JP2016160287A - Active oxygen detection indicator - Google Patents
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- JP2016160287A JP2016160287A JP2015037534A JP2015037534A JP2016160287A JP 2016160287 A JP2016160287 A JP 2016160287A JP 2015037534 A JP2015037534 A JP 2015037534A JP 2015037534 A JP2015037534 A JP 2015037534A JP 2016160287 A JP2016160287 A JP 2016160287A
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- 229910052760 oxygen Inorganic materials 0.000 title claims abstract description 87
- 238000001514 detection method Methods 0.000 title claims abstract description 47
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Landscapes
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
Description
本発明は、気体や液体に含まれる特定の活性酸素を選択的に検知するためのインジケータに関する。 The present invention relates to an indicator for selectively detecting specific active oxygen contained in a gas or liquid.
医療、食品、バイオなどの幅広い分野において、微生物やウイルス等による汚染を防ぐために、対象物を殺菌または滅菌（以下、単に滅菌という）することが必須である。対象物を滅菌する方法として、人工的に生成させた活性酸素を対象物に暴露することにより、対象物の表面に付着していた微生物やウイルスを死滅させる方法が知られている。 In a wide range of fields such as medicine, food, and biotechnology, it is essential to sterilize or sterilize an object (hereinafter simply referred to as sterilization) in order to prevent contamination by microorganisms, viruses, and the like. As a method for sterilizing an object, a method is known in which microorganisms or viruses attached to the surface of the object are killed by exposing the object to artificially generated active oxygen.
こうした現状から、最近では、気体や液体中の活性酸素を検知・モニタリングできる技術の確立が求められている。活性酸素とは、一般的に、大気中の酸素よりも活性化され、高い反応性を有する酸素とその関連分子の総称をいい、細胞に対して酸素障害を引き起こす活性種全般を指す。医療分野などでの活性酸素を利用した滅菌方法では、活性酸素のうち、ある一定以上の反応性を有する活性酸素（例えば、ヒドロキシルラジカルを含む活性酸素（以下、「特定の活性酸素」という））が、対象物の滅菌に有効であると考えられる。 Under these circumstances, recently, establishment of a technique capable of detecting and monitoring active oxygen in gas and liquid is required. Active oxygen is generally a general term for oxygen and its related molecules that are activated and have higher reactivity than oxygen in the atmosphere, and refers to all active species that cause oxygen damage to cells. In the sterilization method using active oxygen in the medical field, etc., active oxygen having a certain reactivity or more (for example, active oxygen containing hydroxyl radical (hereinafter referred to as “specific active oxygen”)) among active oxygen. However, it is considered effective for sterilization of an object.
例えば、特許文献１には、ＱＣＭ（水晶振動子マイクロバランス）法を用いた測定装置を使用して、基板上に配置された水晶振動子の共振周波数を測定することによって、活性酸素をモニタリングすることが開示されている。また、特許文献２には、酸化還元色素の色の変化を利用した酸素インジケータが開示されている。 For example, in Patent Document 1, active oxygen is monitored by measuring a resonance frequency of a crystal resonator disposed on a substrate using a measurement device using a QCM (quartz crystal microbalance) method. It is disclosed. Patent Document 2 discloses an oxygen indicator that utilizes a change in the color of a redox dye.
しかしながら、一般的に、ＱＣＭ法を利用した測定装置は、非常に高価である。さらには、専用のリーダーを用いてセンシング部分をモニタリングしなければならないなど、測定装置の操作が非常に煩雑である。また、引用文献２に開示される酸素インジケータにおいては、反応性の低い、すなわち特定の活性酸素ではないオゾンによっても当該酸化還元色素の色の変化を生じている可能性がある。 However, in general, a measuring apparatus using the QCM method is very expensive. Furthermore, the operation of the measuring apparatus is very complicated, for example, the sensing part must be monitored using a dedicated reader. Further, in the oxygen indicator disclosed in Citation 2, there is a possibility that the color of the redox dye is changed even by ozone having low reactivity, that is, not specific active oxygen.
本発明は、以上のような実情に鑑みてなされたものであり、安価に、かつ簡便な操作で、特定の活性酸素を選択的に検知することができる、インジケータを提供することを目的とするものである。また、本発明のインジケータを有する滅菌容器を提供することも目的とするものである。本発明のインジケータ及び滅菌容器は、医療用のインジケータ及び滅菌容器として用いることができる。 The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an indicator that can selectively detect specific active oxygen at low cost and with a simple operation. Is. It is another object of the present invention to provide a sterilization container having the indicator of the present invention. The indicator and sterilization container of the present invention can be used as a medical indicator and sterilization container.
本発明の一態様によれば、特定の活性酸素を選択的に検知するインジケータであって、該インジケータは、支持体と、該支持体上に形成された検知層とを有し、該検知層は、メチレンブルーと多糖類、好ましくはプルランとを含むことを特徴とするインジケータが提供される。 According to one aspect of the present invention, an indicator that selectively detects specific active oxygen, the indicator having a support and a detection layer formed on the support, the detection layer Provides an indicator characterized in that it comprises methylene blue and a polysaccharide, preferably pullulan.
また、本発明のインジケータを滅菌容器と共に使用することで、特定の活性酸素で対象物を滅菌するための滅菌容器を提供することが可能となる。 Moreover, it becomes possible to provide the sterilization container for sterilizing a target object with specific active oxygen by using the indicator of this invention with a sterilization container.
以下に、本発明のインジケータ及び滅菌容器の実施形態について詳細に説明する。なお、本発明は以下の実施形態に限定されるものではない。 Hereinafter, embodiments of the indicator and the sterilization container of the present invention will be described in detail. In addition, this invention is not limited to the following embodiment.
（１）インジケータ
本発明のインジケータは、気体や液体中の一定以上の反応性を有する特定の活性酸素を、選択的に検知することができる。すなわち、活性酸素のなかでも反応性の低いオゾンには反応せずに、反応性の高いヒドロキシルラジカルなどの特定の活性酸素を選択的に検知することができる。言い換えれば、本発明のインジケータが選択的に検知する特定の活性酸素とは、代表的には、ヒドロキシルラジカル、ヒドロペルオキシド、過酸化水素、スーパーオキシド、一重項酸素等を含み、少なくともオゾンを含まないものである。
(1) Indicator The indicator of the present invention can selectively detect specific active oxygen having a certain level of reactivity in gas or liquid. That is, specific active oxygen such as highly reactive hydroxyl radical can be selectively detected without reacting with ozone having low reactivity among active oxygen. In other words, the specific active oxygen that is selectively detected by the indicator of the present invention typically includes hydroxyl radical, hydroperoxide, hydrogen peroxide, superoxide, singlet oxygen, etc., and at least does not contain ozone. Is.
前記インジケータの検知可能な特定の活性酸素の濃度域は、数十％の高濃度からｐｐｍオーダーの極低濃度まで、非常に広い。例えば、本発明のインジケータを用いることにより、空気中等に含まれる特定の活性酸素を選択的に検知することが可能である。 The specific active oxygen concentration range detectable by the indicator is very wide, from a high concentration of several tens of percent to a very low concentration on the order of ppm. For example, by using the indicator of the present invention, it is possible to selectively detect specific active oxygen contained in air or the like.
本発明のインジケータは、少なくとも、支持体と、支持体上に形成された検知層とを有する。インジケータを構成する検知層は、メチレンブルーと水溶性高分子とを含む。 The indicator of the present invention has at least a support and a detection layer formed on the support. The detection layer constituting the indicator contains methylene blue and a water-soluble polymer.
前記検知層を構成するメチレンブルーと水溶性高分子との組成比は特には限定されないが、活性酸素の検知判断時に行われる目視観察などの分析方法が容易である組成比が好ましい。 The composition ratio of methylene blue and water-soluble polymer constituting the detection layer is not particularly limited, but a composition ratio that facilitates an analysis method such as visual observation performed at the time of detection of active oxygen is preferable.
前記水溶性高分子としては、メチレンブルー水溶液を包接し、特定の活性酸素が透過することができる薄膜を形成できる水溶性高分子であればいかなる水溶性高分子を用いることができるが、具体的には多糖類が好ましい。前記多糖類としては、寒天（アガロース）、ペクチン、グアーガム、カラギーナン、キサンタンガム、タマリンドガム、プルラン、ローカストビーンガム、タラガム、デンプン、デキストリン、デキストラン、グルコマンナン等及びこれらの誘導体を挙げることができ、これらを単独で、または複数を組み合わせて用いてもよい。 As the water-soluble polymer, any water-soluble polymer can be used as long as it is a water-soluble polymer that can form a thin film that can include a methylene blue aqueous solution and allow specific active oxygen to permeate. Is preferably a polysaccharide. Examples of the polysaccharide include agar (agarose), pectin, guar gum, carrageenan, xanthan gum, tamarind gum, pullulan, locust bean gum, tara gum, starch, dextrin, dextran, glucomannan, and derivatives thereof. May be used alone or in combination.
前記多糖類のうち、好ましくは、下記式（１）で表される構造を有するプルラン（ｐｕｌｌｕｌａｎ）が用いられる。 Of the polysaccharides, preferably, pullulan having a structure represented by the following formula (1) is used.
インジケータによる活性酸素の検知の有無の判断には、検知層を構成しているメチレンブルーの色の変化を利用している。特定の活性酸素を検知する前の検知層は、メチレンブルーによる青色を呈している。ここで、インジケータを構成している検知層の表面に特定の活性酸素が吸着すると、メチレンブルーと特定の活性酸素とが反応し、メチレンブルーは分解する。すなわち、検知層の青色が薄くなり、最終的に、検知層は透明になる。メチレンブルーと特定の活性酸素との反応によりメチレンブルーが分解することから、検知層はメチレンブルーと特定の活性酸素との反応量に応じた色消失を示す。このように、本発明のインジケータによる特定の活性酸素の検知の有無は、検知層の色の変化に基づいて判断する。 For the determination of whether or not active oxygen is detected by the indicator, the change in the color of methylene blue constituting the detection layer is used. The detection layer before detecting the specific active oxygen is blue with methylene blue. Here, when specific active oxygen is adsorbed on the surface of the detection layer constituting the indicator, methylene blue reacts with the specific active oxygen, and methylene blue is decomposed. That is, the blue color of the detection layer becomes lighter and finally the detection layer becomes transparent. Since methylene blue is decomposed by the reaction between methylene blue and specific active oxygen, the detection layer shows a color disappearance according to the reaction amount between methylene blue and specific active oxygen. Thus, the presence or absence of the detection of the specific active oxygen by the indicator of this invention is judged based on the change of the color of a detection layer.
本来、下記式（２）で表されるメチレンブルーは、オゾンを含む活性酸素種全般に対して、反応性を有する。一方、本発明のインジケータは、活性酸素のなかでも、オゾンをほとんど検知せずに、反応性の高いヒドロキシルラジカル（ＯＨ＊）などの特定の活性酸素を選択的に検知する。なお、本発明のインジケータが特定の活性酸素を選択的に検知する理由について、完全には明らかになっていないが、本発明者らは以下のように考えている。 Originally, methylene blue represented by the following formula (2) has reactivity with all active oxygen species including ozone. On the other hand, the indicator of the present invention selectively detects specific active oxygen such as highly reactive hydroxyl radical (OH * ) without detecting ozone in active oxygen. Although the reason why the indicator of the present invention selectively detects specific active oxygen is not completely clarified, the present inventors consider as follows.
本発明のインジケータを構成する検知層は、メチレンブルーと多糖類とを含む。検知層内でのメチレンブルーと多糖類との存在状態は、多糖類とメチレンブルーとが反応して新たな化合物が生成されるようなものではなく、おそらくは、メチレンブルーの水溶液が多糖類の構成する高分子ポリマーの網目構造中に存在する、すなわち前記網目構造中に前記水溶液が包接されている状態にあると考えられる。このとき、検知層はメチレンブルーの青色を呈する。 The detection layer constituting the indicator of the present invention contains methylene blue and a polysaccharide. The presence state of methylene blue and polysaccharide in the detection layer is not such that the polysaccharide and methylene blue react to produce a new compound. Perhaps a polymer in which an aqueous solution of methylene blue constitutes the polysaccharide. It is considered that the polymer exists in the polymer network structure, that is, the aqueous solution is included in the network structure. At this time, the detection layer exhibits a blue color of methylene blue.
そして、特定の活性酸素は、前記多糖類の構成する高分子ポリマーの網目を通過し、前記メチレンブルー水溶液中のメチレンブルーを、直接または間接的に分解する。その結果、インジケータの検知層の色が青色から透明に変化する。ここで、間接的なメチレンブルーの分解とは、スーパーオキシドや一重項酸素がメチレンブルーを直接分解せず、例えば、水溶液中の水と反応することで生じたヒドロキシルラジカル等の別の化学種がメチレンブルーを分解することである。メチレンブルーの直接的分解及び間接的分解は、それぞれ別々にまたは同時に進行しても良い。
一方、オゾンは水への溶解度が０．５７ｇ／Ｌ（２０℃）であり、酸素よりも水へ溶解しやすい。また、水へ溶解したオゾンは、塩基性水溶液中であれば速やかに分解され、ヒドロキシルラジカルを生じることが知られている。しかし、本発明のインジケータ中のメチレンブルー水溶液は酸性であるため、メチレンブルー水溶液中でのオゾンの分解によるヒドロキシルラジカルの生成は非常に遅いと考えられる。そのため、生成したヒドロキシルラジカルは、メチレンブルーと反応するよりも早く、何らかの形で還元される（例えば、メチレンブルー水溶液を包接する高分子ポリマー等と反応する）ために、メチレンブルーが分解されず、インジケータの検知層の色が変化しないと推測している。
And specific active oxygen passes the network of the high molecular polymer which the said polysaccharide comprises, and decomposes | disassembles the methylene blue in the said methylene blue aqueous solution directly or indirectly. As a result, the color of the detection layer of the indicator changes from blue to transparent. Here, indirect decomposition of methylene blue means that superoxide or singlet oxygen does not decompose methylene blue directly, but another chemical species such as a hydroxyl radical generated by reacting with water in an aqueous solution does not decompose methylene blue. It is to decompose. The direct and indirect degradation of methylene blue may proceed separately or simultaneously.
On the other hand, ozone has a solubility in water of 0.57 g / L (20 ° C.) and is more soluble in water than oxygen. Further, it is known that ozone dissolved in water is quickly decomposed in a basic aqueous solution to generate hydroxyl radicals. However, since the methylene blue aqueous solution in the indicator of the present invention is acidic, the generation of hydroxyl radicals due to the decomposition of ozone in the methylene blue aqueous solution is considered to be very slow. Therefore, the generated hydroxyl radical is reduced in some form before it reacts with methylene blue (for example, it reacts with a polymer polymer that includes a methylene blue aqueous solution). I guess that the color of the layer does not change.
検知層の色の変化、つまり、特定の活性酸素の検知の有無の判断方法は、特には限定されない。例えば、目視観察や光学的な分析装置を用いて判断することができる。また、特定の活性酸素を検知する前後のインジケータの検知層に含まれるメチレンブルーを所定の溶液に抽出させて得られる抽出溶液について、同様に、目視観察や光学的な分析装置を用いて判断してもよい。検知層からメチレンブルーを抽出させるときに用いられる溶液は、インジケータによる特定の活性酸素の検知の有無の判断に影響を与えなければ限定されず、例えば、エタノールが挙げられる。また、メチレンブルーの抽出方法についても、特には限定されない。一例として、所定の溶液に浸漬させたインジケータに対して、超音波処理等の所定のプロセスを施すことにより、メチレンブルーの抽出を行うことができる。なお、あらかじめインジケータから剥離させた検知層からメチレンブルーを抽出してもよい。 The method for determining the color change of the detection layer, that is, whether or not a specific active oxygen is detected is not particularly limited. For example, it can be judged using visual observation or an optical analyzer. In addition, the extraction solution obtained by extracting methylene blue contained in the detection layer of the indicator before and after detecting specific active oxygen into a predetermined solution is similarly judged using visual observation or an optical analyzer. Also good. The solution used when extracting methylene blue from the detection layer is not limited as long as it does not affect the determination of whether or not specific active oxygen is detected by the indicator, and includes, for example, ethanol. Also, the extraction method of methylene blue is not particularly limited. As an example, methylene blue can be extracted by subjecting an indicator immersed in a predetermined solution to a predetermined process such as ultrasonic treatment. In addition, you may extract methylene blue from the detection layer peeled from the indicator beforehand.
なお、本発明の検知層には、必要に応じて、インジケータの検知特性が劣化されない範囲において、各種添加物が含まれてもよい。各種添加物は、例えば、検知層の成形性や、検知感度の向上、耐久性の向上等の目的として、検知層に含まれる。 In addition, various additives may be contained in the detection layer of this invention in the range which the detection characteristic of an indicator does not deteriorate as needed. Various additives are contained in the detection layer, for example, for the purpose of improving the formability of the detection layer, improving the detection sensitivity, and improving the durability.
本発明における支持体に用いられる材料としては、特定の活性酸素に対して耐久性があり、検知層を担持することができればよく、リジッドな材料でも、フレキシブルな材料でもよい。 The material used for the support in the present invention may be a rigid material or a flexible material as long as it is durable to a specific active oxygen and can carry the detection layer.
本発明のインジケータの製造方法は、支持体上にメチレンブルーと多糖類からなる検知層を形成し、該検知層がメチレンブルー水溶液を包接するようなものであれば、特には限定されない。例えば、メチレンブルーとプルラン等の多糖類との水溶液を支持体上に塗布し、乾燥することにより薄膜の形成を行い、インジケータを製造することができる。 The method for producing an indicator of the present invention is not particularly limited as long as a detection layer made of methylene blue and a polysaccharide is formed on a support and the detection layer encloses an aqueous solution of methylene blue. For example, an indicator can be manufactured by forming a thin film by applying an aqueous solution of methylene blue and a polysaccharide such as pullulan onto a support and drying.
上記薄膜の形成方法は、特には限定されず、目的に応じて適宜選択される。例えば、支持体上の所定の箇所に検知層を形成させたい場合や微小領域に検知層を形成させたい場合には、前記水溶液を支持体上に滴下する方法が選択される。また、大面積の領域に検知層を形成させたい場合には、ディップコーティングやスピンコーティング等が選択される。 The method for forming the thin film is not particularly limited, and is appropriately selected depending on the purpose. For example, when the detection layer is to be formed at a predetermined location on the support or when the detection layer is to be formed in a minute region, a method of dropping the aqueous solution onto the support is selected. Further, when it is desired to form the detection layer in a large area, dip coating or spin coating is selected.
また、薄膜の形成時の乾燥処理については、検知層を構成する各種物質を分解せずに、塗膜を乾燥できる温度であれば、特には限定されない。 Further, the drying process at the time of forming the thin film is not particularly limited as long as it is a temperature at which the coating film can be dried without decomposing various substances constituting the detection layer.
本発明のインジケータは、気体や液体中の特定の活性酸素を、選択的に検知することができる。すなわち、活性酸素のなかでも、反応性の低いオゾンには反応せずに、反応性の高いヒドロキシルラジカル等の特定の活性酸素を選択的に検知することができる。このため、本発明のインジケータは、オゾンとヒドロキシルラジカルとを識別することができる。したがって、本発明のインジケータは、活性酸素を用いて対象物の表面に付着しているウイルス等を死滅させる場合、ヒドロキシルラジカルのような反応性の高い特定の活性酸素を選択的に検知する医療用インジケータとして利用することができる。 The indicator of the present invention can selectively detect specific active oxygen in gas or liquid. That is, among active oxygens, specific active oxygens such as hydroxyl radicals having high reactivity can be selectively detected without reacting with ozone having low reactivity. For this reason, the indicator of the present invention can distinguish ozone from hydroxyl radicals. Therefore, the indicator of the present invention is for medical use that selectively detects a specific reactive oxygen having high reactivity such as a hydroxyl radical when the active oxygen is used to kill a virus or the like attached to the surface of an object. It can be used as an indicator.
また、本発明のインジケータの利用は、上記医療用インジケータとしての利用だけに限られない。例えば本発明のインジケータは、これらに限られるものではないが、集積回路作成の光アッシングによるフォトレジスト除去におけるインジケータや、エンジニアプラスチック等の高分子材料、アルミ、ステンレス等の活性酸素を用いた表面改質におけるインジケータ等の工業用インジケータとしても利用することができる。 The use of the indicator of the present invention is not limited to the use as the medical indicator. For example, the indicator of the present invention is not limited to these, but is an indicator for removing a photoresist by optical ashing for creating an integrated circuit, a polymer material such as engineer plastic, and surface modification using active oxygen such as aluminum or stainless steel. It can also be used as industrial indicators such as quality indicators.
（２）滅菌容器
本発明の別の態様としての滅菌容器は、少なくとも、活性酸素を用いて滅菌させる対象物と、上記のインジケータとを内部に有する。そして、滅菌容器の内部に活性酸素を通気、または滅菌容器の内部で活性酸素を発生して、対象物を活性酸素で滅菌する。このとき、対象物を滅菌する活性酸素が本発明のインジケータで検知可能な特定の活性酸素である場合、滅菌効果の向上、滅菌処理時間の短縮が期待できる。
(2) Sterilization container The sterilization container as another aspect of the present invention has at least an object to be sterilized using active oxygen and the above indicator inside. Then, active oxygen is ventilated inside the sterilization container, or active oxygen is generated inside the sterilization container, and the object is sterilized with active oxygen. At this time, when the active oxygen for sterilizing the object is a specific active oxygen that can be detected by the indicator of the present invention, an improvement in sterilization effect and a reduction in sterilization time can be expected.
本発明の滅菌容器を用いた滅菌処理方法は、特には限定されず、目的に応じて適宜選択される。例えば、本発明のインジケータと対象物とを内部に備える本発明の滅菌容器の少なくとも一部を波長１８５ｎｍ及び波長２５４ｎｍの紫外線を透過する部材から構成し、当該部材から波長１８５ｎｍ及び波長２５４ｎｍの紫外線を照射して滅菌容器の内部に特定の活性酸素、例えばヒドロキシルラジカルを発生し、発生した特定の活性酸素によって対象物を滅菌処理する。このような条件で発生する特定の活性酸素は本発明のインジケータで検知可能であることから、本発明のインジケータを目視で観察するだけで、対象物の滅菌状況が簡易的に瞬時に判断することができることから、本発明の滅菌容器は医療用の滅菌容器としても利用することができる。 The sterilization method using the sterilization container of the present invention is not particularly limited, and is appropriately selected according to the purpose. For example, at least a part of the sterilization container of the present invention having the indicator of the present invention and the object inside is constituted by a member that transmits ultraviolet light having a wavelength of 185 nm and wavelength of 254 nm, and ultraviolet light having a wavelength of 185 nm and wavelength of 254 nm is emitted from the member. Irradiation generates specific active oxygen such as hydroxyl radicals in the sterilization container, and the target is sterilized by the generated specific active oxygen. Since the specific active oxygen generated under such conditions can be detected by the indicator of the present invention, the sterilization status of the object can be easily and instantaneously determined simply by visually observing the indicator of the present invention. Therefore, the sterilization container of the present invention can also be used as a medical sterilization container.
以下に、具体的な実施例を挙げて本発明をさらに詳細に説明する。下記の実施例は、本発明の最良な実施形態の一例であるものの、本発明はこれら実施例により何ら限定されるものではない。 Hereinafter, the present invention will be described in more detail with reference to specific examples. The following examples are examples of the best mode of the present invention, but the present invention is not limited to these examples.
［実施例１］
本実施例に使用される支持体として、ポリスチレン製シャーレを用意した。次に、濃度１００ｇ／Ｌのプルラン水溶液（７００μＬ）及び濃度０．１２ｇ／Ｌのメチレンブルー水溶液（４００μＬ）を純水１３００μＬに添加して、混合水溶液を調製した。前記シャーレに前記混合水溶液を一定量流し込み、乾燥機で４０℃、１２時間以上乾燥させることにより、本発明のインジケータを製造した。
［比較例１及び２］
上記実施例１に示した本発明のインジケータの製造法と同様に、濃度２０ｇ／Ｌのアルギン酸ナトリウム水溶液（５００μＬ）及び濃度０．１２ｇ／Ｌのメチレンブルー水溶液（４００μＬ）を純水１５００μＬに添加して調整した混合水溶液より得た検知層を有するインジケータを比較例１とし、濃度９９ｇ／Ｌのアルギン酸ナトリウム水溶液（５００μＬ）及び濃度０．１２ｇ／Ｌのメチレンブルー水溶液（４００μＬ）を純水１５００μＬに添加して調整した混合水溶液より得た検知層を有するインジケータを比較例２とした。
[Example 1]
A polystyrene petri dish was prepared as a support used in this example. Next, a 100 g / L aqueous pullulan solution (700 μL) and a 0.12 g / L methylene blue aqueous solution (400 μL) were added to 1300 μL of pure water to prepare a mixed aqueous solution. The indicator of the present invention was manufactured by pouring a certain amount of the mixed aqueous solution into the petri dish and drying it at 40 ° C. for 12 hours or more with a dryer.
[Comparative Examples 1 and 2]
Similar to the method for producing the indicator of the present invention shown in Example 1 above, a sodium alginate aqueous solution (500 μL) having a concentration of 20 g / L and a methylene blue aqueous solution (400 μL) having a concentration of 0.12 g / L were added to 1500 μL of pure water. An indicator having a detection layer obtained from the prepared mixed aqueous solution is set as Comparative Example 1, and a 99 g / L sodium alginate aqueous solution (500 μL) and a 0.12 g / L methylene blue aqueous solution (400 μL) are added to 1500 μL pure water. An indicator having a detection layer obtained from the prepared mixed aqueous solution was set as Comparative Example 2.
表１に上記インジケータの各物性値を示す。各インジケータの最大吸収波長及び該最大波長における透過率の測定は、分光光度計（ＵＶ−２４５０、株式会社島津製作所製）により、波長範囲３００ｎｍ〜８００ｎｍ、スリット幅２．０ｎｍで行った。また、検知層の膜厚の変動係数（ＣＶ値）は、前記検知層の平均膜厚を標準偏差で割った値であり、その値が小さいほど検知層の膜厚が均一に近いことを示す。 Table 1 shows the physical property values of the indicators. The maximum absorption wavelength of each indicator and the transmittance at the maximum wavelength were measured with a spectrophotometer (UV-2450, manufactured by Shimadzu Corporation) in a wavelength range of 300 nm to 800 nm and a slit width of 2.0 nm. The variation coefficient (CV value) of the thickness of the detection layer is a value obtained by dividing the average thickness of the detection layer by the standard deviation, and the smaller the value, the closer the thickness of the detection layer is. .
上記で製造したインジケータの検知特性を、以下に示す条件で測定した。すなわち、紫外線（ＵＶ）ランプ（波長１８５／２５４ｎｍ）による紫外線を照射させたときに発生する活性酸素に対する検知特性を調査した。 The detection characteristics of the indicator manufactured above were measured under the following conditions. That is, the detection characteristic for active oxygen generated when ultraviolet rays were irradiated by an ultraviolet ray (UV) lamp (wavelength 185/254 nm) was investigated.
紫外線により発生する活性酸素を利用した洗浄器であるアクティブドライ（登録商標、医療用／器具除染用洗浄器、紫外線ランプ（波長１８５／２５４ｎｍ）、岩崎電気製）内に、上記で製造したインジケータを設置した。図１（ａ）に示すように、３０℃の水でバブリングした気体をアクティブドライの内部に通気させて、アクティブドライ内を高湿度（湿度９０％ＲＨ以上）に制御した。これに対して、図１（ｂ）に示すように、３０℃の湯浴で温められたシリカゲルの詰まった瓶中に気体を通気させて、前記気体をアクティブドライ内に通じることで、アクティブドライ内を低い湿度（湿度２０％ＲＨ以下）に制御した。その後、前記各環境下で、紫外線ランプを点灯することにより、アクティブドライ内においてインジケータに活性酸素を１２０分間暴露した。この際、各インジケータに紫外線が直接照射されないように、各インジケータは不織布で覆われた状態で活性酸素に曝された。 In the active dry (registered trademark, medical / instrument decontamination washer, ultraviolet lamp (wavelength 185/254 nm), manufactured by Iwasaki Electric Co., Ltd.) Was installed. As shown in FIG. 1A, a gas bubbled with 30 ° C. water was passed through the inside of the active dry to control the inside of the active dry to a high humidity (humidity of 90% RH or more). On the other hand, as shown in FIG. 1B, a gas is passed through a bottle filled with silica gel heated in a hot water bath at 30 ° C. The inside was controlled to a low humidity (humidity of 20% RH or less). After that, the active oxygen was exposed to the indicator for 120 minutes in the active dry by turning on the ultraviolet lamp under each environment. At this time, each indicator was exposed to active oxygen in a state of being covered with a nonwoven fabric so that each indicator was not directly irradiated with ultraviolet rays.
活性酸素に曝された各インジケータの透過率を図２に示す。各インジケータの透過率の測定は、分光光度計（ＵＶ−２４５０、株式会社島津製作所製）により、波長範囲３００ｎｍ〜８００ｎｍ、スリット幅２．０ｎｍで行った。 The transmittance of each indicator exposed to active oxygen is shown in FIG. The transmittance of each indicator was measured by a spectrophotometer (UV-2450, manufactured by Shimadzu Corporation) in a wavelength range of 300 nm to 800 nm and a slit width of 2.0 nm.
その結果、図２（ａ）に示すように、実施例１のインジケータは高湿度環境下において、６７０ｎｍの波長光の透過率がおよそ３０％向上し、インジケータ中のメチレンブルーの青色が脱色されたことが明らかとなった。これに対して、実施例１のインジケータは低湿度環境下ではほとんど脱色されなかった。一方、図２（ｂ）に示すように、比較例１のインジケータは高湿度環境下及び低湿度環境下において、５９５ｎｍの波長光の透過率が若干向上した。また、図２（ｃ）に示すように、比較例２のインジケータは高湿度環境下及び低湿度環境下においてほとんど透過率の変化は観測されなかった。 As a result, as shown in FIG. 2A, in the indicator of Example 1, the transmittance of 670 nm wavelength light was improved by about 30% in a high humidity environment, and the blue color of methylene blue in the indicator was decolorized. Became clear. In contrast, the indicator of Example 1 was hardly decolorized in a low humidity environment. On the other hand, as shown in FIG. 2 (b), the indicator of Comparative Example 1 slightly improved the transmittance of 595 nm wavelength light in a high humidity environment and a low humidity environment. Moreover, as shown in FIG.2 (c), the indicator of the comparative example 2 hardly observed the change of the transmittance | permeability in the high humidity environment and the low humidity environment.
発明者らは、低湿度条件下において紫外線照射により活性酸素を発生させた場合、発生する活性酸素種の大部分がオゾンでありヒドロキシルラジカル等の特定の活性酸素がほとんど発生しないのに対して、高湿度条件下において同様に活性酸素を発生させた場合では、ヒドロキシルラジカル等の特定の活性酸素が低湿度条件下よりも高い濃度で発生することを見出している（Oya, K., Watanabe, R., Soga, Y., Ikeda, Y., Nakamura, T., and Iwamori, S. “Effect of humidity conditions on active oxygen species generated under ultraviolet light irradiation and etching characteristics of fluorocarbon polymer” Journal of Photochemistry and Photobiology A: Chemistry 298 (2014) 3339）。そのため、上記実施例１の結果より、本発明のインジケータは、オゾンをほとんど検知せず、ヒドロキシルラジカル等の特定の活性酸素のみを選択的に検知することが示唆された。 When the inventors generate active oxygen by irradiation with ultraviolet rays under low humidity conditions, most of the generated active oxygen species are ozone, while specific active oxygen such as hydroxyl radicals are hardly generated. When active oxygen is similarly generated under high humidity conditions, it has been found that specific active oxygen such as hydroxyl radicals are generated at higher concentrations than under low humidity conditions (Oya, K., Watanabe, R ., Soga, Y., Ikeda, Y., Nakamura, T., and Iwamori, S. “Effect of humidity conditions on active oxygen species generated under ultraviolet light irradiation and etching characteristics of fluorocarbon polymer” Journal of Photochemistry and Photobiology A: Chemistry 298 (2014) 3339). Therefore, from the result of Example 1 above, it was suggested that the indicator of the present invention hardly detects ozone but selectively detects only specific active oxygen such as hydroxyl radical.
［実施例２］
実施例１に示した製造法と同様に、濃度１００ｇ／Ｌのプルラン水溶液（５２５μＬ）及び濃度０．１２ｇ／Ｌのメチレンブルー水溶液（６００μＬ）を純水１４７５μＬに添加して調整した混合水溶液より得た検知層を有するインジケータを実施例２のインジケータとした。
[Example 2]
Similar to the production method shown in Example 1, a pullulan aqueous solution (525 μL) having a concentration of 100 g / L and a methylene blue aqueous solution (600 μL) having a concentration of 0.12 g / L were added to 1475 μL of pure water to obtain a mixed aqueous solution prepared. The indicator having the detection layer was used as the indicator of Example 2.
実施例２のインジケータに対して、高湿度環境下（湿度９０％ＲＨ以上）において活性酸素に暴露した。１０分毎に実施例２のインジケータの６７０ｎｍの波長光での脱色率を図３に示した。前記脱色率は、メチレンブルーを有さないプルランのみの薄膜の６７０ｎｍの波長光の透過率を１００として、該インジケータの透過率を差し引いた値で示した。 The indicator of Example 2 was exposed to active oxygen in a high humidity environment (humidity of 90% RH or higher). The decolorization rate of the indicator of Example 2 with a wavelength of 670 nm every 10 minutes is shown in FIG. The decolorization rate was expressed as a value obtained by subtracting the transmittance of the indicator, assuming that the transmittance of light having a wavelength of 670 nm of a pullulan-only thin film having no methylene blue was 100.
図３の結果より、実施例２のインジケータに対して、特定の活性酸素の暴露時間が１０分〜３０分の間では効果がほとんど表れておらず、暴露時間が５０分〜６０分の間に大きく脱色していることが分かった。 From the result of FIG. 3, the effect of the specific active oxygen exposure time between 10 minutes and 30 minutes is hardly exhibited with respect to the indicator of Example 2, and the exposure time is between 50 minutes and 60 minutes. It turns out that it is greatly decolored.
［実施例３］
実施例１に記載の方法で製造したインジケータに対して、酸素ボンベから酸素を供給したオゾナイザー（ＹＲＧ−２０３Ｎ，レイシー社製)からオゾンを発生させ、３０分間暴露した。この間のオゾン濃度は１５０〜１８０ｐｐｍ程度である。その後、該インジケータの透過率を、実施例１と同様に測定した。
[Example 3]
Ozone was generated from an ozonizer (YRG-203N, manufactured by Lacey Co., Ltd.) supplied with oxygen from an oxygen cylinder and exposed to the indicator produced by the method described in Example 1 for 30 minutes. The ozone concentration during this period is about 150 to 180 ppm. Thereafter, the transmittance of the indicator was measured in the same manner as in Example 1.
上記透過率の測定結果を図４に示す。その結果、本発明のインジケータはオゾンではほとんど脱色されないことが分かった。 The measurement result of the transmittance is shown in FIG. As a result, it was found that the indicator of the present invention is hardly decolorized with ozone.
以上から、本発明のインジケータは、特定の活性酸素を選択的に検知することを見出した。特定の活性酸素とは、高湿度環境（湿度９０％ＲＨ以上）で紫外線ランプ（波長１８５／２５４ｎｍ）による紫外線を照射させたときに発生する一定以上の反応性を有する、オゾンではない活性酸素である。すなわち、本発明のインジケータは、オゾンとそれ以外の活性酸素とを識別することができることから、特定の活性酸素を選択的に検知する色素センサとして応用できることが示唆された。 From the above, it has been found that the indicator of the present invention selectively detects specific active oxygen. The specific active oxygen is an active oxygen that is not ozone and has a reactivity higher than a certain level that is generated when an ultraviolet lamp (wavelength 185/254 nm) is irradiated with ultraviolet rays in a high humidity environment (humidity of 90% RH or more). is there. That is, since the indicator of the present invention can distinguish between ozone and other active oxygen, it was suggested that it can be applied as a dye sensor that selectively detects specific active oxygen.
Claims (7)
該インジケータは、支持体と、該支持体上に形成された検知層とを有し、
該検知層は、メチレンブルー及び多糖類からなることを特徴とするインジケータ。 An indicator that selectively detects specific active oxygen,
The indicator has a support and a sensing layer formed on the support;
The detection layer is composed of methylene blue and a polysaccharide.
該滅菌容器の内部に、請求項１〜５のいずれか１項に記載のインジケータを有することを特徴とする滅菌容器。 A sterilization container for sterilizing an object with specific active oxygen,
A sterilization container comprising the indicator according to any one of claims 1 to 5 inside the sterilization container.
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