JP2021195346A - Composition for sterilization - Google Patents
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- JP2021195346A JP2021195346A JP2020104041A JP2020104041A JP2021195346A JP 2021195346 A JP2021195346 A JP 2021195346A JP 2020104041 A JP2020104041 A JP 2020104041A JP 2020104041 A JP2020104041 A JP 2020104041A JP 2021195346 A JP2021195346 A JP 2021195346A
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- 239000000203 mixture Substances 0.000 title claims abstract description 97
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 67
- 238000004659 sterilization and disinfection Methods 0.000 title abstract description 16
- 150000003839 salts Chemical class 0.000 claims abstract description 43
- 230000002378 acidificating effect Effects 0.000 claims abstract description 37
- 235000013373 food additive Nutrition 0.000 claims abstract description 20
- 239000002778 food additive Substances 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 150000007524 organic acids Chemical class 0.000 claims abstract description 18
- 229940107702 grapefruit seed extract Drugs 0.000 claims description 22
- 229910019142 PO4 Inorganic materials 0.000 claims description 20
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 17
- 239000010452 phosphate Substances 0.000 claims description 17
- 239000000284 extract Substances 0.000 claims description 12
- 235000005985 organic acids Nutrition 0.000 claims description 12
- 230000000694 effects Effects 0.000 abstract description 7
- 238000012360 testing method Methods 0.000 description 42
- 239000000243 solution Substances 0.000 description 40
- 241000700605 Viruses Species 0.000 description 38
- 239000000126 substance Substances 0.000 description 19
- 235000021317 phosphate Nutrition 0.000 description 18
- 238000000034 method Methods 0.000 description 17
- 229910017053 inorganic salt Inorganic materials 0.000 description 15
- 210000004027 cell Anatomy 0.000 description 13
- 230000003833 cell viability Effects 0.000 description 13
- 241000894006 Bacteria Species 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 230000000844 anti-bacterial effect Effects 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- 230000003139 buffering effect Effects 0.000 description 11
- 238000011156 evaluation Methods 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 9
- 239000000645 desinfectant Substances 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 241000714201 Feline calicivirus Species 0.000 description 8
- 230000002779 inactivation Effects 0.000 description 7
- 231100000344 non-irritating Toxicity 0.000 description 7
- 241001263478 Norovirus Species 0.000 description 6
- 206010040880 Skin irritation Diseases 0.000 description 6
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- 231100000135 cytotoxicity Toxicity 0.000 description 6
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- 238000000338 in vitro Methods 0.000 description 5
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- 230000007794 irritation Effects 0.000 description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 5
- 210000003491 skin Anatomy 0.000 description 5
- 239000001488 sodium phosphate Substances 0.000 description 5
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 5
- 229910000406 trisodium phosphate Inorganic materials 0.000 description 5
- 235000019801 trisodium phosphate Nutrition 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 239000005708 Sodium hypochlorite Substances 0.000 description 4
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 230000000120 cytopathologic effect Effects 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 238000013207 serial dilution Methods 0.000 description 4
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 4
- 238000010998 test method Methods 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 241001333951 Escherichia coli O157 Species 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 241000607142 Salmonella Species 0.000 description 3
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- 239000004480 active ingredient Substances 0.000 description 3
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 230000008029 eradication Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 206010015946 Eye irritation Diseases 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241001105894 Murine norovirus Species 0.000 description 2
- 231100000694 OECD Guidelines for the Testing of Chemicals Toxicity 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
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- 210000001339 epidermal cell Anatomy 0.000 description 2
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- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
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- 239000011521 glass Substances 0.000 description 2
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- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000000774 hypoallergenic effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
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- 210000003292 kidney cell Anatomy 0.000 description 2
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- 235000013372 meat Nutrition 0.000 description 2
- 150000007522 mineralic acids Chemical class 0.000 description 2
- 231100000286 mucous membrane, eye irritation or corrosion testing Toxicity 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 244000309711 non-enveloped viruses Species 0.000 description 2
- 230000007505 plaque formation Effects 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 231100000330 serious eye damage Toxicity 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 230000008023 solidification Effects 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 241000712461 unidentified influenza virus Species 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 241000207199 Citrus Species 0.000 description 1
- 240000000560 Citrus x paradisi Species 0.000 description 1
- 241001646719 Escherichia coli O157:H7 Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 235000020971 citrus fruits Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 229940065115 grapefruit extract Drugs 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000002085 irritant Substances 0.000 description 1
- 231100000021 irritant Toxicity 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 229940126601 medicinal product Drugs 0.000 description 1
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical compound [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 239000002444 monopotassium citrate Substances 0.000 description 1
- 235000015861 monopotassium citrate Nutrition 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 239000004745 nonwoven fabric Substances 0.000 description 1
- QVLTXCYWHPZMCA-UHFFFAOYSA-N po4-po4 Chemical compound OP(O)(O)=O.OP(O)(O)=O QVLTXCYWHPZMCA-UHFFFAOYSA-N 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- WKZJASQVARUVAW-UHFFFAOYSA-M potassium;hydron;2-hydroxypropane-1,2,3-tricarboxylate Chemical compound [K+].OC(=O)CC(O)(C(O)=O)CC([O-])=O WKZJASQVARUVAW-UHFFFAOYSA-M 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
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- 239000002994 raw material Substances 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
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- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- KZQSXALQTHVPDQ-UHFFFAOYSA-M sodium;butanedioate;hydron Chemical compound [Na+].OC(=O)CCC([O-])=O KZQSXALQTHVPDQ-UHFFFAOYSA-M 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000021 stimulant Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 229910000404 tripotassium phosphate Inorganic materials 0.000 description 1
- 235000019798 tripotassium phosphate Nutrition 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Description
本発明は除菌用組成物に関し、家庭内やオフィスなどの身の回りの物品に対し、安心安全に除菌を行うための除菌用組成物に関する。 The present invention relates to a sterilizing composition, and relates to a sterilizing composition for safely and safely sterilizing personal items such as homes and offices.
従来から、良好な衛生を保つために、家庭内やオフィスにおける物品の表面を除菌するための除菌用組成物が提案されている。一般的には、次亜塩素酸ナトリウムを有効成分として含む除菌用組成物（殺菌剤）が知られる。しかし、次亜塩素酸ナトリウムを含む除菌用組成物は刺激が強く、誤って人体に取り込まれた場合の有害性が危惧される。また次亜塩素酸ナトリウムと酸性洗剤等とが混合された場合、塩素ガスの発生が問題となる虞がある。
またアルコールを有効成分として含む除菌用組成物も知られている。しかし、アルコールを主成分として含む除菌用組成物は、引火性があり取り扱いに注意を要する必要があり、また独特な臭いがする上、特定のノンエンベロープウイルスに対し効果が低いことが示唆されている。
Conventionally, in order to maintain good hygiene, a sterilizing composition for sterilizing the surface of an article in a home or an office has been proposed. Generally, a disinfectant composition (bactericidal agent) containing sodium hypochlorite as an active ingredient is known. However, the disinfectant composition containing sodium hypochlorite is highly irritating, and there is a concern that it may be harmful if it is accidentally taken into the human body. Further, when sodium hypochlorite and an acidic detergent or the like are mixed, generation of chlorine gas may become a problem.
A sterilizing composition containing alcohol as an active ingredient is also known. However, it is suggested that the disinfectant composition containing alcohol as a main component is flammable and needs to be handled with care, has a peculiar odor, and is less effective against specific non-enveloped viruses. ing.
そこで、水の電気分解により生成された強アルカリ電解水を用いる除菌用組成物が提案されている。たとえば下記特許文献１には、ｐＨ１１以上の強アルカリ電解水と、柑橘類の種子から抽出した成分とを有し、当該成分が濃縮エキス換算で０．００５％以上０．５％未満である除菌用組成物（以下、従来技術１ともいう）が開示されている。特許文献１には、従来技術１によれば、強アルカリ電解水の洗浄効果が発揮されるとともに、上記成分の含有により良好な除菌効果が発揮される旨、記載されている。 Therefore, a composition for sterilization using strongly alkaline electrolyzed water produced by electrolysis of water has been proposed. For example, Patent Document 1 below has a strong alkaline electrolyzed water having a pH of 11 or higher and a component extracted from citrus seeds, and the component is 0.005% or more and less than 0.5% in terms of concentrated extract. A composition for use (hereinafter, also referred to as prior art 1) is disclosed. Patent Document 1 describes that according to the prior art 1, the cleaning effect of strong alkaline electrolyzed water is exhibited, and the inclusion of the above-mentioned components exhibits a good sterilizing effect.
しかしながら、本発明者らの検討によれば、上述する従来技術１は、除菌効果が不十分であることがわかった。
即ち、除菌が求められる汚れた除菌対象面のｐＨは、一般的に中性から酸性の場合が多く、少なくともｐＨ１１以上ということは通常ではありえない。従来技術１は、ｐＨ１１以上の強アルカリに調整されてはいるものの、上述する除菌対象面に対し散布等により接触した場合、当該除菌対象面のｐＨに影響されて短時間でｐＨが１１未満に下がってしまう。そのため、ｐＨ１１以上とすることで期待された除菌性や洗浄性が十分に発揮されないか、あるいは、十分な除菌効果を得るために除菌時間を長く確保する必要があった。すなわち、強アルカリに調整された電解水は、実質的な除菌効果が十分ではなかった。
However, according to the studies by the present inventors, it was found that the above-mentioned prior art 1 has an insufficient sterilizing effect.
That is, the pH of the dirty sterilization target surface for which sterilization is required is generally neutral to acidic in many cases, and it cannot usually be at least pH 11 or higher. Although the prior art 1 is adjusted to a strong alkali having a pH of 11 or higher, when the surface to be sterilized is brought into contact with the above-mentioned surface by spraying or the like, the pH is affected by the pH of the surface to be sterilized and the pH is 11 in a short time. It goes down to less than. Therefore, it is necessary to ensure that the expected sterilizing property and detergency are not sufficiently exhibited by setting the pH to 11 or higher, or that the sterilizing time is long in order to obtain a sufficient sterilizing effect. That is, the electrolyzed water adjusted to a strong alkali did not have a sufficient sterilizing effect.
本発明は上述のような課題に鑑みてなされたものである。即ち、本発明は、人体に対し安全であり、かつ実質的に除菌効果の高い除菌用組成物を提供することを課題とする。 The present invention has been made in view of the above-mentioned problems. That is, it is an object of the present invention to provide a sterilizing composition that is safe for the human body and has a substantially high sterilizing effect.
本発明の除菌用組成物は、食品添加物として認可を受けた塩基性無機塩類と、食品添加物として認可を受けた酸性無機塩類および／または有機酸と、水と、を含み、ｐＨが１０．５以上１１．５以下であることを特徴とする。 The sterilizing composition of the present invention contains basic inorganic salts approved as food additives, acidic inorganic salts and / or organic acids approved as food additives, and water, and has a pH. It is characterized by being 10.5 or more and 11.5 or less.
本発明の除菌用組成物は、塩基性無機塩類と、酸性無機塩類および／または有機酸とを含み、これらによりｐＨが調整されるとともに、緩衝作用が生じ得る。そのため、本発明の除菌用組成物は、除菌対象面と接触した場合、当該除菌対象面が中性あるいは酸性であったとしても、ｐＨが所定範囲（つまりｐＨ１０．５〜ｐＨ１１．５）内に維持されやすい。したがって、本発明の除菌用組成物は、除菌対象面において良好な除菌効果を発揮するとともに、除菌時間を短縮することが可能であり、実質的に除菌効果が高い。
しかも本発明の除菌用組成物に含有される塩基性無機塩類ならびに酸性無機塩類および／または有機酸は、いずれも食品添加物として認可を受けた化合物であるため、誤って人体にかかり、あるいは人体に取り込まれた場合であっても極めて安全である。そのため、日常の様々な物品を除菌することができ、また誰でも危険を伴わずに使用することができる。
The sterilizing composition of the present invention contains basic inorganic salts and acidic inorganic salts and / or organic acids, which can adjust the pH and cause a buffering action. Therefore, when the composition for sterilization of the present invention comes into contact with the surface to be sterilized, the pH is in a predetermined range (that is, pH 10.5 to pH 11.5) even if the surface to be sterilized is neutral or acidic. ) Is easy to maintain. Therefore, the sterilizing composition of the present invention exhibits a good sterilizing effect on the surface to be sterilized, can shorten the sterilizing time, and has a substantially high sterilizing effect.
Moreover, since the basic inorganic salts, the acidic inorganic salts and / or the organic acids contained in the sterilization composition of the present invention are all compounds approved as food additives, they may be erroneously applied to the human body or may be affected by the human body. It is extremely safe even when taken into the human body. Therefore, various daily articles can be sterilized, and anyone can use it without danger.
本発明の除菌用組成物（以下、単に組成物という場合がある）は、食品添加物として認可を受けた塩基性無機塩類と、食品添加物として認可を受けた酸性無機塩類および／または有機酸と、水と、を含む。本発明の組成物のｐＨは、１０．５以上１１．５以下に調整される。
なお、本発明に関し除菌とは、ウイルスおよび／または菌に対し、不活化作用、死滅化作用などを発揮することをいう。すなわち、本発明の組成物は、ウイルスおよび／または菌の除去および／または死滅させるために用いられる。
また本発明において「食品添加物として認可を受けた」とは、具体的には日本国食品衛生法第１０条に基づき厚生労働大臣が使用を認めた、指定添加物及び既存添加物を示す。
The sterilizing composition of the present invention (hereinafter, may be simply referred to as a composition) is a basic inorganic salt approved as a food additive, an acidic inorganic salt approved as a food additive, and / or an organic substance. Contains acid and water. The pH of the composition of the present invention is adjusted to 10.5 or more and 11.5 or less.
In addition, in this invention, sterilization means exerting an inactivating action, a killing action, etc. against a virus and / or a bacterium. That is, the compositions of the present invention are used to eliminate and / or kill viruses and / or fungi.
Further, in the present invention, "approved as a food additive" specifically means a designated additive and an existing additive approved for use by the Minister of Health, Labor and Welfare based on Article 10 of the Food Sanitation Law of Japan.
上述する構成を備える本発明の組成物は、ｐＨ１０．５以上ｐＨ１１．５以下と除菌剤としては低めの範囲に調整される上、食品添加物として認定を受けた化合物によってｐＨが調整されているため、人体に対し安心安全である。
本発明における塩基性無機塩類と、酸性無機塩類および／または有機酸とは、ｐＨ調整作用だけでなく、組成物のｐＨの緩衝剤としても作用する。そのため、本発明の組成物のｐＨは、除菌対象面に接触した際に当該除菌対象面のｐＨに影響を受けて所定範囲を外れにくく、良好な除菌性を維持することができ、また比較的短時間で除菌を実施することが可能である。
またこのように、本発明の組成物は、ｐＨの緩衝作用が働くことから、除菌対象面に接触した際のｐＨの低下を想定してあらかじめ高いアルカリ性に調整する必要がない。そのためｐＨ１０．５以上ｐＨ１１．５以下という、アルカリ性としては低めのｐＨ範囲でも実質的に高い除菌性を発揮することができる。かかるｐＨの範囲であれば、人体に組成物が付着した場合であっても肌荒れを防止し、また刺激性が小さいか、実質的に刺激を感じさせない程度であって、人体に対し極めて安心である。
以下に、本発明の除菌用組成物についてさらに詳細に説明する。
The composition of the present invention having the above-mentioned constitution is adjusted to have a pH of 10.5 or more and a pH of 11.5 or less, which is a low range as a disinfectant, and the pH is adjusted by a compound certified as a food additive. Therefore, it is safe and secure for the human body.
The basic inorganic salts and the acidic inorganic salts and / or organic acids in the present invention act not only as a pH adjusting action but also as a buffering agent for the pH of the composition. Therefore, the pH of the composition of the present invention is affected by the pH of the surface to be sterilized when it comes into contact with the surface to be sterilized, and it is difficult to deviate from a predetermined range, so that good sterilization can be maintained. In addition, it is possible to carry out sterilization in a relatively short time.
Further, as described above, since the composition of the present invention acts as a pH buffering action, it is not necessary to adjust the composition to a high alkalinity in advance assuming a decrease in pH when it comes into contact with the surface to be sterilized. Therefore, it is possible to exhibit substantially high sterilization even in a pH range of 10.5 or more and 11.5 or less, which is a low alkaline pH range. Within such a pH range, even if the composition adheres to the human body, it prevents rough skin, and the irritation is small or practically non-irritating, so it is extremely safe for the human body. be.
Hereinafter, the sterilizing composition of the present invention will be described in more detail.
（塩基性無機塩類）
本発明の組成物は、食品添加物として認可を受けた塩基性無機塩類を含む。上記塩基性無機塩類の例としては、塩基性リン酸塩、炭酸塩、水酸化物などが挙げられる。ここで無機塩類とは、無機酸の水素を置換してできる塩のことをいい、塩基性無機塩類とは、無機塩類を有し水溶液中でアルカリ性を呈する化合物を意味する。本発明の組成物は、塩基性無機塩類を１種または２種以上を含有することができる。
本発明に用いられる塩基性無機塩類は、本発明の組成物のｐＨを塩基性側に調整するｐＨ調整剤であるとともに、後述する酸性無機塩類とともに緩衝作用を発揮することが期待される化合物である。
(Basic inorganic salts)
The composition of the present invention contains basic inorganic salts approved as food additives. Examples of the basic inorganic salts include basic phosphates, carbonates, hydroxides and the like. Here, the inorganic salt means a salt formed by substituting hydrogen of an inorganic acid, and the basic inorganic salt means a compound having inorganic salts and exhibiting alkalinity in an aqueous solution. The composition of the present invention may contain one or more basic inorganic salts.
The basic inorganic salts used in the present invention are pH adjusters that adjust the pH of the composition of the present invention to the basic side, and are compounds that are expected to exert a buffering action together with the acidic inorganic salts described later. be.
上記塩基性リン酸塩としては、リン酸三ナトリウム、リン酸三カリウム、などが挙げられる。
上記炭酸塩としては、炭酸カリウム、炭酸ナトリウムなどが挙げられる。
上記水酸化物としては、水酸化カルシウムなどが挙げられる。
Examples of the basic phosphate include trisodium phosphate, tripotassium phosphate, and the like.
Examples of the carbonate include potassium carbonate and sodium carbonate.
Examples of the hydroxide include calcium hydroxide.
中でも、緩衝効果に優れるとともに、高い電荷を有し化学反応の活性が高いため高い除菌効果（不活化効果）が期待されるという観点から本発明の塩基性無機塩類としては、塩基性リン酸塩が好ましく、特にリン酸三ナトリウムは高い不活化効果を発揮し得るため好ましい。 Among them, the basic inorganic salt of the present invention is a basic phosphate from the viewpoint that it has an excellent buffering effect and is expected to have a high sterilizing effect (inactivating effect) because it has a high charge and a high chemical reaction activity. Salts are preferable, and trisodium phosphate is particularly preferable because it can exert a high inactivating effect.
（酸性無機塩類、有機酸）
本発明の組成物は、食品添加物として認可を受けた酸性無機塩類および／または有機酸を含む。つまり本発明は、除菌作用発揮可能な食品添加物である塩基性無機塩類とともに、酸性無機塩類および／または有機酸を用いることで、緩衝液を構成し、これによって、所定範囲のｐＨを維持することを可能とするものである。これによって本発明の組成物は、人体に有害な影響を与えることなく、たとえばノロウイルスなどのウイルスや種々の菌に対し、望ましい除菌効果を発揮させ得る。
(Acid inorganic salts, organic acids)
The compositions of the present invention contain acidic inorganic salts and / or organic acids approved as food additives. That is, the present invention constitutes a buffer solution by using acidic inorganic salts and / or organic acids together with basic inorganic salts which are food additives capable of exerting a sterilizing effect, thereby maintaining a pH within a predetermined range. It is what makes it possible to do. Thereby, the composition of the present invention can exert a desirable sterilizing effect against a virus such as norovirus and various bacteria without adversely affecting the human body.
したがって、緩衝作用をより有効に発揮するという観点からは、上述する塩基性無機塩類は、強塩基であることが好ましく、酸性無機塩類および／または有機酸は、弱酸であることが好ましい。ここでいう強塩基とは、２％溶液でｐＨ１２以上を示す化合物を指し、弱酸とは、２％溶液でｐＨ５以下を示す化合物を指す。 Therefore, from the viewpoint of more effectively exerting the buffering action, the above-mentioned basic inorganic salts are preferably strong bases, and the acidic inorganic salts and / or organic acids are preferably weak acids. The strong base here refers to a compound exhibiting a pH of 12 or higher in a 2% solution, and the weak acid refers to a compound exhibiting a pH of 5 or lower in a 2% solution.
上記酸性無機塩類の例としては、酸性リン酸塩などが挙げられる。ここで無機塩類とは、無機酸の水素を置換してできる塩のことをいい、酸性無機塩類とは、無機塩類を有し水溶液中で酸性を呈する化合物を意味する。
本発明に用いられる酸性無機塩類は、本発明の組成物のｐＨの調整に関与するとともに、当該組成物において緩衝剤としての役割を担う。
Examples of the acidic inorganic salts include acidic phosphates and the like. Here, the inorganic salt means a salt formed by substituting the hydrogen of the inorganic acid, and the acidic inorganic salt means a compound having an inorganic salt and exhibiting acidity in an aqueous solution.
The acidic inorganic salts used in the present invention are involved in adjusting the pH of the composition of the present invention and play a role as a buffer in the composition.
上記酸性リン酸塩としては、リン酸二水素カリウム、リン酸二水素ナトリウムなどが挙げられる。尚、本発明では、酸性リン酸塩の代わりにリン酸を用いることもできる。 Examples of the acidic phosphate include potassium dihydrogen phosphate and sodium dihydrogen phosphate. In the present invention, phosphoric acid can be used instead of the acidic phosphate.
中でも、本発明の酸性無機塩類としては、酸性リン酸塩が好ましく、特にリン酸二水素カリウムは、緩衝効果および不活化効果に優れるため好ましい。 Among them, as the acidic inorganic salts of the present invention, acidic phosphate is preferable, and potassium dihydrogen phosphate is particularly preferable because it is excellent in buffering effect and inactivating effect.
上記有機酸の例としては、クエン酸、クエン酸一カリウム、コハク酸、コハク酸一ナトリウムなどが挙げられる。
本発明に用いられる有機酸は、本発明の組成物のｐＨの調整に関与するとともに、当該組成物において緩衝剤としての役割を担う。
Examples of the organic acid include citric acid, monopotassium citrate, succinic acid, monosodium succinate and the like.
The organic acid used in the present invention is involved in adjusting the pH of the composition of the present invention and also plays a role as a buffer in the composition.
本発明の組成物は、酸性無機塩類または有機酸を含んでいてもよく、または酸性無機塩類および有機酸の両方を含んでいてもよく、当該組成物に含まれる酸性無機塩類は１種または２種以上であってよく、当該組成物に含まれる有機酸も１種または２種以上であってよい。 The composition of the present invention may contain an acidic inorganic salt or an organic acid, or may contain both an acidic inorganic salt and an organic acid, and the acidic inorganic salt contained in the composition may be one or two. It may be one or more kinds, and the organic acid contained in the composition may be one kind or two or more kinds.
上述する塩基性無機塩類と、酸性無機塩類および／または有機酸とに関し、特に好ましい組み合わせとしては、塩基性無機塩類として塩基性リン酸塩を含み、かつ、酸性無機塩類として酸性リン酸塩を含むことが好ましい。これらリン酸塩は、３価の電荷をもち、リン酸塩を用いた緩衝液は３段階の緩衝領域を示すとともに広いｐＨ領域で価数変化を生ずるため優れた緩衝作用を発揮する。またリン酸塩は、一般的に化学活性が高いため、本発明の組成物にリン酸塩を用いることによって、良好な、抗菌性や抗ウイルス性が期待される上、肌に触れた際の脱水作用が生じにくいので低刺激性である点でも好ましい。またリン酸は、ＤＮＡやＲＮＡを構成する成分として知られる。本発明者らは、菌が増殖し、あるいはウイルスが複製される際に組成物中に含まれるリン酸（リン酸塩）が、これらの増殖や複製に何らかの生化学的作用を与え、これによって効果的な抗菌、抗ウイルス作用が発揮されるのではないかと推察する。
したがって、かかる組み合わせであれば、組成物のｐＨ調整効果および緩衝効果に優れるとともに、非刺激性であって優れた除菌性を発揮可能な除菌用組成物を提供可能である。
より具体的には、本発明の組成物は、たとえば塩基性リン酸塩としてリン酸三ナトリウムを含み、酸性リン酸塩としてリン酸二水素カリウムを含むことが好ましい。
With respect to the above-mentioned basic inorganic salts and acidic inorganic salts and / or organic acids, a particularly preferable combination includes a basic phosphate as a basic inorganic salt and an acidic phosphate as an acidic inorganic salt. Is preferable. These phosphates have a trivalent charge, and the buffer solution using the phosphate exhibits a three-step buffering region and exhibits an excellent buffering action because the valence changes in a wide pH range. In addition, since phosphate generally has high chemical activity, good antibacterial and antiviral properties are expected by using phosphate in the composition of the present invention, and when it comes into contact with the skin, it is expected. It is also preferable in that it is hypoallergenic because it does not easily cause dehydration. Phosphoric acid is also known as a component constituting DNA and RNA. We have found that the phosphoric acid (phosphate) contained in the composition when the fungus grows or the virus replicates causes some biochemical effect on their growth and replication. It is speculated that effective antibacterial and antiviral effects may be exhibited.
Therefore, with such a combination, it is possible to provide a sterilizing composition which is excellent in pH adjusting effect and buffering effect of the composition and is non-irritating and can exhibit excellent sterilizing property.
More specifically, the composition of the present invention preferably contains, for example, trisodium phosphate as a basic phosphate and potassium dihydrogen phosphate as an acidic phosphate.
本発明の組成物に含有される塩基性無機塩類および酸性無機塩類の量は特に限定されないが、それぞれの含有量は、たとえば０．０１質量％以上１０．０質量％以下の範囲で含まれることが好ましく、０．０５質量％以上３．０質量％以下であることがより好ましい。い。上記含有量が０．０１質量％未満であると、好ましい除菌効果が発揮されない場合があるとともに良好なｐＨ緩衝作用が生じない場合がある。一方、上記含有量が１０．０質量％を超えた場合、安定して水に溶解し難くなり、温度や状態の変化（たとえば一度、凍結した後融解する際など）によって成分が析出する場合があり、安定性の点で十分でない場合がある。なお、上記含有量は、本発明の組成物を実際に使用する際の濃度を示すものである。本発明の組成物は安定性も良好であるため、５倍以上２０倍以下の濃縮物組成物として取り扱うこともできる。 The amounts of the basic inorganic salts and the acidic inorganic salts contained in the composition of the present invention are not particularly limited, but the respective contents are contained in the range of, for example, 0.01% by mass or more and 10.0% by mass or less. Is preferable, and more preferably 0.05% by mass or more and 3.0% by mass or less. stomach. If the content is less than 0.01% by mass, a preferable sterilizing effect may not be exhibited and a good pH buffering action may not be produced. On the other hand, when the above content exceeds 10.0% by mass, it becomes difficult to stably dissolve in water, and the components may precipitate due to changes in temperature or state (for example, once frozen and then thawed). Yes, it may not be sufficient in terms of stability. The above-mentioned content indicates the concentration when the composition of the present invention is actually used. Since the composition of the present invention has good stability, it can be handled as a concentrate composition of 5 times or more and 20 times or less.
本発明の組成物のｐＨは、１０．５以上１１．５以下に調整される。ｐＨ１０．５未満であると、除菌性が十分でない場合があり、一方、ｐＨ１１．５を上回ると皮膚に対する刺激性が強くなり安心安全とは言えない場合があるからである。本発明の組成物は、上述の範囲にｐＨが調整されているとともに、組成物自体が緩衝液を構成するため除菌対象面などに接した際や希釈した際に、上記範囲のｐＨが維持されやすいという有利な点を有する。 The pH of the composition of the present invention is adjusted to 10.5 or more and 11.5 or less. This is because if the pH is less than 10.5, the sterilizing property may not be sufficient, while if the pH exceeds 11.5, the irritation to the skin becomes strong and it may not be safe and secure. The pH of the composition of the present invention is adjusted to the above range, and since the composition itself constitutes a buffer solution, the pH in the above range is maintained when it comes into contact with the surface to be sterilized or when diluted. It has the advantage of being easily sterilized.
本発明の組成物は、さらにグレープフルーツ種子抽出物を含むことが好ましい。グレープフルーツ種子抽出物は抗菌性を示すため、より優れた除菌性を示す組成物を提供することができる。すなわち、グレープフルーツ種子抽出物を含む本発明の組成物は、人体に安心であって、かつ優れた抗ウイルス性および抗菌性を示し得る。 The composition of the present invention preferably further comprises a grapefruit seed extract. Since the grapefruit seed extract exhibits antibacterial properties, it is possible to provide a composition exhibiting better sterilizing properties. That is, the composition of the present invention containing grapefruit seed extract can be safe for the human body and exhibit excellent antiviral and antibacterial properties.
グレープフルーツ種子抽出物の主成分は、脂肪酸とフラボノイドであり、製造方法は特に限定されないが、たとえばアルコールや水などで抽出し製造することができる。フラボノイドは脂溶性であるため水よりもアルコールによって良好に抽出されるため、特にアルコール抽出物が好ましい。アルコールで抽出した場合、後工程においてアルコール成分を蒸発させることで実質的にアルコール濃度を０％に調整することが好ましい。また市販品であるグレープフルーツ抽出物を本発明の組成物に用いてもよく、この場合、食品添加物として認定されたグレードの組成物を用いることが好ましい。 The main components of the grapefruit seed extract are fatty acids and flavonoids, and the production method is not particularly limited, but the grapefruit seed extract can be extracted and produced with, for example, alcohol or water. Alcohol extracts are particularly preferred because flavonoids are fat-soluble and are better extracted with alcohol than with water. When extracted with alcohol, it is preferable to adjust the alcohol concentration to 0% by evaporating the alcohol component in the subsequent step. Further, a commercially available grapefruit extract may be used for the composition of the present invention, and in this case, it is preferable to use a composition of a grade certified as a food additive.
グレープフルーツ種子抽出物の含有量は特に限定されないが、一般的に単価の高い原料であるため、抗菌性を担保しつつ含有量を抑えることが好ましい。具体的には、本発明の組成物１００質量％において、グレープフルーツ種子抽出物は、抽出物換算で０．００１質量％以上１．０質量％以下の範囲であることが好ましく、０．００２質量％以上０．５質量％以下の範囲であることがより好ましく、０．００３質量％以上０．１質量％以下であることがさらに好ましく、０．００５質量％以上０．０１質量％以下であることが特に好ましい。上記範囲は、抗菌性を発揮させるために用いる従来のグレープフルーツ種子抽出物としては少ない含有量であるが、本発明の組成物は、上述するとおり、除菌対象面に接触し、あるいは希釈などされた場合にも所定のｐＨの範囲が維持されやすく、実質的に除菌効果が高い。そのため本発明の組成物は、グレープフルーツ種子抽出物の含有量が上記範囲のように少量であっても、高い抗菌性を発揮し得る。
上記グレープフルーツ種子抽出物の含有量が抽出物換算で０．００１質量％未満であると、抗菌効果が十分ではない場合があり、一方、１．０量％を超えると、コストの増大に見合う抗菌性効果の向上が確認し難い。
なお、上記抽出物換算とは、グレープフルーツ種子から抽出した原液であって希釈等していない抽出物に換算した濃度を示す。
The content of the grapefruit seed extract is not particularly limited, but since it is generally a raw material having a high unit price, it is preferable to suppress the content while ensuring antibacterial properties. Specifically, in 100% by mass of the composition of the present invention, the grapefruit seed extract is preferably in the range of 0.001% by mass or more and 1.0% by mass or less in terms of extract, and 0.002% by mass. It is more preferably in the range of 0.5% by mass or more, further preferably 0.003% by mass or more and 0.1% by mass or less, and more preferably 0.005% by mass or more and 0.01% by mass or less. Is particularly preferable. The above range has a small content as a conventional grapefruit seed extract used for exhibiting antibacterial properties, but the composition of the present invention is in contact with or diluted with the surface to be sterilized as described above. Even in this case, the predetermined pH range is easily maintained, and the sterilizing effect is substantially high. Therefore, the composition of the present invention can exhibit high antibacterial properties even if the content of the grapefruit seed extract is as small as the above range.
If the content of the grapefruit seed extract is less than 0.001% by mass in terms of extract, the antibacterial effect may not be sufficient, while if it exceeds 1.0% by mass, the antibacterial effect is commensurate with the increase in cost. It is difficult to confirm the improvement of sexual effect.
In addition, the above-mentioned extract conversion means the concentration converted into the undiluted extract which is the undiluted solution extracted from grapefruit seeds.
本発明の組成物の製造方法は特に限定されないが、たとえば、水にリン酸三ナトリウムなどの塩基性無機塩類を添加し溶解させた後、リン酸二水素カリウムなどの酸性無機塩類を添加して溶解させ、その後、適宜、グレープフルーツ種子抽出物を添加するとよい。最終的に、本発明の特定するｐＨの範囲となるよう、予め、添加する化合物の配合比率を決定しておくとよい。 The method for producing the composition of the present invention is not particularly limited. For example, a basic inorganic salt such as trisodium phosphate is added and dissolved in water, and then an acidic inorganic salt such as potassium dihydrogen phosphate is added. It may be dissolved and then the grapefruit seed extract may be added as appropriate. Finally, it is advisable to determine in advance the compounding ratio of the compound to be added so as to be within the pH range specified by the present invention.
本発明の組成物には、本発明の趣旨を逸脱しない範囲において、さらに異なる化合物を添加してもよい。本発明の組成物の安全性を担保するという観点からは、本発明の組成物に含まれる全ての組成は、食品添加物、食品、または食品から抽出されたものなど、肌に触れ、あるいは誤って体内に取り込まれた場合でも実質的に極めて安全なもののみから構成されることが好ましい。 Further different compounds may be added to the composition of the present invention without departing from the spirit of the present invention. From the viewpoint of ensuring the safety of the composition of the present invention, all the compositions contained in the composition of the present invention may come into contact with the skin such as food additives, foods, or those extracted from foods, or may be mistaken. It is preferable that it is composed of only those that are substantially safe even when taken into the body.
表１に示す組成比率で構成された組成物（実施例１、２）を調整した。具体的には、まず精製水にリン酸三ナトリウムを添加して溶解させ、次いで、リン酸二水素カリウムを添加して溶解させ、最後にグレープフルーツ種子抽出物を添加し、全体で１００質量％とした。組成物のｐＨは、１１．４に調整した。
なお、グレープフルーツ種子抽出物は、株式会社アデプト製の抽出物３．３％液（Ｄｅｓｆａｎ−１０）を用い、組成物においてグレープフルーツ種子抽出物が抽出物換算で実施例１では０．００６６質量％、実施例２では０．００３３質量％となるよう添加した。
The compositions (Examples 1 and 2) composed of the composition ratios shown in Table 1 were prepared. Specifically, trisodium phosphate is first added and dissolved in purified water, then potassium dihydrogen phosphate is added and dissolved, and finally grapefruit seed extract is added to make a total of 100% by mass. did. The pH of the composition was adjusted to 11.4.
As the grapefruit seed extract, an extract 3.3% solution (Desfan-10) manufactured by Adept Co., Ltd. was used, and the grapefruit seed extract was 0.0066% by mass in Example 1 in terms of extract in the composition. In Example 2, it was added so as to be 0.0033% by mass.
（ウイルスの不活化評価１）
国立医薬品食品衛生研究所のホームページに開示される「平成２７年度ノロウイルスの不活化条件に関する調査報告書」（以下、単に調査報告書ともいう）に記載の方法に倣い、本実施例のウイルスの不活化評価を行った。評価結果は表１に示す。
また比較対象として、上記調査報告書に示される塩素系消毒剤（Ｎｏ．Ｄ）を参考例１とし、エタノール系消毒剤（Ｎｏ．Ｎ）を参考例２として、それぞれの評価を参照し表１に併記した。
(Virus inactivation evaluation 1)
Following the method described in the "2015 Survey Report on Norovirus Inactivation Conditions" (hereinafter, also referred to simply as the Survey Report) disclosed on the homepage of the National Institute of Health Sciences, the virus in this example is not used. An activation evaluation was performed. The evaluation results are shown in Table 1.
For comparison, the chlorine-based disinfectant (No. D) shown in the above survey report was used as Reference Example 1, and the ethanol-based disinfectant (No. N) was used as Reference Example 2, and each evaluation was referred to in Table 1. Also described in.
より具体的には、上記調査報告書に記載の方法に倣い、実施例１を用いて、５０％感染終末点法（ｍｅｄｉａｎ ｔｉｓｓｕｅ ｃｕｌｔｕｒｅ ｉｎｆｅｃｔｉｏｕｓ ｄｏｓｅ；ＴＣＩＤ５０／ｍｌ法）によりウイルスの除菌性を評価した。
ウイルス液は、以下の２種類を準備した。
・１０％肉エキス含有組織培養維持液（ＭＥＭ培地）およびネコカリシウイルス（Ｆ−９；接種ウイルス量：７．７ｌｏｇ１０）を１：１の割合で混合したネコカリシウイルス液
・５％肉エキス含有組織培養維持液（ＭＥＭ培地）およびマウスノロウイルス（マウスノロウイルス１（ＭＮＶ−１）ＡＴＣＣ株；接種ウイルス量：７．１ｌｏｇ１０）を１：１の割合で混合したマウスノロウイルス液
More specifically, following the method described in the above investigation report, the virus eradication property was determined by the median tissue culture infectious dose (TCID 50 / ml method) using Example 1. evaluated.
The following two types of virus solutions were prepared.
・ 10% meat extract-containing tissue culture maintenance solution (MEM medium) and feline calicivirus (F-9; inoculum amount: 7.7 log 10) mixed at a ratio of 1: 1 feline calicivirus solution ・ 5% meat extract contained A mouse norovirus solution in which a tissue culture maintenance solution (MEM medium) and a mouse norovirus (mouse norovirus 1 (MNV-1) ATCC strain; inoculum virus amount: 7.1 log 10) are mixed at a ratio of 1: 1.
そして、実施例１の除菌用組成物と各ウイルス液とを、除菌用組成物：ウイルス液＝９：１の割合で、接触させた。接触時間は６０秒とした。除菌用組成物とウイルス液を６０秒間接触させた後、直ちに１０％牛胎児血清加Ｄｕｌｂｅｃｃｏ’ＭＥＭ（ＤＭＥＭ）培地で１０倍段階希釈し反応を停止した。
ウイルス液の反応停止後、以下のとおり、５０％感染終末点法（ＴＣＩＤ５０／ｍｌ法）で生存ウイルス量を定量した。すなわち、上述のとおり反応停止したウイルス混合液（１０倍段階希釈したもの）を、あらかじめ９６ウェルプレートに単層培養したＲＡＷ２６４．７細胞に１ウエルあたり１００μＬ接種し、３７度、ＣＯ２インキュベーター内で１時間培養した。
その後、試験液を除き、新たに３％ＤＭＥＭを１００μＬ各ウエルに加えて３７℃、ＣＯ２インキュベーター内で培養した。
そして培養７日後に倒立顕微鏡下でウイルスによる細胞変性効果（ＣＰＥ）を観察してＢｅｈｒｅｎｎｓ−Ｋａｒｂｅｒ法を用いてウイルス感染価を算出した。ウイルス液の作用停止後の溶液の細胞毒性を顕微鏡下で、ウイルスによる細胞変性効果（ＣＰＥ）と判別した。
なお、ネコカリシウイルス試験では、実施例１の代わりに精製水を用いたものをコントロールとした。またマウスノロウイルス試験では、実施例１の代わりにＰＢＳ（リン酸緩整理食塩水）を用いたものをコントロールとした。
Then, the sterilizing composition of Example 1 and each virus solution were brought into contact with each other at a ratio of sterilizing composition: virus solution = 9: 1. The contact time was 60 seconds. After contacting the disinfectant composition with the virus solution for 60 seconds, the reaction was immediately terminated by 10-fold serial dilution with 10% fetal bovine serum-added Dulbecco'MEM (DMEM) medium.
After the reaction of the virus solution was stopped, the amount of viable virus was quantified by the 50% infection end point method (TCID 50 / ml method) as follows. That is, 100 μL of the virus mixture (10-fold serially diluted) in which the reaction was stopped as described above was inoculated into RAW264.7 cells previously cultured in a single layer on a 96-well plate per well, and at 37 ° C. in a CO 2 incubator. Incubated for 1 hour.
Then, the test solution was removed, 3% DMEM was newly added to each well of 100 μL, and the cells were cultured in a CO 2 incubator at 37 ° C.
Then, 7 days after culturing, the cytopathic effect (CPE) by the virus was observed under an inverted microscope, and the viral infectivity titer was calculated using the Behrenns-Carber method. The cytotoxicity of the solution after the action of the virus solution was stopped was determined to be the cytopathic effect (CPE) by the virus under a microscope.
In the feline calicivirus test, purified water was used as a control instead of Example 1. Moreover, in the mouse norovirus test, the control using PBS (Phosphate Buffered Saline) instead of Example 1 was used.
上記生存ウイルス量の定量の結果、実施例１は、ネコカリシウイルスを用いた評価では、生存ウイルス量は２．７ｌｏｇ１０であり、ウイルスの減少量は５．０ｌｏｇ１０であった。またマウスノロウイルスを用いた評価では、生存ウイルス量は１．４ｌｏｇ１０であり、ウイルスの減少量は５．７ｌｏｇ１０であった。 As a result of the above-mentioned quantification of the viable virus amount, in Example 1, the viable virus amount was 2.7 log10 and the virus reduction amount was 5.0 log10 in the evaluation using feline calicivirus. In the evaluation using murine norovirus, the amount of viable virus was 1.4 log10, and the amount of virus reduction was 5.7 log10.
これに対し、比較対象として示した公知のネコカリシウイルス評価によれば、次亜塩素酸ナトリウムが用いられた参考例１は、同様の試験によってウイルスの減少量は２．０ｌｏｇ１０未満であり、また指定医薬部外品である市販のエタノール系消毒剤を用いた参考例２は、同様の試験によってウイルスの減少量は２．０ｌｏｇ１０以上４．０ｌｏｇ１０未満であった。すなわち、実施例１、２は、上述する参考例１、２のいずれに対しても非常に優れた除菌効果（ウイルスの不活化効果）を示すことがわかった。
また、上記報告書によれば、ウイルスの減少は、４ｌｏｇ１０以上の減少は十分な効果あり、２ｌｏｇ１０以上４ｌｏｇ１０未満の減少は効果あり、２ｌｏｇ１０未満の減少は効果なしという評価基準で評価されており、かかる評価基準に基づいて判断した場合、実施例１は、十分な効果があると判定される。
On the other hand, according to the known feline calicivirus evaluation shown as a comparison target, in Reference Example 1 in which sodium hypochlorite was used, the amount of virus reduction was less than 2.0 log 10 by the same test, and In Reference Example 2 using a commercially available ethanol-based disinfectant, which is a designated non-medicinal product, the amount of virus reduction was 2.0 log 10 or more and less than 4.0 log 10 by the same test. That is, it was found that Examples 1 and 2 showed a very excellent sterilizing effect (virus inactivating effect) with respect to any of the above-mentioned Reference Examples 1 and 2.
In addition, according to the above report, the virus is evaluated based on the evaluation criteria that a decrease of 4log10 or more is sufficiently effective, a decrease of 2log10 or more and less than 4log10 is effective, and a decrease of 2log10 or less is ineffective. When the judgment is made based on the evaluation criteria, it is judged that the first embodiment has a sufficient effect.
（ウイルスの不活化評価２）
以下の方法で、実施例１、２の不活化を評価した。本評価に用いたウイルスと、宿主細胞は以下のとおりである。
・ネコカリシウイルス（Ｆ９）、ＡＴＣＣ ＶＲ−７８２
宿主細胞：ネコ腎臓細胞（ＣＲＦＫ細胞）、ＡＴＣＣ ＣＣＬ−９４
・インフルエンザウイルス（Ｈ１Ｎ１Ａ／ＰＲ８／３／４）、ＡＴＣＣ ＶＲ−１４６９
宿主細胞：イヌ腎細胞（ＭＤＣＫ細胞）、ＡＴＣＣ ＣＣＬ−３４
(Virus inactivation evaluation 2)
The inactivation of Examples 1 and 2 was evaluated by the following method. The viruses and host cells used in this evaluation are as follows.
・ Feline calicivirus (F9), ATCC VR-782
Host cells: cat kidney cells (CRFK cells), ATCC CCL-94
-Influenza virus (H1N1A / PR8 / 3/4), ATCC VR-1469
Host cells: canine kidney cells (MDCK cells), ATCC CCL-34
上述するウイルスを、約１×１０７ｐｆｕ／ｍＬ前後になるよう、滅菌水にて希釈してウイルス液を調製した。そして実施例１、２それぞれの除菌用組成物を用い、除菌用組成物：ウイルス液＝９：１の割合で、接触させた（反応時ウイルス濃度１〜８×１０６ｐｆｕ／ｍＬ）。除菌用組成物とウイルス液とを６０秒間接触させた後、直ちにダルベッコ改変イーグル培地（Ｄ−ＭＥＭ）にて１０倍ずつ希釈し、１０倍段階希釈系を作成した。
事前に６ウェルプレートに播種培養し上層の培地を吸引除去し、ダルベッコリン酸緩衝液（Ｄ−ＰＢＳ（−））２ｍＬで洗浄した宿主細胞に上述のとおり調製した１０倍段階希釈系溶液を１ｍＬずつ添加し、１時間３７℃の感染処理後、細胞上の試験希釈液を吸引し、各２ｍＬのＰＢＳで洗浄吸引後、０．８％オキソイド寒天培地に置換し、３７℃で二日間静置培養した。プラーク形成を確認後、グルタルアルデヒド溶液にて固定し、メチレンブルー染色を行った。形成されたプラーク数を目測によりカウントし、フィルム処理による残存ウイルス力価の変化をプラーク形成率測定によって評価し、不活化率（％）を算出し、表１に示した。尚、除菌用組成物の代わりに滅菌水用い、滅菌水：ウイルス液＝９：１の割合で接触させたものをコントロールとし、接触直後のコントロールを上述と同様に試験し、これを初プラーク数とした。
実施例１、実施例２は、いずれも、ネコカリシウイルスおよびインフルエンザウイルスに対し非常に高い不活化率を示した。
The above-mentioned virus was diluted with sterile water so as to be about 1 × 10 7 pfu / mL to prepare a virus solution. Then, the sterilizing compositions of Examples 1 and 2 were used and brought into contact with each other at a ratio of sterilizing composition: virus solution = 9: 1 (virus concentration at the time of reaction 1 to 8 × 10 6 pfu / mL). .. After contacting the sterilizing composition with the virus solution for 60 seconds, the mixture was immediately diluted 10-fold with Dulbecco's modified Eagle's medium (D-MEM) to prepare a 10-fold serial dilution system.
1 mL of the 10-fold serial diluted solution prepared as described above was added to the host cells which had been seeded and cultured in a 6-well plate in advance, the upper medium was aspirated and removed, and washed with 2 mL of d'Albeccholine acid buffer (D-PBS (-)). Add each, and after infection treatment at 37 ° C for 1 hour, aspirate the test diluted solution on the cells, wash and aspirate with 2 mL of PBS each, replace with 0.8% oxoid agar medium, and leave at 37 ° C for 2 days. It was cultured. After confirming plaque formation, it was fixed with a glutaraldehyde solution and stained with methylene blue. The number of plaques formed was counted by eye, the change in residual virus titer due to film treatment was evaluated by measuring the plaque formation rate, and the inactivation rate (%) was calculated and shown in Table 1. In addition, sterilized water was used instead of the sterilizing composition, and the one contacted with sterilized water: virus solution = 9: 1 was used as the control, and the control immediately after the contact was tested in the same manner as described above, and this was the first plaque. It was a number.
Both Example 1 and Example 2 showed a very high inactivation rate against feline calicivirus and influenza virus.
（抗菌試験）
大腸菌（Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ；ＮＢＲＣ３９７２）、および黄色ブドウ球菌（Ｓｔａｐｈｙｌｏｃｏｃｃｕｓ ａｕｒｅｕｓ；ＮＢＲＣ１２７３２）を用いて以下のとおり試験を行った。
大腸菌および黄色ブドウ球菌はそれぞれ所定の培地で２４〜４８時間培養後、トーマ血球盤により顕微鏡下で濃度を測定し、およそ１０７ｃｆｕ／ｍｌオーダーとなるように滅菌生理食塩水で段階希釈し、被検菌液を調製した。被検菌液の調整は試験当日速やかに行い、直ちに試験に供した。尚、被検菌液は、別途段階希釈法により菌数測定を実施した。
(Antibacterial test)
Escherichia coli (NBRC3972) and Staphylococcus aureus (NBRC12732) were used in the following tests.
After each E. coli and Staphylococcus aureus 24-48 hours at a predetermined medium, the Thoma blood cell board to determine the concentration under a microscope, and serially diluted in sterile saline so as to be approximately 10 7 cfu / ml order A test bacterial solution was prepared. The test bacterial solution was adjusted promptly on the day of the test and immediately subjected to the test. The bacterial count of the test bacterial solution was separately measured by a serial dilution method.
実施例１、２それぞれをガラス製試験管に１０ｍｌずつ分注し、これに上述のとおり調製した被検菌液を０．１ｍｌ添加して反応液を得た。１５秒経過後、直ちに反応液１ｍｌと固化前のＳＣＤ寒天培地とを混合し、混釈培養を行った。コントロールとして、生理食塩水に同様に被検菌液を添加し、接種直後（０秒）に回収した。またネガティブコントロールとして菌を添加しない反応液を用意し、同様に培養（原倍のみ）を行った。そして、３５℃４８時間培養し、出現コロニーを計数した。コロニー数が多い場合には、区画に分けて計数後に全体のコロニー数を算出した。
上述のとおり算出し、コントロールでの菌数を初発菌数とし、初発菌数に対する残存菌数の比率（百分率）を求め、これを除菌率とした。結果は表１に示す。
10 ml of each of Examples 1 and 2 was dispensed into a glass test tube, and 0.1 ml of the test bacterial solution prepared as described above was added thereto to obtain a reaction solution. Immediately after 15 seconds, 1 ml of the reaction solution and the SCD agar medium before solidification were mixed, and pour culture was carried out. As a control, the test bacterial solution was similarly added to physiological saline and recovered immediately after inoculation (0 seconds). In addition, as a negative control, a reaction solution to which no bacteria was added was prepared, and the same culture was performed (only the original size). Then, the cells were cultured at 35 ° C. for 48 hours, and the appearance colonies were counted. When the number of colonies was large, the total number of colonies was calculated after counting by dividing into sections.
Calculated as described above, the number of bacteria in the control was taken as the initial number of bacteria, the ratio (percentage) of the number of residual bacteria to the number of initial bacteria was obtained, and this was used as the eradication rate. The results are shown in Table 1.
腸管出血性大腸菌Ｏ１５７（Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ Ｏ１５７：Ｈ７ ＲＩＭＤ ０５０９９３９）、サルモネラ菌（Ｓａｌｍｏｎｅｌｌａ ｅｎｔｅｒｉｔｉｄｉｓ ＮＢＲＣ ３３１３）、腸炎ビブリオ菌（Ｖｉｂｒｉｏ ｐａｒａｈａｅｍｏｌｙｔｉｃｕｓ ＮＢＲＣ １２７１１）、緑膿菌（Ｐｓｅｕｄｏｍｏｎａｓ ａｅｒｕｇｉｎｏｓａ ＮＢＲＣ １３２７５）に対する抗菌性について以下のとおり試験を行った。
腸管出血性大腸菌Ｏ１５７、サルモネラ菌、腸炎ビブリオ菌、緑膿菌はそれぞれ所定の培地で２４〜４８時間培養後、トーマ血球盤により顕微鏡下で濃度を測定し、およそ１０７ｃｆｕ／ｍｌオーダーとなるように滅菌生理食塩水で段階希釈し、被検菌液を調製した。被検菌液の調整は試験当日速やかに行い、直ちに試験に供した。尚、被検菌液は、別途段階希釈法により菌数測定を実施した。
Intestinal hemorrhagic Escherichia coli O157 (Escherichia coli O157: H7 RIMD 0509939), Salmonella enteridisis NBRC 3313, Vibrio parahaemolyticus bacillus (Vibrio parahaemolyticus NBRC 127) Was done.
Enterohemorrhagic Escherichia coli O157, Salmonella, Vibrio parahaemolyticus, after each Pseudomonas aeruginosa 24-48 hours at a predetermined medium, the Thoma blood cell board to determine the concentration under a microscope, so as to be approximately 10 7 cfu / ml order The test bacterial solution was prepared by serially diluting with sterile physiological saline. The test bacterial solution was adjusted promptly on the day of the test and immediately subjected to the test. The bacterial count of the test bacterial solution was separately measured by a serial dilution method.
実施例１、２それぞれをガラス製試験管に１０ｍｌずつ分注し、これに上述のとおり調製した被検菌液を０．１ｍｌ添加して反応液を得た。３０秒経過後、直ちに反応液に２倍濃度ＮＢ培地０．１ｍｌを添加し、十分に攪拌した後回収して滅菌生理食塩水による１０倍から１０４倍までの希釈を速やかに行った。得られた原倍、１０２倍、１０３倍および１０４倍希釈液１ｍｌと固化前のＳＣＤ寒天培地とを混合し、混釈培養を行った。コントロールとして、生理食塩水に同様に被検菌液を添加し、接種直後（０秒）および３０秒後に回収した。またネガティブコントロールとして菌を添加しない反応液を用意し、同様に培養（原倍のみ）を行った。そして、３５℃４８時間培養し、出現コロニーを計数した。コロニー数が多い場合には、区画に分けて計数後に全体のコロニー数を算出した。
上述のとおり算出し、コントロールでの菌数を初発菌数とし、初発菌数に対する残存菌数の比率（百分率）を求め、これを除菌率とした。結果は表１に示す。
10 ml of each of Examples 1 and 2 was dispensed into a glass test tube, and 0.1 ml of the test bacterial solution prepared as described above was added thereto to obtain a reaction solution. After 30 seconds, it was added immediately reaction solution 2 fold concentration NB medium 0.1 ml, was carried out quickly enough dilution from 10 times by recovering and sterile saline was stirred until 104 times. Obtained original magnification, 10 twice, before solidification and 10 3-fold and 10 4 fold dilution 1ml an SCD agar culture medium were mixed and subjected to pour culture. As a control, the test bacterial solution was similarly added to physiological saline and recovered immediately after inoculation (0 seconds) and 30 seconds later. In addition, as a negative control, a reaction solution to which no bacteria was added was prepared, and the same culture was performed (only the original size). Then, the cells were cultured at 35 ° C. for 48 hours, and the appearance colonies were counted. When the number of colonies was large, the total number of colonies was calculated after counting by dividing into sections.
Calculated as described above, the number of bacteria in the control was taken as the initial number of bacteria, the ratio (percentage) of the number of residual bacteria to the number of initial bacteria was obtained, and this was used as the eradication rate. The results are shown in Table 1.
上記抗菌試験の結果、実施例１、２により、大腸菌、黄色ブドウ球菌は、１５秒後には９９．９９９％超除菌されたことが確認された。
また、同様に実施例１、２により、腸管出血性大腸菌Ｏ１５７、サルモネラ属菌、腸炎ビブリオ菌、緑膿菌は、３０秒後には９９．９９％超除菌されたことが確認された。
As a result of the above antibacterial test, it was confirmed by Examples 1 and 2 that Escherichia coli and Staphylococcus aureus were eradicated by more than 99.999% after 15 seconds.
Similarly, from Examples 1 and 2, it was confirmed that enterohemorrhagic Escherichia coli O157, Salmonella spp., Vibrio parahaemolyticus, and Pseudomonas aeruginosa were eradicated by more than 99.99% after 30 seconds.
（Ｉｎ Ｖｉｔｒｏ皮膚一次刺激性試験）
実施例１の組成物を用い、皮膚に対する刺激性試験を行った。
外来性物質（刺激物）と表皮細胞が接触して細胞傷害が誘導される。これが一次刺激性接触皮膚炎(Ｉｒｒｉｔａｎｔ Ｃｏｎｔａｃｔ Ｄｅｒｍａｔｉｔｉｓ，ＩＣＤ)の原因と考えられている。そこで本試験では、被験物質（実施例１）と表皮細胞との接触による細胞傷害を指標としたＩｎ Ｖｉｔｒｏ再生ヒト表皮試験法によって皮膚一次刺激性を評価するものである。すなわち、１５時間培養したＲｅｃｏｎｓｔｒｕｃｔｅｄ Ｈｕｍａｎ Ｅｐｉｄｅｒｍｉｓ（ＲｈＥ）モデルに被験物質の組成物を１５分間曝露した後、３−（４，５−ｄｉｍｅｔｈｙｌｔｈｉａｚｏｌ−２−ｕｌ）−２，５−ｄｉｐｈｅｎｙｌｔｅｔｒａｚｏｌｉｕｍ ｂｒｏｍｉｄｅ（ＭＴＴ，ＣＡＳ Ｎｏ．２９８−９３−１，Ｓｉｇｍａ−Ａｌｄｒｉｃｈ）の取り込み量を基にしたＭＴＴ還元法によって細胞生存率を測定し、被験物質の皮膚一次刺激性を評価した。
(In vitro primary skin irritation test)
The skin irritation test was performed using the composition of Example 1.
External substances (stimulants) come into contact with epidermal cells to induce cytotoxicity. This is believed to be the cause of primary irritant contact dermatitis (ICD). Therefore, in this test, the primary skin irritation is evaluated by the In vitro regenerated human epidermal test method using the cytotoxicity due to the contact between the test substance (Example 1) and the epidermal cells as an index. That is, after exposing the composition of the test substance to a Reconstructed Human Epidermis (RhE) model cultured for 15 minutes for 15 minutes, then 3- (4,5-dimethylthialzol-2-ul) -2,5-diphenyltellazolium bromide (MTT, CAS). The cell viability was measured by the MTT reduction method based on the uptake amount of No. 298-93-1, Sigma-Aldrich), and the primary skin irritation of the test substance was evaluated.
具体的には、ヒト三次元培養表皮を用いたＩｎ Ｖｉｔｒｏ再生ヒト表皮試験法による皮膚一次刺激性試験代替法により実施例１の刺激性を評価した。上記Ｉｎ Ｖｉｔｒｏ再生ヒト表皮試験法は、ＯＥＣＤテストガイドライン４３９［ＯＥＣＤ Ｇｕｉｄｅｌｉｎｅｓ ｆｏｒ ｔｈｅ Ｔｅｓｔｉｎｇ ｏｆ Ｃｈｅｍｉｃａｌｓ， Ｉｎ Ｖｉｔｒｏ Ｓｋｉｎ Ｉｒｒｉｔａｔｉｏｎ：Ｒｅｃｏｎｓｔｒｕｃｔｅｄ Ｈｕｍａｎ Ｅｐｉｄｅｒｍｉｓ Ｔｅｓｔ Ｍｅｔｈｏｄ（Ａｄｏｐｔｅｄ ２６ Ｊｕｌｙ ２０１３）］に則って実施した。 Specifically, the irritation of Example 1 was evaluated by an alternative method of the primary skin irritation test by the In vitro regenerated human epidermis test method using a human three-dimensional cultured epidermis. The above-mentioned In Viro regenerated human epidermis test method was carried out according to the OECD test guideline 439 [OECD Guidelines for the Testing of Chemicals, In Viro Skin Irritation: Reconstructed Human Epidermis Test 26] Test 26.
上述する所定の方法で上記細胞生存率を測定した結果、実施例１の細胞生存率は５０％を超えた。ＯＥＣＤテストガイドライン４３９の判定基準は、細胞生存率≦５０％で刺激性と判定され、細胞生存率＞５０％で非刺激性と判定される。かかる判定基準から、実施例１は非刺激性と判定された。 As a result of measuring the cell viability by the above-mentioned predetermined method, the cell viability of Example 1 exceeded 50%. The criteria of the OECD test guideline 439 is that cell viability ≤ 50% is determined to be irritating, and cell viability> 50% is determined to be non-irritating. From such a criterion, Example 1 was determined to be non-irritating.
（Ｉｎ Ｖｉｔｒｏ眼刺激性試験）
実施例１の組成物を用い、眼に対する刺激性試験を行った。
目に異物が入った場合、眼への刺激性は最表面の角膜上皮細胞に対する細胞傷害から始まる。そこで本試験では、角膜上皮細胞に対する細胞毒性を指標として眼刺激性を評価するＳＴＥ法を用いた。すなわち、コンフルエントに単層培養したウサギ角膜由来細胞株（Ｓｔａｔｅｎｓ Ｓｅｒｕｍｉｎｓｔｉｔｕｔ Ｒａｂｂｉｔ Ｃｏｒｎｅａ， ＳＩＲＣ細胞）に、０．０５％（ｖ／ｖ）、５．０％（ｖ／ｖ）に希釈した被験物質希釈液（実施例１希釈液）を５分間曝露した。次に３−（４，５−ｄｉｍｅｔｈｙｌｔｈｉａｚｏｌ−２−ｕｌ）−２，５−ｄｉｐｈｅｎｙｌｔｅｔｒａｚｏｌｉｕｍ ｂｒｏｍｉｄｅ （ＭＴＴ，ＣＡＳ Ｎｏ．２９８−９３−１，Ｓｉｇｍａ−Ａｌｄｒｉｃｈ，ＵＳＡ）の取り込み量をもとにしたＭＴＴ還元法によって細胞生存率を測定し、被験物質の眼刺激性を評価した。
尚、眼に入った異物の大部分がヒトでは１〜２分で眼内から排出され、ウサギでは３〜４分で８０％が排出されると報告されている。これら実際の曝露条件を考慮して、ＳＴＥ法では通常の細胞毒性と比べて短時間曝露の試験として設計されている。
(In vitro eye irritation test)
An eye irritation test was performed using the composition of Example 1.
When a foreign body enters the eye, irritation to the eye begins with cytotoxicity to the outermost corneal epithelial cells. Therefore, in this test, we used the STE method to evaluate ocular irritation using cytotoxicity to corneal epithelial cells as an index. That is, a diluted solution of the test substance diluted to 0.05% (v / v) and 5.0% (v / v) in a rabbit corneal-derived cell line (Statens Seruminstat Rabbit Cornea, SIRC cell) cultured in a confluent monolayer. (Example 1 diluted solution) was exposed for 5 minutes. Next, MTT reduction based on the uptake amount of 3- (4,5-dimethylthialzol-2-ul) -2,5-diphenyltetrazolium bromide (MTT, CAS No. 298-93-1, Sigma-Aldrich, USA). Cell viability was measured by the method and the eye irritation of the test substance was evaluated.
It has been reported that most of the foreign substances that have entered the eye are excreted from the eye in 1 to 2 minutes in humans and 80% in 3 to 4 minutes in rabbits. Considering these actual exposure conditions, the STE method is designed as a short-term exposure test compared to normal cytotoxicity.
上記ＳＴＥ法として、ＯＥＣＤテストガイドライン４９１［ＯＥＣＤ Ｇｕｉｄｅｌｉｎｅｓ ｆｏｒ ｔｈｅ Ｔｅｓｔｉｎｇ ｏｆ Ｃｈｅｍｉｃａｌｓ，Ｓｈｏｒｔ Ｔｉｍｅ Ｅｘｐｏｓｕｒｅ Ｉｎ Ｖｉｔｒｏ Ｔｅｓｔ Ｍｅｔｈｏｄ ｆｏｒ Ｉｄｅｎｔｉｆｙｉｎｇ Ｃｈｅｍｉｃａｌｓ Ｉｎｄｕｃｉｎｇ Ｓｅｒｉｏｕｓ Ｅｙｅ Ｄａｍａｇｅ ａｎｄ Ｃｈｅｍｉｃａｌｓ Ｎｏｔ Ｒｅｑｕｉｒｉｎｇ Ｃｌａｓｓｉｆｉｃａｔｉｏｎ ｆｏｒ Ｅｙｅ Ｉｒｒｉｔａｔｉｏｎ ｏｒ Ｓｅｒｉｏｕｓ Ｅｙｅ Ｄａｍａｇｅ（Ａｄｏｐｔｅｄ ２８ Ｊｕｌｙ ２０１５） ]を参考に実施した。 As the STE method, OECD Test Guideline 491 [OECD Guidelines for the Testing of Chemicals, Short Time Exposure In Vitro Test Method for Identifying Chemicals Inducing Serious Eye Damage and Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (Adopted 28 July 2015) ] Was carried out with reference to.
上述する所定の方法で上記細胞生存率を測定した結果、被験物質である実施例１への曝露後の細胞生存率の平均値±標準偏差は、０．０５％曝露条件において８０．６±９．１％、５．０％曝露条件において８５．３±８．３％を示し、いずれの条件下でも細胞生存率が７０％を超えた。ＯＥＣＤテストガイドライン４９１では、目に対する重篤な損傷性を有する物質および目刺激性物質と分類されるか否かは、以下のとおり判定される。以下の判定基準に基づき、実施例１は「区分外（非刺激性）」と判定された。
＜非験物質希釈濃度０．０５％＞
平均細胞生存率＞７０％・・・区分外（非刺激性）
平均細胞生存率≦７０％・・・区分１（目に対する重篤な損傷性物質）
＜非験物質希釈濃度５．０％＞
平均細胞生存率＞７０％・・・区分外（非刺激性）
平均細胞生存率≦７０％・・・区分１（目に対する重篤な損傷性物質）
As a result of measuring the cell viability by the above-mentioned predetermined method, the average value ± standard deviation of the cell viability after exposure to Example 1 which is a test substance is 80.6 ± 9 under 0.05% exposure condition. It showed 85.3 ± 8.3% under the exposure conditions of .1% and 5.0%, and the cell viability exceeded 70% under both conditions. In OECD Test Guidelines 491, whether or not a substance is classified as a substance having serious damage to the eye and an eye irritant is determined as follows. Based on the following criteria, Example 1 was determined to be "out of category (non-irritating)".
<Dilution concentration of non-experimental substance 0.05%>
Average cell viability> 70% ・ ・ ・ Not classified (non-irritating)
Average cell viability ≤ 70% ・ ・ ・ Category 1 (Seriously damaging substances to the eyes)
<Dilution concentration of non-experimental substance 5.0%>
Average cell viability> 70% ・ ・ ・ Not classified (non-irritating)
Average cell viability ≤ 70% ・ ・ ・ Category 1 (Seriously damaging substances to the eyes)
本発明の除菌用組成物は、実質的に、有効成分として食品添加物、および食品類などの人体に無害な組成から構成される。そのため、誤って体内に入り、あるいは肌に触れても低刺激性であるため実質的に害がなく、日常において誰でも安心安全に使用することができる。かかる本発明の除菌用組成物は、台所用品、家庭内の家具、子供の玩具などのより安全性が求められる被除菌物の除菌にも好適である。特にグレープフルーツ種子抽出物を含む態様の本発明の除菌用組成物は、上述する本実施例の結果からも明らかなとおり、抗菌作用に優れる上、ネコカリシウイルスやマウスノロウイルスなどのノンエンベロープウイルスだけでなく、インフルエンザウイルスのようなエンベロープウイルスに対しても短時間で非常に優れた不活化を示す。グレープフルーツ種子抽出物、特には食品添加物として使用し得るグレープフルーツ種子抽出物は高価であるところ、本発明の除菌用組成物は非常に少量のグレープフルーツ種子抽出物を含むことで優れた除菌性を示すことから、経済的優位性にも優れる。
また、本発明の除菌用組成物は、安心安全を重視するばかりではなく、除菌効果も高い。これは、塩基性無機塩類と、酸性無機塩類および／または有機酸と、によりｐＨを１０．５以上１１．５以下に調整することによって、適度なアルカリ性を呈するとともに、このｐＨを緩衝作用により維持することを可能としたことによる。
上述する本発明は、たとえば、任意の除菌対象面に噴霧可能なスプレーボトルタイプ、あるいは、布や不織布などに浸漬させて拭き取るタイプなど広範な使用態様に適用できる上、希釈してもｐＨが変わり難いので、濃縮液としても取引可能である。
The sterilizing composition of the present invention is substantially composed of a food additive as an active ingredient and a composition that is harmless to the human body such as foods. Therefore, even if it accidentally enters the body or touches the skin, it is hypoallergenic and therefore substantially harmless, and anyone can use it safely and securely in daily life. The sterilizing composition of the present invention is also suitable for sterilizing sterilized substances that require more safety, such as kitchen utensils, household furniture, and children's toys. In particular, the sterilizing composition of the present invention, which comprises a grapefruit seed extract, has an excellent antibacterial effect and is only a non-enveloped virus such as feline calicivirus or murine norovirus, as is clear from the results of the above-mentioned Examples. Not only, it shows very good inactivation in a short time against enveloped viruses such as influenza virus. Whereas grapefruit seed extract, especially grapefruit seed extract that can be used as a food additive, is expensive, the disinfectant composition of the present invention contains a very small amount of grapefruit seed extract and thus has excellent sterilization properties. Therefore, it is also excellent in economic superiority.
Further, the sterilizing composition of the present invention not only emphasizes safety and security, but also has a high sterilizing effect. By adjusting the pH to 10.5 or more and 11.5 or less with basic inorganic salts, acidic inorganic salts and / or organic acids, it exhibits appropriate alkalinity and maintains this pH by buffering action. By making it possible to do.
The present invention described above can be applied to a wide range of uses such as a spray bottle type that can be sprayed on an arbitrary surface to be sterilized, or a type that is dipped in a cloth or non-woven fabric and wiped off, and the pH changes even when diluted. Since it is difficult, it can be traded as a concentrate.
上述する本発明は、下記の技術的思想を包含する。
（１）食品添加物として認可を受けた塩基性無機塩類と、
食品添加物として認可を受けた酸性無機塩類および／または有機酸と、
水と、を含み、
ｐＨが１０．５以上１１．５以下であることを特徴とする除菌用組成物。
（２）グレープフルーツ種子抽出物を含む上記（１）に記載の除菌用組成物。
（３）前記グレープフルーツ種子抽出物が、抽出物換算で０．００１質量％以上１．０質量％以下の範囲で含まれる上記（２）に記載の除菌用組成物。
（４）前記塩基性無機塩類として塩基性リン酸塩を含む、かつ、
前記酸性無機塩類がとして酸性リン酸塩を含む上記（１）から（３）のいずれか一項に記載の除菌用組成物。
The present invention described above includes the following technical ideas.
(1) Basic inorganic salts approved as food additives and
Acidic inorganic salts and / or organic acids approved as food additives,
Including water
A sterilizing composition having a pH of 10.5 or more and 11.5 or less.
(2) The sterilizing composition according to (1) above, which contains a grapefruit seed extract.
(3) The sterilizing composition according to (2) above, wherein the grapefruit seed extract is contained in the range of 0.001% by mass or more and 1.0% by mass or less in terms of extract.
(4) The basic inorganic salts contain a basic phosphate and have a basic phosphate.
The composition for sterilization according to any one of (1) to (3) above, wherein the acidic inorganic salts contain an acidic phosphate.
Claims (4)
食品添加物として認可を受けた酸性無機塩類および／または有機酸と、
水と、を含み、
ｐＨが１０．５以上１１．５以下であることを特徴とする除菌用組成物。 Basic inorganic salts approved as food additives and
Acidic inorganic salts and / or organic acids approved as food additives,
Including water
A sterilizing composition having a pH of 10.5 or more and 11.5 or less.
前記酸性無機塩類がとして酸性リン酸塩を含む請求項１から３のいずれか一項に記載の除菌用組成物。 The basic inorganic salts include basic phosphate and
The sterilizing composition according to any one of claims 1 to 3, wherein the acidic inorganic salts contain an acidic phosphate.
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