KR100468380B1 - Antimicrobial composition containing the calcium powder of starfish - Google Patents

Abstract

The present invention relates to a calcium powder having an antimicrobial activity prepared by high-temperature heating the starfish, the calcium powder according to the present invention exhibits a strong antimicrobial activity against a wide range of bacteria, the antimicrobial composition containing it has an antimicrobial action It can be usefully used as a pharmaceutical and food composition.

Description

ANTIMICROBIAL COMPOSITION CONTAINING THE CALCIUM POWDER OF STARFISH}

The present invention relates to an antimicrobial composition containing calcium powder obtained by subjecting a starfish to high temperature heat treatment.

In recent years, the importance of hygiene management is increasing with the introduction of a new concept of HACCP (Hazard Analysis and Critical Control) in relation to food manufacturing and distribution. Until now, hygiene management during the manufacturing process has played an important role as a main medium for heat treatment or disinfection sterilization, but this method of treatment has fundamental problems. In particular, due to the demands of the times where sufficient consideration for the preservation of the environment is required, it is time for a means to drastically replace the hygiene measures in food manufacturing that have been used until now. In addition, recent cases of food poisoning tend to be enlarged at home and abroad.

At this point, research on the development of a natural substance that is harmless to the human body and has an antimicrobial effect has been conducted in various fields, but the development of a natural antibacterial agent having industrial value has not been made yet. In particular, there is no substance suitable for the use of natural antimicrobial agents to extend the shelf life of traditionally processed fish foods such as salted fish and cod roe. In addition, in the case of farmed fish, a variety of bacterial diseases are a problem, and since there is no appropriate antimicrobial agent, antibiotics used in humans are used as it is without a test, which is very likely to be a serious problem.

Meanwhile, starfish have been caught as marine pirates, and several researchers have attempted to prepare feeds or fertilizers, and have applied for patents as fertilizers. However, in this series of studies, economics are not being seen as they are combined with economics.

Accordingly, the present inventors have become a raw material for the production of natural substances, and attempted to develop an antibacterial calcium agent from the most natural starfish of the aquaculture, so that the antimicrobial-activated calcium agent could be manufactured in a very simple method, and thus the patent application for this technology. .

An object of the present invention is to provide an antimicrobial composition containing calcium powder obtained by high temperature heating (heating) the starfish.

In accordance with the above object, the present invention provides an antimicrobial composition containing calcium powder obtained from starfish by high temperature heat treatment.

Hereinafter, the present invention will be described in more detail.

Kinds of starfish used in the raw material of the present invention include starfish ( Asterina pectinifera, Asterina batheri, Asterina carinifera ) and Amurbula ( Asterias amurensis et al.) And the like, and the present invention includes all the starfish species.

In order to obtain calcium powder from the starfish, lipid components are first removed from the starfish and dried, preferably vacuum freeze-dried, by a method such as hot air drying, vacuum lyophilization, or solar drying. The dried product is preheated at a heating temperature of 80 to 120 ° C., preferably at about 100 ° C. for 20 to 40 minutes, and then continuously at a high temperature heating temperature of about 400 to 1500 ° C., preferably 600 to 1100 ° C., most preferably The high temperature heat treatment is performed at 800 to 900 ° C. for 2 to 6 hours, preferably 4 hours. High-temperature heated starfish After cooling the high-temperature heated material, a starch calcium powder is obtained through a grinding process.

Calcium preparations prepared contain 70 to 80% by weight of calcium, in addition to magnesium, iron, strontium and the like. In addition, trace amounts of metals such as nickel, sodium, zinc and copper are contained.

The hot-heated calcium preparation isolated from the starfish according to the present invention is Escherichia coli , Bacillus subtilis , Staphylococcus aureus , Pseudomonas aeruginosa , Krebsiella aerogenes , Proteus mirabilis , nutrient agar medium, yeast is Candia solani , Saccharomyces cerevisiae , Zygosaccharomyces rouxii , and fungus is Aspergillus. Since it exhibits strong antimicrobial activity against a wide variety of fungi including yeasts such as Aspergillus niger ) and penicillium chrysogenum , the preparation can be added as a food preservative to foods.

In addition, the calcium preparation isolated from the starfish according to the present invention does not show any toxicity, such as acute toxicity in the test using mice, and does not show side effects on liver function.

The calcium preparation isolated from the starfish of the present invention may be mixed with a suitable pharmaceutically acceptable carrier or excipient or diluted with a diluent according to conventional methods to prepare a pharmaceutical composition having an antimicrobial action. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like. Pharmaceutical compositions of the invention can be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.

The pharmaceutical compositions of the invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular. Typical daily dosages of the starfish calcium powder of the present invention range from 1 to 1,000 mg / kg body weight, and can be administered once or in divided doses. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the patient, and therefore the dosage may be determined in any aspect of the invention. It does not limit the scope of.

The present invention also provides a food or beverage composition exhibiting an antimicrobial effect, comprising a calcium preparation isolated from an effective amount of starfish. Examples of the food to which the calcium preparation isolated from the starfish of the present invention can be added to exhibit an antimicrobial effect include various foods, beverages, gums, teas, vitamin complexes, and health supplements.

Calcium preparation isolated from the starfish of the present invention can be added to the raw material during food manufacturing or properly mixed with the cooked food to produce a food having an antimicrobial effect or to impart antimicrobial to the food, in this case the final food prepared Or the content of the calcium preparation isolated from the starfish in the beverage is in the range of 0.001 to 10% by weight, preferably 0.01 to 0.2% by weight.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only for illustrating the present invention, the present invention is not limited to these.

In addition, in the examples below, the following percentages for solids and solid mixtures, liquids and liquids, and liquids and solids are based on weight / weight, volume / volume and weight / volume, respectively, and all reactions are carried out at room temperature unless otherwise noted. It was.

Example 1 Preparation and Analysis of Starfish Calcium Prepared by High Temperature Heat Treatment

(1) Component Analysis of Starfish Raw Materials

Starfish and Amur starfish from the east and south coasts were collected as raw materials, and the ingredients were analyzed according to the AOAC method (Refer to Official methods of analysis, 15th ed., Association of Official Analytical Chemists, Arlington, 1990). It is shown in Table 1 below.

General Ingredients of Starfish and Amur Starfish content(%) Starfish Amur Buddha Starfish moisture 1.6-1.8 1.9-2.1 Crude fat 2.6-4.6 2.2-3.2 Crude protein 17.4-20.3 18.5-18.9 View minutes 63.0-64.8 65.2-67.9 Sugar 10.5-13.2 8.5-9.0

* Ingredient analysis: moisture-atmospheric drying method, crude fat-Soxhlet method, crude protein-Kjeldahl method, ash-dry painting method, sugar- {100- (water + crude fat + crude protein + ash)}

Starfish had a water content of 1.6-1.8%, crude fats 2.6-4.6%, crude protein contents 17.4-20.3%, sugar content 10.5-13.2%, and ash content 63.0-64.8%. On the other hand, Amur vulgaris were characterized by higher ash content than starfish, and crude protein or crude fat was slightly lower than starfish.

(2) Yield of Calcium Preparation Heat-Treated from Starfish

Starfish and Amur starfish were subjected to vacuum lyophilization (Freezone 12, Labconco Co.) to obtain 2.5 kg of lyophilisate, denitrification by solvent extraction, and preheating at 100 ° C. for 2 hours, followed by Among them, the high temperature heating treatment temperature and time were judged to be the most important factors, and the calcium preparation was prepared while varying the treatment temperature and time. Table 2 shows the results of confirming the yield of the calcium preparation prepared for each condition.

Yield Analysis of Calcium Products by Heating (High Temperature Heating) Treatment Conditions Processing conditions Yield (%) Treatment temperature (℃) Processing time (hours) Starfish Amur Buddha Starfish 600 2 70.6 72.2 4 65.1 68.4 6 64.3 65.8 700 2 67.2 68.9 4 64.9 66.5 6 62.0 62.2 800 2 47.0 48.4 4 41.4 43.2 6 40.6 42.7 900 2 43.8 44.5 4 41.6 43.1 6 41.2 41.9 1000 2 41.9 43.0 4 41.7 42.8 6 40.8 41.6 1100 2 41.0 42.9 4 39.8 41.1 6 40.2 41.3

The production yield of the obtained calcium preparation was calculated as (dry weight of the hot heated formulation / dry weight of the raw materials) × 100.

As a result of the experiment, it was confirmed that the treatment temperature was significantly influenced regardless of the type of starfish. Particularly, the yield was more than 60% up to the level of 700 ° C. These results indicate that part of the form of calcium carbonate (CaCO ₃ ) is changed to the form of calcium oxide (CaO) at a specific temperature or more. It was confirmed that it had to be heated (see Isshiki, 1994, Nippon Shokuhin Kogyo Gakkaishi, 41 (2), 135-140). In addition, since no change in yield was observed at temperatures above 1000 ° C., it was determined that there was no change in form of calcium formulation at a specific temperature. The yield was slightly different depending on the heat treatment time, but no significant change in yield was observed with the treatment for 4 hours or more, and the treatment time was determined to be 4 hours or less at the time of calcium formulation. On the other hand, there was almost no difference in yield according to the sample type of starfish or Amur starfish, which could conclude that the two starfish are similar in form of calcium.

(3) Elemental analysis result of starfish calcium preparation

In the method of (2), elemental analysis was performed using an atomic absorption spectrometer (A Analyst 300 device) in order to analyze the components of the high-temperature-calculated calcium preparation at 800 or 900 ° C for 4 hours.

Elemental analysis results are shown in Table 3.

Elemental Components of Antibacterial Calcium Agents Isolated from Starfish element content(%) Starfish Amur Buddha Starfish 800 ℃ 900 ℃ 800 ℃ 900 ℃ Ca 75.9 77.4 78.2 81.6 Mg 0.16 0.21 0.25 0.20 Fe 0.07 0.03 0.11 0.08 Sr 0.01 0.04 0.01 0.01 Ni (ppm) 17 21 26 19 Na (ppm) 21 27 31 24 Zn (ppm) 19 14 12 18 Cu (ppm) 11 9 20 17

As a result of elemental analysis, calcium was 70-80%, mostly magnesium, iron, and trace amounts of metals such as sodium, nickel, and copper.

Example 2: Antimicrobial Experiments of Calcium Preparations Prepared from Starfish

The calcium preparation prepared from the starfish obtained in Example 1 was re-dissolved in distilled water at a concentration of 300 µg / ml and used as an antimicrobial test as a sample. The strains used for the antimicrobial test were Escherichia coli KCTC 1924, Bacillus subtilis KCTC 1021, Pseudomonas aeruginosa KCTC 1636, and Klebsiella aerogenes KCTC 2619 Villi. ( Proteus mirabilis KCTC 2566) and nutrient agar was used as a medium. Candida solani KCTC 7185, Saccharomyces cerevisiae KCTC 7914, Zygosaccharomyces rouxii KCTC 7966, and the mold is Aspergillus niger KCTC 6985, Penicillium chrysogenum KCTC 6447) and YM agar or PDA, respectively. The antimicrobial experiment was carried out in accordance with the paper disk method (refer to Documents Tokyo Research Institute, Microbial Experiment Manual, Society Publication Center, p. 80, 1987) as follows.

A plate was prepared by injecting 15 mL of sterile medium into a sterile petri dish (9 cm in diameter), and then, 0.1 mL of a test strain shaken at 37 ° C. for 16 hours was added to a sterile medium plate and uniformed with a conradi stick. Spread and preincubated at 20 ° C. for 2 hours. Then, put a paper disk (8mm, Advantec Toyo, Japan) on it, and inject (40μL) a sample of a certain concentration (calcium preparation in water at 1% concentration), and leave it at 20 ℃ for 1 hour, and then at 37 ℃. 48 hours of incubation measured antimicrobial activity by the diameter of the clear zone around the paper disk. First, the antimicrobial activity was confirmed by measuring the absorbance by liquid culture.

As a result of testing the antimicrobial activity of the calcium preparation obtained by treating for 4 hours and 6 hours at each treatment temperature condition, as shown in Tables 4 to 13 below, there were some differences depending on the type of test bacteria, but 800-900 It was confirmed that the calcium preparation prepared at ℃ is particularly excellent antibacterial effect and the experimental results for the antimicrobial effect is shown as follows.

In the following, " + " indicates activity in which the transparent disk size of the paper disk is 1 cm or more, " +-" indicates activity in which the transparent ring size of the paper disk is 0.85 cm or more, and "-+ " Is more than 0.8 cm and represents an active size of less than 0.85 cm, "-" means that the transparent ring size of the paper disk represents an active size of 0.8 cm or less.

Antimicrobial Effects on Escherichia Coli Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hours) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 + + 6 +- +- 900 4 + +- 6 +- +- 1000 4 +- +- 6 +- - 1100 4 +- +- 6 - -

Antibacterial effect against Pseudomonas bacteria Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 +- +- 700 4 - - 6 - - 800 4 + +- 6 +- +- 900 4 +- + 6 +- +- 1000 4 +- +- 6 +- +- 1100 4 - - 6 - -

Antimicrobial Effects on Creepsiella Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 +- - 800 4 +- +- 6 +- + 900 4 +- +- 6 +- +- 1000 4 - - 6 +- - 1100 4 - - 6 - -

Antimicrobial Effect Against Proteus Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 +- - 700 4 - - 6 - - 800 4 + +- 6 + +- 900 4 + + 6 +- +- 1000 4 - - 6 - - 1100 4 - - 6 - -

Antibacterial effect against Bacillus bacteria Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 +- +- 6 +- +- 900 4 + + 6 +- +- 1000 4 +- +- 6 +- +- 1100 4 - - 6 - -

Antifungal Effects on Candida Fungi Processing conditions Antifungal effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - +- 800 4 +- +- 6 +- +- 900 4 +- +- 6 +- +- 1000 4 +- +- 6 +- +- 1100 4 +- +- 6 - -

Antifungal effect on Saccharomyces fungi Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 -+ -+ 6 +- -+ 900 4 +- +- 6 -+ -+ 1000 4 -+ -+ 6 -+ -+ 1100 4 -+ -+ 6 -+ -+

Antifungal Effect on Shima Infected Caromyces Fungi Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 -+ -+ 6 -+ -+ 900 4 -+ -+ 6 -+ -+ 1000 4 - - 6 - - 1100 4 - - 6 - -

Antifungal Effect on Aspergillus Fungi Processing conditions Antifungal effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 - - 6 -+ -+ 900 4 -+ - 6 - - 1000 4 - - 6 - - 1100 4 - - 6 - -

Antifungal Effect on Penicillium Fungi Processing conditions Antimicrobial effect Treatment temperature (℃) Processing time (hr) Starfish Amur Buddha Starfish 600 4 - - 6 - - 700 4 - - 6 - - 800 4 -+ -+ 6 -+ +- 900 4 -+ -+ 6 -+ -+ 1000 4 -+ -+ 6 -+ - 1100 4 - - 6 - -

As shown in the results of Tables 4 to 13, the effect on the mold was judged to be rather low, and the effect on the mold was not confirmed even in the results confirmed in the liquid culture.

(2) Determination of minimum growth inhibition concentration and determination of effective amount

In addition, as a result of confirming the minimum growth inhibitory concentrations for the various strains of the present calcium preparations (see Table 14), it was confirmed that the antimicrobial effect of the calcium formulation of about 0.5%, although slightly different depending on the test strain.

Test strain Minimum growth concentration (%) Starfish Amur Buddha Starfish NaOH Escherichia coli 0.42 0.40 0.15 Staphylococcus aureus 0.78 0.65 0.15 Bacillus subtilis 0.42 0.48 0.12 Candida Solani 0.85 0.72 0.18

(3) Application and Antibacterial Effect of Starfish Calcium Formula to Cod Salted Salted Fish

The starfish calcium preparation prepared by heating at 800 to 900 ° C. for 6 hours was added at a level of 0.5% and 1.0% for spicy cod roe, and the bacterium of salted cod roe was stored during storage, and the experimental results are shown in Table 15.

Sample storage days Total bacteria (cells / g-sample) 0 One 3 5 7 9 Spicy cod roe 1.1 × 10 ⁴ 5.42 × 10 ⁴ 4.84 × 10 ⁷ 3.12 × 10 ⁹ 6.26 × 10 ⁹ 1.24 × 10 ¹¹ 800 ℃ 0.5% 1.1 × 10 ⁴ 2.50 × 10 ⁴ 4.55 × 10 ⁴ 1.94 × 10 ⁶ 5.74 × 10 ⁶ 7.14 × 10 ⁷ 1.0% " 1.61 × 10 ⁴ 3.18 × 10 ⁴ 9.22 × 10 ⁵ 2.96 × 10 ⁶ 1.56 × 10 ⁷ 900 ℃ 0.5% " 1.29 × 10 ⁴ 8.61 × 10 ⁴ 3.75 × 10 ⁶ 8.42 × 10 ⁷ 1.29 × 10 ⁸ 1.0% " 8.09 × 10 ³ 7.35 × 10 ⁴ 1.91 × 10 ⁶ 3.88 × 10 ⁶ 7.72 × 10 ⁷

As a result of the experiment, it was confirmed that the growth of bacteria was significantly suppressed compared to the sample without addition of the calcium preparation by the 9th day of storage.

Formulation Example: Preparation of Pharmaceutical Composition Containing High Temperature Heated Calcium Powder from Starfish of the Present Invention

A pharmaceutical composition was prepared comprising the following ingredients.

High Temperature Heated Calcium Powder from Starfish 200 mg

Vitamin C 50 mg

Lactose (carrier) 150 mg

400 mg total

The ingredients were mixed well and filled into hard gelatin capsules to a total of 400 mg per capsule. By ingesting the prepared capsules 1 to 10 per day can bring about an antibacterial effect. In addition to vitamin C, the compositions may further include other healthful ingredients, and various oral preparations, such as tablets, granules, powders, microcapsules, drinks, suspensions, emulsions, syrups, and other liquids, according to conventional general preparations. It can be prepared as.

Calcium formulation of the present invention prepared by heating the starfish at an appropriate temperature using a simple high temperature heating process exhibits strong antimicrobial activity against a wide range of bacteria, and the antimicrobial composition containing the same is useful as a pharmaceutical and food composition having an antimicrobial effect. Can be used. In addition, this application has the effect of producing high value-added natural material from starfish, which is a waste of expensive treatment.

Claims (11)

delete delete The starfish was vacuum lyophilized, the dried product was preheated at 80 to 120 ° C. for 20 to 40 minutes, and subsequently subjected to high temperature heat treatment at 800 to 890 ° C. for 2 to 6 hours. Method for producing an antibacterial calcium powder prepared by cooling and grinding. delete delete delete delete An antimicrobial composition containing calcium powder prepared by the method of claim 3. The method of claim 8, An antimicrobial composition, which is in the form of a pharmaceutical composition. The method of claim 8, An antimicrobial composition, which is in the form of a food composition. The method of claim 10, Antimicrobial composition, characterized in that the form of dietary supplements.