JP7058490B2 - Antibacterial composition that suppresses viruses, bacteria and fungi - Google Patents
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本発明は、ウイルスを不活化し、かつ細菌および真菌を殺菌するエタノール製剤に関する。 The present invention relates to ethanol formulations that inactivate viruses and kill bacteria and fungi.
ノロウイルスによる急性胃腸炎、大腸菌や黄色ブドウ球菌等による食中毒が往々にして問題になっている。また真菌類の中には、食品中で増殖する過程で人にとって有害な代謝産物を作るものもある。
これらの微生物やその産生する有害物質が、人体に取り込まれる場合、その経路はほとんど経口である。そのため、食品の調理環境における微生物のコントロール、すなわち調理器具の表面や、調理室内の空気におけるウイルス、細菌および真菌の低減は、食品衛生上重要である。
Acute gastroenteritis due to norovirus and food poisoning due to Escherichia coli and Staphylococcus aureus are often problems. Some fungi also produce metabolites that are harmful to humans as they grow in food.
When these microorganisms and the harmful substances produced by them are taken up by the human body, the route is almost oral. Therefore, control of microorganisms in the cooking environment of food, that is, reduction of viruses, bacteria and fungi in the surface of cooking utensils and the air in the cooking room is important for food hygiene.
厚生労働省によると、ノロウイルスを失活化させるために用いる薬剤としては、消毒用エタノールや逆性石鹸は効果が無く、次亜塩素酸ナトリウム水溶液が有効であるとされている。また、次亜塩素酸ナトリウムは、芽胞菌以外の細菌にも有効であることが知られている。
しかし一方で、次亜塩素酸ナトリウムは特有の強い臭気を有する、皮膚腐食性がある、衣服等を脱色する、酸と反応して塩素ガスを発生させるなど、調理環境で使用するには難しい物性を有している。
According to the Ministry of Health, Labor and Welfare, disinfectant ethanol and inverted soap are ineffective as chemicals used to inactivate norovirus, and sodium hypochlorite aqueous solution is effective. In addition, sodium hypochlorite is known to be effective against bacteria other than spore-forming bacteria.
However, on the other hand, sodium hypochlorite has a peculiar strong odor, is corrosive to the skin, decolorizes clothes, etc., reacts with acid to generate chlorine gas, and has physical characteristics that are difficult to use in a cooking environment. have.
そのため、食品周りの微生物コントロールに用いる薬剤は、通常の飲食品に含まれている成分からなる、より安全性の高い薬剤であることが望ましい。
エタノールは酒類等の食品に含まれる成分のため安全性が高いが、上記のとおり単独ではノロウイルスをはじめとするノンエンベロープウイルスを不活化する効果は期待できない。
Therefore, it is desirable that the drug used for controlling microorganisms around foods is a drug having higher safety, which is composed of the components contained in ordinary foods and drinks.
Ethanol is highly safe because it is a component contained in foods such as alcoholic beverages, but as mentioned above, it cannot be expected to have the effect of inactivating non-enveloped viruses such as norovirus by itself.
本発明者らはこれまでに、エタノール、リン酸、グリセリン脂肪酸エステルを含みｐＨ２．５～５である水溶液を用いることにより、カリシウイルスを不活化できることを開示している（特許文献１）。さらに、エタノール、乳酸、クエン酸、それらの塩を所定の比率で含み、ｐＨ３～４であるエタノール製剤は、高い抗ウイルス活性を有するとともに、噴霧・塗布後の白残り、液残りが無いということを見出した（特許文献２）。 The present inventors have previously disclosed that calicivirus can be inactivated by using an aqueous solution containing ethanol, phosphoric acid, and glycerin fatty acid ester and having a pH of 2.5 to 5. (Patent Document 1). Furthermore, ethanol preparations containing ethanol, lactic acid, citric acid, and salts thereof in a predetermined ratio and having a pH of 3 to 4 have high antiviral activity, and have no white residue or liquid residue after spraying / coating. (Patent Document 2).
本発明の課題は、ウイルスの不活化だけでなく、細菌や真菌の殺菌もできる薬剤を提供することである。原料として用いるものは、食品周りで用いても問題の無いよう、臭いや腐食性、刺激性が少なく、一般の食品に含有されている成分であることが望ましい。 An object of the present invention is to provide a drug capable of not only inactivating viruses but also killing bacteria and fungi. It is desirable that the raw material used is a component contained in general foods, which has little odor, corrosiveness, and irritation so that there is no problem even if it is used around foods.
本発明者らは、特定の成分組成を有するエタノール製剤が、ウイルス、細菌および真菌を、不活化または殺菌できることを見出した。すなわち、本発明は以下の（１）～（３）に関する。 The present inventors have found that ethanol preparations having a specific composition can inactivate or kill viruses, bacteria and fungi. That is, the present invention relates to the following (1) to (3).
（１）各成分の含有量及びｐＨが以下（ａ）～（ｅ）を満たす水溶液である、抗ウイルス、抗細菌および抗真菌用の薬剤。
（ａ）エタノール濃度が６５～７５重量％である
（ｂ）グリセリン脂肪酸エステルの含有量が０．０３～０．１５重量％である
（ｃ）乳酸またはその塩の含有量が、乳酸として１．０～１．８重量％である
（ｄ）クエン酸またはその塩の含有量が、クエン酸として０．２～０．５重量％である
（ｅ）ｐＨ３～４である
（２）前記薬剤において、乳酸またはその塩とクエン酸またはその塩の含量比（重量比）が、乳酸とクエン酸として３：１～５：１である、前記（１）に記載の薬剤。
（３）前記（１）または（２）に記載の薬剤を対象物に接触させる、対象物表面のウイルス、細菌および真菌を、不活化または殺菌する方法。
(1) An antiviral, antibacterial and antifungal agent which is an aqueous solution in which the content and pH of each component satisfy the following (a) to (e).
(A) The ethanol concentration is 65 to 75% by weight, (b) the content of glycerin fatty acid ester is 0.03 to 0.15% by weight, and (c) the content of lactic acid or a salt thereof is 1. The content of (d) citric acid or a salt thereof, which is 0 to 1.8% by weight, is 0.2 to 0.5% by weight as citric acid, (e) pH is 3 to 4, and (2) in the above-mentioned drug. , The agent according to (1) above, wherein the content ratio (weight ratio) of lactic acid or a salt thereof and citric acid or a salt thereof is 3: 1 to 5: 1 as lactic acid and citric acid.
(3) A method for inactivating or sterilizing viruses, bacteria and fungi on the surface of an object, in which the agent according to (1) or (2) above is brought into contact with the object.
本発明の薬剤（エタノール製剤）は、対象物に噴霧、塗布、混合等をすることにより、対象物表面のウイルス、細菌および真菌といった幅広い種類の微生物を、効率的に不活化させたり殺菌したりすることが出来るものである。本製剤を使用した際、長時間残る臭いは無く、人体への悪影響も無いため、食品周りの消毒に好適に用いることができる。また本発明の製剤は、金属への腐食性がほとんど無く、さらに、乳酸とクエン酸を適切な比率にすれば、使用後に固形分の残存による白残り、液残りも無いため、噴霧剤として簡便に用いることができる。 The agent (ethanol preparation) of the present invention efficiently inactivates or sterilizes a wide variety of microorganisms such as viruses, bacteria and fungi on the surface of an object by spraying, applying, mixing, etc. on the object. It is something that can be done. When this product is used, it has no odor that remains for a long time and has no adverse effect on the human body, so that it can be suitably used for disinfecting foods. Further, the pharmaceutical product of the present invention has almost no corrosiveness to metals, and if lactic acid and citric acid are used in an appropriate ratio, there is no white residue or liquid residue due to residual solid content after use, so that it is convenient as a spraying agent. Can be used for.
本発明のエタノール製剤は、エタノール濃度が６５～７５重量％である。この範囲内であれば、安全でかつ十分な抗ウイルス、抗菌効果が得られる。６５重量％未満になると、抗ウイルス、抗菌効果がやや弱くなる。 The ethanol preparation of the present invention has an ethanol concentration of 65 to 75% by weight. Within this range, safe and sufficient antiviral and antibacterial effects can be obtained. If it is less than 65% by weight, the antiviral and antibacterial effects are slightly weakened.
本発明のエタノール製剤は、グリセリン脂肪酸エステルを０．０３～０．１５重量％含有する。グリセリン脂肪酸エステルは、具体的にはモノラウリン酸モノグリセリド、モノカプリル酸モノグリセリド等が挙げられ、抗細菌活性、抗真菌活性を高める効果がある。 The ethanol preparation of the present invention contains 0.03 to 0.15% by weight of glycerin fatty acid ester. Specific examples of the glycerin fatty acid ester include monolauric acid monoglyceride and monocaprylic acid monoglyceride, which have an effect of enhancing antibacterial activity and antifungal activity.
本発明のエタノール製剤は、乳酸またはその塩の含有量が、乳酸として１．０～１．８重量％である。かつ、クエン酸またはその塩の含有量が、クエン酸として０．２～０．５重量％である。この範囲内であれば、十分なウイルス不活化効果を発揮する。
さらに、乳酸またはその塩とクエン酸またはその塩の含量比（重量比）が、乳酸とクエン酸として３：１～５：１であれば、噴霧後や塗布後に白残りや液残りをしにくいため、なお望ましい。
The ethanol preparation of the present invention has a content of lactic acid or a salt thereof as lactic acid of 1.0 to 1.8% by weight. Moreover, the content of citric acid or a salt thereof is 0.2 to 0.5% by weight as citric acid. Within this range, a sufficient virus inactivating effect is exhibited.
Furthermore, if the content ratio (weight ratio) of lactic acid or its salt to citric acid or its salt is 3: 1 to 5: 1 as lactic acid and citric acid, it is difficult to leave white residue or liquid residue after spraying or coating. Therefore, it is still desirable.
本発明のエタノール製剤は、ｐＨ３～４である。ｐＨは低いほど薬効は強くなるが、一方で金属への使用の際に腐食が起こりやすくなるため、バランスを考慮すると、ｐＨ３．４～３．７が最も望ましい。 The ethanol preparation of the present invention has a pH of 3 to 4. The lower the pH, the stronger the medicinal effect, but on the other hand, corrosion is more likely to occur when used on metals. Therefore, considering the balance, pH 3.4 to 3.7 is the most desirable.
有効成分の含有量及びｐＨが上記条件を満たすエタノール製剤は、十分な抗ウイルス、抗細菌および抗真菌活性を示す。
さらに、薬効を妨げない範囲で、ポリグリセリン脂肪酸エステル、ヒアルロン酸、グリセリン等の保湿剤、色素、香料等を含有させても良い。
ポリグリセリン脂肪酸エステルは、具体的にはラウリン酸テトラグリセリル、ラウリン酸ペンタグリセリル、ラウリン酸ヘキサグリセリル等があり、これらはエタノール製剤の表面張力を低下させるため、対象物への接触面積を大きくする効果がある。
Ethanol preparations in which the content and pH of the active ingredient satisfy the above conditions show sufficient antiviral, antibacterial and antifungal activities.
Further, a moisturizer such as polyglycerin fatty acid ester, hyaluronic acid and glycerin, a pigment, a fragrance and the like may be contained within a range that does not interfere with the medicinal effect.
Specific examples of the polyglycerin fatty acid ester include tetraglyceryl laurate, pentaglyceryl laurate, hexaglyceryl laurate, etc., which reduce the surface tension of the ethanol preparation and thus have the effect of increasing the contact area with the object. There is.
本発明のエタノール製剤は、例えば乳酸またはその塩およびクエン酸またはその塩、および必要に応じてポリグリセリン脂肪酸エステル等を上記濃度となるようにエタノール水に溶解させ、エタノール濃度およびｐＨを上記濃度となるように、酸やアルカリにより調整することにより調製することができるが、各工程の順はこれに限られない。 In the ethanol preparation of the present invention, for example, lactic acid or a salt thereof, citric acid or a salt thereof, and if necessary, a polyglycerin fatty acid ester or the like are dissolved in ethanol water so as to have the above concentrations, and the ethanol concentration and pH are adjusted to the above concentrations. It can be prepared by adjusting with an acid or an alkali so as to be, but the order of each step is not limited to this.
本発明のエタノール製剤によりウイルス、細菌および真菌を不活化または殺菌する方法は、エタノール製剤とこれらの微生物とを十分な時間、接触させることである。
必要な接触時間は、防除する微生物の種類、その密度や状況により変わってくるが、細菌や真菌を殺菌したい場合は、通常３０秒間の接触で概ね十分である。ウイルスを不活化したい場合は１分間以上、望ましくは５分間以上の接触をさせる。
A method of inactivating or sterilizing a virus, bacterium and fungus with the ethanol preparation of the present invention is to bring the ethanol preparation and these microorganisms into contact for a sufficient time.
The required contact time varies depending on the type of microorganism to be controlled, its density and situation, but if it is desired to kill bacteria and fungi, contact for 30 seconds is usually sufficient. If you want to inactivate the virus, contact it for 1 minute or longer, preferably 5 minutes or longer.
具体的には、これらの微生物の存在が疑われる対象物にエタノール製剤を噴霧、塗布、混合などする。対象物をエタノール製剤に浸漬してもよい。対象物の例としては、食器類、調理機械器具、冷蔵庫、調理者の衣服等がある。
以下に本発明の実施例を挙げるが、本発明はこれらの実施例に限定されるものではない。
Specifically, an ethanol preparation is sprayed, applied, mixed, or the like on an object suspected of having these microorganisms. The object may be immersed in an ethanol preparation. Examples of objects include tableware, cooking utensils, refrigerators, and cooks' clothes.
Examples of the present invention are given below, but the present invention is not limited to these examples.
表１記載の組成のエタノール製剤（実施例としてサンプル１～３、比較例としてサンプル４、５）を調製した。また、サンプル６として次亜塩素酸Ｎａを２００ｐｐｍ含有する製剤を用いた。 Ethanol preparations having the compositions shown in Table 1 (Samples 1 to 3 as Examples and Samples 4 and 5 as Comparative Examples) were prepared. In addition, a pharmaceutical product containing 200 ppm of sodium hypochlorous acid was used as sample 6.
＜実施例１＞ ネコカリシウイルス不活化効果
ネコカリシウイルス Ｆ９株／ネコ腎臓細胞（ＣｒＦＫ）を用い、以下の方法によりウイルス感染価を測定し、ウイルス活性を評価した。
<Example 1> Feline calicivirus inactivating effect Using feline calicivirus F9 strain / feline kidney cells (CrFK), the virus infectivity titer was measured by the following method, and the virus activity was evaluated.
サンプル液１００μｌとウイルス液を１００μｌずつ等量混合し、室温にて３０秒間または６０秒間作用させた。なお、コントロールとして、サンプル液の代わりにリン酸緩衝液を用いた。作用後ただちに混合液１００μｌを２％ウシ胎児血清含有ＭＢＭ培地９００μｌに加え、１０倍に希釈した。
１０倍希釈した混合液を、ＭＢＭ培地にてさらに１０段階に１０倍希釈した。
６穴プレート上に３日間培養させたＣＲＦＫ細胞の細胞増殖用培地（ウシ胎児血清１０％含むＭＥＭ培地）をアスピレーターにて取り除き、各階段希釈した混合液について２穴の細胞を使用し、それぞれに１００μｌ接種した（Ｎ＝２）。
３４℃、ＣＯ２培養器内で１時間ウイルス吸着を行った後、寒天培地で重層し５％ＣＯ２インキュベーター内で３４℃、６０時間培養した。
培養終了後、１０％ホルマリン固定、メチレンブルー染色により形成されたプラーク数を数え、感染価を測定した。結果を表２に示す。
An equal amount of 100 μl of the sample solution and 100 μl of the virus solution were mixed and allowed to act at room temperature for 30 seconds or 60 seconds. As a control, a phosphate buffer solution was used instead of the sample solution. Immediately after the action, 100 μl of the mixture was added to 900 μl of MBM medium containing 2% fetal bovine serum and diluted 10-fold.
The 10-fold diluted mixture was further diluted 10-fold in MBM medium in 10 steps.
The cell proliferation medium (MEM medium containing 10% fetal bovine serum) of CRFK cells cultured on a 6-well plate for 3 days was removed with an aspirator, and 2-well cells were used for each step-diluted mixed solution. 100 μl was inoculated (N = 2).
After virus adsorption at 34 ° C. in a CO 2 incubator for 1 hour, the cells were layered on an agar medium and cultured at 34 ° C. for 60 hours in a 5% CO 2 incubator.
After completion of the culture, the number of plaques formed by 10% formalin fixation and methylene blue staining was counted, and the infectious titer was measured. The results are shown in Table 2.
表２に示すとおり、本発明のエタノール製剤に該当するサンプル１および２は、作用時間３０秒、６０秒でいずれも感染価が低く、ネコカリシウイルスに対して高い不活化効果を示した。 As shown in Table 2, Samples 1 and 2 corresponding to the ethanol preparation of the present invention had a low infectious titer at an action time of 30 seconds and 60 seconds, and showed a high inactivating effect on feline calicivirus.
＜実施例２＞ 細菌に対する殺菌効果
大腸菌（Escherichia coli）、黄色ブドウ球菌（Staphylococcus aureus）および乳酸菌（Leuconostoc mesenteroides）をそれぞれ用い、サンプル１～５の原液、２倍または３倍希釈液の殺菌効果を評価した。なお、大腸菌は標準寒天培地にて３０℃、２４時間培養したものを、黄色ブドウ球菌は標準寒天培地にて３０℃、４８時間培養したものを、乳酸菌はＭＲＳ寒天培地にて３０℃、４８時間の嫌気培養したものを、それぞれ用いた。
<Example 2> Bacterial effect against bacteria Escherichia coli, Staphylococcus aureus and Leuconostoc mesenteroides were used to obtain the bactericidal effect of the stock solution of Samples 1 to 5 and the 2- or 3-fold diluted solution, respectively. evaluated. Escherichia coli was cultured on a standard agar medium at 30 ° C. for 24 hours, yellow staphylococcus was cultured on a standard agar medium at 30 ° C. for 48 hours, and lactic acid bacteria were cultured on an MRS agar medium at 30 ° C. for 48 hours. The anaerobic cultures of the above were used respectively.
大腸菌、黄色ブドウ球菌および乳酸菌に対する殺菌効果は、以下の方法により評価した。
各サンプル３ｍｌに菌液（濃度１０６ｃｆｕ／ｍｌ）をそれぞれ０．０５ｍｌずつ投入し撹拌後、０、１０、３０秒後に処理液を直ちに１００倍量の滅菌水で希釈することで処理を停止し、その０．０５ｍｌを各培地に塗布後、培養し、発生したコロニー数から生残菌数を求めた。大腸菌、黄色ブドウ球菌および乳酸菌に関する結果をそれぞれ表３、表４および表５に示す。
The bactericidal effect on Escherichia coli, Staphylococcus aureus and lactic acid bacteria was evaluated by the following method.
Add 0.05 ml each of the bacterial solution (concentration 106 cfu / ml) to 3 ml of each sample, stir, and after 0, 10, and 30 seconds, immediately dilute the treatment solution with 100 times the amount of sterile water to stop the treatment. Then, 0.05 ml of the mixture was applied to each medium and then cultured, and the number of surviving bacteria was determined from the number of colonies generated. Results for Escherichia coli, Staphylococcus aureus and Lactic acid bacteria are shown in Table 3, Table 4 and Table 5, respectively.
表３、表４および表５に示すとおり、本発明のアルコール製剤サンプル１～３は、大腸菌、黄色ブドウ球菌および乳酸菌のいずれに対しても、顕著な殺菌効果を示した。また、本発明の製剤は、２倍または３倍に希釈しても、その殺菌効果が比較サンプルに比べて失われにくいことから、対象物が水に濡れたような状況下で使用しても、優れた殺菌効果を期待でき、また高度に汚染された対象物に対しても有効であることが示唆された。 As shown in Tables 3, 4 and 5, the alcohol preparation samples 1 to 3 of the present invention showed a remarkable bactericidal effect against any of Escherichia coli, Staphylococcus aureus and lactic acid bacteria. Further, since the bactericidal effect of the pharmaceutical product of the present invention is less likely to be lost even when diluted 2-fold or 3-fold, the bactericidal effect is less likely to be lost as compared with the comparative sample. It was suggested that an excellent bactericidal effect can be expected and that it is also effective for highly contaminated objects.
＜実施例３＞ 真菌に対する殺菌効果
食品を変敗させる酵母様真菌（Pichia anomala）を用い、サンプル１～５の原液、２倍または３倍希釈液の殺菌効果を評価した。なお、酵母様真菌はポテトデキストロース寒天培地にて２５℃４８時間培養したものを用いた。
<Example 3> Bactericidal effect on fungi Using yeast-like fungi (Pichia anomala) that alters food, the bactericidal effect of the stock solution of Samples 1 to 5 and the 2-fold or 3-fold diluted solution was evaluated. The yeast-like fungus used was cultured in a potato dextrose agar medium at 25 ° C. for 48 hours.
酵母様真菌に対する殺菌効果は、以下の方法により評価した。 The bactericidal effect on yeast-like fungi was evaluated by the following method.
各サンプル３ｍｌに菌液（濃度１０６ｃｆｕ／ｍｌ）を０．０５ｍｌずつ投入し撹拌後、０、１０、３０秒後に処理液を直ちに１００倍量の滅菌水で希釈することで処理を停止し、その０．０５ｍｌを各培地に塗布後、培養し、発生したコロニー数から生残菌数を求めた。結果を表６に示す。 0.05 ml of the bacterial solution (concentration 106 cfu / ml) was added to 3 ml of each sample, and after stirring, the treatment solution was immediately diluted with 100 times the amount of sterile water after 0, 10 and 30 seconds to stop the treatment. After applying 0.05 ml thereof to each medium, the cells were cultured, and the number of surviving bacteria was determined from the number of colonies generated. The results are shown in Table 6.
表６に示すとおり、本発明のエタノール製剤サンプル１～３は、酵母様真菌に対しても、優れた殺菌効果を示した。また、本発明の製剤は、２倍または３倍に希釈しても、その殺菌効果が比較サンプルに比べて失われにくいことから、対象物が水に濡れたような状況下で使用しても、殺菌効果を期待でき、また高度に汚染された対象物に対しても有効であることが示唆された。 As shown in Table 6, the ethanol-prepared samples 1 to 3 of the present invention also showed an excellent bactericidal effect against yeast-like fungi. Further, since the bactericidal effect of the pharmaceutical product of the present invention is less likely to be lost even when diluted 2-fold or 3-fold, the bactericidal effect is less likely to be lost as compared with the comparative sample. It was suggested that it can be expected to have a bactericidal effect and is also effective for highly contaminated objects.
以上のことより、本発明のエタノール製剤は、ウイルス、細菌および真菌といった幅広い種類の微生物に対して、不活化や殺菌の効果があることが示された。 From the above, it was shown that the ethanol preparation of the present invention has an inactivating and bactericidal effect on a wide variety of microorganisms such as viruses, bacteria and fungi.
本発明のエタノール製剤は、あらゆる微生物を対象として消毒に用いることができる。また長時間残存する臭気や固形物も無いため、特に食品周り、具体的には食器類、調理機械器具、冷蔵庫、調理者の衣服等の消毒に好適に用いることができる。
The ethanol preparation of the present invention can be used for disinfection of any microorganism. Further, since there is no odor or solid matter remaining for a long time, it can be suitably used for disinfecting foods, specifically tableware, cooking machinery, refrigerators, clothes of cooks, and the like.
Claims (3)
（ａ）エタノール濃度が６５～７５重量％である
（ｂ）モノラウリン酸モノグリセリドの含有量が０．０３～０．１５重量％である
（ｃ）乳酸またはその塩の含有量が、乳酸として１．０～１．８重量％である
（ｄ）クエン酸またはその塩の含有量が、クエン酸として０．２～０．５重量％である
（ｅ）ｐＨ３～４である
（ｆ）ラウリン酸ペンタグリセリドを含む An antiviral, antibacterial and antifungal agent which is an aqueous solution in which the content and pH of each component satisfy the following (a) to ( f ).
(A) The ethanol concentration is 65 to 75% by weight, (b) the content of monolauric acid monoglyceride is 0.03 to 0.15% by weight, and (c) the content of lactic acid or a salt thereof is 1. The content of (d) citric acid or a salt thereof, which is 0 to 1.8% by weight, is 0.2 to 0.5% by weight as citric acid, and (e) pH is 3 to 4.
(F) Contains lauric acid pentaglyceride
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