gas molecules with polymers to isolate unstable substances such as perfumes and nutrients from the external environment such as light, oxygen, and moisture and conceal toxicity, smell and taste. It is used for the purpose of simplifying the handling and adjusting the dissolution rate of the contents by solidifying.

Specific microencapsulation methods include spray drying method, flow coating method, and emulsion solvent evaporation method.

On the other hand, in order to microencapsulate the useful components, physical and chemical properties and physiological characteristics are different according to each useful component, so conditions such as the amount of the useful components encapsulated in the polymer, the type and amount of the polymer used, the particle size, and the degree of release of the useful component, etc. Should be optimized.

Therefore, while the present inventors are studying a method that can be masked the strong irritation and high volatility of wasabi extract, to prepare a microcapsule containing the wasabi extract by microencapsulating the concentrate of the wasabi extract under optimized conditions and Using the microcapsules, the present invention was completed by confirming that the stimulating activity and high volatility of wasabi may be maintained while maintaining the physiological activity of wasabi.

Therefore, an object of the present invention is to provide a method for preparing alginate microcapsules containing horseradish extract.

Still another object of the present invention is to provide an alginate microcapsules containing horseradish extract prepared by the above method.

Still another object of the present invention is to provide an antimicrobial composition comprising alginate microcapsules containing the wasabi extract.

In order to achieve the above object, the present invention provides a method for preparing alginate microcapsules containing wasabi extract.

Another object of the present invention to provide an alginate microcapsules encapsulated wasabi extract prepared by the above method.

Another object of the present invention provides an antimicrobial composition comprising alginate microcapsules containing the wasabi extract.

Hereinafter, the present invention will be described in detail.

Preparation method of alginate microcapsules containing wasabi extract of the present invention

(a) preparing wasabi water extract;

(b) concentrating wasabi extract of step (a);

(c) mixing the wasabi extract and the alginate solution of step (b);

(d) spraying the mixed solution of step (c) into the polyvalent cation solution through a nozzle; And

(e) preferably washing and drying the microcapsules formed in step (d).

 The manufacturing method of the polymer beads according to the present invention will be described in detail for each step as follows.

(a) Preparation of Wasabi Water Extract

In order to prepare a microcapsule containing the horseradish extract of the present invention, it is preferable to use horseradish water extract. This is because the water extract is safer than the organic solvent extract.

Wasabi is not particularly limited, and Wasabia japonica or Wasabia japonica or Cocholearia armoracia may be used. In addition, wasabi may be selected from wasabi, roots, stems and leaves of wasabi.

The manufacturing method of the water extract of wasabi is not particularly limited and can be prepared by adding water to wasabi. Preferably, water was added to horseradish at a weight ratio of 3 to 7 times and extracted at 35 to 37 ° C. for 1 to 2 hours. At this time, in order to increase the extraction efficiency during the extraction can be used by stirring with a shaker (shaker) or finely cut wasabi.

In addition, the horseradish water extract is further subjected to the step of standing wasabi for 1 to 2 hours at 35-37 ℃ before extraction, while isothiocyanate glycosides are present in the wasabi cells to the isothiocyanate It is possible to maximize the activity of degrading myrosinase (myrosinase).

(b) Concentration of Wasabi Water Extract

This step is the step of concentrating the wasabi water extract of step (a) isothiocyanates, preferably alkyl isothiocyanate, 6-methylsulfinylhexyl isothiocyanate contained in the wasabi water extract (6-MeS-NCS), isothiocyanates such as allyl isothiocyanate, and most preferably, can be concentrated until the concentration of allyl isothiocyanate reaches 2,000 to 8,000 ppm. In order to make the concentration of allylisothiocyanate fall within the above range, it is preferable to concentrate the horseradish water extract of step (a) 5 to 15 times. The concentration can be easily carried out using a method known in the art, such as a rotary evaporator concentration method.

On the other hand, the content range of the preferred allyl isothiocyanate contained in the concentrate of the horseradish water extract is optimized so that the microcapsules prepared by the method of the present invention is a suitable amount to exhibit the biological activity of wasabi. That is, when the concentration of allyl isothiocyanate in the concentrate of the wasabi extract is less than 2,000ppm, the microcapsules prepared by using it were insufficient to show the physiological activity of wasabi and the concentration of allylisothiocyanate was 8,000ppm. When exceeded, it was shown that the biological activity is no longer increased (see Table 1).

(c) mixing the concentrate and alginate solution of horseradish water extract

This step is to mix the wasabi extract and the alginate solution. In the present invention, the alginate may be in the form of a salt of alginic acid, which is preferably linear polyglucuronic acid, and more preferably, sodium (Na), calcium (Ca), or magnesium of alginic acid. (Mg) in salt form. Most preferably, of the water soluble alginic acid Sodium salt form. The alginate salt of the present invention can be extracted from brown algae such as kelp and seaweed, or a commercially available product can be used. In the present invention, the alginate solution may be prepared by dissolving alginate in a suitable solvent. For example, it may be prepared by adding alginate to the purified water to dissolve it. Preferably, the alginate may be prepared by dissolving it in purified water to 1.0 to 3.0% (w / v). Most preferably, the alginate solution may be prepared by adding alginic acid to purified water at a concentration of 1.8% (w / v). If the concentration of alginate is less than 1.0% (w / v), there is a problem that gel formation does not occur. If the concentration exceeds 3.0% (w / v), the gel is too hard and the useful components inside the gel are free. There is no problem.

In addition, the alginate solution preferably has a viscosity of 2,500 to 3,500 cps.

Meanwhile, the mixing ratio of the useful ingredient of the microcapsules and the alginate solution, which is a capsule material, has an important effect on the density, yield and stability of the microcapsules. Accordingly, the present inventors prepared microcapsules by varying the ratio of the concentrate and alginate solution of horseradish water extract, and then investigated the number, yield, irritation and antimicrobial activity of the microcapsules according to the mixing ratio. As a result, when the mixing ratio of wasabi extract and alginate solution was less than 1: 4, the number of microcapsules was small, so that empty spaces were observed, and strong irritation of wasabi extract was detected. On the other hand, when the mixing ratio exceeded 1:10, a lot of microcapsules were formed, but the yield of the microcapsules including the concentrate of horseradish water extract was decreased and the amount of the microcapsules containing alginic acid was increased. In addition, the microcapsules when the mixing ratio exceeded 1:10 was shown to decrease the antimicrobial activity due to the low concentration of wasabi extract concentration containing (see Table 2).

Therefore, it is preferable to mix the wasabi extract and the alginate solution in the step (c) in a volume ratio of 1: 4 to 10.

In addition, it is preferable that the mixture of the wasabi water extract and the alginate solution are mixed at a temperature of 35 to 37 ° C.

(d) spraying the mixed liquid

This step is a step of preparing the alginate microcapsules by curing by crosslinking the polyvalent metal cation and alginate by spraying the mixed solution of step (c) through the nozzle to the polyvalent cation solution.

The present inventors made the alginate microcapsules encapsulated with wasabi water extract nanoscale to further extend the scope of application. That is, the microcapsules made of nano-sizes can be prepared in the form of a suspension, which has the advantage of being sprayed or applied to the surface of the product to be applied. In the present invention, the nano size is preferably 0.1 to 1.0nm.

The present inventors have optimized the injection conditions such as nozzle size, injected air pressure and liquid injection pressure so that the horseradish water extract encapsulated microcapsules can be produced in the preferred size of the nano-sized range. Specific spray conditions include nozzle size 0.5 to 2.0 mm, sprayed air pressure of 0.1 to 2.0 kgf / cm ² and liquid spray pressure of 0.1 to 2.0 kgf / cm ² .

In the above polyvalent metal cation solution, calcium chloride (CaCl ₂ ), barium chloride (BaCl ₂ ), strontium chloride (SrCl ₂ ), magnesium chloride (MgCl ₂ ), and zinc chloride (ZnCl), which are compounds having an ionic group that can be crosslinked with alginate ₂ ) and the like, and preferably a solution of calcium chloride (CaCl ₂ ). The concentration of the polyvalent metal cation used may be 0.05 to 0.5M, preferably 0.5M. The reaction time may be 10 to 40 minutes, preferably 30 minutes.

(e) washing and drying of microcapsules

This step is to wash and dry the microcapsules formed in step (d). In the washing, it is preferable to use distilled water, and the drying may be air drying, freeze drying or heat drying.

The method of the present invention described above has the effect of obtaining alginate microcapsules encapsulated with wasabi extract in a high yield of 97% or more.

Accordingly, the present invention provides alginate microcapsules encapsulated with wasabi extract prepared according to the above method. The microcapsules exhibit strong antimicrobial activity, but also have strong stimulation and high volatility.

The microcapsules include, but are not limited to, Staphylococcus sp. Bacteria, Lactobacillus sp. Bacteria, cholera bacteria, typhoid bacteria, Helicobacter sp. Bacteria, pathogenic E. coli ( E. coli ), Vibrio sp. bacteria and Bacillus sp. bacteria.

In one experiment of the present invention the microcapsules according to the invention Helicobacter pylori (Helicobacter pylori, NCTC 11637), Escherichia coli (E. coli O157: H7, ATCC 43889), Staphylococcus aureus (Staphylococcus aureus, ATCC 25923 ), for enteritis non brioche bacteria (Vibrio parahaemolyticus, ATCC 2210001), Salmonella typhimurium (Salmonella typhimurium, KCTC 2058), Bacillus cereus (Bacillus cereus, KCTC 1012) and Bacillus subtilis (Bacillus subtilis, ATCC 6633) It was confirmed that it has a strong antibacterial activity (see Table 1 and Table 3).

In addition, the present invention provides an antimicrobial composition comprising alginate microcapsules encapsulated with wasabi water extract of the present invention. More specifically, the antimicrobial composition may be used to inhibit or kill the growth of harmful bacteria and may be formulated in various forms. The formulations include, but are not limited to, cosmetics, disinfectants, sterile detergents and food preservatives.

In the above cosmetics may be easily prepared according to known methods in the art, including one or more excipients and additives generally used in the manufacture of cosmetic compositions with the microcapsules of the present invention. More specifically, the cosmetics, such as face lotion, cream, essence, cleansing foam and cleansing water, basic cosmetics such as packs, body oil, etc., color cosmetics such as foundation, lipstick, mascara and makeup base, shampoo, rinse, hair conditioner And hair products such as hair gels, soaps, and the like.

The disinfectant may be prepared by diluting the microcapsules of the present invention with a dermatologically and pharmacologically acceptable diluent or solvent. The disinfectant may be used on the surface of the organism, preferably on the skin of a mammal, most preferably on the skin of a human. The disinfectant may also be used on inanimate surfaces, such as wood, metal, glass, ceramics, plastics, paper and cloth. The disinfectant may be treated on the surface of the living or non-living object using a method including dipping, swab, spraying and brushing. Preferably, the disinfectant may be used in hospitals and homes for the purpose of wound surface, pre-surgical skin disinfection, surgical instrument disinfection, conduit disinfection and the like.

In the case of the sterilizing detergent, in addition to the microcapsules according to the present invention, one or more excipients and additives generally used in the preparation of the detergent may be easily prepared according to methods known in the art. The disinfectant detergent may be used for kitchen or food, and the disinfectant detergent for kitchen may further include anionic surfactant, nonionic surfactant, amphoteric surfactant, etc. in addition to the microcapsules of the present invention. Examples of anionic surfactants include alkylbenzenesulfonates, alphaolefin sulfates, sodium or ammonium lauryl ether sulfates, and examples of nonionic surfactants include fatty acid amides, alkylpolyglucosides, ethoxylated fatty alcohols, and ethoxylates. And cylated nonylphenol. Examples of amphoteric surfactants include betaine-based surfactants, amine oxides, and the like. In addition to the above surfactants, diluents such as flavors, pigments and water may be optionally added. In the case of food sterilizing detergent, it can be prepared by further adding a food acceptable solvent or diluent to the microcapsules according to the present invention and diluting to an appropriate concentration. The food sterilizing detergent may be used for sterilization and washing of fruits, fish and meat, for example.

Food preservatives are added to the microcapsules according to the present invention in addition to the microcapsules according to the present invention in order to maintain the freshness of the food by preventing the decay of food products by rancidosis, especially for the purpose of enhancing the preservation of foods. Can be prepared. The food preservative of the present invention may be added together with the raw materials in the manufacturing process of the food or prepared into a separate water-soluble suspension. In addition, the food preservative according to the present invention may be used to immerse the target food or spray the food preservative of the present invention onto the target food. Furthermore, the food preservative of the present invention can prevent deterioration and corruption such as rancidity, cloudiness, and swelling of bags due to the proliferation of rancid microorganisms, thereby extending its shelf life when used in packaged foods.

For example, by adding an antimicrobial composition comprising alginate microcapsules encapsulated wasabi extract of the present invention to the immersion liquid of the tofu to inhibit the propagation of microorganisms during the storage period it is possible to enable long-term storage.

On the other hand, the antimicrobial composition comprising the alginate microcapsules encapsulated wasabi extract of the present invention as described above is excellent in antimicrobial effect, but almost no smell or skin irritation can be used safely in the human body.

In the experimental example of the present invention, the antimicrobial activity and odor or skin irritation of the soap containing the microcapsules according to the present invention was tested, it was confirmed that the antibacterial activity is very excellent compared to the general soap, but almost no smell or skin irritation. (See Table 4 and Table 6).

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1>

Preparation of Wasabi Extract

After cutting 100 g of wasabi from Japan, 300 ml of water was added thereto and left at 35 ° C. for 2 hours. Then it was extracted for 2 hours at 37 ℃ under atmospheric pressure to give 1690ml horseradish water extract.

<Example 2>

Preparation of Alginate Microcapsules Enclosed with Wasabi Water Extract According to the Method of the Present Invention

The horseradish water extract of Example 1 was concentrated 10 times on a volume basis using a rotary evaporator so that the concentration of allyl isothiocyanate contained was 8000 ppm to obtain 169 ml of the wasabi extract. It was.

5.4 g of sodium alginate (Sigma Aldrich Co., Ltd) was added to 127.5 ml of purified water to prepare a 1.8% (w / v) sodium alginate solution, and 169 ml of the concentrate of the wasabi water extract was added to prepare a total of 300 ml of the mixture. . Next, 2.4 L of purified water was added to 43.2 g of calcium chloride (CaCl ₂ ) to prepare 2.4 L of a 0.5 M calcium chloride solution. The mixture of the sodium alginate solution and the concentrate of the wasabi extract was sprayed onto the calcium chloride solution using an air atomizer (FIG. 1). That is, the sodium alginate solution and wasabi extract were placed in the tank 1 of the air atomizer and sprayed onto the calcium chloride solution 3 at a liquid spray pressure of 0.6 kgf / cm ² through a nozzle 2 of 0.9 mm. At this time, the air sterilized through the air filter 4 was to be injected together at a pressure of 0.6kgf / cm ² . Thereafter, the mixture was stirred at 25 ° C. for 30 minutes, and the obtained microcapsules were washed with distilled water and then dried. The prepared microcapsules were examined under a microscope and the diameter was measured using a hematocytometer (Paul Marienfeld GmbH & Co. KG, Lauda-Koenigshofen). Microencapsulation yield was calculated by representing the total volume of microcapsules produced as a percentage of the total volume of the samples used (aqueous sodium alginate, wasabi extract and calcium chloride solution).

As a result, it was confirmed that a bead of beads in the range 0.1 ~ 0.4mm in diameter was formed (Fig. 2), the yield was confirmed to be 97%.

Experimental Example 1

Determination of the Preferred Concentration Range of Wasabi Extract

<1-1> Preparation of Microcapsules Containing Wasabi Water Extract Concentrated at Various Concentrations

In order to determine the preferred concentration range of wasabi water extract for the preparation of microcapsules, the wasabi extract of Example 1 was used in a rotary vacuum evaporator to allyl isothiocyanate at various concentrations (1,000, 2,000, 4,000, 6,000, 8,000, 10,000 ppm).

The concentration of allylisothiocyanate was measured using gas chromatography (model HP6890; Hewlett Packard Co., Palo Alto, CA, USA). For gas chromatography analysis, HP-INNOWax capillary column (30m × 0.32mm id, 0.5㎛ film thickness; Agilent Technologies, Inc., Palo Alto, CA, USA) was used, and the sample injection volume was 1 μl. 14.2 psi silver, helium was used as the carrier gas. The oven temperature was raised at a rate of 15 ° C./min from 50 to 250 ° C. and the injector and flame ionization detectors were at 280 ° C. and 270 ° C., respectively. Allyl isothiocyanate (purity 98% or more, Fluka Chemical Co., Buchs SG, Swizerland) was used as a standard. The microcapsules were prepared in the same manner as in Example 2 using the concentrate of horseradish water extract prepared to include allyl isothiocyanate at various concentrations.

<1-2> Antimicrobial Activity of Microcapsules According to Concentration of Allyl Isothiocyanate in Wasabi Extract

By measuring the antimicrobial activity of the microcapsules of Experimental Example <1-1> by disk diffusion method, the preferred concentration range of wasabi water extract for preparing microcapsules was determined. Is Helicobacter pylori (Helicobacter pylori, NCTC 11637) as test organism, Escherichia coli (E. coli O157: H7, ATCC 43889), Staphylococcus aureus (Staphylococcus aureus, ATCC 25923), Vibrio parahaemolyticus (Vibrio parahaemolyticus, ATCC to 2,210,001), Salmonella typhimurium (Salmonella typhimurium, KCTC 2058), Bacillus cereus (Bacillus cereus, KCTC 1012) and Bacillus subtilis (Bacillus subtilis, ATCC 6633) were used.

More specifically, each of the pre-cultured pathogenic bacteria was dispensed by dispensing 10 μl into a plate medium for measuring the disk diffusion method and a sterilized paper disk of 8 mm diameter was attached to the plated plate medium. Herein, the microcapsules containing the wasabi extract prepared above were dispensed on the paper disk, and the plate medium was incubated at 37 ° C. for 24 hours, and then the size of the transparent ring (growth block) was measured.

As a result, it was found that the concentration of allyl isothiocyanate contained in horseradish water extract for preparing microcapsules was 2,000 to 8,000 ppm. That is, when the concentration of allyl isothiocyanate contained in wasabi extract is less than 1,000ppm, the microcapsules prepared using the same showed low antimicrobial activity, and when the concentration of allyl isothiocyanate was 10,000ppm or more It was found that there was no difference with the activity of 8,000 ppm (Table 1).

Antimicrobial Activity of Microcapsules According to the Concentration of Allyl Isothiocyanate in Wasabi Extract Allylisothiocyanate concentration Antimicrobial Activity (Growth Base, mm) Helicobacter pylori Escherichia coli Enteritis vibrio Salmonella Typhimurium Staphylococcus aureus Bacillus cereus Bacillus subtilus 1,000 ppm 8 15 28 18 2 4 9 2,000 ppm 15 23 27 29 11 17 14 4,000ppm 25 23 28 29 21 26 28 6,000 ppm 26 24 28 29 28 25 29 8,000 ppm 26 24 28 29 32 26 29 10,000 ppm 26 24 28 29 32 26 29

Experimental Example 2

Determination of the Preferred Mixing Ratio of Wasabi Water Extract Concentrate and Alginate Solution

In order to determine the desired mixing ratio of the wasabi extract and the alginate solution, the concentrate and sodium alginate solution of wasabi extract prepared in the same manner as in Example 2 were 1: 3, 1: 4, 1: 5, 1:10 and 1:11, respectively, and mixed in volume ratios. Next, the microcapsules were prepared in the same manner as in Example 2, and the microcapsules were examined under a microscope. And the like. The particle count of the microcapsules was analyzed by microscopic examination of the prepared microcapsules and the diameter of the microcapsules (Paul Marienfeld GmbH & Co. KG, Lauda-Koenigshofen). Absorb to the gauze and attach it to the upper arm of the panel, remove the band-aid after 24 or 48 hours and observe the smell and skin irritation. It was measured by the method. Yield was calculated by expressing the total volume of microcapsules containing the concentrate of wasabi water extract relative to the total volume of the sample used (aqueous sodium alginate solution, wasabi extract and calcium chloride solution) as a percentage. Antimicrobial activity was measured in the same manner as in Experimental Example <1-2> using Helicobacter pylori as the target strain.

As a result, the lower the addition ratio of the alginate solution, the smaller the number of particles, and the empty spaces were observed, and the strong irritation of wasabi extract was detected. The higher the addition ratio of the alginate solution, the more microcapsules were formed, but the yield of the microcapsules containing the wasabi extract was lowered and the antibacterial activity was lower due to the lower concentration of the wasabi extract (Table 2). Therefore, in consideration of all the above experimental results, the preferred mixing ratio of wasabi extract and alginate solution was determined to be 1: 4 to 10 by volume ratio.

Number, irritation, yield, and antimicrobial activity of wasabi extracts according to the mixing ratio of the concentrate and alginate solution Mixing ratio Particle count pepper yield Antimicrobial Activity (Helicobacter Pylori) 1: 3 + +++ ++ ++ 1: 4 ++ ++ +++ +++ 1.5 ++ + +++ +++ 1:10 ++ + ++ ++ 1:11 +++ + + +

Experimental Example 3

Determination of Minimum Growth Concentration and Minimum Sterilization Concentration for Various Strains of Microcapsules Containing Wasabi Water Extract Concentrate According to the Present Invention

Minimum inhibitory concentration (MIC) and minimum sterilization concentration (Minimum inhibitory concentration, MIC) for various pathogenic strains of microcapsules containing the concentrate of horseradish water extract prepared in Example 2 (allyl isothiocyanate concentration 8000 ppm) bactericidal concentration (MBC) was measured.

Is Helicobacter pylori (Helicobacter pylori, NCTC 11637) as test organism, Escherichia coli (E. coli O157: H7, ATCC 43889), Staphylococcus aureus (Staphylococcus aureus, ATCC 25923), Vibrio parahaemolyticus (Vibrio parahaemolyticus, ATCC to 2,210,001), Salmonella typhimurium (Salmonella typhimurium, KCTC 2058), Bacillus cereus (Bacillus cereus, KCTC 1012) and Bacillus subtilis (Bacillus subtilis, ATCC 6633) were used.

Minimum growth inhibitory concentrations were determined by liquid media dilution. That is, 0.1 ml of the diluted solution was added to 9.8 ml of Mueller hinton broth so that the microcapsules prepared in Example 2 had a constant concentration, and the final concentration of each strain was incubated for 18 to 24 hours. After inoculating 0.1 ml at a rate of 10 ⁵ CFU / ml and incubating for 48 hours at 37 ° C., bacterial growth was measured by absorbance (660 nm).

The minimum sterilization concentration is in the Mueller hinton agar containing the microcapsules prepared in Example 2 in which the platinum culture medium in which the bacteria were not grown in the minimum growth inhibitory concentration measurement experiment was diluted to a constant concentration of 1 platinum each. After inoculation and incubation at 37 ° C. for 24 hours, the minimum concentration that failed to form colonies was measured.

As a result, the minimum growth inhibition concentration of wasabi extract was 450-5,500ppm, and the minimum sterilization concentration was about 660 ± 220-8,500 ± 3,000ppm (Table 3).

Minimum Growth Concentration and Minimum Sterilization Concentration of Wasabi Extract Test strain MIC concentration (mg of dry weight / ℓ: ppm) MBC concentration (mg of dry weight / ℓ: ppm) Helicobacter pylori 2,000 3,130 ± 1,125 * Escherichia coli 1,000 1,600 ± 900 Enteritis vibrio 450 660 ± 220 Salonella Typhimurium 1,000 1,375 ± 360 Staphylococcus aureus 5,500 8,500 ± 3,000 Bacillus cereus 3,000 4,690 ± 1,350 Bacillus subtilus 4,500 7,500 ± 2,900

^* Mean ± standard deviation (n = 3)

<Manufacture example 1>

Preparation of skin cleansing agent comprising microcapsules according to the present invention

0.5 g of microcapsules containing horseradish water extract prepared in Example 2 were added to 12 g of a cleanser base containing 6 g of glycerin, 2.0 g of monoalkyl phosphate, 0.5 g of sodium hydroxide solution, 1.5 g of myristic acid, and a trace amount of fragrance. The mixture was stirred in a homo mixer, heated at 60 ° C. for 3 minutes, degassed, and cooled to 37 ° C. to prepare a face wash composition.

<Manufacture example 2>

Preparation of soap containing microcapsules according to the present invention

99.5% by weight of the soap base (including water) and 0.5% or 0.3% by weight of the microcapsules containing the horseradish water extract prepared in Example 2 were mixed well with a mixer, and then extruded, cut and molded in a soap maker to obtain a solid. A soap composition was prepared.

<Manufacture example 3>

Preparation of kitchen cleaners containing microcapsules according to the present invention

20% by weight alkylethersulfonate, 4% by weight alkylpolyglucoside, 4% by weight lauryldimethylamine oxide, 7% by weight of Cocamidopropyl Betaine, ethoxy The cleaning agent was prepared by adding 5% by weight of lauryl alcohol, 0.3% by weight of EDTA, a small amount of pigment, 0.5% by weight of flavor, 0.5% by weight of microcapsules encapsulated with wasabi water extract of Example 2 and the remaining amount of purified water.

<Manufacture example 4>

Preparation of toothpaste comprising microcapsules according to the present invention

40% by weight of monohydrogen phosphate, 30% by weight of sorbitol, 1.50% by weight of sodium alkyl sulfate, 0.1% by weight of sodium saccharin, 0.2% by weight of paraoxybenzoic acid ester, 1.0% by weight of carrageenan, 1.0% by weight of perfume, horseradish extract of Example 2 Toothpaste was prepared by adding 1.0 wt% of the encapsulated microcapsules and the remaining amount of purified water.

Experimental Example 4

Evaluation of antimicrobial effect and skin irritation of soap containing microcapsules according to the present invention

<4-1> Antimicrobial Effect Evaluation

The antimicrobial effect and skin irritation degree of the soap prepared by the method of Preparation Example 2 were evaluated by the following method. First, 15 panelists selected as the test subjects were divided into three groups of five. That is, divided into a general soap use group, a soap use group containing 0.3% microcapsules according to the present invention and a soap use group containing 0.5% by weight of microcapsules according to the present invention, for measuring general bacteria without washing hands The plate was plated on a plate count agar. Then wash the hands with a normal soap, soap containing 0.3% by weight of microcapsules according to the present invention, and soap containing 0.5% by weight of microcapsules according to the present invention. After time incubation, the growth of the bacteria was measured.

As a result, bacteria were detected in all groups before hand washing. On the other hand, even in the case of washing hands with normal soap, bacteria were detected in all 15 hands. On the other hand, in the soap containing 0.3% by weight of microcapsules according to the present invention, bacteria were detected in one hand, and in the soap containing 0.5% by weight, no bacteria were detected in 15 hands. 4).

Antibacterial Effect of Soap Containing Microcapsules According to the Present Invention group panel Unwashed Plain Soap 0.3 wt% soap 0.5 wt% soap Primary Secondary Primary Secondary Primary Secondary A One Fungus detection Fungus detection Fungus detection Fungus detection 2 Fungus detection Fungus detection Fungus detection Fungus detection 3 Fungus detection Fungus detection Fungus detection Fungus detection 4 Fungus detection Fungus detection Fungus detection Fungus detection 5 Fungus detection Fungus detection Fungus detection Fungus detection B One Fungus detection Fungus detection No bacteria detected No bacteria detected 2 Fungus detection Fungus detection No bacteria detected No bacteria detected 3 Fungus detection Fungus detection No bacteria detected No bacteria detected 4 Fungus detection Fungus detection No bacteria detected No bacteria detected 5 Fungus detection Fungus detection Fungus detection Fungus detection C One Fungus detection Fungus detection No bacteria detected No bacteria detected 2 Fungus detection Fungus detection No bacteria detected No bacteria detected 3 Fungus detection Fungus detection No bacteria detected No bacteria detected 4 Fungus detection Fungus detection No bacteria detected No bacteria detected 5 Fungus detection Fungus detection No bacteria detected No bacteria detected

<4-2> Odor and Skin Irritation

A 10% aqueous solution was prepared by dissolving the soap containing the microcapsules according to the present invention prepared in Preparation Example 2 in water. 1 ml of the aqueous solution was absorbed into the gauze portion of the adhesive test bandage and attached to the upper arm of the panel. After 24 hours or 48 hours, the band-aid was removed and the smell and skin irritation were observed. Fifteen participants participated in the experiment, and odor and skin irritation were evaluated in 4 grades (Table 5).

Odor and skin irritation evaluation criteria Rating Odor Stimulation Skin irritation One No No response at all 2 It smells a bit Partial skin erythema 3 Smells overall Skin erythema generally appears. 4 Smell bad Skin erythema appears and the skin swells.

As a result of the experiment, the smell and skin irritation were found to be little overall. However, the odor and skin irritation tended to be slightly higher as the time of applying the bandage for irritation test was observed, but it was found that the degree of safe use (Table 6).

Odor and skin irritation of the soap containing the microcapsules according to the present invention division Odor Stimulation Skin irritation Immediately after attachment 24 hours later 48 hours later Immediately after attachment 24 hours later 48 hours later One One One One 2 2 3 2 One 2 2 One One One 3 2 2 2 3 3 3 4 One One 2 One One One 5 One One One One One 2 6 2 2 2 One One One 7 One 2 2 One One One 8 One 2 2 One One One 9 One One One One One One 10 2 2 3 2 2 2 11 2 2 2 2 2 2 12 One One One One One 2 13 One One One One 2 2 14 One One One 2 2 2 15 2 2 2 2 2 2 system 1.33 1.53 1.67 1.47 1.53 1.73

As described above, the alginate microcapsules of the present invention have a strong antimicrobial activity by capturing wasabi extract at a high concentration while shielding the strong irritation and high volatility of wasabi extract to improve the usability of wasabi extract in various fields It can be usefully used.

Claims (9)

(a) preparing wasabi water extract; (b) concentrating the wasabi water extract of step (a) 5 to 15 times to concentrate the concentration of allylisothiocyanate to 2000 to 8000 ppm; (c) mixing the wasabi extract and the alginate solution of step (b) in a volume ratio of 1: 4 to 10; (d) spraying the mixed solution of step (c) under a pressure of 0.1 to 2.0 kgf / cm ² while supplying air at 0.1 to 2.0 kgf / cm ² using a 0.5 to 2.0 mm nozzle; And (e) A method for preparing alginate microcapsules containing horseradish extract containing washing and drying the microcapsules formed in step (d). The wasabi extract of step (a) is prepared by adding water to a fine wasabi at a weight ratio of 3 to 7 times and extracting at 35 to 37 ° C. for 1 to 2 hours. The manufacturing method to make. delete delete delete The method of claim 1, wherein the polyvalent cation solution of step (d) is calcium chloride (CaCl ₂ ), barium chloride (BaCl ₂ ), strontium chloride (SrCl ₂ ), magnesium chloride (MgCl ₂ ) and zinc chloride (ZnCl ₂ ) The manufacturing method characterized in that it is selected from the group consisting of. Alginate microcapsules prepared by the method of any one of claims 1, 2 and 6 and containing wasabi extract. 8. The alginate microcapsules according to claim 7, wherein the alginate microcapsules are 0.1 to 1.0 nm in diameter. An antimicrobial composition comprising alginate microcapsules containing horseradish extract of claim 7.

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