JP2016113373A - Water-soluble composition having antibacterial and antiviral properties - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a uniform acidic alcohol aqueous solution having antibacterial and antiviral properties which can express excellent antibacterial and antiviral effects and is composed of ε-polylysine and grape seed extract.SOLUTION: A water-soluble composition having antibacterial and antiviral properties is composed of an acidic alcohol aqueous solution comprising ε-polylysine and grape seed extract and having a pH of 3.40 or more and 5.90 or less.SELECTED DRAWING: None

Description

The present invention relates to an aqueous solution composition having antibacterial and antiviral properties, which comprises an acidic alcohol aqueous solution, containing ε-polylysine, grape seed extract, and alcohol.

In general, in order to improve the preservability of food, a method of subjecting food to heat treatment has been adopted. For foods and foods that cannot be heat-treated, adjust the method of directly killing bacteria, storage temperature, water activity, pH, etc. by treating with hypochlorite, chlorite, ozone, etc. Method, Lysozyme, Protamine, Polylysine, Glycine, Sodium acetate, Citric acid, Lactic acid, Tea extract, Pepper extract, Hop extract, Rosemary extract, Chitosan and other substances having bacteriostatic effect The method etc. which suppress propagation of bacteria by doing are taken.

By the way, bacteria and molds that cause food corruption and deterioration are often derived from bacteria originally attached to food materials used as food ingredients. Ordinary bacteria present in the skin, etc., or falling bacteria present on the surface of food production machinery / equipment may cause food to decay or deteriorate.

Therefore, those who are engaged in food production not only sterilize fingers etc. with alcohol sterilization preparations before entering the food processing plant, but also apply alcohol sterilization preparations to the outer parts of food production machinery and equipment. It is common to prevent contamination by bacteria.

In addition, recently, Clostridium botulinum, pathogenic E. coli typified by O-157, food poisoning microorganisms such as Norovirus, avian influenza virus, hepatitis virus (HV), human immunodeficiency virus (HIV) The threat of pathogenic microbes causing disease is increasing.

As a method for sterilizing and disinfecting pathogenic microorganisms, an alcohol preparation is generally used (see, for example, Patent Document 1).

In addition, methods using chemical agents such as hypochlorous acid, hydrogen peroxide, and ozone used for food poisoning bacteria, and physical and chemical sterilization methods such as ultraviolet irradiation, radiation irradiation, and electrochemical treatment have been devised. ing. In general, antibacterial means to prevent the growth of prokaryotic and eukaryotic protists, and the antibacterial target does not include viruses, and here the term “bactericidal” means to kill microorganisms and viruses. The term “disinfection (inactivation)” is used to eliminate the pathogenicity of microorganisms and viruses.

Up to now, antiviral agents containing proanthocyanidins from alcohol and grape seed extract have been disclosed as mentioning antiviral effects against non-enveloped viruses such as Norovirus (see, for example, Patent Document 2). Yes.

However, these are intended to inactivate viruses having an envelope, and 70% to 80% by volume of alcohol is necessary to achieve a sufficient inactivation effect. Depending on the material, undesired effects such as alteration may be observed, and the composition may be colored to adversely affect the object to be disinfected.

As described above, as an alcohol sterilizing preparation used for the purpose of sterilization, a hydrous alcohol in which alcohol is contained in water at a high concentration of 70% by volume or more is widely used.

Since alcohol (ethanol) has a function of degreasing oil on the surface of the hand, there is a problem that if the hydrated alcohol is frequently used for sterilization of fingers, the hand is degreased and eventually the hand becomes rough.

Moreover, there are cases where the sterilizing effect is insufficient only with the above-mentioned hydrous alcohol, and those having a stronger sterilizing effect are desired. There is an example of combining grape seed extract with anti-norovirus effect for food use, glycerin fatty acid ester, ethanol, and water (for example, see Patent Document 3), but it is expanded to sterilizer for food by combining ε-polylysine There are no examples.

Although grape seed extract has been reported as having a useful anti-norovirus effect, there has been no example in which ε-polylysine has been combined to expand into food sanitizers. (For example, refer to Patent Document 4 and Non-Patent Document 5).
JP 2004-269410 A JP 2008-214297 A JP2013-47196A JP2008-214297 Applied and Environmental Microbiology (vol. 78 (21), 7572 (2012))

As described above, as an alcohol sterilization preparation used for the purpose of sterilization, a hydrous alcohol in which alcohol is contained in water at a high concentration of about 70% is widely used. Since there is an action to degrease the oil on the skin and the surface of the hand, if the water-containing alcohol is frequently used for sterilization of fingers, there is a problem that the hand is degreased and eventually the hand becomes rough.

Moreover, there are cases where the sterilizing effect is insufficient only with the above-mentioned hydrous alcohol, and those having a stronger sterilizing effect are desired.

In view of the above, the present inventors considered that a disinfectant that is widely effective against bacteria and norovirus can be achieved by mixing both alcohol and grape seed extract.

However, it was confirmed that when both were mixed in an aqueous solution or water-containing ethanol, a precipitate was formed and a precipitate was formed to form a heterogeneous system, resulting in spraying difficulty such as nozzle clogging due to the precipitate.

In such a case, even if it is applied, the surface becomes uneven due to the precipitate, the effect is lowered, and the coated surface becomes dirty due to the precipitate, and cannot be put to practical use.

The present invention solves the above-mentioned problems, and contains ε-polylysine and grape seed extract as a disinfectant that is widely effective against bacteria and norovirus, and has a pH of 3.40 or more to 5.90. The present invention provides a water-soluble composition comprising the following acidic alcohol aqueous solution.

As a result of intensive studies for achieving the above-mentioned problems, the present inventors have adjusted the pH to 3.40 or more and 5.90 or less with respect to an aqueous solution containing ε-polylysine, grape seed extract, and alcohol. The present inventors have found that the acidic alcohol aqueous solution can maintain the uniformity without generating precipitates and have completed the present invention.

As described above, according to the present invention, the aqueous solution composition of the present invention has high uniformity, and can provide an antibacterial / antiviral aqueous solution composition having antibacterial / antiviral performance against bacteria and viruses. Furthermore, the influence on the human body and the environment can be suppressed, and the cost can be reduced.

When ε-polylysine and grape seed extract are mixed with water or hydrous alcohol, precipitation occurs. When trying to spray, the nozzle is clogged and spraying becomes difficult. Moreover, when mixed with food, it is difficult to mix evenly, and ε-polylysine and grape seed extract precipitate, resulting in sterilization and a decrease in antiviral effect. This precipitation is a fatal problem in homogenization.

The aqueous solution composition according to the present invention preferably has a pH of 3.40 or more and 5.90 or less, more preferably a pH of 3.40 or more and a pH of 5.70 or less.

In the present embodiment, the ε-polylysine concentration range is preferably 0.02 wt% or more and 1.0 wt% or less, more preferably 0.03 wt% or more and 0.98 wt% or less. is there.

Moreover, in this embodiment, it is preferable that the density | concentration range of grape seed extract is 0.002 to 0.30 weight%, More preferably, it is 0.005 to 0.25 weight%. is there.

In this embodiment, the alcohol concentration range of the acidic alcohol aqueous solution is preferably 20% by weight to 70% by weight, more preferably 40% by weight to 69% by weight.

In the present embodiment, the solvent for dissolving ε-polylysine and grape seed extract is water and alcohol. The alcohol that can be used in the present invention is preferably ethyl alcohol or isopropyl alcohol. Furthermore, it is also possible to add antioxidants which are food additives.

Moreover, in this embodiment, the acid for adjusting pH other than a solvent is added. The acid is not particularly limited as long as it can be adjusted to the desired pH and is soluble in water. However, inorganic acids such as hydrochloric acid and phosphoric acid, lactic acid, acetic acid, malic acid, tartaric acid, Examples thereof include organic acids such as citric acid, fumaric acid, and succinic acid.

In this embodiment, a salt having a buffering ability can be added as a substance for adjusting pH other than acid.

Examples of the salt having pH buffering ability include salts of the above-mentioned acids and metals, specifically, lactic acid metal salts, acetic acid metal salts, tartaric acid metal salts, phosphoric acid metal salts, and the like.

In this case, the metal is not particularly limited, and examples thereof include sodium and potassium.

As described above, according to the present embodiment, it is possible to provide an antibacterial / antiviral agent having antibacterial / antiviral performance against bacteria and viruses, which is highly uniform and highly effective. As a result, the influence on the human body and the environment can be suppressed, and the cost can be reduced.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated in detail, the aspect of this invention is not limited to this. In the following text, “part” means “part by mass”.

Here, the water-soluble composition according to the above embodiment was actually produced and examined. This will be specifically described below.

First, 100 g of a solution prepared by adding a predetermined amount of grape seed extract (produced by Kikkoman Biochemifa), ε-polylysine (produced by JNC) and ethanol, respectively, and adding lactic acid to the solution to adjust the pH. It adjusted and it checked whether the precipitate based on a grape seed extract and (epsilon) -polylysine melt | dissolved. The results are shown in Table 1. In the table below, ε-polylysine weight% and grape extract weight% indicate the weight% in the solution at the end pH (final pH at which dissolution could be confirmed or dissolution could not be confirmed). In Table 1, isopropyl alcohol was used only for sample 4, ethyl alcohol was used for the other samples, and sample 5 was prepared using citric acid for pH adjustment.

From the above results, the aqueous solution composition of the present invention having a final solution pH of 3.40 or more and 5.90 or less becomes a uniform solution depending on the concentration of ε-polylysine, grape extract and alcohol. And good dissolution uniformity.

To measure the antibacterial performance, 0.25 ml of a solution containing Staphylococcus aureus (a solution containing 1 × 10 ³ S. aureus / ml) was adsorbed on a 16 cm ² sterilized gauze. Add 1.0 ml of each comparative sample and leave at room temperature for 0.5 hour.

Next, in order to detect Staphylococcus aureus, each sample was brought into contact with Petan Check ® 25 egg yolk mannitol salt medium (Eiken Chemical Co., Ltd.) and cultured at a temperature of 33 ° C. for 24 hours. The presence or absence of fungal inactivation was determined based on whether the medium changed from vermilion to yellowish white derived from Staphylococcus aureus. In the same manner as in Example 1, Comparative Sample 7 was prepared by removing ε-polylysine from Sample 11. The results are shown in Table 2.

From the above results, Comparative Sample 5 had precipitates and did not show uniform antibacterial properties, and the growth of the bacteria was partially confirmed. Comparative Samples 1, 2, 3, 6, and 7 had antibacterial properties. However, all the samples of the present invention were uniform solutions and sufficient antibacterial properties were confirmed.

The feline calicivirus established in the culture system was used as a substitute for norovirus, and the virus infection suppression effect was evaluated by the 50% cultured cell infection concentration method (hereinafter TCID ₅₀ method).

The feline calicivirus (hereinafter referred to as FCV) used was the FCV-F9 strain, and the test cells were feline kidney cells (hereinafter referred to as CRFK cells). CRFK cells are cultured using a medium (D-MEN F10) obtained by adding 10% fetal bovine serum (FBS) to Dulbecco's modified Eagle medium (D-MEM) so that the final concentration of FBS exceeds 5%. The medium was appropriately adjusted according to the number.

Moreover, the maintenance culture for a test was performed using the medium (D-MEM F1) which added 1% of FBS to the D-MEM basal medium.

The virus infection inhibitory effect was evaluated under the condition in which a sample prepared in the same manner as in Example 1 or a comparative sample and a test virus solution were allowed to act at an equivalent amount of 1: 1 for 5 minutes.

Comparative sample 8 was prepared by removing the grape seed extract from sample 8 in the same manner as in Example 1. In addition, as a test virus solution, a suspension prepared by adjusting FCTC-F9 strain with phosphate buffered saline (PBS) to a log TCID ₅₀ / ml of 6.0 was used.

In order to determine the infectious titer, the working solution of the test virus and sample was serially diluted 10-fold with D-MEM F1, and 5 CRFK cells cultured with D-MEM F1 were placed in each well of a 96-well microplate. Inoculate cells at 10 ⁵ cfu / well, inoculate 4 working wells of each working solution at each dilution stage, and culture in a CO ₂ incubator at 36 ° C. and 5% CO ₂ for 5-7 days. The effect (CPE) was observed under a microscope, and the value of TCID ₅₀ / ml was calculated by the Reed-Munch method.

As a control, the infectivity titer was determined in the same manner except that PBS was used instead of the sample solution. Calculated infectivity logarithm (logTCID ₅₀ / ml) infectivity logarithm of virus reacted with a sample solution from (logTCID ₅₀ / ml) obtained by subtracting the value of the control (ΔlogTCID ₅₀ / ml), this reduction value When it exceeded 3, it was judged that there was sufficient infection suppression effect. Table 3 shows the evaluation results of the viral infection suppression effect by the TCID ₅₀ method.

From the above results, in each composition of the present invention, Δlog TCI of 3.0 or more even with an action for 5 minutes.

The present invention has industrial applicability as an antibacterial / antiviral agent. A D ₅₀ / ml value was obtained, and a good virus infection suppressing effect was observed.

First, 100 g of a solution prepared by adding a predetermined amount of grape seed extract (produced by Kikkoman Biochemifa), ε-polylysine (produced by JNC) and ethanol, respectively, and adding lactic acid to the solution to adjust the pH. It adjusted and it checked whether the precipitate based on a grape seed extract and (epsilon) -polylysine melt | dissolved. The results are shown in Table 1. In the table below, ε-polylysine weight% and grape extract weight% indicate the weight% in the solution at the end pH (final pH at which dissolution could be confirmed or dissolution could not be confirmed). In Table 1, isopropyl alcohol was used only for sample 4, ethyl alcohol was used for the other samples, and sample 5 was prepared using citric acid for pH adjustment.

Next, in order to detect Staphylococcus aureus, each sample was brought into contact with Petan Check ® 25 egg yolk mannitol salt medium (Eiken Chemical Co., Ltd.) and cultured at a temperature of 33 ° C. for 24 hours. The presence or absence of fungal inactivation was determined based on whether the medium changed from vermilion to yellowish white derived from Staphylococcus aureus. In the same manner as in Example 1, Comparative Sample 7 was prepared by removing ε-polylysine from Sample 11. The results are shown in Table 2.

Claims (4)

A water-soluble composition having antibacterial and antiviral properties, comprising ε-polylysine and a grape seed extract, and comprising an acidic alcohol aqueous solution having a pH of 3.40 to 5.90. The water-soluble composition according to claim 1, wherein the ε-polylysine concentration range is 0.02 wt% or more and 1.0 wt% or less. The water-soluble composition according to claim 1 or 2, wherein a concentration range of the grape seed extract is 0.002 wt% or more and 0.30 wt% or less. The water-soluble composition according to any one of claims 1 to 3, wherein an alcohol concentration range of the acidic alcohol aqueous solution is 20% by weight or more and 70% by weight or less.