JP2020096542A - Evaluation method and evaluation sample of antibacterial antifungal performance of cleaning composition - Google Patents
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Abstract
Description
本発明は、洗浄用組成物の抗菌抗カビ性能の評価方法及び評価用試料に関する。 The present invention relates to a method for evaluating antibacterial and antifungal performance of a cleaning composition and a sample for evaluation.
住環境設備には、便器、浴槽、洗面台、壁面等に使用されるセラミックス、プラスチック、タイル、金属等の硬質材料が多く存在する。これら硬質材料の表面（以下において、「硬質表面」という。）には、細菌類や真菌類（カビ等）など様々な菌種が付着する。これらの菌種の繁殖は衛生上問題となるだけではなく、黒ずみや赤色汚れなど、微生物に由来する汚れの発生を招くものである。硬質表面に付着する菌種の繁殖やこれに伴う汚れを防止するために、種々多様な抗菌抗カビ剤を含有する洗浄用組成物や、抗菌製品が開発されている。 In living environment facilities, there are many hard materials such as ceramics, plastics, tiles and metals used for toilets, bathtubs, washbasins, wall surfaces and the like. Various bacterial species such as bacteria and fungi (molds, etc.) adhere to the surface of these hard materials (hereinafter referred to as “hard surface”). Propagation of these bacterial species is not only a sanitary problem, but also causes the generation of stains derived from microorganisms such as darkening and red stains. In order to prevent the propagation of bacterial species adhering to a hard surface and the accompanying stains, cleaning compositions and antibacterial products containing various antibacterial and antifungal agents have been developed.
洗浄用組成物や抗菌製品の開発においては、その抗菌抗カビ性能（抗菌抗カビ効果）が評価される。その評価方法として、例えば、特許文献１では、カビ胞子を塗布した濾紙に洗浄用組成物を塗布して数分間放置した後、濾紙を水洗し、この濾紙をポテトデキストロース寒天培地（Potato Dextrose Agar培地；ＰＤＡ培地）上で培養した後における黒色部分の発生状態を観察することで、抗菌抗カビ性能を評価している。 In the development of cleaning compositions and antibacterial products, their antibacterial and antifungal performance (antibacterial and antifungal effect) is evaluated. As an evaluation method thereof, for example, in Patent Document 1, after applying a cleaning composition to a filter paper coated with mold spores and leaving it for a few minutes, the filter paper is washed with water, and the filter paper is potato dextrose agar medium (Potato Dextrose Agar medium). The antibacterial and antifungal performance is evaluated by observing the generation state of the black portion after culturing on PDA medium).
また、特許文献２では、シャーレ内の寒天培地に黒カビを培養し、上から円筒形状のガラスチューブを嵌め込み、そのガラスチューブ内に漂白剤をたらし、３０分放置後の黒カビ漂白の程度を評価している。 Further, in Patent Document 2, black mold is cultivated on an agar medium in a petri dish, a cylindrical glass tube is fitted from above, a bleaching agent is poured into the glass tube, and the degree of black mold bleaching after leaving for 30 minutes is evaluated. doing.
また、特許文献３〜５では、汚れの原因菌液に洗浄用組成物を添加後、シャーレ等に固化させた培地上で培養し、発生したコロニー数をカウントすることにより、初発菌数からの減少菌数を算出することで評価している。 In addition, in Patent Documents 3 to 5, after adding a cleaning composition to a stain-causing bacterial solution, it is cultured on a medium solidified in a petri dish or the like, and the number of colonies generated is counted to obtain the number from the initial number of bacteria. It is evaluated by calculating the reduced number of bacteria.
また、特許文献６では、便器等の衛生陶器表面における抗菌抗カビ性能の評価方法として、衛生陶器表面に寒天培地を形成し、この寒天培地に形成されたコロニーを目視にて観察することで衛生陶器の抗菌抗カビ性能を評価している。 Further, in Patent Document 6, as an evaluation method of antibacterial and antifungal performance on the surface of sanitary ware such as a toilet bowl, an agar medium is formed on the surface of the sanitary ware, and colonies formed on the agar medium are visually observed to be hygienic. The antibacterial and antifungal performance of pottery is evaluated.
一方、上記硬質表面の洗浄用組成物として、洗浄対象である硬質表面に泡状に付着するよう洗浄用組成物を塗布することにより、硬質表面をこすることなく水で洗い流すタイプが近年増加している。例えば、「モコ泡わ（登録商標）トイレクリーナー」（エステー株式会社製）等のトイレ用洗剤、「バスマジックリン（登録商標）」（花王株式会社製）等の風呂用洗剤などの各種製品が挙げられる。 On the other hand, as the cleaning composition for the hard surface, by applying the cleaning composition so that it adheres to the hard surface to be cleaned in a foamy manner, the number of types in which the hard surface is washed off with water without rubbing has increased in recent years. ing. For example, various products such as toilet detergents such as "Moko Awawa (registered trademark) Toilet Cleaner" (manufactured by ST Co., Ltd.) and bath detergents such as "Bath Magic Lynn (registered trademark)" (manufactured by Kao Corporation) are available. Can be mentioned.
これら洗浄用組成物は、泡の状態で被洗浄面に付着することにより、分離した汚れが洗浄対象物に再度付着することを防止するほか、洗浄対象物上に洗浄用組成物が滞留する時間を延ばし、汚れ成分に対する洗浄成分の接触時間を長くするといった効果が期待できる。このため、こするなどの機械的な力を加えることなく、所定時間放置した後、水で洗い流すことで容易に洗浄することができるという利点を有する。また、菌又はカビに由来する汚れに対しても同様に、抗菌抗カビ成分の接触時間を長くすることができるため、菌又はカビに由来する汚れの発生を防止することができるものである。 These cleaning compositions adhere to the surface to be cleaned in the form of bubbles to prevent the separated dirt from re-adhering to the object to be cleaned, and the time for which the cleaning composition stays on the object to be cleaned. And the effect of extending the contact time of the cleaning component with the dirt component can be expected. For this reason, there is an advantage that it can be easily washed by leaving it for a predetermined time and then rinsing it with water without applying mechanical force such as rubbing. Similarly, for stains derived from fungi or mold, the contact time of the antibacterial/antifungal component can be prolonged, so that the generation of stains derived from fungi or mold can be prevented.
上記の通り、洗浄用組成物において、硬質表面などの洗浄対象表面に泡状に付着することにより、機械的な力を加えることなく、所定時間放置した後に水で洗い流すタイプの洗浄用組成物が近年増加している。このため、洗浄用組成物における抗菌抗カビ性能を、正確且つ簡便に評価することの重要性が高まっている。 As described above, in the cleaning composition, by adhering to the surface to be cleaned such as a hard surface in a foam form, a cleaning composition of the type that is rinsed with water after being left for a predetermined time without applying mechanical force. It has been increasing in recent years. Therefore, the importance of accurately and simply evaluating the antibacterial and antifungal performance in the cleaning composition is increasing.
しかしながら、例えば、特許文献１〜５に示される従来の抗菌抗カビ性能の評価方法は、洗浄用組成物が実際に使用される環境とはかけ離れた環境下でなされるものである。このため、評価での培養段階における菌種の生育状況が実態と異なり、実環境における菌又はカビに由来する汚れの抑制が可能であるかを確認できるものではない。また、特許文献６のように評価試料として洗浄対象物そのものを使用する場合、評価環境は洗浄用組成物が使用される実環境に近いものの、多数のサンプルの評価や有効成分のスクリーニングが困難であり、簡便な評価方法とは言い難い。 However, for example, the conventional methods for evaluating antibacterial and antifungal performance shown in Patent Documents 1 to 5 are performed in an environment far from the environment in which the cleaning composition is actually used. Therefore, the growth status of the bacterial species at the culture stage in the evaluation is different from the actual state, and it is not possible to confirm whether it is possible to suppress stains derived from the bacteria or mold in the actual environment. Further, when the cleaning object itself is used as the evaluation sample as in Patent Document 6, the evaluation environment is close to the actual environment in which the cleaning composition is used, but it is difficult to evaluate a large number of samples and screen active ingredients. There is no simple evaluation method.
本発明は、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果を簡便に視認評価することを可能とする評価方法を提供することを目的とする。また、本発明は、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果の簡便な視認評価を可能とする評価用試料を提供することを目的とする。 It is an object of the present invention to provide an evaluation method that enables simple and visual evaluation of the effect of suppressing stains derived from fungi or mold based on the antibacterial and antifungal performance of the cleaning composition. It is another object of the present invention to provide an evaluation sample that enables simple visual evaluation of the effect of suppressing stains derived from bacteria or mold based on the antibacterial and antifungal properties of the cleaning composition.
本発明者等は、評価環境を洗浄用組成物が実際に使用される環境に近づけるという観点から鋭意研究を重ねた。その結果、表面の少なくとも一部が、洗浄用組成物の洗浄対象表面を構成する材料と同一材料から構成される基板を、評価用試料に用いる基板として使用することにより、洗浄用組成物が実際に使用される環境を模擬した簡便な評価を実施することができることを見出し、本発明を完成するに至った。 The present inventors have conducted intensive studies from the viewpoint of bringing the evaluation environment closer to the environment in which the cleaning composition is actually used. As a result, the cleaning composition is actually used by using a substrate in which at least a part of the surface is composed of the same material as the material forming the surface to be cleaned of the cleaning composition, as the substrate used for the evaluation sample. The inventors have found that a simple evaluation that simulates the environment used for can be performed, and have completed the present invention.
すなわち、本発明の一側面によると、洗浄用組成物の抗菌抗カビ性能の評価方法であって、基板として、表面の少なくとも一部が、前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料から構成される基板を選択すること、前記基板上に、液状の培地と菌懸濁液又はカビ胞子液の混合物を塗布すること、前記菌懸濁液又は前記カビ胞子液を含む前記培地を固化すること、前記培地表面に前記洗浄用組成物を接触させること、前記培地表面に不活化剤含有水溶液を接触させること、及び、所定時間経過後に前記培地におけるコロニーの生育状況を目視により観察することをこの順序で含む第一の評価方法が提供される。 That is, according to one aspect of the present invention, there is provided a method for evaluating antibacterial and antifungal performance of a cleaning composition, wherein at least a part of the surface of the substrate is a material constituting a surface to be cleaned by the cleaning composition. Selecting a substrate composed of the same material, applying a mixture of a liquid medium and a bacterial suspension or fungal spore solution onto the substrate, the medium containing the bacterial suspension or the fungal spore solution Solidifying, contacting the cleaning composition with the culture medium surface, contacting an inactivating agent-containing aqueous solution with the culture medium surface, and visually observing the growth state of colonies in the culture medium after a predetermined period of time. A first evaluation method is provided that includes doing in this order.
また、本発明の他の側面によると、洗浄用組成物の抗菌抗カビ性能の評価方法であって、基板として、表面の少なくとも一部が、前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料から構成される基板を選択すること、前記基板上に固体状の培地を形成すること、前記培地上に菌懸濁液又はカビ胞子液を塗布すること、前記培地表面に前記洗浄用組成物を接触させること、前記培地表面に不活化剤含有水溶液を接触させること、及び、所定時間経過後に前記培地におけるコロニーの生育状況を目視により観察することをこの順序で含む第二の評価方法が提供される。 Further, according to another aspect of the present invention, there is provided a method for evaluating antibacterial and antifungal performance of a cleaning composition, wherein at least a part of the surface of the substrate is a material constituting a surface to be cleaned by the cleaning composition. A substrate composed of the same material as above, forming a solid medium on the substrate, applying a bacterial suspension or fungal spore solution on the medium, and cleaning the surface of the medium with A second evaluation method comprising contacting a composition, contacting an inactivating agent-containing aqueous solution with the surface of the medium, and visually observing the growth state of colonies in the medium after a lapse of a predetermined time in this order. Will be provided.
上記第一の評価方法及び第二の評価方法は、一形態において、上記培地の厚さが３ｍｍ以下であってよく、１ｍｍ以下であってよい。 In one aspect of the first evaluation method and the second evaluation method, the thickness of the medium may be 3 mm or less, or 1 mm or less.
また、上記第一の評価方法及び第二の評価方法は、他の形態において、上記培地が寒天培地であってよい。 Further, in the first evaluation method and the second evaluation method, in another embodiment, the medium may be an agar medium.
また、上記第一の評価方法及び第二の評価方法は、他の形態において、上記洗浄用組成物が硬質材料の表面を洗浄対象とする硬質表面洗浄用組成物であってよい。 Moreover, the said 1st evaluation method and the 2nd evaluation method may be a hard surface cleaning composition in which the said cleaning composition makes the surface of a hard material a cleaning object in another form.
また、上記第一の評価方法及び第二の評価方法は、他の形態において、上記基板の表面の少なくとも一部を構成する上記材料が硬質材料であってよい。 Moreover, the said 1st evaluation method and the 2nd evaluation method WHEREIN: The said material which comprises at least one part of the surface of the said board|substrate may be a hard material in another form.
また、上記第一の評価方法及び第二の評価方法は、他の形態において、上記硬質材料が、セラミックス、プラスチック、タイル又は金属であってよい。 Moreover, in the said 1st evaluation method and the 2nd evaluation method, the said hard material may be ceramics, a plastic, a tile, or a metal in another form.
また、上記第一の評価方法及び第二の評価方法は、他の形態において、上記培地表面と上記洗浄用組成物との接触が、上記培地表面を泡状で覆うように上記洗浄用組成物を付着させることによりなされる。 Further, the first evaluation method and the second evaluation method, in another form, the contact between the medium surface and the cleaning composition, the cleaning composition so that the medium surface is covered with foam It is made by attaching.
また、本発明の他の側面によると、洗浄用組成物の抗菌抗カビ性能評価に使用する評価用試料であって、表面の少なくとも一部が前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料からなる基板と、前記基板上に菌又はカビを含む培地とを備える第一の評価用試料が提供される。 Further, according to another aspect of the present invention, an evaluation sample used for the evaluation of antibacterial and antifungal performance of a cleaning composition, wherein at least a part of the surface is a material constituting a surface to be cleaned by the cleaning composition. There is provided a first evaluation sample including a substrate made of the same material as above and a medium containing fungi or mold on the substrate.
また、本発明の他の側面によると、洗浄用組成物の抗菌抗カビ性能評価に使用する評価用試料であって、表面の少なくとも一部が前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料からなる基板と、前記基板上に培地とを備える第二の評価用試料が提供される。 Further, according to another aspect of the present invention, an evaluation sample used for the evaluation of antibacterial and antifungal performance of a cleaning composition, wherein at least a part of the surface is a material constituting a surface to be cleaned by the cleaning composition. A second evaluation sample including a substrate made of the same material as above and a culture medium on the substrate is provided.
上記第一の評価用試料及び第二の評価用資料は、一形態において、上記培地の厚さが３ｍｍ以下であってよく、１ｍｍ以下であってよい。 In one form of the first evaluation sample and the second evaluation material, the thickness of the medium may be 3 mm or less, or 1 mm or less.
また、上記第一の評価用試料及び第二の評価用試料は、他の形態において、上記培地が寒天培地であってよい。 Further, in the above-mentioned first evaluation sample and second evaluation sample, in another embodiment, the medium may be an agar medium.
また、上記第一の評価用試料及び第二の評価用試料は、他の形態において、上記洗浄用組成物が、硬質材料の表面を洗浄対象とする硬質表面洗浄用組成物であってよい。 Moreover, the said 1st evaluation sample and the 2nd evaluation sample WHEREIN: In the other form, the said cleaning composition may be a hard surface cleaning composition which makes the surface of a hard material a cleaning target.
また、上記第一の評価用試料及び第二の評価用試料は、他の形態において、上記基板の表面の少なくとも一部を構成する上記材料が硬質材料であってよい。 In addition, in the first evaluation sample and the second evaluation sample, in another form, the material forming at least a part of the surface of the substrate may be a hard material.
また、上記第一の評価用試料及び第二の評価用試料は、他の形態において、上記硬質材料が、セラミックス、プラスチック、タイル又は金属であってよい。 In addition, in another form of the first evaluation sample and the second evaluation sample, the hard material may be ceramics, plastic, tile, or metal.
また、上記第一の評価用試料及び第二の評価用試料は、他の形態において、上記洗浄用組成物は、洗浄対象表面へ泡状に付着するように塗布される用途に用いられる洗浄用組成物であってよい。 Further, the first evaluation sample and the second evaluation sample are, in another form, the cleaning composition, which is used for cleaning to be applied so as to adhere to the surface to be cleaned in a foamy manner. It may be a composition.
本発明により、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果を簡便に視認評価する方法を提供することが可能となる。また、本発明により、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果の簡便な視認評価を可能とする評価用試料を提供することが可能となる。 According to the present invention, it is possible to provide a method for easily visually evaluating the effect of suppressing stains derived from bacteria or mold based on the antibacterial and antifungal properties of the cleaning composition. Further, according to the present invention, it is possible to provide an evaluation sample that enables simple visual evaluation of the effect of suppressing stains derived from bacteria or mold based on the antibacterial and antifungal properties of the cleaning composition.
以下、本発明の実施形態に係る洗浄用組成物の抗菌抗カビ性能の評価方法（以下、「実施形態に係る評価方法」という。）と、そこで使用される評価用試料について説明する。 Hereinafter, a method for evaluating antibacterial and antifungal performance of a cleaning composition according to an embodiment of the present invention (hereinafter, referred to as “evaluation method according to an embodiment”) and an evaluation sample used therein will be described.
＜基板の選択工程＞
実施形態に係る評価方法は、培地が形成される評価用基板として、表面の少なくとも一部が、洗浄用組成物による洗浄対象表面を構成する材料と同一材料から構成される基板を選択する工程を含むことを特徴の一つとする。評価用基板として上記基板を選択することにより、洗浄用組成物が実際に使用される環境を模擬することができ、評価での培養段階における菌種の生育状況が実態に近づくため、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果の簡便な視認評価が可能となる。
<Substrate selection process>
The evaluation method according to the embodiment includes a step of selecting a substrate, at least a part of the surface of which is formed of the same material as the material forming the surface to be cleaned by the cleaning composition, as the evaluation substrate on which the medium is formed. Including is one of the features. By selecting the above substrate as the evaluation substrate, the environment in which the cleaning composition is actually used can be simulated, and the growth situation of the bacterial species at the culture stage in the evaluation approaches the actual condition. A simple visual evaluation of the effect of suppressing stains derived from bacteria or mold based on the antibacterial and antifungal properties of a product becomes possible.
実施形態に係る評価方法により抗菌抗カビ性能が評価される洗浄用組成物は、一形態において、硬質表面を洗浄対象とする硬質表面洗浄用組成物であってよい。ここで硬質表面は、特に限定されるものではないが、一例を挙げると、トイレ、浴室、台所、洗面台などの、壁や床、器具、機器等に設けられる表面であり、硬質材料の具体例として、例えば、セラミックス、プラスチック、金属、タイル等が挙げられる。 The cleaning composition whose antibacterial and antifungal performance is evaluated by the evaluation method according to the embodiment may be, in one form, a hard surface cleaning composition in which a hard surface is a cleaning target. Here, the hard surface is not particularly limited, but, for example, a surface provided on a wall or floor such as a toilet, a bathroom, a kitchen, and a washbasin, an appliance, a device, etc., and a specific example of a hard material. Examples include ceramics, plastics, metals, tiles, and the like.
したがって、実施形態に係る評価方法は、一形態において、硬質表面洗浄用組成物の抗菌抗カビ性能の評価方法であり、評価用基板を選択する工程において選択される基板は、表面の少なくとも一部が、硬質表面洗浄用組成物による洗浄対象である硬質表面を構成する材料と同一材料から構成される基板である。例えば、表面の少なくとも一部がセラミックス、プラスチック、金属、タイル等の硬質表面から構成される基板が挙げられる。実施形態に係る評価方法において、滅菌後の基板が使用される。 Therefore, the evaluation method according to the embodiment is, in one form, an evaluation method of the antibacterial and antifungal performance of the hard surface cleaning composition, and the substrate selected in the step of selecting the evaluation substrate is at least a part of the surface. Is a substrate composed of the same material as that of the hard surface which is the object of cleaning by the hard surface cleaning composition. For example, there may be mentioned a substrate in which at least a part of the surface is made of a hard surface such as ceramics, plastic, metal or tile. In the evaluation method according to the embodiment, the sterilized substrate is used.
＜培地の形成と、培地に対する菌／カビの仕込み工程＞
実施形態に係る評価方法は、培地に対する菌又はカビの仕込み方の相違により、第一の評価方法と、第二の評価方法に分けることができる。ここで、培地としては、特に限定されるものではなく、寒天培地などであってよい。寒天培地としては、例えば、標準寒天培地、ポテトデキストロース寒天培地、Ｒ２Ａ寒天培地、ＸＭ−Ｇ寒天培地、デゾキシコレート寒天培地等が挙げられ、一形態においてポテトデキストロース寒天培地が好ましい。
<Formation of medium and process of preparing bacteria/mold on medium>
The evaluation method according to the embodiment can be divided into a first evaluation method and a second evaluation method, depending on the difference in the method of charging the culture medium with fungi or mold. Here, the medium is not particularly limited, and may be an agar medium or the like. Examples of the agar medium include standard agar medium, potato dextrose agar medium, R2A agar medium, XM-G agar medium, dezoxycholate agar medium, and the like, and in one form, potato dextrose agar medium is preferable.
第一の評価方法では、上述した評価用基板の選択工程で選択した基板上に、液状の培地と菌懸濁液又はカビ胞子液との混合物を塗布することにより、菌又はカビを含有する培地が形成される。次いで、菌懸濁液又はカビ胞子液を含む培地は固化される。このとき得られる試料が後述する第一の評価用試料である。培地が寒天培地である場合、一形態において、所定温度で加熱溶解した寒天培地に、評価を希望する菌種を含む液体（例えば、カビ胞子液等）を所定濃度で混合し、得られる混合物を評価用基板に滴下、あるいはスプレー等により塗布し、これを例えばクリーンベンチ内で培地が固化するまで放置する。 In the first evaluation method, on the substrate selected in the selection step of the evaluation substrate described above, by applying a mixture of a liquid medium and a bacterial suspension or fungal spore solution, a medium containing fungi or mold. Is formed. Then, the medium containing the bacterial suspension or the mold spore fluid is solidified. The sample obtained at this time is a first evaluation sample described later. When the medium is an agar medium, in one embodiment, a liquid containing a bacterial species desired to be evaluated (for example, mold spore fluid) is mixed at a predetermined concentration with an agar medium heated and dissolved at a predetermined temperature, and the resulting mixture is The evaluation substrate is dropped or applied by spraying or the like, and is left in a clean bench, for example, until the medium is solidified.
第二の評価方法では、まず、上述した評価用基板の選択工程で選択した基板上に、液状の培地が塗布され、固化される。このとき得られる試料が後述する第二の評価用試料である。次いで、この固体状の培地上に菌懸濁液又はカビ胞子液が塗布されることにより、培地上に菌又はカビ含有層が形成される。培地が寒天培地である場合、一形態において、所定温度で加熱溶解した寒天培地を評価用基板に滴下、あるいはスプレー等により塗布し、これを例えばクリーンベンチ内で放置し培地を固化させる。次いで、評価を希望する菌種を含む液体（例えば、カビ胞子液等）を上記培地上に均一に塗布することにより菌又はカビ含有層が形成される。 In the second evaluation method, first, a liquid medium is applied and solidified on the substrate selected in the above-described evaluation substrate selecting step. The sample obtained at this time is a second evaluation sample described later. Then, a bacterial suspension or fungal spore solution is applied onto the solid medium to form a fungal or fungal-containing layer on the medium. When the medium is an agar medium, in one embodiment, the agar medium heated and melted at a predetermined temperature is dropped or applied on the evaluation substrate by spraying or the like, and this is left in, for example, a clean bench to solidify the medium. Next, a bacterium- or mold-containing layer is formed by uniformly applying a liquid containing the bacterium species to be evaluated (for example, mold spore solution or the like) onto the medium.
菌糸は、培地に対し下方に成長することがあり、培地の厚みが厚い場合、例えば便器等の硬質表面上での菌糸の生育の実態と異なってしまうため、正確な抗菌抗カビ性能の評価が困難となる。この観点から培地の厚みは適宜設定することが好ましい。第一の評価方法（第一の評価用試料）及び第二の評価方法（第二の評価用試料）では、上記培地の厚みは３ｍｍ以下であってよく、１ｍｍ以下であってよい。 Mycelia may grow downward with respect to the medium, and if the medium is thick, for example, it will differ from the actual state of mycelium growth on a hard surface such as a toilet bowl, so an accurate evaluation of antibacterial and antifungal performance is possible. It will be difficult. From this viewpoint, it is preferable to set the thickness of the medium appropriately. In the first evaluation method (first evaluation sample) and the second evaluation method (second evaluation sample), the thickness of the medium may be 3 mm or less, or 1 mm or less.
本発明の実施形態に係る洗浄用組成物の抗菌抗カビ性能評価に使用する評価用試料（以下において、「実施形態に係る評価用試料」という。）は、実施形態に係る評価方法における上述した工程を経て得られる試料である。 The evaluation sample used for the antibacterial/antifungal performance evaluation of the cleaning composition according to the embodiment of the present invention (hereinafter, referred to as “evaluation sample according to the embodiment”) is as described above in the evaluation method according to the embodiment. It is a sample obtained through the steps.
すなわち、第一の評価方法において作製され、使用される第一の評価用試料は、表面の少なくとも一部が上述した洗浄用組成物による洗浄対象表面を構成する材料と同一材料からなる基板と、この基板上に菌又はカビを含む培地を備える。 That is, the first evaluation sample that is produced in the first evaluation method and is used, at least a part of the surface is a substrate made of the same material as the material forming the surface to be cleaned by the cleaning composition described above, A medium containing fungi or mold is provided on this substrate.
第二の評価方法において作製され、使用される第二の評価用試料は、表面の少なくとも一部が上述した洗浄用組成物による洗浄対象表面を構成する材料と同一材料からなる基板と、この基板上に培地を備える。第二の評価用試料は、培地上に更に菌又はカビ含有層を更に備えていてもよい。 The second evaluation sample produced and used in the second evaluation method is a substrate in which at least a part of the surface is made of the same material as the material forming the surface to be cleaned by the cleaning composition described above, and this substrate. Provide the medium on top. The second evaluation sample may further include a fungus- or mold-containing layer on the medium.
＜洗浄用組成物との接触工程＞
実施形態に係る評価方法は、培地が形成され、菌又はカビが仕込まれた実施形態に係る上記評価用試料（第一の評価試料又は第二の評価試料）に対し、評価対象である任意の洗浄用組成物を接触させる工程を含む。接触の形態は、特に限定されるものではない。例えば、評価対象である洗浄用組成物が、洗浄対象表面へ泡状に付着するように塗布される用途に用いられるものである場合、当該接触は、上記評価用試料を泡で覆うように洗浄用組成物を付着させることによりなされることが好ましい。その手段として、例えば、内容物を泡状にして噴霧又は塗布することができるスプレー容器又はエアゾール容器を用いて作製される泡を評価用試料に付着させることによりなされる。
<Step of contact with cleaning composition>
The evaluation method according to the embodiment is an arbitrary evaluation target for the evaluation sample (first evaluation sample or second evaluation sample) according to the embodiment in which a medium is formed and fungi or mold are charged. The step of contacting the cleaning composition is included. The form of contact is not particularly limited. For example, when the cleaning composition to be evaluated is used for the purpose of being applied so as to adhere to the surface to be cleaned in a foamy manner, the contact is performed so that the evaluation sample is covered with foam. Preferably, it is done by depositing a composition for use. As a means for this, for example, a foam produced by using a spray container or an aerosol container capable of spraying or applying the content in the form of foam is attached to the evaluation sample.
＜不活化剤含有水溶液との接触工程（抗菌抗カビ活性の不活化工程）＞
実施形態に係る評価方法は、洗浄用組成物に接触させた後の上記評価用試料表面に対し、不活化剤含有水溶液を接触させる工程を含む。これは、洗浄用組成物における抗菌抗カビ活性を停止させるための工程であり、不活化剤含有水溶液としてレシチン・ポリソルベート８０（ＬＰ）希釈液を用いて上記評価用試料表面を洗浄することにより実施することができる。
<Step of contact with aqueous solution containing inactivating agent (step of inactivating antibacterial and antifungal activity)>
The evaluation method according to the embodiment includes a step of bringing an inactivating agent-containing aqueous solution into contact with the surface of the evaluation sample after being brought into contact with the cleaning composition. This is a step for stopping the antibacterial and antifungal activity in the cleaning composition, and is carried out by cleaning the surface of the sample for evaluation with a diluting solution of lecithin polysorbate 80 (LP) as an aqueous solution containing an inactivating agent. can do.
実施形態に係る評価方法は、不活化剤含有水溶液を接触させる工程の後、更に水（好ましくは滅菌水）を上記評価用試料表面に接触させ、洗浄する工程を更に含んでもよい。 The evaluation method according to the embodiment may further include a step of bringing water (preferably sterilized water) into contact with the surface of the sample for evaluation and washing after the step of contacting with the aqueous solution containing the inactivating agent.
＜培養及び目視による観察工程＞
実施形態に係る評価方法は、上記評価用試料表面を洗浄した後、培養のために所定時間評価用試料を放置し、培地におけるコロニーの生育状況を目視により観察する工程を含む。
<Culture and visual observation step>
The evaluation method according to the embodiment includes a step of washing the surface of the evaluation sample, allowing the evaluation sample to stand for a predetermined time for culturing, and visually observing the growth state of colonies in the medium.
実施形態に係る評価方法により評価することができる菌種は、増殖した際、視認できる菌又はカビであればよい。具体的には、カビの一例として、クロカワカビ（Ｃｌａｄｏｓｐｏｒｉｕｍ ｃｌａｄｏｓｐｏｒｉｏｉｄｅｓ）等のクラドスポリウム（Ｃｌａｄｏｓｐｏｒｉｕｍ）属、アオカビ（Ｐｅｎｉｃｉｌｌｉｕｍ ｃｉｔｒｉｎｕｍ）等のペニシリウム（Ｐｅｎｉｃｉｌｌｉｕｍ）属、コウジカビ（Ａｓｐｅｒｇｉｌｌｕｓ ｂｒａｓｉｌｉｅｎｓｉｓ）等のアスペルギルス（Ａｓｐｅｒｇｉｌｌｕｓ）属、ススカビ（Ａｌｔｅｒｎａｒｉａ ａｌｔｅｒｎａｔａ）等のアルテルナリア（Ａｌｔｅｒｎａｒｉａ）属、アカカビ（Ｆｕｓａｒｉｕｍ ｓｏｌａｎｉ）等のフザリウム（Ｆｕｓａｒｉｕｍ）属、Ｅｕｒｏｔｉｕｍ ｈｅｒｂａｒｉｏｒｕｍ等のユーロチウム（Ｅｕｒｏｔｉｕｍ）属、ロドトルラ属（Ｒｈｏｄｏｔｏｒｕｌａ ｍｕｃｉｌａｇｉｎｏｓａ）等の赤色酵母類、アウレオバシジウム（Ａｕｒｅｏｂａｓｉｄｉｕｍ）属、エキソフィアラ（Ｅｘｏｐｈｉａｌａ）属等の黒色酵母類、フォーマ（Ｐｈｏｍａ）属、カンジダ（Ｃａｎｄｉｄａ）属、サッカロマイセス（Ｓａｃｃｈａｒｏｍｙｃｅｓ）属等が挙げられる。 The bacterium species that can be evaluated by the evaluation method according to the embodiment may be any bacterium or mold that can be visually recognized when grown. Specifically, as an example of mold, the genus Cladosporus such as Cladosporium such as Cladosporium cladosporioides, and the genus Penicillus aspergillus of Aspergillus sp. , Alternaria genus such as Alternaria alternata, Fusarium genus such as Fusarium solani, genus Eurotium such as Eurotium herbarium orum, and Rhodotorula vulgaris such as Eurotium aurora ulta, Rhodotorula (Rhodrora) Black yeasts such as Aureobasidium genus and Exophiala genus, Phoma genus, Candida genus, Saccharomyces genus and the like can be mentioned.
菌の一例としては、ミクロバクテリウム（Ｍｉｃｒｏｂａｃｔｅｒｉｕｍ）属、ミクロコッカス（Ｍｉｃｒｏｃｏｃｃｕｓ）属等、黄色ブドウ球菌等のブドウ球菌（Ｓｔａｐｈｙｌｏｃｏｃｃｕｓ）属、枯草菌等のバシラス（Ｂａｃｉｌｌｕｓ）属等のグラム陽性菌、緑膿菌等のシュードモナス（Ｐｓｅｕｄｏｍｏｎａｓ）属、メチロバクテリウム（Ｍｅｔｈｙｌｏｂａｃｔｅｒｉｕｍ）属、肺炎桿菌等のクラブシエラ（Ｋｌｅｂｓｉｅｌｌａ）属、サルモネラ（Ｓａｌｍｏｎｅｌｌａ）属、モラクセラ（Ｍｏｒａｘｅｌｌａ）属等のグラム陰性菌が挙げられる。 Examples of the bacterium include genus Microbacterium, genus Micrococcus, genus Staphylococcus such as Staphylococcus aureus, gram-positive bacterium such as genus Bacillus such as Bacillus subtilis, and green Gram-negative bacteria such as Pseudomonas genus such as Pseudomonas aeruginosa, Methylobacterium genus, Klebsiella genus such as Klebsiella pneumoniae, Salmonella genus, Moraxella genus and the like can be mentioned.
培養は、一形態において、蓋つきの透明容器に評価用試料を配置し、外部からの菌の混入を防ぐために蓋を閉めた状態で、菌種の育成状況の経過を容器外から観察しながら行うことができる。培養時間は菌種により異なるが、例えば３日〜４日、室温（（例えば、２０〜２５℃）で培養することができる。菌種の生育に適した湿度を維持するために、容器内には水を設置することが好ましい。また、評価用試料以外の器具はエタノール消毒をした上で使用することが好ましい。 In one form, the culture is performed while arranging the sample for evaluation in a transparent container with a lid and closing the lid to prevent contamination of bacteria from the outside while observing the progress of the growth of the bacterial species from the outside of the container. be able to. Although the culturing time varies depending on the bacterial species, it can be carried out at room temperature ((eg, 20 to 25° C.)) for 3 to 4 days. In order to maintain the humidity suitable for the growth of the bacterial species, It is preferable to install water, and it is preferable to use instruments other than the sample for evaluation after sterilizing with ethanol.
次に本発明を以下の実施例によりさらに詳細に説明するが、本発明はこれに限定されるものではない。 Next, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.
＜評価用試料の作製＞
基板として、滅菌したセラミック製タイル（サイズ：２０ｃｍ×２０ｃｍ）を用いた。このセラミック製タイルの表面に、ポテトデキストロース寒天培地を厚さ１ｍｍになるよう均一に塗布した後、クリーンベンチ内で寒天培地が固まるまで放置し、寒天培地タイルを得た。
<Preparation of evaluation sample>
A sterilized ceramic tile (size: 20 cm×20 cm) was used as a substrate. A potato dextrose agar medium was evenly applied to the surface of the ceramic tile to a thickness of 1 mm, and then left in a clean bench until the agar medium was solidified to obtain an agar medium tile.
菌種として、下記に示すカビおよび菌を使用し、３種のカビ胞子液または菌懸濁液を調製した。
・例１：Ｃｌａｄｏｓｐｏｒｉｕｍ ｓｐ．（環境分離カビ）
・例２：Ｅｘｏｐｈｉａｌａ ｓｐ．（環境分離カビ）
・例３：Ｍｅｔｈｙｌｏｂａｃｔｅｒｉｕｍ ｓｐ．（環境分離菌）
The following fungi and fungi were used as bacterial species, and three types of fungal spore solution or fungal suspension were prepared.
-Example 1: Cladosporium sp. (Environmental separation mold)
-Example 2: Exophiala sp. (Environmental separation mold)
-Example 3: Methylobacterium sp. (Environmentally isolated bacteria)
各寒天培地タイルの上に、上記カビ胞子液又は菌懸濁液の各々を０．１μＬ滴下し、コーンラージ棒を用いて均一に塗り広げ、３種の評価用試料を得た。 0.1 μL of each of the above mold spore solution or bacterial suspension was dropped on each agar medium tile and spread uniformly using a cone large stick to obtain three kinds of evaluation samples.
＜洗浄用組成物の抗菌抗カビ性能の評価＞
トイレ用洗剤「モコ泡わトイレクリーナー」（エステー株式会社製）を、１００ｍＬビーカー内に噴霧し、泡状の洗剤でビーカー内を満たした。この泡状洗剤を、上記評価用試料各々の表面にコーンラージ棒を用いて均一に満遍なく塗り広げた。１分間放置した後、ＬＰ（レシチン・ポリソルベート８０）希釈液１０ｍＬで各評価用試料の表面を洗浄し、次いで滅菌水１０ｍＬで洗浄した。
なお、ブランク用として、カビ胞子懸濁の各々を塗布後の各評価用試料に、上記泡状洗剤を塗布しない試料３種を用意した。
<Evaluation of antibacterial and antifungal performance of cleaning composition>
Toilet detergent "Mokowawa Toilet Cleaner" (manufactured by ST Co., Ltd.) was sprayed into a 100 mL beaker, and the beaker was filled with a foamy detergent. This foamy detergent was uniformly and evenly spread on the surface of each of the evaluation samples by using a cone large stick. After standing for 1 minute, the surface of each evaluation sample was washed with 10 mL of LP (lecithin polysorbate 80) diluted solution, and then washed with 10 mL of sterilized water.
As blanks, three kinds of samples to which the above foamy detergent was not applied were prepared for each evaluation sample after applying each of the mold spore suspensions.
各評価用試料及び各ブランク用試料を、縦２０ｃｍ×横２８ｃｍ×高さ１０ｃｍの蓋付透明プラスチック容器にそれぞれ別々に設置し、２５℃にて４日間培養した。この際、培地の乾燥を防ぐために、滅菌水２０ｍＬを入れた５０ｍＬビーカーを上記蓋付透明プラスチック容器に入れた。培養後、各試料に形成されたコロニーを目視により観察し、ブランクとの差を確認した。結果を表１に示す。 Each evaluation sample and each blank sample were separately installed in a transparent plastic container with a lid of 20 cm in length×28 cm in width×10 cm in height and cultured at 25° C. for 4 days. At this time, in order to prevent the culture medium from drying, a 50 mL beaker containing 20 mL of sterilized water was placed in the transparent plastic container with a lid. After culturing, the colonies formed in each sample were visually observed to confirm the difference from the blank. The results are shown in Table 1.
表１の評価結果から、本発明の実施形態にかかる評価方法では、洗浄用組成物の抗菌抗カビ性能に基づく菌又はカビに由来する汚れの抑制効果を簡便に視認評価することが可能であることがわかる。 From the evaluation results in Table 1, in the evaluation method according to the embodiment of the present invention, the effect of suppressing stains derived from bacteria or mold based on the antibacterial and antifungal performance of the cleaning composition can be easily visually evaluated. I understand.
Claims (17)
基板として、表面の少なくとも一部が、前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料から構成される基板を選択すること、
前記基板上に、液状の培地と菌懸濁液又はカビ胞子液の混合物を塗布すること、
前記菌懸濁液又は前記カビ胞子液を含む前記培地を固化すること、
前記培地表面に前記洗浄用組成物を接触させること、
前記培地表面に不活化剤含有水溶液を接触させること、及び
所定時間経過後に前記培地におけるコロニーの生育状況を目視により観察すること
をこの順序で含む評価方法。 A method for evaluating antibacterial and antifungal performance of a cleaning composition,
As the substrate, selecting a substrate in which at least a part of the surface is composed of the same material as the material constituting the surface to be cleaned by the cleaning composition,
On the substrate, applying a mixture of a liquid medium and a bacterial suspension or fungal spore liquid,
Solidifying the medium containing the bacterial suspension or the mold spore fluid,
Contacting the cleaning composition with the medium surface,
An evaluation method comprising contacting an aqueous solution containing an inactivating agent with the surface of the medium and visually observing the growth state of colonies in the medium after a predetermined time has passed in this order.
基板として、表面の少なくとも一部が、前記洗浄用組成物による洗浄対象表面を構成する材料と同一材料から構成される基板を選択すること、
前記基板上に固体状の培地を形成すること、
前記培地上に菌懸濁液又はカビ胞子液を塗布すること、
前記培地表面に前記洗浄用組成物を接触させること、
前記培地表面に不活化剤含有水溶液を接触させること、及び
所定時間経過後に前記培地ににおけるコロニーの生育状況を目視により観察すること
をこの順序で含む評価方法。 A method for evaluating antibacterial and antifungal performance of a cleaning composition,
As the substrate, selecting a substrate in which at least a part of the surface is composed of the same material as the material constituting the surface to be cleaned by the cleaning composition,
Forming a solid medium on the substrate,
Applying a bacterial suspension or fungal spore solution on the medium,
Contacting the cleaning composition with the medium surface,
An evaluation method comprising contacting an inactivating agent-containing aqueous solution with the surface of the medium and visually observing the growth state of colonies on the medium after a predetermined time has passed in this order.
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