JP5158352B2 - Antiviral agent - Google Patents

Antiviral agent Download PDF

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JP5158352B2
JP5158352B2 JP2008018773A JP2008018773A JP5158352B2 JP 5158352 B2 JP5158352 B2 JP 5158352B2 JP 2008018773 A JP2008018773 A JP 2008018773A JP 2008018773 A JP2008018773 A JP 2008018773A JP 5158352 B2 JP5158352 B2 JP 5158352B2
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antiviral agent
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ethanol
norovirus
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JP2009179577A (en
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範子 伊藤
優 鍋田
麗 石崎
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Lion Hygiene Co Ltd
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本発明は、ウイルス不活化効果を有する、エタノール含有抗ウイルス剤に関するものである。   The present invention relates to an ethanol-containing antiviral agent having a virus inactivating effect.

アルコールを主剤とするアルコール製剤は、食品へ直接使用できることや、調理器具、食器、厨房スペース等で使用できる等、使用性が極めて高い点から、除菌剤として広く使用されている。一方、急性胃腸炎を引き起こすノロウイルスは、近年猛威をふるっており、昨今の食中毒・感染症の主流となりつつある。しかしながら、従来のアルコール製剤では、ノロウイルス(ネコカリシウイルスによる代替実験による)に対して効果が不十分であるという問題があった。以上のことから、特に、ノロウイルスに対応できるアルコール製剤が望まれていた。   Alcohol preparations based on alcohol are widely used as disinfectants because they can be used directly in foods and can be used in cooking utensils, tableware, kitchen spaces, and the like. On the other hand, norovirus causing acute gastroenteritis has been on the rise in recent years and is becoming the mainstream of recent food poisoning and infectious diseases. However, the conventional alcohol preparation has a problem that its effect on Norovirus (by a substitute experiment with feline calicivirus) is insufficient. From the above, an alcohol preparation capable of dealing with norovirus has been particularly desired.

また、アルコール製剤は、食品へ直接使用する点から食品への影響が少ないことが求められ、組成物の安定性に優れることが望まれていた。さらに、アルコール製剤が使用される対象には、金属性調理器具等があるため、金属の腐食が防止できるアルコール製剤が望まれていた。   In addition, the alcohol preparation is required to have little influence on food because it is used directly on food, and it is desired that the composition has excellent stability. Furthermore, since the subject to which the alcohol preparation is used includes a metal cooking utensil or the like, an alcohol preparation capable of preventing metal corrosion has been desired.

特公昭58−2668号公報Japanese Patent Publication No.58-2668 特公平2−31946号公報JP-B-2-31946

本発明は上記事情に鑑みなされたもので、ウイルス、特にノロウイルス(ネコカリシウイルスによる代替実験による)不活化効果を有し、かつ食品への影響が少なく、金属の腐食が防止され、組成物の安定性に優れる抗ウイルス剤を提供することを目的とする。   The present invention has been made in view of the above circumstances, has an inactivating effect on viruses, particularly norovirus (by a substitute experiment with feline calicivirus), has little influence on food, prevents metal corrosion, An object is to provide an antiviral agent having excellent stability.

本発明者らは、上記目的を達成するため鋭意検討した結果、(A)エタノールと、(B)有機酸塩と、(C)ユーカリ抽出物とをそれぞれ特定量で配合することにより、ウイルス、特にノロウイルス(ネコカリシウイルスによる代替実験による)不活化効果を有し、かつ食品への影響が少なく、金属の腐食が防止され、組成物の安定性に優れることを知見し、本発明をなすに至ったものである。   As a result of intensive studies to achieve the above object, the present inventors have formulated (A) ethanol, (B) organic acid salt, and (C) eucalyptus extract in specific amounts, respectively. In particular, it has been found that it has an inactivating effect on norovirus (by alternative experiment with feline calicivirus), has little influence on food, prevents corrosion of metal, and has excellent stability of the composition. It has come.

従って、本発明は、(A)エタノール40〜60質量%と、(B)有機酸塩0.05〜0.5質量%と、(C)ユーカリ抽出物0.05〜0.5質量%とを含有し、pH(15℃)が8〜9である、ノロウイルス又はネコカリシウイルスに対する抗ウイルス剤を提供する。 Therefore, the present invention comprises (A) ethanol 40-60% by mass, (B) organic acid salt 0.05-0.5% by mass, (C) eucalyptus extract 0.05-0.5% by mass, And an antiviral agent against norovirus or feline calicivirus having a pH (15 ° C.) of 8-9 .

本発明によれば、ウイルス、特にノロウイルス(ネコカリシウイルスによる代替実験による)不活化効果を有し、かつ食品への影響が少なく、金属の腐食が防止され、組成物の安定性に優れる抗ウイルス剤を提供することができる。   ADVANTAGE OF THE INVENTION According to this invention, the anti-virus which has the inactivation effect of viruses, especially Norovirus (by the alternative experiment by a feline calicivirus), has little influence on food, prevents metal corrosion, and has excellent composition stability. An agent can be provided.

本発明の抗ウイルス剤は、(A)エタノール40〜60質量%と、(B)有機酸塩0.05〜0.5質量%と、(C)ユーカリ抽出物0.05〜0.5質量%とを含有するものである。   The antiviral agent of the present invention comprises (A) ethanol 40 to 60% by mass, (B) organic acid salt 0.05 to 0.5% by mass, and (C) eucalyptus extract 0.05 to 0.5% by mass. %.

(A)エタノール
エタノールとしては、発酵エタノール、合成エタノール等が挙げられ、いずれも用いることができ、1種単独で又は2種以上を適宜組み合わせて用いることができる。エタノールは殺菌性能を有するものであり、その配合量は、抗ウイルス剤全量に対して40〜60質量%である。通常、70〜80質量%の高濃度エタノール溶液が、殺菌性能が高いとされているが、配合量を40〜60質量%、好ましくは45〜58質量%にすることで、特にノロウイルス不活化効果が高くなる。一方、配合量が40質量%未満だと、ウイルス不活化効果が不十分となり、60質量%を超えると、安定性(25℃)が悪くなる。
(A) Ethanol Examples of ethanol include fermented ethanol, synthetic ethanol, and the like, and any of them can be used alone or in combination of two or more. Ethanol has bactericidal performance, and its blending amount is 40 to 60% by mass with respect to the total amount of the antiviral agent. Usually, a high-concentration ethanol solution of 70 to 80% by mass is considered to have high bactericidal performance. However, it is particularly effective to inactivate norovirus by setting the blending amount to 40 to 60% by mass, preferably 45 to 58% by mass. Becomes higher. On the other hand, when the blending amount is less than 40% by mass, the virus inactivating effect is insufficient, and when it exceeds 60% by mass, the stability (25 ° C.) is deteriorated.

(B)有機酸塩
有機酸塩を配合することで、ウイルス不活化効果をより向上させることができ、金属性の調理器具の腐食の心配もない。有機酸塩としては、例えば、クエン酸塩、乳酸塩、リンゴ酸塩、酒石酸塩、シュウ酸塩、酢酸塩、ギ酸塩、コハク酸塩、フマル酸塩、グルコンサン塩等が挙げられ、1種単独で又は2種以上を適宜組み合わせて用いることができる。塩としては、アルカリ金属塩等が挙げられ、水和物であってもよい。この中でも、クエン酸塩が好ましく、クエン酸3ナトリウム・2水和物がより好ましい。
(B) Organic acid salt By adding an organic acid salt, the virus inactivating effect can be further improved, and there is no concern about corrosion of metallic cookware. Examples of the organic acid salt include citrate, lactate, malate, tartrate, oxalate, acetate, formate, succinate, fumarate, gluconate salt and the like. It can use individually or in combination of 2 or more types as appropriate. Examples of the salt include alkali metal salts and the like, and may be hydrates. Among these, citrate is preferable, and trisodium citrate dihydrate is more preferable.

(B)成分の配合量は、抗ウイルス剤全量に対して0.05〜0.5質量%であり、0.1〜0.3質量%が好ましい。配合量が0.05質量%未満だと、(B)成分配合の効果が得られず、0.5質量%を超えると、安定性が悪くなり、結晶が析出する。   (B) The compounding quantity of a component is 0.05-0.5 mass% with respect to antiviral agent whole quantity, and 0.1-0.3 mass% is preferable. When the blending amount is less than 0.05% by mass, the effect of component (B) blending cannot be obtained, and when it exceeds 0.5% by mass, the stability is deteriorated and crystals are precipitated.

(C)ユーカリ抽出物
ユーカリ抽出物とは、フトモモ科ユーカリ属の植物からの抽出物をいう。ユーカリ抽出物としては、公知の抽出方法で得られたもの、市販品等を用いることができる。抽出部位は特に限定されないが、葉が好ましい。抽出方法は特に限定されず、溶媒抽出、水蒸気蒸留、超臨界抽出等の公知の抽出方法を採用することができる。例えば、溶媒抽出の場合、ユーカリを生のまま、又は乾燥した後に適当な大きさに切断・加工し、抽出溶媒に浸漬、撹拌することによって得ることができる。抽出は必要に応じて加温してもよく、抽出時間は適宜選定され、抽出pHは、極端な酸性又はアルカリ性でなければ特に制限はない。上記抽出方法に用いる溶媒としては、水;メタノール、エタノール、プロパノール、ブタノール等のアルコール類;プロピレングリコール、ブチレングリコール等の多価アルコール類等が挙げられ、これらを1種単独で又は2種以上を適宜組み合わせて用いることができる。抽出後ろ過を行い、適宜公知の精製を行ってもよい。
(C) Eucalyptus extract Eucalyptus extract refers to an extract from a plant belonging to the genus Eucalyptus. As a eucalyptus extract, what was obtained by the well-known extraction method, a commercial item, etc. can be used. The extraction site is not particularly limited, but leaves are preferred. The extraction method is not particularly limited, and a known extraction method such as solvent extraction, steam distillation, or supercritical extraction can be employed. For example, in the case of solvent extraction, it can be obtained by cutting and processing eucalyptus raw or after drying to a suitable size, and immersing and stirring in the extraction solvent. The extraction may be heated as necessary, the extraction time is appropriately selected, and the extraction pH is not particularly limited unless it is extremely acidic or alkaline. Examples of the solvent used in the extraction method include water; alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as propylene glycol and butylene glycol, and the like. They can be used in appropriate combinations. After extraction, filtration may be performed to appropriately perform known purification.

ユーカリ抽出物の形状は特に限定されず、上記処理により得られたものをそのまま、その希釈液、濃縮液等の液状、ペースト状、凍結乾燥等による乾燥粉末物等が挙げられる。   The shape of the eucalyptus extract is not particularly limited, and examples thereof include those obtained by the above-described treatment as they are, such as liquids such as dilutions and concentrates, pastes, and dry powders by freeze drying.

(C)成分の配合量は、抗ウイルス剤全量に対して0.05〜0.5質量%であり、0.1〜0.4質量%が好ましい。配合量が0.5質量%を超えると、食品に用いた場合、苦味を生じる等の悪影響があり、抗ウイルス剤の色味が濃くなる。   (C) The compounding quantity of a component is 0.05-0.5 mass% with respect to antiviral agent whole quantity, and 0.1-0.4 mass% is preferable. When the blending amount exceeds 0.5% by mass, when used in food, there is an adverse effect such as causing bitterness, and the color of the antiviral agent becomes deep.

本発明の抗ウイルス剤には、本発明の効果を損なわない範囲で、アルコール製剤に配合する成分を1種単独で又は2種以上を適宜組み合わせて、適量用いることができる。このような成分としては、防腐剤、酸化防止剤、pH調整剤、界面活性剤、色素、香料、水等が挙げられる。   In the antiviral agent of the present invention, an appropriate amount can be used alone or in combination of two or more of the components to be blended in the alcohol preparation within a range not impairing the effects of the present invention. Examples of such components include preservatives, antioxidants, pH adjusters, surfactants, pigments, fragrances, and water.

本発明の抗ウイルス剤は、例えば、上記(A)〜(C)成分、必要に応じて任意成分及び水(残部)を混合し、常法に基づいて調製することができる。   The antiviral agent of the present invention can be prepared, for example, by mixing the above components (A) to (C), optional components and water (remainder) as necessary, based on a conventional method.

本発明の抗ウイルス剤のpH(15℃)は8〜9が好ましい。pHが8未満であると、ウイルス不活化効果が低下するおそれがあり、ユーカリ抽出物の安定性や金属腐食を防ぐ点から、pHが9を超えないことが望ましい。   The pH (15 ° C.) of the antiviral agent of the present invention is preferably 8-9. If the pH is less than 8, the virus inactivating effect may be lowered, and it is desirable that the pH does not exceed 9 from the viewpoint of preventing the stability of the eucalyptus extract and metal corrosion.

本発明の抗ウイルス剤は、特にノロウイルス(ネコカリシウイルスによる代替実験による)不活化効果が顕著であり、ノロウイルス又はネコカリシウイルスに対する抗ウイルス剤として好適である。   The antiviral agent of the present invention is particularly effective as a norovirus (by alternative experiment with feline calicivirus) inactivation effect, and is suitable as an antiviral agent against norovirus or feline calicivirus.

本発明の抗ウイルス剤は、食品、手等の皮膚、包丁、鍋等の調理器具、皿等の食器類、厨房の調理台、流し台(三角コーナー)、厨房の床、壁等の厨房スペース等、冷蔵庫、トイレ、バス、洗面所、その他ウイルスの付着のおそれがある部分に適用することができる。   The antiviral agent of the present invention is food, skin such as hands, kitchen utensils, cooking utensils such as pans, dishes such as dishes, kitchen countertops, sinks (triangular corners), kitchen floors, kitchen spaces such as walls, etc. It can be applied to refrigerators, toilets, baths, toilets, and other areas where there is a risk of virus attachment.

本発明の抗ウイルス剤の使用方法としては特に限定されないが、例えば、対象物を抗ウイルス剤に浸漬、対象物に抗ウイルス剤を塗布又は噴霧(例えば、トリガースプレー、ディスペンサースプレー、エアゾール等)し、そのまま自然乾燥してもよいし、風乾、拭き取り等をして使用することができる。対象物に塗布又は噴霧する量としては、対象物にまんべんなく塗布又は噴霧できればよく、その量は特に限定されないが、約1〜10mLである。   The method of using the antiviral agent of the present invention is not particularly limited. For example, the object is immersed in the antiviral agent, and the antiviral agent is applied to or sprayed on the object (for example, trigger spray, dispenser spray, aerosol, etc.). It can be naturally dried as it is, or it can be used after being air-dried or wiped off. The amount to be applied or sprayed onto the object is not particularly limited as long as it can be applied or sprayed evenly onto the object, and the amount is about 1 to 10 mL.

以下、実施例及び比較例を示し、本発明を具体的に説明するが、本発明は下記の実施例に制限されるものではない。なお、下記の例において特に明記のない場合は、組成の「%」は質量%、表中の各成分の量は純分の量である。   EXAMPLES Hereinafter, although an Example and a comparative example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to the following Example. In the following examples, unless otherwise specified, “%” of the composition is mass%, and the amount of each component in the table is a pure amount.

[実施例1〜7、比較例1〜6]
精製水に、クエン酸3ナトリウム・2水和物、エタノール、ユーカリ抽出物を順に添加し、最後に精製水で量を調整して、表2,3に示す組成の抗ウイルス剤を得た。得られた抗ウイルス剤について下記評価を行った。結果を表中に併記する。
[Examples 1-7, Comparative Examples 1-6]
Trisodium citrate dihydrate, ethanol, and eucalyptus extract were sequentially added to purified water, and finally the amount was adjusted with purified water to obtain antiviral agents having the compositions shown in Tables 2 and 3. The following evaluation was performed about the obtained antiviral agent. The results are also shown in the table.

[ウイルス不活性化]
ネコカリシウイルスを用いて調べた。ノロウイルスは培養系が確立されていないため、ノロウイルスの代替ウイルスとして、同じカリシ科に属するネコカリシウイルスを用いて、ウイルス感染力を確認する方法が一般的に採用されている。ネコカリシウイルスは、ネコカリシウイルス感染症というネコの感染症の病原体で、ノロウイルスと構造が似ており、似た性質を持つものである。
[Virus inactivation]
It investigated using the feline calicivirus. Since norovirus culture systems have not been established, a method for confirming virus infectivity using feline calicivirus belonging to the same caliciidae as a substitute virus for norovirus is generally employed. Feline calicivirus is a pathogen of feline infectious disease called feline calicivirus infection, which is similar in structure to norovirus and has similar properties.

細胞にウイルスを感染させ、限界希釈法により、50%培養細胞感染濃度(TCID50)に基づき、ウイルス感染価を求めた。
供試ウイルス:ネコカリシウイルス(Feline calicivirus)、ATCC VR−782
供試細胞:CRFK細胞:ネコ腎臓細胞(CRFK:Crandell−Reese Feline Kidney)、ATCC CCL−94
The cells were infected with virus, and the virus infection titer was determined based on the 50% cultured cell infection concentration (TCID 50 ) by the limiting dilution method.
Test virus: Feline calicivirus, ATCC VR-782
Test cells: CRFK cells: Cat kidney cells (CRFK: Crandell-Reese Feline Kidney), ATCC CCL-94

試験管内で試料900μLにネコカリシウイルス液100μLを接種し、ボルテックスでよく混合し、室温で60秒作用させた(原液)。その後、この混合液をPBS(−):Phospate−Buffered Saline、品名:ダルベッコリン酸バッファー(invitrogen株式会社)で10倍に希釈中和した。さらに、ウイルス感染価を求めるため、PBS(−)を用いて10倍で段階希釈した。
96穴ウエルプレートに10%FCS(+)MEMを用いて単層培養したCRFK細胞に、上記希釈品を適宜25μL、それぞれ4ウェル接種し、ネコカリシウイルスとCRFK細胞とを接触させた。さらに、0.2%MEM FCS(+)を100μL重層し、37℃のCO2ガスフラン器内で3〜5日間培養した。
顕微鏡下で観察を行い、4ウェル中半数のウェル(50%)に、ウイルスにより引き起こされた細胞変性が見られた希釈倍数を求めた。この希釈倍数の対数値をウイルス感染価とした。
ブランクは、試料の代わりにPBS(−)溶液を用いる以外は同様に行い、ブランクのウイルス感染価が4のウイルスを用いた。ブランクに対する試料のウイルス感染価の減少値(ブランクの感染価−試料のウイルス感染価)を求めた。このウイルス感染価の減少値に基づき、ウイルス不活性化効果を下記評価基準で示した。
<評価基準>
◎:ウイルス感染価の減少値4
○:ウイルス感染価の減少値3
△:ウイルス感染価の減少値2〜1
×:ウイルス感染価の減少値0
10%FCS(+)MEM組成を下記に示す。
In a test tube, 900 μL of a sample was inoculated with 100 μL of feline calicivirus solution, mixed well by vortexing, and allowed to act at room temperature for 60 seconds (stock solution). Then, this mixed solution was diluted and neutralized 10 times with PBS (-): Phospate-Buffered Saline, product name: Dulbecco's phosphate buffer (Invitrogen). Furthermore, in order to determine the virus infectivity titer, serial dilution was performed 10 times with PBS (-).
CRFK cells monolayer cultured in 96-well plates using 10% FCS (+) MEM were inoculated with 25 μL of the diluted product in 4 wells, respectively, and feline calicivirus and CRFK cells were contacted. Furthermore, 100 μL of 0.2% MEM FCS (+) was overlaid and cultured in a CO 2 gas furan vessel at 37 ° C. for 3 to 5 days.
Observation was carried out under a microscope, and the dilution factor at which cytopathy caused by the virus was observed in half of the four wells (50%) was determined. The logarithm of the dilution factor was used as the virus infectivity value.
Blank was performed in the same manner except that a PBS (−) solution was used instead of the sample, and a virus having a virus infectivity of 4 was used. The decrease value of the virus infectivity value of the sample with respect to the blank (blank infectivity value-virus infectivity value of the sample) was determined. Based on the decreased value of the virus infectivity, the virus inactivation effect was shown by the following evaluation criteria.
<Evaluation criteria>
A: Decreased value of virus infection value 4
○: Decrease value of virus infection value 3
Δ: Decrease value of virus infection value 2-1
×: Reduced value of virus infection value 0
The 10% FCS (+) MEM composition is shown below.

Figure 0005158352
Figure 0005158352

[金属腐食防止]
アルミ板(A1100P、厚さ×幅×長さ=1.0×25×75mm)に、試料0.5mLを滴下し、24時間後に表面を拭き取った。金属表面の腐食の有無を、試料未滴下の金属板と比較し、目視にて確認した。結果を下記評価基準で示す。
<評価基準>
○:腐食していない
×:腐食している
[Metal corrosion prevention]
0.5 mL of the sample was dropped onto an aluminum plate (A1100P, thickness × width × length = 1.0 × 25 × 75 mm), and the surface was wiped off after 24 hours. The presence or absence of corrosion on the metal surface was compared with a metal plate not dripped with the sample and confirmed visually. A result is shown by the following evaluation criteria.
<Evaluation criteria>
○: Not corroded ×: Corroded

[食品への影響]
赤みの刺身(マグロ)に対して、試料約1.2mLをスプレーヤーにて噴霧した。冷蔵で2時間放置後、刺身をそのまま食した時の味への影響を評価した。
<評価基準>
○:変化なし
△:若干影響あり
×:影響あり
(○、△までは、食品に使用しても問題はないと判断する。)
[Impact on food]
About 1.2 mL of the sample was sprayed with a sprayer on reddish sashimi (tuna). After leaving for 2 hours in the refrigerator, the effect on taste when eating sashimi as it was was evaluated.
<Evaluation criteria>
○: No change △: Slightly affected ×: Influenced (Up to ○ and △, it is judged that there is no problem even if used for food)

[安定性]
試料(50mL)を50mLのバイアル瓶に充填し、常温で30日保存した。保存後、外観を下記評価基準に基づき評価した。
<評価基準>
○:均一透明
×:結晶析出
[Stability]
The sample (50 mL) was filled in a 50 mL vial and stored at room temperature for 30 days. After storage, the appearance was evaluated based on the following evaluation criteria.
<Evaluation criteria>
○: Uniform and transparent ×: Crystal precipitation

Figure 0005158352
Figure 0005158352

Figure 0005158352
Figure 0005158352

上記例で用いた原料を下記に示す。
エタノール:発酵エタノールトレーサブル95 1級(日本アルコール販売株式会社製)に、食品香料No.10(フレーバーH−No.13)(長谷川香料株式会社製)を1kg/200Lの割合で添加する。アルコール有効成分91.88%とし、純分換算して配合した。
クエン酸3ナトリウム・2水和物:クエン酸三ナトリウム(食品添加物グレード)(純正化学株式会社製)
ユーカリ抽出物:ユーカリエキス、50%エタノール抽出物(王子木材緑化株式会社製)
精製水:正起薬品工業株式会社製
The raw materials used in the above examples are shown below.
Ethanol: Fermented ethanol traceable 95 grade 1 (manufactured by Nippon Alcohol Sales Co., Ltd.) 10 (flavor H-No. 13) (manufactured by Hasegawa Fragrance Co., Ltd.) is added at a rate of 1 kg / 200 L. The alcohol active ingredient was 91.88%, and it was blended in terms of pure content.
Trisodium citrate dihydrate: Trisodium citrate (food additive grade) (manufactured by Junsei Co., Ltd.)
Eucalyptus extract: Eucalyptus extract, 50% ethanol extract (Oji Wood Greening Co., Ltd.)
Purified water: manufactured by Seiki Pharmaceutical Co., Ltd.

Claims (3)

(A)エタノール40〜60質量%と、(B)有機酸塩0.05〜0.5質量%と、(C)ユーカリ抽出物0.05〜0.5質量%とを含有し、pH(15℃)が8〜9である、ノロウイルス又はネコカリシウイルスに対する抗ウイルス剤。 (A) ethanol 40-60 mass%, (B) organic acid salt 0.05-0.5 mass%, and (C) eucalyptus extract 0.05-0.5 mass%, pH ( An antiviral agent against norovirus or feline calicivirus, wherein 15 ° C) is 8-9 . (B)成分が、クエン酸塩、乳酸塩、リンゴ酸塩、酒石酸塩、シュウ酸塩、酢酸塩、ギ酸塩、コハク酸塩、フマル酸塩及びグルコンサン塩から選ばれる請求項1記載の抗ウイルス剤。The antibacterial composition according to claim 1, wherein the component (B) is selected from citrate, lactate, malate, tartrate, oxalate, acetate, formate, succinate, fumarate and glucosan salt. Viral agent. (B)成分が、クエン酸塩である請求項2記載の抗ウイルス剤。The antiviral agent according to claim 2, wherein the component (B) is citrate.
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