JP6708715B2 - 次亜塩素酸を含む抗微生物剤 - Google Patents
次亜塩素酸を含む抗微生物剤 Download PDFInfo
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- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 title claims description 39
- 239000004599 antimicrobial Substances 0.000 title claims 2
- 239000007864 aqueous solution Substances 0.000 claims description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 33
- 230000000844 anti-bacterial effect Effects 0.000 claims description 24
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 24
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 23
- 241000894006 Bacteria Species 0.000 claims description 15
- 239000008213 purified water Substances 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 7
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 6
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 6
- 241000588724 Escherichia coli Species 0.000 claims description 6
- 239000000460 chlorine Substances 0.000 claims description 6
- 229910052801 chlorine Inorganic materials 0.000 claims description 6
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims description 5
- 239000003242 anti bacterial agent Substances 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 claims description 3
- 239000002778 food additive Substances 0.000 claims description 3
- 235000013373 food additive Nutrition 0.000 claims description 3
- 239000000243 solution Substances 0.000 description 14
- 239000002609 medium Substances 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 9
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 5
- 235000019345 sodium thiosulphate Nutrition 0.000 description 5
- 238000010998 test method Methods 0.000 description 4
- 241000193755 Bacillus cereus Species 0.000 description 3
- 241000194108 Bacillus licheniformis Species 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 241000607142 Salmonella Species 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000001965 potato dextrose agar Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000001974 tryptic soy broth Substances 0.000 description 3
- 108010050327 trypticase-soy broth Proteins 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 description 2
- 230000007794 irritation Effects 0.000 description 2
- 229940127554 medical product Drugs 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 206010029803 Nosocomial infection Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002070 germicidal effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002350 laparotomy Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003330 sporicidal effect Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
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Description
次亜塩素酸ナトリウムを、0.018〜0.026W/V%、好ましくは0.026W/V%となるように秤量し、精製水と混合希釈する。混合希釈により生成した次亜塩素酸ナトリウム水溶液を、希塩酸(約9.5〜10.5W/V%)でpH6.0〜6.5となるように調整し、攪拌混合する。これは、次亜塩素酸ナトリウムを、有効塩素濃度で150〜260ppm、好ましくは220ppm含むようにしてもよい。例えば、得られる混合液の全量を100%とした場合、精製水を99.9%以上とし、残りの0.1%未満を、ほぼ同量の次亜塩素酸ナトリウム水溶液(有効塩素12%のもの)と希塩酸水溶液(濃度約10%のもの)とすることで、得ることができる。
以下に次亜塩素酸水溶液の成分分析表を示す。
(1)性状
無色透明の液体で、においはない。
(2)純度
有機体炭素試験を行うとき、0.50mg/L以下である。
(3)導電性
次の方法により試験を行うとき、導電率(25℃)は、2.1μS/cm以下である。
精製水の適当量をビーカーにとり、かき混ぜる。温度を25±1℃に調節し、強くかき混ぜながら、一定時間ごとにこの液の導電率の測定を行う。5分当たりの導電率変化が0.1μS/cm以下となったときの導電率を精製水の導電率(25℃)とする。
(1)試験方法
1)供試菌
次亜塩素酸水溶液の殺菌効果の確認を以下の供試菌を用いて行った。
Escherichia coli(大腸菌)
Salmonella Enteritidis(サルモネラ)
Candida sp.(カンジダ)
Pseudomonas aeruginosa(緑膿菌)
次亜塩素酸水溶液を、それぞれ有効塩素濃度が、200、20、5、2、1、0.5ppmとなるように希釈し、試験の試料とした。これらの試料を、それぞれ20ml容試験管に5ml分注した。また、次亜塩素酸水溶液を含まない滅菌純水を対照とした。
大腸菌及びサルモネラについては、供試菌をTSB(Tryptic Soy Broth)で35℃、20〜24時間静置培養し、試験に供した。供試した菌液濃度は、培養液を滅菌純水で希釈して調製した。菌数は、大腸菌が1.2×106/ml、サルモネラが1.7×106/mlであった。
緑膿菌については、TSBで25℃、44〜48時間静置培養し、培養液を滅菌純水で希釈して調製した。菌数は2.3×106/mlであった。
各濃度の試料に菌液を0.2ml接種し、混合した。0.5分、5分、及び、10分経過後のそれぞれにおいて、各試料から0.2mlを取り出し、1.8mlの1mg/mlチオ硫酸ナトリウム含有滅菌純水に懸濁した。この懸濁液と、さらに1mg/mlチオ硫酸ナトリウム含有滅菌純水を用いて10倍希釈した液とを、細菌の場合はSA培地に、酵母の場合はPDA培地に0.1ml塗抹した。Controlについては、チオ硫酸ナトリウム含有滅菌純水の替わりに滅菌純水を用いた。培地を培養した後、平板上に出現したコロニーを計測した。
試料の濃度別に、出現コロニー数を処理時間毎に生残菌数を計測し、殺菌効果を判定した。
以下の表2〜表5、及び図1〜4に示すように、本試験に使用したすべての微生物は、次亜塩素酸水溶液の有効塩素濃度5ppmにおいて、0.5分間作用させることにより死滅した。図1〜4では、0.5ppmで処理した場合を一点鎖線、1ppmで処理した場合を破線、2ppmで処理した場合を二点鎖線で、それぞれ示した。
(1)試験に使用した試料
次亜塩素酸水溶液及び次亜塩素酸ナトリウム水を以下の表6のように調製した。
1)供試菌
本発明の一実施形態に係る次亜塩素酸水溶液と次亜塩素酸ナトリウム水との殺菌効果の比較試験を以下の供試菌を用いて行った。
Bacillus subtilis NBRC 13719
Bacillus cereus NBRC 13494
Bacillus licheniformis NBRC 12200
供試した3種のBacillus属の菌体を保存スラント上からそれぞれ釣菌し、滅菌純水にそれぞれ懸濁したあと80℃で15分間加熱した。各懸濁液をNA(普通寒天)平板培地上に塗抹して35℃で3日間培養してコロニーを形成させた。芽胞の多いコロニーを選んで釣菌し、滅菌純水に懸濁したあと80℃で15分間加熱した。懸濁液をNA培地上に塗抹して35℃で4〜6日間培養した。生育したコロニーから菌体を釣菌して滅菌純水に懸濁し、80℃で15分間加熱した菌液を供試芽胞菌液とした。3株の供試芽胞菌液の濃度は、1〜4×106/mlであった。
各濃度の試料4.5mlに芽胞菌液を0.5ml接種し、混合した。0.5分、1分、2分、4分経過後のそれぞれにおいて各試料から20μlを取り出し、2mlの1mg/mlチオ硫酸ナトリウム含有滅菌純水に懸濁した。さらに、同じチオ硫酸ナトリウム含有滅菌純水を用いて10倍希釈し、NA培地に塗抹した。Controlについては滅菌純水を用い、NA培地に塗抹した。上記平板培地を35℃で2日間培養した後、各平板培地上に出現したコロニーを計測した。
試料濃度別に、処理時間毎に生存芽胞菌数を計測し、殺芽胞効果を判定した。
1)Bacillus subtilis における殺芽胞試験結果
以下の表7及び図6に示すように、試料A及び試料Bを用いた場合は、2分間処理すると死滅したが、試料Cを用いた場合は、4分間処理ではControlと比較して芽胞が約1/2生残していた。次亜塩素酸ナトリウム水を用いた場合は、対照D、対照E、及び、対照Fの全てにおいて4分間の処理で芽胞が約1/5〜1/2生残していた。図6中、次亜塩素酸水溶液で処理した場合を白抜きのマークと実線で示し、次亜塩素酸ナトリウム水で処理した場合を、二点鎖線、一点鎖線及び破線でそれぞれ示した。
以下の表8に示すように、試料A及び試料Bを用いた場合は、それぞれ1分間処理、4分間処理において死滅した。試料Cを用いた場合は、4分間の処理においてControlと比較して芽胞がかなり生残していた。次亜塩素酸ナトリウム水を用いた場合は、対照D及び対照Eにおいて4分間処理で死滅した。対照Fにおいては4分間の処理で芽胞が約1/4〜1/2生残していた。図7中、次亜塩素酸水溶液で処理した場合を白抜きのマークと実線で示し、次亜塩素酸ナトリウム水で処理した場合を、二点鎖線、一点鎖線及び破線でそれぞれ示した。
以下の表9に示すように、試料A及び試料Bを用いた場合は、2分間処理すると死滅したが、試料Cを用いた場合は、4分間の処理においてもControlと比較して芽胞が約1/2生残していた。次亜塩素酸ナトリウム水を用いた場合は、対照D、対照E、及び、対照Fの全てにおいて4分間の処理で芽胞が約1/3〜1/2生残していた。図8中、次亜塩素酸水溶液で処理した場合を白抜きのマークと実線で示し、次亜塩素酸ナトリウム水で処理した場合を、二点鎖線、一点鎖線及び破線でそれぞれ示した。
Claims (3)
- 食品添加物用次亜塩素酸ナトリウムと、日本薬局方精製水と、日本薬局方希塩酸のみとを含むpH6.0〜6.5の次亜塩素酸水溶液であって、前記水溶液中の有効塩素濃度が0.5〜5ppmであり、サルモネラ・エンテロティディス、大腸菌、カンジダ及び緑膿菌からなる群から選ばれる菌を、10分処理で死滅させる、抗菌剤。
- 前記次亜塩素酸水溶液のpHが6.4であることを特徴とする、請求項1に記載の抗菌剤。
- 請求項1又は2に記載の抗菌剤中で、抗菌処理の対象となる部材を5〜10分間処理することを特徴とする、抗菌処理方法。
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