᪂つヨ㦂ἲᥦ᱌᭩             ᖹᡂ 29 ᖺ 10 ᭶ 1 ᪥                                 No. 2017-03⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ࡟㛵ࡍࡿᥦ᱌ ᖹᡂ 29 ᖺ 7 ᭶ 25 ᪥࡟ᮾிࠊᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ࡟࡚㛤ദࡉࢀࡓ᪂つヨ㦂ἲホ౯఍㆟㸦㏻⛠㸸JaCVAM ホ౯఍㆟㸧࡟࠾࠸࡚௨ୗࡢᥦ᱌ࡀ࡞ࡉࢀࡓࠋ  ᥦ᱌ෆᐜ㸸 ⤒⓶㟁Ẽ᢬ᢠヨ㦂ࡣࠊ✀ࠎࡢ໬Ꮫ≀㉁࡟㐺⏝࡛ࡁࠊ⭉㣗ᛶ࠾ࡼࡧ㠀⭉㣗ᛶࢆศ㢮ࡍࡿࡓࡵࡢ௦᭰ἲ࡜ࡋ࡚ࠊ⾜ᨻⓗ฼⏝ࡀྍ⬟࡛࠶ࡿ࡜⪃࠼ࡿࠋࡓࡔࡋࠊEU ࡛ࡣື≀ᐇ㦂ࢆ⾜ࡗࡓ໬⢝ရ➼ࡢ㈍኎ࢆ⚗Ṇࡋ࡚࠸ࡿࡇ࡜࠿ࡽࠊ໬⢝ရ➼ࡢᏳ඲ᛶホ౯࡬ࡢᮏヨ㦂᪉ἲࡢ㐺⏝ࡣၥ㢟ࡀ⏕ࡌࡿᠱᛕࡀ࠶ࡿࠋ
ࡇࡢᥦ᱌᭩ࡣࠊOrganisation for Economic Co-operation and Development (OECD) Test Guideline(TG) 430, In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER)ࢆࡶ࡜࡟ࠊ
⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾࡲ࡜ࡵࡽࢀࡓᩥ᭩ࢆ⏝࠸࡚ࠊJaCVAMホ౯఍㆟ࡀホ౯࠾ࡼࡧ᳨ウࡋࡓ⤖ᯝࠊࡑࡢ᭷⏝ᛶࡀ☜ㄆࡉࢀࡓࡇ࡜࠿ࡽసᡂࡉࢀࡓࠋ ௨ୖࡢ⌮⏤࡟ࡼࡾࠊ⾜ᨻᙜᒁࡢᏳ඲ᛶホ౯᪉ἲ࡜ࡋ࡚⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢ౑⏝ࢆᥦ᱌ࡍࡿࡶࡢ࡛࠶ࡿࠋ 㔝Ὀ㞝                      すᕝ⛅ె኱㔝Ὀ㞝
JaCVAM ホ౯఍㆟ ㆟㛗               JaCVAM 㐠Ⴀጤဨ఍ ጤဨ㛗

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JaCVAM ホ౯఍㆟኱ 㔝 Ὀ 㞝 㸦බ┈㈈ᅋἲே ᮌཎグᛕᶓ὾⏕࿨⛉Ꮫ᣺⯆㈈ᅋ㸧㸸ᗙ㛗㣤 ሯ ᑦ ᩥ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧*஬༑ᔒⰋ᫂ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ ⏕ά⾨⏕໬Ꮫ㒊㸧▼ ஭ 㞝 ஧ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧஭ ୖ ᬛ ᙲ 㸦᪥ᮏච␿ẘᛶᏛ఍㸧௒ ஭ ᩍ Ᏻ 㸦᪥ᮏື≀ᐇ㦂௦᭰ἲᏛ఍㸧ᒾ℩⿱⨾Ꮚ 㸦᪥ᮏ〇⸆ᕤᴗ༠఍㸧⠛ ⏣ ࿴ ಇ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ᮡᒣ┿⌮Ꮚ 㸦᪥ᮏ໬⢝ရᕤᴗ㐃ྜ఍㸧௰ ஭ ಇ ྖ 㸦᪥ᮏ໬Ꮫᕤᴗ༠఍㸧୰ᮧࡿࡾࡇ 㸦⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ㸧す ᕝ ⛅ ె 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧἟ ⃝  ⪽ 㸦᪥ᮏẘᛶᏛ఍㸧㔝 ཱྀ ┿ ᕼ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧**᳃ ⏣  ೺ 㸦᪥ᮏ⎔ቃኚ␗ཎᏛ఍㸧ᶓ 㛵 ༤ 㞝 㸦᪥ᮏ⓶⭵࢔ࣞࣝࢠ࣮࣭᥋ゐ⓶⭵⅖Ꮫ఍㸧                ௵ᮇ㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥*㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 29 ᖺ 3 ᭶ 31 ᪥**㸸ᖹᡂ 29 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥
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JaCVAM 㐠Ⴀጤဨ఍す ᕝ ⛅ ె㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧㸸ጤဨ㛗ᕝ す  ᚭ㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ㸧኱ ཎ  ᣅ㸦ཌ⏕ປാ┬ ་⸆࣭⏕ά⾨⏕ᒁ ་⸆ရᑂᰝ⟶⌮ㄢ㸧ᑠᕝஂ⨾Ꮚ㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ ⑓⌮㒊㸧ຍ ⸨  ⠜㸦ᅜ❧ឤᰁ⑕◊✲ᡤ㸧ㅋ ⏣ Ὀ ᡂ㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ ⸆⌮㒊㸧ᑠụ⣫୍㑻㸦ཌ⏕ປാ┬ ་⸆࣭⏕ά⾨⏕ᒁ ་⸆ရᑂᰝ⟶⌮ㄢ ໬Ꮫ≀㉁Ᏻ඲ᑐ⟇ᐊ㸧⠛ ⏣ ࿴ ಇ㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧㧗 ᮌ ⠜ ஓ㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ ẘᛶ㒊 ື≀⟶⌮ᐊ㸧᮰ 㔝 ṇ ᫂㸦ཌ⏕ປാ┬ ་⸆࣭⏕ά⾨⏕ᒁ ་⸆ရᑂᰝ⟶⌮ㄢ ໬Ꮫ≀㉁Ᏻ඲ᑐ⟇ᐊ㸧ᖹ ᯘ ᐜ Ꮚ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ ẘᛶ㒊㸧ᗈ ℩ ᫂ ᙪ㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ Ᏻ඲ᛶண ホ౯㒊㸧ᘅ ⏣ ග ᜨ㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ῲ ᒸ  Ꮫ㸦ཌ⏕ປാ┬ ་⸆࣭⏕ά⾨⏕ᒁ ་⸆ရᑂᰝ⟶⌮ㄢ ໬Ꮫ≀㉁Ᏻ඲ᑐ⟇ᐊ㸧ᮏ 㛫 ṇ ඘ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ ኚ␗㑇ఏ㒊㸧Ώ 㑔 ఙ ୍㸦ཌ⏕ປാ┬ ་⸆࣭⏕ά⾨⏕ᒁ ་⸆ရᑂᰝ⟶⌮ㄢ㸧ᑠ ᓥ  ⫕ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮ Ᏻ඲ᛶணホ౯㒊➨஧ᐊ㸧㸸஦ົᒁ

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JaCVAM Statement on the
In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method(TER)
At a meeting held on 25 July 2017 at the National Institute of Health Sciences (NIHS) in
Tokyo, Japan, the Japanese Center for the Validation of Alternative Methods (JaCVAM)
Regulatory Acceptance Board unanimously endorsed the following statement: 
Proposal: We consider the Transcutaneous Electrical Resistance Test Method (TER) to besuitable for predicting the skin corrosion potential of a test chemical in a
regulatory context. We are concerned, however, that the use of isolated rat skin inthis test method for the safety evaluation of cosmetic ingredients and products
could be problematic due to the prohibition on animal testing for cosmetics thatare sold in the EU.
This statement was prepared following a review of the Organisation for Economic
Co-operation and Development (OECD) Test Guideline (TG) 430 In Vitro Skin Corrosion:
TER Test Method together with other materials prepared by the Skin Corrosion Testing
JaCVAM Editorial Committee to acknowledge that the results of a review and study by the
JaCVAM Regulatory Acceptance Board have confirmed the usefulness of this assay.
Based on the above, we propose the TER test method as a useful means for assessing skincorrosion potential during safety assessments by regulatory agencies.

Yasuo
suo Ohno
Chairperson
JaCVAM Regulatory Acceptance Board

Akiyoshi Nishikawa
Chairperson
JaCVAM Steering Committee

October 1, 2017v

The JaCVAM Regulatory Acceptance Board was established by the JaCVAM Steering
Committee, and is composed of nominees from the industry and academia.
This statement was endorsed by the following members of the JaCVAM Regulatory
Acceptance Board:

Mr. Yasuo Ohno (Kihara Memorial Yokohama Foundation for the Advancement ofLife Sciences) : Chairperson
Mr. Naofumi Iizuka (Pharmaceuticals and Medical Devices Agency)*
Mr. Yoshiaki Ikarashi (National Institute of Health Sciences: NIHS)
Mr. Noriyasu Imai (Japanese Society for Alternatives to Animal Experiments)
Mr. Tomoaki Inoue (Japanese Society of Immunotoxicology)
Mr. Yuji Ishii (Biological Safety Research Center: BSRC, NIHS)
Ms. Yumiko Iwase (Japan Pharmaceutical Manufacturers Association)
Mr. Takeshi Morita (Japanese Environmental Mutagen Society)
Mr. Shunji Nakai (Japan Chemical Industry Association)
Ms. Ruriko Nakamura (National Institute of Technology and Evaluation)
Mr. Akiyoshi Nishikawa (BSRC, NIHS)
Ms. Maki Noguchi (Pharmaceuticals and Medical Devices Agency)**
Mr. Satoshi Numazawa (Japanese Society of Toxicology)
Mr. Kazutoshi Shinoda (Pharmaceuticals and Medical Devices Agency)
Ms. Mariko Sugiyama (Japan Cosmetic Industry Association)
Mr. Hiroo Yokozeki (Japanese Society for Dermatoallergology and Contact Dermatitis)
Term: From 1st April 2016 to 31st March 2018*: From 1st April 2016 to 31st March 2017**: From 1st April 2017 to 31st March 2018vi

This statement was endorsed by the following members of the JaCVAM Steering Committee
after receiving the report from JaCVAM Regulatory Acceptance Board:

Mr. Akiyoshi Nishikawa (BSRC, NIHS): Chairperson
Mr. Toru Kawanishi (NIHS)
Mr. Manabu Fuchioka (Ministry of Health, Labour and Welfare)
Ms. Yoko Hirabayashi (Division of Toxicology, BSRC, NIHS)
Mr. Akihiko Hirose (Division of Risk Assessment, BSRC, NIHS)
Ms. Mitsue Hirota (Pharmaceutical & Medical Devices Agency)
Mr. Masamitsu Honma (Division of Genetics and Mutagenesis, BSRC, NIHS)
Mr. Yasunari Kanda (Division of Pharmacology, BSRC, NIHS)
Mr. Atsushi Kato (National Institute of Infectious Diseases)
Mr. Kouichirou Koike (Ministry of Health, Labour and Welfare)
Ms. Kumiko Ogawa (Division of Pathology, BSRC, NIHS)
Mr. Taku Oohara (Ministry of Health, Labour and Welfare)
Mr. Kazutoshi Shinoda (Pharmaceuticals and Medical Devices Agency)
Mr. Atsuya Takagi (Animal Management Section of the Division of Toxicology, BSRC,
NIHS)
Mr. Masaaki Tsukano (Ministry of Health, Labour and Welfare)
Mr. Shinichi Watanabe (Ministry of Health, Labour and Welfare)Mr. Hajime Kojima (Division of Risk Assessment, BSRC, NIHS): Secretary

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⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ┠  ḟ
ホ౯఍㆟ሗ࿌᭩ ------------------------------------------------------------------------------------ 1
ホ౯ሗ࿌᭩ ------------------------------------------------------------------------------------------ 7
OECD GUIDELINE FOR THE TESTING OF CHEMICALS 430,
In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER) ----------- 21ixxホ౯఍㆟ሗ࿌᭩

⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ

JaCVAM ホ౯఍㆟

ᖹᡂ 29 ᖺ㸦2017 ᖺ㸧7 ᭶ 25 ᪥

1
1

JaCVAM ホ౯఍㆟኱ 㔝 Ὀ 㞝 㸦බ┈㈈ᅋἲே ᮌཎグᛕᶓ὾⏕࿨⛉Ꮫ᣺⯆㈈ᅋ㸧㸸ᗙ㛗㣤 ሯ ᑦ ᩥ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧*஬༑ᔒⰋ᫂ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ ⏕ά⾨⏕໬Ꮫ㒊㸧▼ ஭ 㞝 ஧ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧஭ ୖ ᬛ ᙲ 㸦᪥ᮏච␿ẘᛶᏛ఍㸧௒ ஭ ᩍ Ᏻ 㸦᪥ᮏື≀ᐇ㦂௦᭰ἲᏛ఍㸧ᒾ℩⿱⨾Ꮚ 㸦᪥ᮏ〇⸆ᕤᴗ༠఍㸧⠛ ⏣ ࿴ ಇ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ᮡᒣ┿⌮Ꮚ 㸦᪥ᮏ໬⢝ရᕤᴗ㐃ྜ఍㸧௰ ஭ ಇ ྖ 㸦᪥ᮏ໬Ꮫᕤᴗ༠఍㸧୰ᮧࡿࡾࡇ 㸦⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ㸧す ᕝ ⛅ ె 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧἟ ⃝  ⪽ 㸦᪥ᮏẘᛶᏛ఍㸧㔝 ཱྀ ┿ ᕼ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧**᳃ ⏣  ೺ 㸦᪥ᮏ⎔ቃኚ␗ཎᏛ఍㸧ᶓ 㛵 ༤ 㞝 㸦᪥ᮏ⓶⭵࢔ࣞࣝࢠ࣮࣭᥋ゐ⓶⭵⅖Ꮫ఍㸧                ௵ᮇ㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥*㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 29 ᖺ 3 ᭶ 31 ᪥**㸸ᖹᡂ 29 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥

22ࣛࢵࢺ᦬ฟ⓶⭵ࢆ⏝࠸ࡿ⤒⓶㟁Ẽ᢬ᢠヨ㦂㸦TER: Transcutaneous Electrical Resistance㸧ࡣࠊ࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ࡢ௦᭰࡜ࡋ࡚㛤Ⓨࡉࢀࡓヨ㦂ἲ࡛࠶ࡿࠋᮏヨ㦂ἲ࡛ࡣࠊ⭉㣗ᛶ≀㉁ࡀゅ㉁ᒙ࡟྾཰
ࡉࢀࡓᚋᣑᩓࡋࠊୗᒙࡢ⾲⓶⣽⬊ࢆയᐖࡍࡿ࡜࠸࠺⪃࠼ࢆࡶ࡜࡟ࠊ⿕㦂≀㉁᭚㟢ᚋࡢ㟁Ẽ᢬ᢠ್ࢆᣦᶆ
࡜ࡋ࡚⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࠋᮏヨ㦂ἲ࡟ࡘ࠸࡚ࡣࠊECVAM㸦European Centre for the Validation of
Alternative Methods㸸Ḣᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮㸧࡛ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࠊESAC㸦ECVAM
Scientific Advisory Committee㸸ECVAM ⛉Ꮫㅎၥ఍㆟㸧ࡸ 1,2,3) ICCVAM㸦Interagency Coordinating Committee
on the Validation of Alternative Methods㸸⡿ᅜ௦᭰ἲ࡟㛵ࡍࡿ┬ᗇ㛫㐃⤡఍㆟㸧4,5)࡟࠾࠸࡚ᮏヨ㦂ἲࡢಙ㢗ᛶ➼ࡀ☜ㄆࡉࢀࡓᚋࠊOECD㸦Organisation for Economic Co-operation and Development㸸⤒῭༠ຊ㛤Ⓨᶵᵓ㸧࡟࡚ࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥ㸦TG㸧430 ࡜ࡋ࡚ 2004 ᖺ࡟ᢎㄆࡉࢀ࡚࠸ࡿࠋࡇࡢ TG ࡣఱᗘ࠿ᨵゞࡉࢀ࡚࠾ࡾࠊ⌧ᅾࡣ 2015 ᖺ∧࡜࡞ࡗ࡚࠸ࡿ 6)ࠋ
JaCVAM ホ౯఍㆟ࡣࠊ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾసᡂࡉࢀࡓࠕ⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩ࠖ7)ࢆ⏝࠸࡚ࠊᮏヨ㦂ἲࡢጇᙜᛶ࡟ࡘ࠸᳨࡚ウࡋࡓࠋ㸯㸬ヨ㦂ἲࡢᐃ⩏ྡ⛠㸸 ⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ௦᭰ࡍࡿᑐ㇟ẘᛶヨ㦂㸸 ࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ヨ㦂ἲࡢᴫ␎㸸ᮏヨ㦂ἲ࡛ࡣࠊࣛࢵࢺ㸦28-30 ᪥㱋࡯࡝㸧࠿ࡽษ㝖ࡋࡓ 20 mm ෇┙≧ࡢ⓶⭵ࢆヨ㦂ࢩࢫࢸ࣒࡜ࡋ࡚౑⏝ࡍࡿࠋ⿕㦂≀㉁ࢆ⾲⓶㠃࡟୍ᐃ᫬㛫㐺⏝ࡋࠊゅ㉁ᒙ࡟྾཰ࡉࢀࡓᚋ
࡟ᣑᩓࡋࠊୗᒙࡢ⾲⓶⣽⬊࡟ᑐࡋയᐖࡍࡿ⛬ᗘࢆࠊ⤒⓶㟁Ẽ᢬ᢠࢆᣦᶆ࡜ࡋ࡚

ᐃࡍࡿࡇ࡜࡟ࡼࡾ⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࠋࡉࡽ࡟ࠊ㟁Ẽ᢬ᢠࡀ 5 kȍ ௨ୗࡢሙྜ࡟ࡣࠊ10㸣ࢫ࣮ࣝ࣍ࣟࢲ࣑ࣥ㹀ᰁⰍᾮ࡟࡚ᰁⰍࡋࠊ⓶⭵࡟྾╔ࡉࢀࡓⰍ⣲ࡢ㔞ࢆ྾ගᗘ࠿ࡽ⟬ฟࡍࡿࠋࡇࡢ⤖ᯝࠊ⤒⓶㟁Ẽ᢬ᢠࡀ 5㹩Ȑࡼࡾᑠࡉࡃࠊ᫂ࡽ࠿࡟ᦆയࡢ࠶ࡿሙྜ㸦౛࠼ࡤ✸Ꮝ㸧ࠊࡲࡓࡣࠊ᫂ࡽ࠿࡞ᦆയࡀ࡞ࡃ࡚ࡶⰍ⣲㔞ࡀ㝧ᛶᑐ↷≀㉁㸦10㹋ሷ㓟㸧ࡢ⤖ᯝ௨ୖࡢሙྜࠊ⓶⭵⭉㣗ᛶ࡜ุᐃࡍࡿࠋ㸰㸬ホ౯࡟⏝࠸ࡓ㈨ᩱཬࡧホ౯ෆᐜࡢ⛉Ꮫⓗጇᙜᛶᮏヨ㦂ἲࡣࠊECVAM ࡟ࡼࡿᗈ࠸⠊ᅖࡢ⿕㦂≀㉁ࢆ⏝࠸ࡓࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡟ࡼࡾࠊጇᙜᛶࡀ᳨ドࡉࢀࠊESAC ࡟ࡼࡾಙ㢗ᛶ࡜෌⌧ᛶࡣ㧗࠸࡜ホ౯ࡉࢀ࡚࠸ࡿ 1,2,3)ࠋࡇࡢ⤖ㄽࡣࠊICCVAM ࡟࠾࠸࡚ࡶ☜ㄆࡉࢀ࡚࠸ࡿࠋࡇࢀࡽࡢ⤖ᯝࡣබ⾲ࡉࢀ࡚࠾ࡾࠊ㏱࡛᫂⊂❧࡞⛉Ꮫⓗホ౯ࡀ⾜ࢃࢀ࡚࠸ࡿ࡜⪃࠼ࡿ 4,5)ࠋ
ᮏヨ㦂ἲࡣࠊOECD ࡟ࡼࡾヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ࡟᥇ᢥࡉࢀࡓ㸦TG 430㸧6)ࠋᮏ㑥࡟࠾࠸࡚ࡣࠊJaCVAM ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍ࡀࠊࡇࢀࡽࡢ㈨ᩱࢆ⏝࠸࡚ᮏヨ㦂ἲࢆホ౯ࡋ࡚࠸ࡿ 7)ࠋࡲࡓࠊᮏヨ㦂ἲࡣࠊ⭉㣗ᛶ≀㉁ࡀゅ㉁ᒙࢆ㏱㐣࣭ᣑᩓࡋࡓࡢࡕ࡟⾲⓶⣽⬊ࢆയᐖࡋࠊ⓶⭵ࡢࣂࣜ࢔ᶵ⬟ࢆയᐖࡍࡿ࡜࠸࠺⓶33⭵യᐖᶵᵓࢆᶍೌࡋ࡚࠾ࡾࠊ⓶⭵ࣂࣜ࢔ᶵ⬟ࡢᣦᶆ࡜ࡋ࡚⤒⓶㟁Ẽ᢬ᢠࢆホ౯ࡋ࡚࠾ࡾࠊ⓶⭵⭉㣗ᛶⓎ⌧ᶵᗎࡀ⪃៖ࡉࢀ࡚࠸ࡿࠋ
௨ୖࡢⅬ࠿ࡽࠊᮏヨ㦂ἲࡢホ౯࡟⏝࠸ࡓ㈨ᩱ࡞ࡽࡧ࡟ホ౯ෆᐜ࡟ࡘ࠸࡚ࡣࠊ⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿ᪉ἲ࡜ࡋ࡚⛉Ꮫⓗጇᙜᛶࡀ࠶ࡿ࡜⪃࠼ࡿࠋ㸱㸬ᮏヨ㦂ἲࡢ᭷⏝ᛶ࡜㐺⏝㝈⏺
ᮏヨ㦂ἲ࡟ᑐࡍࡿ Botham1,8)࡜ Fentem3)ࡢண

ᛶࡢ᳨ウ⤖ᯝ࠾ࡼࡧ ICCVAM ࡟ࡼࡿ㞟ィ⤖ᯝࢆ⾲㸯࡟♧ࡋࡓࠋ୧⪅ࡢ⤖ᯝࢆྜࢃࡏࡓ⤖ᯝ࡜ࡑࢀࡒࢀࡢㄽᩥࡢṇ☜ᗘࠊឤᗘࠊ≉␗ᗘ࡟኱ࡁ࡞ᕪࡣ࡞ࡃࠊṇ☜ᗘࡣ⣙ 81㸣ࠊឤᗘࡣ⣙ 94㸣ࠊ≉␗ᗘࡣ⣙ 71㸣࡛࠶ࡗࡓࠋࡇࡢ୰࡛ࠊഇ㝜ᛶࡣ 3 ≀㉁ (6㸣) ࡟ࡘ࠸࡚ㄆ
ࡵࡽࢀ࡚࠸ࡿࡶࡢࡢࠊࡇࢀࡽ 3 ≀㉁ࡢ⭉㣗ᛶࡢ UN GHS1⣽ศ㢮ࡣ࠸ࡎࢀࡶᙉᗘࡀᙅ࠸ഃࡢࣞ࣋ࣝ࡟࠶ࡿ
(UN GHS ༊ศ 1A ࡟ࡣ┦ᙜࡋ࡞࠸)࡜᥎ᐹࡉࢀࡿࡓࡵࠊࠕ⭉㣗ᛶࡣᙉࡃ࡞࠸࡜ุ᩿࡛ࡁࡿ 7)ࠖ࡜࠸࠺㈨ᩱ
⦅⧩ጤဨ఍ࡢุ᩿ࡣᨭᣢ࡛ࡁࡿࠋ

⾲㸯. TER ࡢணᛶICCVAM ࡟ࡼࡿ㞟

Botham et al. (1992 andFentem et al. (1998)

ィ⤖ᯝ

1996)

ឤᗘ

94% (51/54)

96% (27/28)

93% (25/27)≉␗ᗘ

71% (48/68)

70% (26/37)

71% (22/31)ṇ☜ᗘ

81% (99/122)

82% (53/65)

81% (47/58)

OECD ࡢ in vitro ⭉㣗ᛶヨ㦂࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ࠊ௚࡟ࠕTG431 ࣄࢺ⾲⓶ࣔࢹࣝヨ㦂ࠖ࠾ࡼࡧࠕTG435
in vitro ⭷ࣂࣜ࢔ヨ㦂ࠖࡀ࠶ࡾࠊICCVAM ࡛ࡣᮏヨ㦂ἲ࡜ࡇࢀࡽࡢヨ㦂ἲ (EpiSkinτ, EpiDermτ࠾ࡼࡧ
Corrositex®)࡟ࡘ࠸࡚≉␗ᛶࠊឤᗘࠊṇ☜ᛶ➼ࡢࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀẚ㍑ࡉࢀ࡚࠸ࡿ 5,9,10)ࠋࡇࡢ⤖ᯝࠊᮏヨ㦂ἲࡣ௚ࡢヨ㦂ἲ࡜ྠ➼ࡢணᛶࢆ᭷ࡍࡿ࡜ホ౯ࡉࢀ࡚࠸ࡿࠋ

௨ୖࡢⅬ࠿ࡽࠊᮏヨ㦂ἲࡣ✀ࠎࡢ໬Ꮫ≀㉁࡟ᑐࡋ࡚㐺⏝࡛ࡁࠊࡍ࡭࡚ࡢࢱ࢖ࣉࡢ⭉㣗ᛶ࠾ࡼࡧ㠀⭉㣗ᛶࢆศ㢮ࡍࡿࡓࡵࡢ௦᭰ἲ࡜ࡋ࡚᭷⏝࡛࠶ࡿ࡜⪃࠼ࡿࠋࡓࡔࡋࠊࣉࣟࢺࢥࣝ࠿ࡽ㢮᥎ࡋ࡚ࠊ㞴Ỉ⁐ᛶࡢᅛయࡸ⢓ᛶࡢ㧗࠸≀㉁ࡣ㐺ษ࡟ホ౯࡛ࡁ࡞࠸ྍ⬟ᛶࡀ࠶ࡿࠋࡲࡓࠊUN GHS ࡟ࡼࡿ⭉㣗ᛶࡢ⣽ศ㢮࡬ࡢ㐺⏝࡟ࡣ᳨ウࡀ୙༑ศ࡛࠶ࡾࠊ᭷⏝ᛶࢆホ౯࡛ࡁ࡞࠸ࠋ࡞࠾ࠊᮏ㑥࡟࠾ࡅࡿ⾜ᨻⓗ࡞ཷࡅධࢀ࡟࠾࠸࡚
ࡣࠊUN GHS ༊ศ㸯ࡢ⣽༊ศࢆồࡵࡽࢀ࡚࠸࡞࠸ࠋ

1ᅜ㐃 ໬Ꮫရࡢศ㢮࠾ࡼࡧ⾲♧࡟㛵ࡍࡿୡ⏺ㄪ࿴ࢩࢫࢸ࣒㸦UN GHS㸸United Nations Globally
Harmonized System of Classification and Labelling of Chemicals㸧44㸲㸬┠ⓗ࡜ࡍࡿ≀㉁ཪࡣ〇ရࡢẘᛶࢆホ౯ࡍࡿヨ㦂ἲ࡜ࡋ࡚ࡢࠊ♫఍ⓗཷࡅධࢀᛶཬࡧ⾜ᨻୖࡢ฼⏝ࡢྍ⬟ᛶ ♫఍ⓗཷࡅධࢀᛶ㸸ᮏヨ㦂ἲࡣࠊࣛࢵࢺࡢ᦬ฟ⓶⭵ࢆ⏝࠸࡚࠸ࡿࡓࡵ᏶඲࡞⨨ࡁ᥮࠼ヨ㦂࡛ࡣ࡞࠸ࠋࡋ࠿ࡋ࡞ࡀࡽࠊᮏヨ㦂ἲࢆ⏝࠸ࡿࡇ࡜࡛ື≀ᩘࡢ๐ῶࡀྍ⬟࡛࠶ࡾࠊࡲࡓࠊ⭉㣗ᛶ≀㉁࡟ࡼࡿື≀࡬ࡢࢫࢺࣞࢫࢆ୚࠼ࡿࡇ
࡜ࡣ↓࠸ࡇ࡜࠿ࡽࠊື≀ᐇ㦂௦᭰ἲࡢ㸱Rs ࡢཎ๎࡟㐺ࡗࡓヨ㦂ἲ࡛࠶ࡿࠋᮏヨ㦂ἲࡣࠊᚲせ࡞ᢏ⾡ࡀ」㞧࡛ࡣ࡞ࡃࠊ㐺ษ࡞カ⦎࡟ࡼࡗ࡚ᐜ᫆࡟⩦ᚓ࡛ࡁࡿ࡜⪃࠼ࡽࢀࡿࠋ
௨ୖࡢほⅬ࠿ࡽࠊᮏヨ㦂ἲࡣ♫఍ⓗ࡟ཷࡅධࢀࡽࢀࡿࡶࡢ࡛࠶ࡿ࡜ุ᩿ࡍࡿࠋ ⾜ᨻୖࡢ฼⏝ᛶ㸸ᮏヨ㦂ἲࡣࠊ✀ࠎࡢ໬Ꮫ≀㉁࡟㐺⏝࡛ࡁࠊ⭉㣗ᛶ࠾ࡼࡧ㠀⭉㣗ᛶࢆศ㢮ࡍࡿࡓࡵࡢ௦᭰ἲ࡜ࡋ࡚ࠊ⾜ᨻⓗ฼⏝ࡀྍ⬟࡛࠶ࡿ࡜⪃࠼ࡿࠋࡓࡔࡋࠊEU ࡛ࡣື≀ᐇ㦂ࢆ⾜ࡗࡓ໬⢝ရ➼ࡢ㈍኎ࢆ⚗Ṇࡋ࡚࠸ࡿࡇ
࡜࠿ࡽࠊ໬⢝ရ➼ࡢホ౯࡬ࡢᮏヨ㦂᪉ἲࡢ㐺⏝ࡣၥ㢟ࡀ⏕ࡌࡿᠱᛕࡀ࠶ࡿࠋ
ཧ⪃ᩥ⊩1)Botham, P.A. et al. (1996) A prevalidation study on in vitro skin corrosivity testing: The report andrecommendations of ECVAM workshop 6. ATLA, 23, 219-255

2)

Barratt, M.D. et al. (1998) The ECVAM international validation study on in vitro tests for skin corrosivity. 1.
Selection and distribution of the test chemicals. Toxicology in Vitro, 12, 471-482

3)Fentem, J.H. et al. (1998) The ECVAM International Validation Study on in vitro tests for skin corrosivity. 2.
Results and evaluation by the management team. Toxicology in Vitro, 12, 483-524

4)

ICCVAM (1999) NIH Publication No.99-4495. Corrositex: An in vitro test method for assessing dermal
corrosivity potential of chemicals.

5)

ICCVAM (2002) NIH Publication No.02-4502. ICCVAM Evaluation of EPISKIN, and EpiDerm (EPI-200)
and rat skin transcutaneous electrical resistance (TER) assay: in vitro test method for assessing dermal
corrosivity potential of chemicals.

6)OECD (2015) Guideline for the testing of chemicals. 430, In Vitro Skin Corrosion: Transcutaneous Electrical
Resistance Test Method (TER).

7)

JaCVAM ⓶⭵⭉㣗ᛶ㈨ᩱ⦅⧩ጤဨ఍㸸⓶⭵⭉㣗ᛶヨ㦂ホ౯ሗ࿌᭩ ⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩㸦2017 ᖺ 2 ᭶ 24 ᪥㸧㸬

8)Botham, P.A. et al.㸦1992㸧The skin corrosivity test in vitro. Results of an inter-laboratory trial. Toxicol In
Vitro. 6(3):191-4

9)ICCVAM (1999) NIH Publication No.99-4495. Corrositex: An in vitro test method for assessing dermal
corrosivity potential of chemicals10) JaCVAM ⓶⭵⭉㣗ᛶ㈨ᩱ⦅⧩ጤဨ఍㸸⓶⭵⭉㣗ᛶヨ㦂ホ౯ሗ࿌᭩ ࣄࢺ⾲⓶ࣔࢹࣝࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩㸦2017 ᖺ 2 ᭶ 24 ᪥㸧㸬

5
5

6

ホ౯ሗ࿌᭩

⤒⓶㟁Ẽ᢬ᢠヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍

ᖹᡂ 29 ᖺ㸦2017 ᖺ㸧2 ᭶ 24 ᪥

17ホ౯ጤဨ㧘ᶫ ♸ḟ

ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ ẘᛶ㒊୰ᮧࡿࡾࡇ⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ ໬Ꮫ≀㉁⟶⌮ࢭࣥࢱ࣮㡲᪉ ╩ኵ

ఫ཭໬Ꮫᰴᘧ఍♫ ࣞࢫ࣏ࣥࢩࣈࣝࢣ࢔㒊㸦໬ᏛရᏳ඲ࢢ࣮ࣝࣉ㸧㸭᪥ᮏ໬Ꮫᕤᴗ༠఍ᑠᓥ ⫕ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶணホ౯㒊┠ḟ
1. ヨ㦂ἲࡢ⛉Ꮫⓗ࠾ࡼࡧつไ㠃࠿ࡽࡢጇᙜᛶ
2. ヨ㦂ࣉࣟࢺࢥࣝᵓᡂࡢጇᙜᛶ
3. 㛤Ⓨ࠾ࡼࡧホ౯࡟౑ࢃࢀࡓ≀㉁ࡢศ㢮ࠊ㑅ᢥ⌮⏤ࡢጇᙜᛶ
4. ヨ㦂ἲࡢṇ☜ᛶࢆホ౯ࡍࡿࡓࡵ࡟⏝࠸ࡽࢀࡓ≀㉁ࡢ in vitro ࠾ࡼࡧཧ↷ࢹ࣮ࢱࡢ᭷↓5. ヨ㦂ἲࡢṇ☜ᛶ㸦෌⌧ᛶ㸧
6. ヨ㦂ἲࡢಙ㢗ᛶ
7. ௚ࡢ⛉Ꮫⓗ࡞ሗ࿌࡜ࡢẚ㍑ࡢ᭷↓8. 㸱Rs ཎ๎࡬ࡢ㛵୚㸦ື≀⚟♴㠃࠿ࡽࡢጇᙜᛶ㸧
9. ヨ㦂ἲࡢ᭷⏝ᛶ࡜㝈⏺㸦ࢥࢫࢺࠊ᫬㛫࠿ࡽࡢጇᙜᛶ࡞࡝㸧10. ࡑࡢ௚㸦≉チࡢ᭷↓࡞࡝㸧
11. ⤖ㄽ
12. ᩥ⊩
Annex
Appendix
OECD Guidelines for the Testing of Chemicals, Test No. 430: In Vitro Skin Corrosion: Transcutaneous
Electrical Resistance Test Method (TER)

2
8

せ᪨
࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ࡢື≀ᐇ㦂௦᭰ἲ㸦௦᭰ἲ㸧࡜ࡋ࡚ࠊ⤒῭༠ຊ㛤Ⓨᶵᵓ㸦OECD㸸
Organisation for Economic Co-operation and Development㸧࡟࡚ヨ㦂࢞࢖ࢻࣛ࢖ࣥ㸦TG: Test
Guideline㸧430࡜ࡋ࡚ᢎㄆࡉࢀࡓࣛࢵࢺ᦬ฟ⓶⭵ࢆ⏝࠸ࡿ⤒⓶㟁Ẽ᢬ᢠヨ㦂㸦TER: Transcutaneous
Electrical Resistance㸧ࡢ᭷⏝ᛶࢆホ౯ࡋࡓࠋಙ㢗ᛶ࡜ጇᙜᛶ࡜࠸࠺どⅬ࡟࠾࠸࡚ࠊTERࢆホ౯ࡋࡓ⤖ᯝࠊ㐺⏝⠊ᅖ࡟㝈⏺ࡣ࠶ࡿࡶࡢࡢࠊᮏヨ㦂ἲ࡟ࡼࡾ⭉㣗ᛶࡢ᭷↓ࢆホ౯࡛ࡁࡿ࡜⪃࠼ࡽࢀࡓࠋ
ࡋ࠿ࡋࠊ⏕ࡁ࡚࠸ࡿࣛࢵࢺ࠿ࡽ᥇ྲྀࡋࡓ⓶⭵ࢆ⏝࠸ࡿࡇ࡜࠿ࡽ໬⢝ရ➼ࡢホ౯࡬ࡢᛂ⏝࡟ࡣၥ㢟
ࡀ࠶ࡿࠋ

3
9

1. ヨ㦂ἲࡢ⛉Ꮫⓗ࠾ࡼࡧつไ㠃࠿ࡽࡢጇᙜᛶ
⓶⭵⭉㣗ᛶヨ㦂ࡣ⓶⭵่⃭ᛶヨ㦂ࡢ୍⎔࡜ࡋ࡚⾜ࢃࢀࠊ࢞࢖ࢻࣛ࢖࡛ࣥࡣ Draize ࡽ࡟ࡼࡾᥦၐ
ࡉࢀࡓ࢘ࢧࢠࢆ⏝࠸ࡿ᪉ἲࡀ᥎ዡࡉࢀ࡚࠸ࡿ 1)ࠋࡇࡢ᪉ἲࡣ⿕㦂≀㉁ࡢ่⃭ᛶࡸ⭉㣗ᛶࢆ᳨ฟࡍ
ࡿヨ㦂ἲ࡜ࡋ࡚㛗ࡃ⏝࠸ࡽࢀ࡚ࡁࡓࡶࡢࡢࠊุᐃࢆ⫗║࡛⾜࠺ࡓࡵᐈほᛶ࡟ஈࡋࡃᐇ㦂㛫ࡸ᪋タ㛫࡛ࡢ෌⌧ᛶࡀஈࡋ࠸ࠊ᭦࡟ື≀࡟⃭ࡋ࠸③ࡳ࡜ࢫࢺࣞࢫࢆ୚࠼ࡿࡇ࡜ࡀ♫఍ⓗ࡟ၥ㢟࡜࡞ࡾࠊ௨๓ࡼࡾື≀ࢆ౑⏝ࡋ࡞࠸௦᭰ἲࡢ㛤Ⓨࡀษᮃࡉࢀ࡚࠸ࡓࠋࡇࡢື≀ᐇ㦂௦᭰ἲ㸦௨ୗࠊ௦᭰ἲ
࡜グࡍ㸧࡜ࡋ࡚ࠊ⤒῭༠ຊ㛤Ⓨᶵᵓ㸦OECD㸸Organisation for Economic Co-operation and Development㸧࡟࡚ᢎㄆࡉࢀࡓヨ㦂࢞࢖ࢻࣛ࢖ࣥ㸦TG: Test Guideline㸧430 ࡟ࡣࠊ⤒⓶㟁Ẽ᢬ᢠヨ㦂㸦TER:
Transcutaneous Electrical Resistance)ࡀグ㍕ࡉࢀ࡚࠸ࡿ 2㸧ࠋࡇࡢヨ㦂ἲࡣࠊ⭉㣗ᛶ≀㉁ࡀゅ㉁ᒙ࡟྾
཰ࡉࢀࡓᚋᣑᩓࡋࠊୗᒙࡢ⾲⓶⣽⬊ࢆയᐖࡍࡿࡇ࡜࡟ࡼࡾ⓶⭵ࡢࣂࣜ࢔࣮ᶵ⬟ࢆയᐖࡍࡿ࡜࠸࠺
⪃࠼ࢆࡶ࡜࡟ࠊ⿕㦂≀㉁᭚㟢ᚋࡢ㟁Ẽ᢬ᢠ್ࢆᣦᶆ࡟⓶⭵⭉㣗ᛶࢆホ౯ࡋ࡚࠸ࡿࠋᮏヨ㦂ἲࡣḢᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮(ECVAM㸸European Centre for the Validation of Alternative Methods㸧࡟ࡼࡿࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࢆ⤒࡚ࠊḢᕞ࡛ࡣ໬Ꮫ≀㉁ࡢ⓶⭵⭉㣗ᛶホ౯ࢆ┠ⓗ࡜ࡋ࡚ᢎㄆࡉࢀࠊ໬Ꮫ≀㉁ࡢࣜࢫࢡ⾲♧ࡸ㆑ู➼࡟฼⏝ࡉࢀ࡚࠸ࡿࠋ≉࡟᫖௒࡛ࡣᅜ㐃໬Ꮫရࡢศ㢮࠾ࡼࡧ⾲♧࡟㛵ࡍࡿୡ⏺ㄪ࿴ࢩࢫࢸ࣒㸦UN GHS㸸United Nations Globally Harmonized System of Classification and Labellingof Chemicals㸧ศ㢮࡟ᚑࡗ࡚ホ౯ࡉࢀࡿࢣ࣮ࢫࡀቑ࠼࡚࠸ࡿࠋ
ᡃࡀᅜ࡛᪤Ꮡࡢ໬Ꮫ≀㉁ࢆホ౯ࡍࡿሙྜࠊOECD ࡛ᢎㄆࡉࢀࡓヨ㦂᪉ἲ࡟ࡼࡿ⤖ᯝࢆ฼⏝ࡍࡿࡇ࡜ࡣྍ⬟࡛࠶ࡿࡀࠊ⌧ᅾࡲ࡛௦᭰ἲ࡛ࡢ⤖ᯝࢆࡶ࡜࡟⭉㣗ᛶࢆホ౯ࡉࢀࡓ౛ࡣከࡃ࡞࠸ࠋᏳ඲ᛶホ౯࡟࠾ࡅࡿ௦᭰ἲࡢᬑཬࡀษᮃࡉࢀ࡚࠸ࡿ⌧≧࡟࠾࠸࡚ࠊᡃࡀᅜ࡛ࡶ✚ᴟⓗ࡟ཷࡅධࢀࡿࡇ࡜ࡀᚲせ࡜࡞ࡗ࡚࠸ࡿࠋ
ࡇࢀࡽࡢ≧ἣ࡟㚷ࡳࠊᮏホ౯᭩࡛ࡣࠊOECD TG430 ࡟ᥖ㍕ࡉࢀࡓ TER ࡢ⭉㣗ᛶホ౯࡟࠾ࡅࡿ
᭷⏝ᛶࢆホ౯ࡋࡓ 2㸧ࠋ
2. ヨ㦂ࣉࣟࢺࢥࣝᵓᡂࡢጇᙜᛶ 2㸧1) ཎ⌮   ษ㝖ࡋࡓࣛࢵࢺ⓶⭵ࢆヨ㦂ࢩࢫࢸ࣒࡜ࡋ࡚౑⏝ࡋࠊࡑࡢ㟁Ẽ᢬ᢠࡢኚ໬ࢆ⓶⭵യᐖᛶࡢᣦᶆ࡜ࡍࡿࠋ
2㸧᪉ἲ   ᦬ฟࡋࡓ┤ᚄ20 mm෇┙≧ࡢ⓶⭵㸦ࣛࢵࢺࠊ28-30᪥㱋࡯࡝㸧ࢆᅗ㸯࡟♧ࡍࡼ࠺࡟ࠊ
ࢸࣇࣟࣥ㸦࣏ࣜࢸࢺࣛࣇ࢚ࣝ࢜ࣟࢳࣞࣥ㸸PTFE㸧ࢳ࣮ࣗࣈࡢୗ㒊࡟⾲⓶㠃ࢆୖ࡟ࡋ࡚࠶࡚ࠊࢦ࣒〇ࡢOࣜࣥࢢ࡛ࡣࡉࡴࠋ⓶⭵ࡢࡘ࠸ࡓPTFEࢳ࣮ࣗࣈࢆ◲㓟࣐ࢢࢿࢩ࣒࢘㸦154mM㸧⁐ᾮ࡟ࡘࡅࠊ⿕㦂≀㉁⁐ᾮ150 ȝLࢆ⾲⓶㠃࡟ࠊ30Υ࡛24᫬㛫㐺⏝ࡍࡿࠋWheatstone㟁ᶫ⿦⨨࡟ప㟁ᅽࢆ࠿ࡅࠊ100 Hz࡟࠾ࡅࡿ㟁Ẽ᢬ᢠᗘࢆᐃࡍࡿࠋ     ࡉࡽ࡟ࠊ㟁Ẽ᢬ᢠࡀ 5 kȍ ௨ୗࡢሙྜ࡟ࡣࠊ10㸣ࢫ࣮ࣝ࣍ࣟࢲ࣑ࣥ㹀ᰁⰍᾮ࡟࡚ 2 ᫬㛫ᰁⰍࡋࠊ⓶⭵࡟྾╔ࡉࢀࡓⰍ⣲ࡢ㔞ࢆ྾ගᗘ࠿ࡽ⟬ฟࡍࡿࠋ     㟁Ẽ᢬ᢠᗘࡀ 5 kȍ ࡼࡾᑠࡉࡃࠊ᫂ࡽ࠿࡟ᦆയࡢ࠶ࡿሙྜࠊࡲࡓࡣᦆയࡀ࡞ࡃ࡚ࡶⰍ
⣲㔞ࡀ㝧ᛶᑐ↷≀㉁࡛࠶ࡿ 10 M ሷ㓟௨ୖࡢሙྜࠊ⭉㣗ᛶ࡜ุ᩿ࡍࡿࠋ

410ᅗ㸯㸬TER ᴫせᅗ 2㸧
3. 㛤Ⓨ࠾ࡼࡧホ౯࡟౑ࢃࢀࡓ≀㉁ࡢศ㢮ࠊ㑅ᢥ⌮⏤ࡢጇᙜᛶ
Botham ࡽ࡟ࡼࡾ TER ࡟ࡘ࠸࡚ࡣࠊ20 ࡢ⿕㦂≀㉁࡟ࡼࡾ᪋タෆ෌⌧ᛶ 3㸧ࠊ࠾ࡼࡧ 50 ࡢ⿕㦂≀㉁࡟ࡼࡾ᪋タ㛫෌⌧ᛶ࠾ࡼࡧண

ᛶࡀㄪ࡭ࡽࢀ࡚࠸ࡿ 4㸧ࠋࡑࢀࡽࡢ≀㉁ࣜࢫࢺ࡜ࡑࢀࡽࡢ≀⌮࣭໬ᏛᏛⓗᛶ≧ࢆ ANNEX 1 ࠾ࡼࡧ 2 ࡟♧ࡍࠋࡲࡓ Fentem ࡽ࡟ࡼࡾࠊ60 ≀㉁ࢆ⏝࠸࡚ࠊ෌⌧ᛶ࠾ࡼࡧணᛶࡀㄪ࡭ࡽࢀ࡚࠸ࡿ 5㸧ࠋࡑࢀࡽࡢ≀㉁ࣜࢫࢺࢆ⭉㣗ᛶࡸ pH, つไ≧ἣ࡜࡜ࡶ࡟ ANNEX 3࡟♧ࡍࠋࡑࢀࡽ࡛ࢃ࠿ࡿࡼ࠺࡟ࠊᮏヨ㦂ἲࡣᖜᗈ࠸ᛶ≧ࡢ໬Ꮫ≀㉁᳨࡛ウࡉࢀ࡚࠸ࡿࠋ
4. ヨ㦂ἲࡢṇ☜ᛶࢆホ౯ࡍࡿࡓࡵ࡟⏝࠸ࡽࢀࡓ≀㉁ࡢ in vitro ࠾ࡼࡧཧ↷ࢹ࣮ࢱࡢ᭷↓⿕㦂≀㉁ࡢከࡃࡣ ECVAM ࡢ⓶⭵⭉㣗ᛶヨ㦂ࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑⏝ࡉࢀࡓ≀㉁࡛࠶ࡾ 6)ࠊࡑࢀ
ࡽࡢ⭉㣗ᛶ࡟㛵ࡍࡿཧ↷ࢹ࣮ࢱࡀධᡭྍ⬟࡛࠶ࡿ
5. ヨ㦂ἲࡢṇ☜ᛶ㸦෌⌧ᛶ㸧 TER ࡢ᪋タෆ෌⌧ᛶࡣ 20 ࡢ⿕㦂≀㉁㸦6 ⭉㣗ᛶ≀㉁࠾ࡼࡧ 14 㠀⭉㣗ᛶ≀㉁㸧ࢆ⏝࠸ࠊ3 ᪋タ
511㛫࡛ ANOVA ゎᯒࢆ⏝࠸ࠊ᪋タ㛫࡟᭷ពᕪࡀ࡞࠸࡜ࡉࢀ࡚࠸ࡿ 3㸧ࠋ ୍᪉ࠊ᪋タ㛫෌⌧ᛶ࡛ࡣࠊ2 ᪋タࡀ 50 ࡢ⿕㦂≀㉁㸦25 ⭉㣗ᛶ≀㉁࠾ࡼࡧ 25 㠀⭉㣗ᛶ≀㉁㸧ࢆ⏝࠸ࠊ92㸣㸦࠸ࡎࢀࡢ≀㉁⩌ࡶ 23 ≀㉁࡛ࡢ⤖ᯝࡀ୍⮴ࡋࡓ㸧࡛࠶ࡿ࡜ࡉࢀ࡚࠸ࡿ 4㸧ࠋ௨ୖࡢ⤖ᯝ࠿ࡽࠊ෌⌧ᛶࡣ㧗࠸࡜ุ᩿ࡉࢀ࡚࠸ࡿࠋ
60 ≀㉁ࢆ⏝࠸ࡓ 3 ᪋タ࡛ࡢ 2 way ࡢ ANOVA ゎᯒ࡟࠾࠸࡚ࠊ᪋タෆ࠾ࡼࡧ᪋タ㛫ࡢኚື࡟᭷ព
࡞ᕪࡣ࡞࠸࡜ุ᩿ࡉࢀࡓ 5)ࠋࡲࡓࠊ60 ≀㉁㸦27 ⭉㣗ᛶ≀㉁࠾ࡼࡧ 33 㠀⭉㣗ᛶ≀㉁㸧ࡢ࠺ࡕࠊ37≀㉁ࡣ 3 ᪋タ࡜ࡶ᪋タෆ࠾ࡼࡧ᪋タ㛫෌⌧ᛶࡀㄆࡵࡽࢀࡓࠋṧࡿ 23 ≀㉁ࡢ࠺ࡕ 10 ≀㉁࡛᪋タෆ෌⌧ᛶࡀᝏ࠿ࡗࡓࠋࡇࡢࡼ࠺࡟ࠊ࠸ࡃࡘ࠿ࡢ≀㉁࡛෌⌧ᛶࡢ⤖ᯝࡀ␗࡞ࡗ࡚࠸ࡓࡀࠊECVAM ࡣᮏヨ㦂ἲࡢಙ㢗ᛶ࡜෌⌧ᛶࡣ㧗࠸࡜ุ᩿ࡋࡓࠋࡇࡢ⤖ㄽࡣࠊECVAM ⛉Ꮫㅎၥ఍㆟ (ESAC:ECVAM Scientific Advisory Committee)7㸧࠾ࡼࡧ⡿ᅜࡢ௦᭰ἲ࡟㛵ࡍࡿ┬ᗇ㛫㐃⤡఍㆟ (ICCVAM㸸
Interagency Coordinating Committee on the Validation of Alternative Methods)8㸧࡛ࡢホ౯࡟࠾࠸࡚ࡶ☜ㄆࡉࢀࡓࠋ
6. ヨ㦂ἲࡢಙ㢗ᛶ Botham et al (1996)࡜Fentem et al (1998)ࡢ᳨ウ⤖ᯝ࠾ࡼࡧࡑࢀࡽࢆྜ⟬ࡋࡓICCVAMࡢ㞟ィ⤖ᯝ6)ࢆ⾲㸯࡟♧ࡍࠋࡑࡢ⤖ᯝ࡜ࡑࢀࡒࢀࡢㄽᩥࡢṇ☜ᗘࠊឤᗘࠊ≉␗ᗘ࡟኱ࡁ࡞ᕪࡣ࡞ࡃࠊṇ☜ᗘࡣ
⣙81㸣ࠊឤᗘࡣ⣙94㸣ࠊ≉␗ᗘࡣ⣙71㸣࡛࠶ࡗࡓࠋഇ㝜ᛶࡣ6㸣ࠊ3≀㉁ㄆࡵࡽࢀࠊࡇࢀࡽࡣ
MethacroleinࠊGlycol bromoacetate (85%)࠾ࡼࡧ1,2-Benzisothiazolin-3-one (33%) in aqueous propylene
glycol࡛࠶ࡗࡓࠋMethacrolein࠾ࡼࡧGlycol bromoacetate (85%)ࡣUNࡢ⭉㣗ᛶࡢ⣽ศ㢮࡛࠸ࡎࢀࡶᙉ
ࡃ࡞࠸ࡶࡢ(1B/1C)࡟ᒓࡍࡿࡶࡢ࡛࠶ࡾࠊ1,2-Benzisothiazolin-3-one (19.3%)࡟࠾࠸࡚ࡣ⓶⭵่⃭ᛶࡶ࡞࠸࡜໬Ꮫ≀㉁➼Ᏻ඲ࢹ࣮ࢱࢩ࣮ࢺ (MSDS㸸Material Safety Data Sheet) ࡟グ㍕ࡀ࠶ࡿ㸦ᮍῧ௜㈨ᩱ㸧ࠋ௨ୖࡢ⤖ᯝ࠿ࡽࠊୖグഇ㝜ᛶ࡜࡞ࡗࡓ3≀㉁ࡢ⭉㣗ᛶࡣᙉࡃ࡞࠸࡜ุ᩿࡛ࡁࡿࠋ
✀ࠎࡢ໬Ꮫ≀㉁ศ㢮࡟ࡼࡿᕪࡣ࡞ࡃࠊTER ࡣ⭉㣗ᛶ࠾ࡼࡧ㠀⭉㣗ᛶࢆศ㢮ࡍࡿࡓࡵࡢ௦᭰ἲ࡜
ࡋ࡚᭷⏝࡛࠶ࡿࠋࡓࡔࡋࠊࡑࢀ௨ୖࡢ⣽ศ㢮࡟ࡘ࠸࡚ࡣ᳨ウࡀ୙༑ศ࡛࠶ࡿ࡜ࡉࢀ࡚࠸ࡿࠋ
⾲㸯. TER ࡢணᛶFentem et al. (1998) 5㸧

ICCVAM (1999) ࡢBotham et al. (1992 and⥲ィ 6㸧1996)3,4㸧ṇ☜ᗘ

81% (99/122)

82% (53/65)

81% (47/58)

ឤᗘ

94%(51/54)

96%(27/28)

93%(25/27)≉␗ᗘ71%(48/68)

70%(26/37)

71%(22/31)

7. ௚ࡢ⛉Ꮫⓗ࡞ሗ࿌࡜ࡢẚ㍑ࡢ᭷↓ OECD ࡢ⭉㣗ᛶヨ㦂௦᭰ἲ࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ࠊ௚࡟ࠕTG431 ࣄࢺ⾲⓶ࣔࢹࣝヨ㦂ࠖ9)࠾ࡼࡧ
ࠕTG435 in vitro ⭷ࣂࣜ࢔ヨ㦂ࠖ10㸧ࡀᢎㄆࡉࢀ࡚࠸ࡿࠋࡇࢀࡽࡣ࠸ࡎࢀࡶࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࠊICCVAM ࡣࡇࢀࡽࡢヨ㦂ἲ(Rat Skin TER, EpiSkin™, EpiDerm™࠾ࡼࡧ Corrositex®)ࡢṇ☜ᗘࠊឤᗘࠊ≉␗ᗘࠊ࡟ࡘ࠸࡚ẚ㍑ࡋࡓ⤖ᯝࢆ⾲ 2 ࡟♧ࡍࠋヨ㦂≀㉁ࡢᩘ㔞ࡸ㑅ᢥ≀㉁ࡢ✀㢮ࡀ␗࡞ࡗ࡚࠸ࡿࡓࡵ⤖ᯝࡢุᐃࡔࡅࢆࡶࡗ࡚ࠊ༢⣧࡟ᙜヱヨ㦂ἲࡢඃ㉺ᛶࢆホ౯ࡍࡿࡇ࡜ࡣᅔ㞴࡛࠶ࡿࡀࠊᙜヱヨ㦂ἲࡣ௚ࡢヨ㦂ἲ࡜ྠ➼ࡢண࡜ᛮࢃࢀࡿࠋ
12

6

ᛶࢆ᭷ࡍࡿ

⾲ 2㸬ヨ㦂ἲࡢẚ㍑⤖ᯝ 6,8㸧TER

EpiSkin

EpiDerm

Corrositex

≀㉁ᩘ

122

60

24

163

ṇ☜ᗘ

81% (99/122)

83% (50/60)

92% (22/24)

79% (128/163)

ឤᗘ

94%(51/54)

82%(23/28)

92%(11/12)

85%(76/89)≉␗ᗘ71%(48/68)

84%(27/32)

83%(10/12)

72%(52/74)

8. 㸱Rs ཎ๎࡬ࡢ㛵୚㸦ື≀⚟♴㠃࠿ࡽࡢጇᙜᛶ㸧 ᮏヨ㦂ἲࡣࠊࣛࢵࢺࡢ᦬ฟ⓶⭵ࢆ⏝࠸࡚࠸ࡿࡓࡵ᏶඲࡞⨨ࡁ᥮࠼ヨ㦂࡜ࡣ࡞ࡽ࡞࠸ࠋࡋ࠿ࡋࠊ
Ᏻᴦẅ࡟ࡼࡾ⓶⭵ࢆ᦬ฟࡍࡿࡇ࡜ࠊ1 ༉࠿ࡽᚓࡽࢀࡿ⓶⭵࡛」ᩘࡢヨ㦂ࡀᐇ᪋ྍ⬟࡛࠶ࡿࠋࡋࡓ
ࡀࡗ࡚ࠊື≀ᩘࡢ๐ῶࡀྍ⬟࡛࠶ࡾࠊࡲࡓࠊ⭉㣗ᛶ≀㉁࡟ࡼࡿື≀࡬ࡢࢫࢺࣞࢫࢆ୚࠼ࡿࡇ࡜ࡣ
↓ࡃࠊື≀ᐇ㦂௦᭰ἲ࡜ࡋ࡚㸱Rs ࡢཎ๎࡟㐺ࡗࡓヨ㦂ἲ࡛࠶ࡿࠋ
9. ヨ㦂ἲࡢ᭷⏝ᛶ࡜㝈⏺
 ᮏヨ㦂ἲࡣ✀ࠎࡢ✀㢮ࡢ໬Ꮫ≀㉁࡟ᑐࡋ࡚㐺⏝࡛ࡁࡿࡀࠊࣉࣟࢺࢥࣝ࠿ࡽ㢮᥎ࡋ࡚ࠊ㞴Ỉ⁐ᛶࡢᅛయࡸ⢓ᛶࡢ㧗࠸≀㉁ࡣ㐺ษ࡟ホ౯࡛ࡁ࡞࠸ྍ⬟ᛶࡀ࠶ࡿࠋ⭉㣗ᛶࡢ⣽ศ㢮㸦UN GHS ࡟ࡼࡿ㸧࡟ࡣ౑⏝࡛ࡁ࡞࠸࡜ゝࢃࢀ࡚࠸ࡿ 2㸧ࠋ⾲ 3 ࡟♧ࡍ⩦⇍ᗘ☜ㄆ≀㉁ࢆ⏝࠸ࡿࡇ࡜࡟ࡼࡾᑓ㛛ᢏ⾡ࡢ⩦⇍࡟ࡘ࠸࡚☜ㄆࡍࡿࡇ࡜ࡀ࡛ࡁࡿ 2)ࠋ
10. ࡑࡢ௚㸦≉チࡢ᭷↓࡞࡝㸧 ≉チ࡟ࡘ࠸࡚ࡣ♧ࡉࢀ࡚࠸࡞࠸ࠋ
11. ⤖ㄽ
ಙ㢗ᛶ࡜ጇᙜᛶ࡜࠸࠺どⅬ࡟࠾࠸࡚ࠊࣛࢵࢺ᦬ฟ⓶⭵ࢆ⏝࠸ࡿ⤒⓶㟁Ẽ᢬ᢠヨ㦂㸦TER㸧ࢆホ౯ࡋࡓ⤖ᯝࠊᮏጤဨ఍ࡣᙜヱヨ㦂ἲࢆ⏝࠸ࢀࡤࠊ⭉㣗ᛶࡢ᭷↓ࢆホ౯࡛ࡁࡿ࡜⪃࠼ࡓࠋ

713

⾲ 3 ⩦⇍ᗘ☜ㄆ୍ぴ⾲ 2)
໬Ꮫ≀㉁

1

CASRN

UN GHS ༊ศ 3

VRM ༊ศ4

In vivo ヨ㦂⤖ᯝ࡟ᇶ࡙ࡃIn vivo ࡟࠾ࡅࡿ⭉㣗ᛶ≀㉁

In vitro ヨ㦂⤖ᯝ࡟ᇶ࡙ࡃ

໬Ꮫ≀㉁
2
ࢡࣛࢫ

≀⌮
ⓗ≧
ែ

pH5

N,N’- ࢪ࣓ࢳ ࣝ ࢪ ࣉࣟࣆࣞࣥࢺࣜ࢔࣑ࣥ

10563-29-8

᭷ᶵሷᇶ

1A

6×⭉㣗ᛶ

ᾮయ

8.3

1,2-ࢪ࢔࣑ࣀࣉࣟࣃࣥ

78-90-0

᭷ᶵሷᇶ

1A

6×⭉㣗ᛶ

ᾮయ

8.3

◲㓟㸦10%㸧

7664-93-9↓ᶵ㓟(1A/)1B/1C

5×⭉㣗ᛶ
1×㠀⭉㣗ᛶ

ᾮయ

1.2

1310-58-3↓ᶵሷᇶ(1A/)1B/1C

6×⭉㣗ᛶ

ᾮయ

13.2

124-07-2

᭷ᶵ㓟

1B/1C

4×⭉㣗ᛶ
2×㠀⭉㣗ᛶ

ᾮయ

3.6

88-18-6

ࣇ࢙ࣀ࣮ࣝ

1B/1C

4×⭉㣗ᛶ
2×㠀⭉㣗ᛶ

ᾮయ

3.9Ỉ㓟໬࣒࢝ࣜ࢘
㸦10㸣aq㸧㸧࢜ࢡࢱࣥ㓟㸦࢝ࣉࣜࣝ 㓟㸧2-tert- ࣈࢳ ࣝ ࣇ࢙ ࣀ࣮ࣝIn vivo ࡟࠾ࡅࡿ㠀⭉㣗ᛶ≀㉁࢖ࢯࢫࢸ࢔ࣜࣥ㓟2724-58-5

᭷ᶵ㓟

㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᾮయ

3.6

4- ࢔࣑ ࣀ -1,2,4- ࢺࣜ࢔ ࢰ࣮ࣝ

584-13-4

᭷ᶵሷᇶ

㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᅛయ

5.5⮯໬ࣇ࢙ࢿࢳࣝ

103-63-9

ồ㟁Ꮚ≀㉁

㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᾮయ

3.64-㸦࣓ࢳࣝࢳ࢜㸧-࣋ࣥࢬ࢔ࣝࢹࣄࢻ

3446-89-7 ồ㟁Ꮚ≀㉁㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᾮయ

6.8

1,9-ࢹ࢝ࢪ࢚ࣥ

1647-16-1

୰ᛶ᭷ᶵ
≀ ㉁

㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᾮయ

3.9ࢸࢺࣛࢡ࢚ࣟࣟࢳࣞࣥ

127-18-4

୰ᛶ᭷ᶵ
≀ ㉁

㠀⭉㣗ᛶ

6×㠀⭉㣗ᛶ

ᾮయ

4.5

␎ㄒ㸸aq.=Ỉ⁐ᾮࠊCASRN=CAS Ⓩ㘓␒ྕࠊUN GHS=ᅜ㐃໬Ꮫရࡢศ㢮࠾ࡼࡧ⾲♧࡟㛵ࡍࡿୡ⏺ㄪ࿴ࢩࢫࢸ࣒(1)ࠊ VRM=ࣂࣜࢹ࣮ࢩࣙࣥ῭ࡳཧ↷ヨ㦂ἲ1㸸ࡇࢀࡽࡢ≀㉁ࡣࠊ᭱ึ࡟⭉㣗ᛶ≀㉁࡜㠀⭉㣗ᛶ≀㉁ࠊḟ࡟⭉㣗ᛶࡢ⣽༊ศࠊࡉࡽ࡟໬Ꮫ≀㉁ࢡࣛࢫ
ࡢ㡰࡛ศ㢮ࡉࢀࠊḢᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮㸦ECVAM㸧࡟ࡼࡿࣛࢵࢺ TER

ᐃἲࡢࣂࣜࢹ࣮ࢩࣙࣥ

11㸧

ヨ㦂࡟⏝࠸ࡽࢀࡿ≀㉁࠿ࡽ㑅ᢥࡉࢀࡓࡶࡢ࡛࠶ࡿ㸦Barratt et al 1998

, Fentem et al 19985㸧㸧ࠋ≉࡟᩿ࡾࡀ࡞࠸㝈ࡾࠊᥖ㍕ࡋࡓ≀㉁ࡣᕷ㈍ရ㉎ධ᫬ࡢ⣧ᗘ࡛ヨ㦂ࢆ⾜ࡗࡓ㸦Barratt et al 199811㸧㸧ࠋྍ⬟࡞㝈ࡾࠊḟࡢᇶ‽ࢆ‶ࡓࡍ≀㉁ࢆ㑅ᢥࡋࡓࠋ(i)VRM ࡟ࡼࡿᐃࡲࡓࡣணࢆ⾜࠼ࡿ⭉㣗ᛶ཯ᛂࡢ⠊ᅖࢆ௦⾲ࡍࡿ≀㉁࡛࠶ࡿࡇ࡜㸦౛࠼ࡤࠊ㠀⭉㣗ᛶ≀㉁ࠊ⭉㣗ᛶ≀㉁ࡢ࠺ࡕ཯ᛂࡀᙅ࠸≀㉁࠿ࡽᙉ࠸≀㉁ࡲ࡛㸧
ࠋ(ii)ࣂࣜࢹ࣮ࢩࣙࣥヨ㦂࡟⏝࠸ࡽࢀࡓ໬Ꮫ≀㉁ࢡࣛࢫࢆ௦⾲ࡍࡿ≀㉁࡛࠶ࡿࡇ࡜ࠋ(iii)VRM ࡢᛶ⬟≉ᛶࢆ཯ᫎࡍࡿ≀㉁࡛࠶ࡿࡇ࡜ࠋ(iv)᫂☜࡟ᐃ⩏ࡉࢀࡓ໬Ꮫᵓ㐀ࢆᣢࡘ≀㉁࡛࠶ࡿࡇ࡜ࠋ(v) In
vivo ࡟࠾ࡅࡿཧ↷ヨ㦂ἲ࡛᫂☜࡞⤖ᯝࢆᑟࡃ≀㉁࡛࠶ࡿࡇ࡜ࠋ(vi)ᕷ㈍ရ࡛࠶ࡿࡇ࡜ࠋ(vii)ᴟ➃࡟㧗㢠࡞ᗫᲠ㈝⏝ࢆకࢃ࡞࠸≀㉁࡛࠶ࡿࡇ࡜ࠋ
2㸸Barratt et al㸦199811㸧㸧࡟ࡼࡾᣦᐃࡉࢀࡓ໬Ꮫ≀㉁ࢡࣛࢫ࡜ࡋࡓࠋ
3㸸UN GHS ࡢ 1Aࠊ1Bࠊ1C ࡟┦ᙜࡍࡿᅜ㐃ᐜჾ➼⣭ࡣࡑࢀࡒࢀ IࠊIIࠊIII ࡛࠶ࡿࠋ4㸸ࣂࣜࢹ࣮ࢩࣙࣥ࡟࡚᪋タ㛫෌⌧ᛶ⤖ᯝࢆグ㍕ࡋ࡚࠸ࡿࠋ⅊Ⰽ㒊ศࡣ 6 ᐇ㦂ࡢุᐃ⤖ᯝࡀ␗࡞ࡿࡇ࡜

148

ࢆព࿡ࡍࡿࠋ
5㸸pH ್ࡣ Fentem et al5㸧 ࠾ࡼࡧ Barratt et al11㸧 ࡢᩥ⊩࠿ࡽᚓࡓࠋ

12.ᩥ⊩1) OECD (2015) Guideline for the testing of chemicals. 404, Acute Dermal Irritation/Corrosion.
2) OECD (2015) Guideline for the testing of chemicals. 430, in vitro Skin Corrosion: Transcutaneouselectrical resistance test (TER).
3) Botham, P.A. et al.㸦1992㸧The skin corrosivity test in vitro. Results of an inter-laboratory trial. Toxicol
In Vitro, 6:191-4
4) Botham, P.A. et al. (1996) A prevalidation study on in vitro skin corrosivity testing: The report andrecommendations of ECVAM workshop 6. ATLA, 23, 219-255
5) Fentem, J.H. et al. (1998) The ECVAM International Validation Study on in vitro tests for skincorrosivity. 2. Results and evaluation by the management team. Toxicology in Vitro, 12, 483-524
6) ICCVAM (1999) NIH Publication No.99-4495. Corrositex: An in vitro test method for assessing dermal
corrosivity potential of chemicals.
7) EC-ECVAM (1998) Statement on the Scientific Validity of the Rat Skin Transcutaneos Electrical
Resistance (TER) Test (an In Vitro Test for Skin Corrosivity), Issued by the ECVAM Scientific
Advisory Committee (ESAC10), 3 April 1998. Available at: [http://www.ecvam.jrc.ec.europa.eu.html].
8) ICCVAM (2002) NIH Publication No.02-4502. ICCVAM Evaluation of EPISKIN, and EpiDerm(EPI-200) and rat skin transcutaneous electrical resistance (TER) assay: in vitro test method forassessing dermal corrosivity potential of chemicals.
9) OECD (2016) Guideline for the testing of chemicals. 431, in vitro Skin Corrosion: Human skin model
test.
10) OECD (2015) Guideline for the testing of chemicals. 435, in vitro membrane barrier test method for
Skin Corrosion.
11) Barratt, M.D. et al. (1998) The ECVAM international validation study on in vitro tests for skincorrosivity. 1. Selection and distribution of the test chemicals. Toxicology in Vitro, 12, 471-482

9

15

ANNEX1: Botham 1992 ࡢࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑ࢃࢀࡓ 20 ≀㉁   Code No. 1㹼6 ࡀ⭉㣗ᛶ≀㉁ࠊࡑࡢ௚ࡣ㠀⭉㣗ᛶ≀㉁

10

16

ANNEX 2: Botham 1996 ࡢࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑ࢃࢀࡓ 50 ≀㉁

11

17

12
18

13
19

ANNEX 3: Fentem 1998 ࡢࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑ࢃࢀࡓ 60 ≀㉁20

14

OECD/OCDE

430
Adopted:28 July 2015

OECD GUIDELINE FOR THE TESTING OF CHEMICALSIn Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER)

INTRODUCTION
1.
Skin corrosion refers to the production of irreversible damage to the skin manifested as visible
necrosis through the epidermis and into the dermis, following the application of a test chemical [as definedby the United Nations (UN) Globally Harmonized System of Classification and Labelling of Chemicals(GHS)] (1). This updated Test Guideline 430 provides an in vitro procedure allowing the identification of
non-corrosive and corrosive substances and mixtures in accordance with UN GHS (1).
2.
The assessment of skin corrosivity has typically involved the use of laboratory animals(OECD Test Guideline 404 (TG 404); originally adopted in 1981, and revised in 1992, 2002 and 2015) (2).
In addition to the present TG 430, other in vitro test methods for testing of skin corrosion potential ofchemicals have been validated and adopted as OECD Test Guidelines 431 (3) and 435 (4), that are alsoable to identify sub-categories of corrosive chemicals when required. Several validated in vitro test
methods have been adopted as OECD TG 439 (5), to be used for the testing of skin irritation. A document
on Integrated Approaches to Testing and Assessment (IATA) for Skin Corrosion and Irritation describes
several modules which group various information sources and analysis tools and provides guidance on (i)how to integrate and use existing testing and non-testing data for the assessment of skin irritation and skincorrosion potentials of chemicals and (ii) proposes an approach when further testing is needed (6).
3.
This Test Guideline addresses the human health endpoint skin corrosion. It is based on the rat
skin transcutaneous electrical resistance (TER) test method, which utilizes skin discs to identify corrosivesby their ability to produce a loss of normal stratum corneum integrity and barrier function. This Test
Guideline was originally adopted in 2004 and updated in 2015 to refer to the IATA guidance document.
4.
In order to evaluate in vitro skin corrosion testing for regulatory purposes, pre-validation studies(7) followed by a formal validation study of the rat skin TER test method for assessing skin corrosion
were conducted (8) (9) (10) (11). The outcome of these studies led to the recommendation that the TER
test method (designated the Validated Reference Method – VRM) could be used for regulatory purposes
for the assessment of in vivo skin corrosivity (12) (13) (14).
5.
Before a proposed similar or modified in vitro TER test method for skin corrosion other than the
VRM can be used for regulatory purposes, its reliability, relevance (accuracy), and limitations for its
proposed use should be determined to ensure its similarity to the VRM, in accordance with therequirements of the Performance Standards (15). The Mutual Acceptance of Data will only be guaranteed
after any proposed new or updated test method following the PS of this Test Guideline have been reviewed
and included in this Test Guideline.
DEFINITIONS
6.

Definitions used are provided in Annex 1.
1© OECD, (2015)
You are free to use this material for personal, non-commercial purposes without seeking prior consent
from the OECD, provided the source is duly mentioned. Any commercial use of this material is subject towritten permission from the OECD.

21

430

OECD/OCDEINITIAL CONSIDERATIONS
7.
A validation study (10) and other published studies (16) (17) have reported that the rat skin TER
test method is able to discriminate between known skin corrosives and non-corrosives with an overallsensitivity of 94% (51/54) and specificity of 71% (48/68) for a database of 122 substances.
8.
This Test Guideline addresses in vitro skin corrosion. It allows the identification of noncorrosive and corrosive test chemicals in accordance with the UN GHS (1). A limitation of this Test
Guideline, as demonstrated by the validation studies (8) (9) (10) (11), is that it does not allow the subcategorization of corrosive substances and mixtures in accordance with the UN GHS (1). The regulatory
framework in member countries will decide how this Test Guideline will be used. While this Test
Guideline does not provide adequate information on skin irritation, it should be noted that OECD TG 439
specifically addresses the health effect skin irritation in vitro (5). For a full evaluation of local skin effects
after a single dermal exposure, the Guidance Document n. 203 on Integrated Approaches for Testing
Assessment should be consulted (6).
9.
A wide range of chemicals representing mainly substances has been tested in the validation
underlying this Test Guideline and the empirical database of the validation study amounted to
60 substances covering a wide range of chemical classes (8) (9). On the basis of the overall data available,
the Test Guideline is applicable to a wide range of chemical classes and physical states including liquids,
semi-solids, solids and waxes. However, since for specific physical states test items with suitable reference
data are not readily available, it should be noted that a comparably small number of waxes and corrosive
solids were assessed during validation. The liquids may be aqueous or non-aqueous; solids may be soluble
or insoluble in water. In cases where evidence can be demonstrated on the non-applicability of the Test
Guideline to a specific category of substances, the Test Guideline should not be used for that specificcategory of substances. In addition, this Test Guideline is assumed to be applicable to mixtures as anextension of its applicability to substances. However, due to the fact that mixtures cover a wide spectrumof categories and composition, and that only limited information is currently available on the testing of
mixtures, in cases where evidence can be demonstrated on the non-applicability of the Test Guideline to a
specific category of mixtures (e.g. following a strategy as proposed by Eskes et al., 2012) (18), the Test
Guideline should not be used for that specific category of mixtures. Before use of the Test Guideline on a
mixture for generating data for an intended regulatory purpose, it should be considered whether, and if sowhy, it may provide adequate results for that purpose. Such considerations are not needed, when there is aregulatory requirement for testing of the mixture. Gases and aerosols have not been assessed yet invalidation studies (8) (9). While it is conceivable that these can be tested using the TER test method, thecurrent Test Guideline does not allow testing of gases and aerosols.
PRINCIPLE OF THE TEST10.
The test chemical is applied for up to 24 hours to the epidermal surfaces of skin discs in a twocompartment test system in which the skin discs function as the separation between the compartments.
The skin discs are taken from humanely killed rats aged 28-30 days. Corrosive chemicals are identified bytheir ability to produce a loss of normal stratum corneum integrity and barrier function, which is measured
as a reduction in the TER below a threshold level (16) (see paragraph 32). For rat skin TER, a cut-off valueof 5k: has been selected based on extensive data for a wide range of substances where the vast majority ofvalues were either clearly well above (often > 10 k:), or well below (often < 3 k:) this value (16).
Generally, test chemicals that are non-corrosive in animals but are irritant or non-irritant do not reduce the
TER below this cut-off value. Furthermore, use of other skin preparations or other equipment may alter the
cut-off value, necessitating further validation.

2© OECD, (2015)22

OECD/OCDE

430

11.
A dye-binding step is incorporated into the test procedure for confirmation testing of positive
results in the TER including values around 5 k:. The dye-binding step determines if the increase in ionicpermeability is due to physical destruction of the stratum corneum. The TER method utilizing rat skin has
shown to be predictive of in vivo corrosivity in the rabbit assessed under OECD guideline 404 (2).
DEMONSTRATION OF PROFICIENCY
12.
Prior to routine use of the rat skin TER test method that adheres to this Test Guideline,
laboratories should demonstrate technical proficiency by correctly classifying the twelve Proficiency
Substances recommended in Table 1. In situations where a listed substance is unavailable or where
justifiable, another substance for which adequate in vivo and in vitro reference data are available may be
used (e.g. from the list of reference chemicals (16)) provided that the same selection criteria as described inTable 1 is applied.
Table 1: List of Proficiency Substances1
UN GHS
Cat. Basedon In Vivo
Results3
In Vivo Corrosives

VRM
Cat. Basedon In Vitro
Results

PhysicalState

Substance

CASRN

ChemicalClass2

N,N’-Dimethyl
dipropylenetriamine

10563-29-8

organic base

1A

6xC

L

8.3

1,2-Diaminopropane

78-90-0

organic base

1A

6xC

L

8.3

Sulfuric acid (10%)

7664-93-9

inorganic acid(1A/)1B/1C

5xC
1x NC

L

1.2

1310-58-3

inorganic base(1A/)1B/1C

6xC

L

13.2

124-07-2

organic acid

1B/1C

L

3.6

88-18-6

phenol

1B/1C

L

3.9

Potassium hydroxide(10% aq.)
Octanoic (Caprylic)acid
2-tert-Butylphenol

4xC
2 x NC
4xC
2 x NC

pH4

In Vivo Non-corrosives
Isostearic acid2724-58-5

organic acidNC

6 x NCL

3.6

4-Amino-1,2,4triazole

584-13-4organic baseNC

6 x NCS

5.5

Phenethyl bromide

103-63-9

electrophileNC

6 x NCL

3.6

4-(Methylthio)benzaldehyde

3446-89-7

electrophileNC

6 x NCL

6.8

1,9-Decadiene

1647-16-1

neutral organicNC

6 x NC

L

3.9

Tetrachloroethylene

127-18-4

neutral organicNC

6 x NCL

4.5

Abbreviations: aq = aqueous; CASRN = Chemical Abstracts Service Registry Number; UN GHS = United
Nations Globally Harmonised System (1); VRM = Validated Reference Method; ND = Not Determined.

3© OECD, (2015)

23

430

OECD/OCDE

1

The proficiency substances, sorted first by corrosives versus non-corrosives, then by corrosive
subcategory and then by chemical class, were selected from the substances used in the ECVAM validation
study of the rat skin TER test method (8) (9). Unless otherwise indicated, the substances were tested at the
purity level obtained when purchased from a commercial source (8). The selection included, to the extent
possible, substances that: (i) are representative of the range of corrosivity responses (e.g. non-corrosives;
weak to strong corrosives) that the VRM is capable of measuring or predicting; (ii) are representative of
the chemical classes used in the validation study; (iii) reflect the performance characteristics of the VRM;(iv) have chemical structures that are well-defined; (v) induce definitive results in the in vivo reference test
method; (vi) are commercially available; and (vii) are not associated with prohibitive disposal costs.
2
Chemical class assigned by Barratt et al. (8).
3The corresponding UN Packing groups are I, II and III, respectively, for the UN GHS 1A, 1B and 1C.
4
The pH values were obtained from Fentem et al. (9) and Barratt et al. (8).
PROCEDURE
13.
Standard Operating Procedures (SOP) for the rat skin TER skin corrosion test method areavailable (19). The rat skin TER test methods covered by this Test Guideline should comply with thefollowing conditions:
Animals
14.
Rats should be used because the sensitivity of their skin to substances in this test method has
been previously demonstrated (12) and is the only skin source that has been formally validated (8) (9). The
age (when the skin is collected) and strain of the rat is particularly important to ensure that the hair follicles
are in the dormant phase before adult hair growth begins.
15.
The dorsal and flank hair from young, approximately 22 day-old, male or female rats (Wistarderived or a comparable strain), is carefully removed with small clippers. Then, the animals are washed bycareful wiping, whilst submerging the clipped area in antibiotic solution (containing, for example,
streptomycin, penicillin, chloramphenicol, and amphotericin, at concentrations effective in inhibiting
bacterial growth). Animals are washed with antibiotics again on the third or fourth day after the first wash
and are used within 3 days of the second wash, when the stratum corneum has recovered from the hairremoval.
Preparation of the skin discs
16.
Animals are humanely killed when 28-30 days old; this age is critical. The dorso-lateral skin ofeach animal is then removed and stripped of excess subcutaneous fat by carefully peeling it away from theskin. Skin discs, with a diameter of approximately 20-mm each, are removed. The skin may be stored
before discs are used where it is shown that positive and negative control data are equivalent to thatobtained with fresh skin.
17.
Each skin disc is placed over one of the ends of a PTFE (polytetrafluoroethylene) tube, ensuringthat the epidermal surface is in contact with the tube. A rubber ‘O’ ring is press-fitted over the end of the
tube to hold the skin in place and excess tissue is trimmed away. The rubber ‘O’ ring is then carefully
sealed to the end of the PTFE tube with petroleum jelly. The tube is supported by a spring clip inside a
receptor chamber containing MgSO4 solution (154 mM) (Figure 1). The skin disc should be fully
submerged in the MgSO4 solution. As many as 10-15 skin discs can be obtained from a single rat skin.
Tube and ‘O’ ring dimensions are shown in Figure 2.

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18.
Before testing begins, the TER of two skin discs are measured as a quality control procedure foreach animal skin. Both discs should give electrical resistance values greater than 10 k: for the remainderof the discs to be used for the test method. If the resistance value is less than 10 k:, the remaining discs
from that skin should be discarded.
Application of the test chemical and control substances
19.
Concurrent positive and negative controls should be used for each run (experiment) to ensure
adequate performance of the experimental model. Skin discs from a single animal should be used in eachrun (experiment). The suggested positive and negative control test chemicals are 10M hydrochloric acidand distilled water, respectively.
20.
Liquid test chemicals (150 PL) are applied uniformly to the epidermal surface inside the tube.
When testing solid materials, a sufficient amount of the solid is applied evenly to the disc to ensure that thewhole surface of the epidermis is covered. Deionised water (150 PL) is added on top of the solid and thetube is gently agitated. In order to achieve maximum contact with the skin, solids may need to be warmed
to 300 C to melt or soften the test chemical, or ground to produce a granular material or powder.
21.
Three skin discs are used for each test and control chemical in each testing run (experiment). Test
chemicals are applied for 24 hours at 20-230 C. The test chemical is removed by washing with a jet of tap
water at up to room temperature until no further material can be removed.
TER measurements
22.
The skin impedance is measured as TER by using a low-voltage, alternating current Wheatstonebridge (18). General specifications of the bridge are 1-3 Volt operating voltage, a sinus or rectangularshaped alternating current of 50 - 1000 Hz, and a measuring range of at least 0.1 -30 k:. The databridge
used in the validation study measured inductance, capacitance and resistance up to values of 2000H,
2000 PF, and 2 M:, respectively at frequencies of 100Hz or 1kHz, using series or parallel values. For the
purposes of the TER corrosivity assay measurements are recorded in resistance, at a frequency of 100 Hz
and using series values. Prior to measuring the electrical resistance, the surface tension of the skin is
reduced by adding a sufficient volume of 70% ethanol to cover the epidermis. After a few seconds, theethanol is removed from the tube and the tissue is then hydrated by the addition of 3 mL MgSO4 solution(154mM). The databridge electrodes are placed on either side of the skin disc to measure the resistance ink:/skin disc (Figure 1). Electrode dimensions and the length of the electrode exposed below the crocodile
clips are shown in Figure 2. The clip attached to the inner electrode is rested on the top of the PTFE tube
during resistance measurement to ensure that a consistent length of electrode is submerged in the MgSO 4
solution. The outer electrode is positioned inside the receptor chamber so that it rests on the bottom of thechamber. The distance between the spring clip and the bottom of the PTFE tube is maintained as a constant(Figure 2), because this distance affects the resistance value obtained. Consequently, the distance between
the inner electrode and the skin disc should be constant and minimal (1-2 mm).
23.
If the measured resistance value is greater than 20 k:, this may be due to the remains of the test
chemical coating the epidermal surface of the skin disc. Further removal of this coating can be attempted,
for example, by sealing the PTFE tube with a gloved thumb and shaking it for approximately 10 seconds;
the MgSO4 solution is discarded and the resistance measurement is repeated with fresh MgSO4.
24.
The properties and dimensions of the test apparatus and the experimental procedure used may
influence the TER values obtained. The 5 k: corrosive threshold was developed from data obtained withthe specific apparatus and procedure described in this Test Guideline. Different threshold and control
values may apply if the test conditions are altered or a different apparatus is used. Therefore, it is necessary

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to calibrate the methodology and resistance threshold values by testing a series of Proficiency Substanceschosen from the substances used in the validation study (8) (9), or from similar chemical classes to thesubstances being investigated. A set of suitable Proficiency Substances is identified in Table 1.
Dye Binding Methods
25.
Exposure of certain non-corrosive materials can result in a reduction of resistance below the cutoff of 5 k: allowing the passage of ions through the stratum corneum, thereby reducing the electricalresistance (9). For example, neutral organics and substances that have surface-active properties (including
detergents, emulsifiers and other surfactants) can remove skin lipids making the barrier more permeable toions. Thus, if TER values produced by such chemicals are less than or around 5 k: in the absence ofvisually perceptible damage of the skin discs, an assessment of dye penetration should be carried out on thecontrol and treated tissues to determine if the TER values obtained were the result of increased skinpermeability, or skin corrosion (7) (9). In case of the latter where the stratum corneum is disrupted, the dye
sulforhodamine B, when applied to the skin surface rapidly penetrates and stains the underlying tissue.
This particular dye is stable to a wide range of substances and is not affected by the extraction proceduredescribed below.
Sulforhodamine B dye application and removal
26.
Following TER assessment, the magnesium sulphate is discarded from the tube and the skin iscarefully examined for obvious damage. If there is no obvious major damage (e.g. perforation), 150 PL ofa 10% (w/v) dilution in distilled water of the dye sulforhodamine B (Acid Red 52; C.I. 45100; CAS
number 3520-42-1), is applied to the epidermal surface of each skin disc for 2 hours. These skin discs arethen washed with tap water at up to room temperature for approximately 10 seconds to remove any
excess/unbound dye. Each skin disc is carefully removed from the PTFE tube and placed in a vial
(e.g. a 20-mL glass scintillation vial) containing deionised water (8 mL). The vials are agitated gently for
5 minutes to remove any additional unbound dye. This rinsing procedure is then repeated, after which theskin discs are removed and placed into vials containing 5ml of 30% (w/v) sodium dodecyl sulphate (SDS)
in distilled water and are incubated overnight at 600 C.
27.
After incubation, each skin disc is removed and discarded and the remaining solution iscentrifuged for 8 minutes at 210 C (relative centrifugal force ~175 x g). A 1mL sample of the supernatant isdiluted 1 in 5 (v/v) [i.e. 1mL + 4mL] with 30% (w/v) SDS in distilled water. The optical density (OD) of
the solution is measured at 565 nm.
Calculation of dye content
28.
The sulforhodamine B dye content per disc is calculated from the OD values (9) (sulforhodamine
B dye molar extinction coefficient at 565nm = 8.7 x l04; molecular weight = 580). The dye content isdetermined for each skin disc by the use of an appropriate calibration curve and mean dye content is thencalculated for the replicates.
Acceptability Criteria
29.
The mean TER results are accepted if the concurrent positive and negative control values fallwithin the acceptable ranges for the method in the testing laboratory. The acceptable resistance ranges forthe methodology and apparatus described above are given in the following table:

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Control
Positive
Negative

Substance
10M Hydrochloric acid
Distilled water

430

Resistance range (k::)
0.5 - 1.0
10 - 25

30.
The mean dye binding results are accepted on condition that concurrent control values fall within
the acceptable ranges for the method. Suggested acceptable dye content ranges for the control substances
for the methodology and apparatus described above are given in the following table:
Control
Positive
Negative

Dye content range (PPg/disc)40 - 100
15 - 35

Substance
10M Hydrochloric acid
Distilled waterInterpretation of results
31.
The cut-off TER value distinguishing corrosive from non-corrosive test chemicals wasestablished during test method optimization, tested during a pre-validation phase, and confirmed in aformal validation study.
32.
The prediction model for rat skin TER skin corrosion test method (9) (19), associated with the
UN GHS (1) classification system, is given below:
The test chemical is considered to be non-corrosive to skin:i) if the mean TER value obtained for the test chemical is greater than (>) 5 k:, orii) the mean TER value obtained for the test chemical is less than or equal to (≤) 5 k:, andx the skin discs show no obvious damage(e.g. perforation), andx the mean disc dye content is less than (<) the mean disc dye content of the 10M HCl
positive control obtained concurrently (see paragraph 30 for positive control values).
The test chemical is considered to be corrosive to skin:if the mean TER value obtained for the test chemical is less than or equal to (≤) 5 k: and the skindiscs are obviously damaged(e.g. perforated), or
the mean TER value obtained for the test chemical is less than or equal to (≤) 5 k:, andx the skin discs show no obvious damage(e.g. perforation), but
x the mean disc dye content is greater than or equal to (≥) the mean disc dye content of the
10M HCl positive control obtained concurrently (see paragraph 30 for positive control
values).
33.
A testing run (experiment) composed of at least three replicate skin discs should be sufficient fora test chemical when the classification is unequivocal. However, in cases of borderline results, such as nonconcordant replicate measurements and/or mean TER equal to 5 ± 0.5 kΩ, a second independent testingrun (experiment) should be considered, as well as a third one in case of discordant results between the first
two testing runs (experiments).

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OECD/OCDEDATA AND REPORTING
Data
34.
Resistance values (k:) and dye content values (μg/disc), where appropriate, for the test chemical,
as well as for positive and negative controls should be reported in tabular form, including data for eachindividual replicate disc in each testing run (experiment) and mean values ± SD. All repeat experiments
should be reported. Observed damage in the skin discs should be reported for each test chemical.
Test report
35.

The test report should include the following information:
Test Chemical and Control Substances: Mono-constituent substance: chemical identification, such as IUPAC or CAS name,
CAS number, SMILES or InChI code, structural formula, purity, chemical identity of
impurities as appropriate and practically feasible, etc;
 Multi-constituent substance, UVCB and mixture: characterised as far as possible bychemical identity (see above), quantitative occurrence and relevant physico-chemical
properties of the constituents;
 Physical appearance, water solubility, and additional relevant physico-chemical
properties;
 Source, lot number if available;
 Treatment of the test chemical/control substance prior to testing, if applicable (e.g.
warming, grinding);
 Stability of the test chemical, limit date for use, or date for re-analysis if known;
 Storage conditions.
Test Animals:
 Strain and sex used; Age of the animals when used as donor animals;
 Source, housing condition, diet, etc.;
 Details of the skin preparation.
Test Conditions: Calibration curves for test apparatus;
 Calibration curves for dye binding test performance, band pass used for measuring
OD values, and OD linearity range of measuring device (e.g. spectrophotometer), if
appropriate; Details of the test procedure used for TER measurements;
 Details of the test procedure used for the dye binding assessment, if appropriate; Test doses used, duration of exposure period(s) and temperature(s) of exposure; Details on washing procedure used after the exposure period;
 Number of replicate skin discs used per test chemical and controls (positive andnegative control); Description of any modification of the test procedure;– Reference to historical data of the model. This should include, but is not limited to;i) Acceptability of the positive and negative control TER values (in kΩ) withreference to positive and negative control resistance ranges

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430ii) Acceptability of the positive and negative control dye content values (inμg/disc) with reference to positive and negative control dye content rangesiii) Acceptability of the test results with reference to historical variability between
skin disc replicates Description of decision criteria/prediction model applied.
Results:

 Tabulation of data from the TER and dye binding assays (if appropriate) for
individual test chemicals and controls, for each testing run (experiment) and eachskin disc replicate (individual animals and individual skin samples), means, SDs and
CVs; Description of any effects observed;
 The derived classification with reference to the prediction model/decision criteria
used.

Discussion of the results
Conclusions

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OECD/OCDELITERATURE(1)

United Nations (UN). (2013). Globally Harmonized System of Classification and Labelling of Chemicals(GHS), Second Revised Edition, UN New York and Geneva, 2013. Available at:[http://www.unece.org/trans/danger/publi/ghs/ghs_rev05/05files_e.html].

(2)OECD. (2015). Guideline for Testing of Chemicals. (No. 404.): Acute Dermal Irritation, Organisation for
Economic Cooperation and Development, Paris..

(3)OECD. (2015). Guideline for the Testing of Chemicals (No. 431.): In Vitro Skin Model, Organisation
for Economic Cooperation and Development, Paris.

(4)OECD. (2015). Guideline for the Testing of Chemicals (No. 435.) In Vitro Membrane Barrier Test
Method, Organisation for Economic Cooperation and Development, Paris.

(5)

OECD. (2015). Guideline for the Testing of Chemicals (No. 439.) In Vitro Skin Irritation: Reconstructed
Human Epidermis Test Method.

(6)

OECD. (2014). Guidance document on Integrated Approached to Testing and Assessment for Skin
Irritation/Corrosion. Environment, Health and Safety Publications, Series on Testing and Assessment,(No. 203), Organisation for Economic Cooperation and Development, Paris.

(7)

Botham P.A., Chamberlain M., Barratt M.D., Curren R.D., Esdaile D.J., Gardner J.R., Gordon V.C.,
Hildebrand B., Lewis R.W., Liebsch M., Logemann P., Osborne R., Ponec M., Regnier J.F., Steiling W.,
Walker A.P., and Balls M. (1995). A Prevalidation Study on In Vitro Skin Corrosivity Testing. The
Report and Recommendations of ECVAM Workshop 6.ATLA 23, 219-255.

(8)

Barratt M.D., Brantom P.G., Fentem J.H., Gerner I., Walker A.P., and Worth A.P. (1998). The ECVAMInternational Validation Study on In Vitro Tests for Skin Corrosivity. 1. Selection and Distribution of the
Test Chemicals. Toxic.In Vitro 12, 471-482.

(9)

Fentem J.H., Archer G.E.B., Balls M., Botham P.A., Curren R.D., Earl L.K., Esdaile D.J., Holzhütter H.G., and Liebsch M. (1998). The ECVAM International Validation Study on In Vitro Tests For Skin
Corrosivity. 2. Results and Evaluation by the Management Team. Toxic.In Vitro12, 483- 524.

(10) Balls M., Blaauboer B.J., Fentem J.H., Bruner L., Combes R.D., Ekwall B., Fielder R.J., Guillouzo A.,
Lewis R.W., Lovell D.P., Reinhardt C.A., Repetto G., Sladowski D., Spielmann H., and Zucco F. (1995).
Practical Aspects of the Validation of Toxicity Test Procedures. The Report and Recommendations of
ECVAM Workshops.ATLA23, 129-147.
(11) ICCVAM (Interagency Coordinating Committee on the Validation of Alternative Methods). (1997).
Validation and Regulatory Acceptance of Toxicological Test Methods. NIH Publication No. 97-3981.
National Institute of Environmental Health Sciences, Research Triangle Park, NC, USA. Available at:[http://www.iccvam.niehs.nih.gov/docs/guidelines/validate.pdf].

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430(12) EC-ECVAM. (1998). Statement on the Scientific Validity of the Rat Skin Transcutaneos Electrical
Resistance (TER) Test (an In Vitro Test for Skin Corrosivity), Issued by the ECVAM Scientific
Advisory Committee (ESAC10), 3 April 1998. Available at: [http://www.ecvam.jrc.ec.europa.eu.html].
(13) ECVAM. (1998). ECVAM News & Views. ATLA 26, 275-280.
(14) ICCVAM (Interagency Coordinating Committee on the Validation of Alternative Methods). (2002).
ICCVAM Evaluation of EpiDermTM (EPI-200), EPISKINTM (SM), and the Rat Skin Transcutaneous
Electrical Resistance (TER) Assay: In Vitro Test Methods for Assessing Dermal Corrosivity Potential ofChemicals. NIH Publication No. 02-4502. National Toxicology Program Interagency Center for the
Evaluation of Alternative Toxicological Methods, National Institute of Environmental Health Sciences,
Research Triangle Park, NC, USA. Available at:[http://www.iccvam.niehs.nih.gov/methods/epiddocs/epis_brd.pdf].
(15) OECD. (2015). Performance Standards for the Assessment of Proposed Similar or Modified In Vitro
Transcutaneous Electrical Resistance (TER) Test Method for Skin Corrosion in Relation to TG 430.
Environmental Health and Safety Publications, Series on Testing and Assessment No. 218. Organisation
for Economic Cooperation and Development, Paris.
(16) Oliver G.J.A., Pemberton M.A., and Rhodes C. (1986). An In Vitro Skin Corrosivity Test -Modifications
and Validation. Fd. Chem. Toxicol.24, 507-512.
(17) Botham P.A., Hall T.J., Dennett R., McCall J.C., Basketter D.A., Whittle E., Cheeseman M., Esdaile
D.J., and Gardner J. (1992). The Skin Corrosivity Test In Vitro: Results of an Interlaboratory Trial.
Toxicol.In Vitro6,191-194.
(18) Eskes C., Detappe V., Koëter H., Kreysa J., Liebsch M., Zuang V., Amcoff P., Barroso J., Cotovio J.,
Guest R., Hermann M., Hoffmann S., Masson P., Alépée N., Arce L.A., Brüschweiler B., Catone T.,
Cihak R., Clouzeau J., D'Abrosca F., Delveaux C., Derouette J.P., Engelking O., Facchini D., Fröhlicher
M., Hofmann M., Hopf N., Molinari J., Oberli A., Ott M., Peter R., Sá-Rocha V.M., Schenk D., Tomicic
C., Vanparys P., Verdon B., Wallenhorst T., Winkler G.C. and Depallens O. (2012). Regulatory
Assessment of In Vitro Skin Corrosion and Irritation Data Within the European Framework: Workshop
Recommendations. Regul.Toxicol.Pharmacol. 62, 393-403.
(19) TER SOP. (December 2008). INVITTOX Protocol (No. 115.) Rat Skin Transcutaneous Electrical
Resistance (TER) Test. Available at: [http://www.ecvam-dbalm.jrc.ec.europa.eu/].
(20) OECD. (2005). Guidance Document on the Validation and International Acceptance of New or Updated
Test Methods for Hazard Assessment. Environment, Health and Safety Publications, Series on Testingand Assessment (No. 34.), Organisation for Economic Cooperation and Devlopment, Paris.

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Figure 1: Apparatus for the rat skin TER assay

Crocodile clip

Inner (thick) electrode
PTFE Tube
Crocodile clip

Outer (thin) electrodeSpring clip

Receptor chamber(disposable tube)

Magnesium sulphate(154mM)

Epidermis of skin disc

Rubber 'O' ring

Magnesium sulphate(154mM)
Dermis of skin disc12
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Figure 2: Dimensions of the polytetrafluoroethylene (PTFE) and receptor tubes and electrodes used
16mm

10mm

95mm

Position of 'O' ring

3mm
0.5mm

1mm

1mm
13mm
0.5mm

Crocodile Clip
81mm
78mm

75mm

1mm

3mm

Critical factors of the apparatus shown above:––

The inner diameter of the PTFE tube,–

The amount of MgSO4 solution in the receptor tube should give a depth of liquid, relative to the level in the
PTFE tube, as shown in Figure 1,–

The skin disc should be fixed well enough to the PTFE tube, such that the electrical resistance is a truemeasure of the skin properties.

The length of the electrodes relative to the PTFE tube and receptor tube, such that the skin disc should not betouched by the electrodes and that a standard length of electrode is in contact with the MgSO4 solution,

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OECD/OCDEANNEX 1DEFINITIONS

Accuracy: The closeness of agreement between test method results and accepted reference values. It is ameasure of test method performance and one aspect of relevance. The term is often used interchangeably
with “concordance” to mean the proportion of correct outcomes of a test method (20).
C: Corrosive.
Chemical: means a substance or a mixture.
Concordance: This is a measure of test method performance for test methods that give a categorical result,and is one aspect of relevance. The term is sometimes used interchangeably with accuracy, and is definedas the proportion of all chemicals tested that are correctly classified as positive or negative. Concordance ishighly dependent on the prevalence of positives in the types of test chemical being examined (20).
GHS (Globally Harmonized System of Classification and Labelling of Chemicals (UN)): A system
proposing the classification of chemicals (substances and mixtures) according to standardized types andlevels of physical, health and environmental hazards, and addressing corresponding communication
elements, such as pictograms, signal words, hazard statements, precautionary statements and safety data
sheets, so that to convey information on their adverse effects with a view to protect people (including
employers, workers, transporters, consumers and emergency responders) and the environment (1).
IATA: Integrated Approach on Testing and Assessment.
Mixture: means as a mixture or solution composed of two or more substances in which they do not react.
Mono-constituent substance: A substance, defined by its quantitative composition, in which one main
constituent is present to at least 80% (w/w).
Multi-constituent substance: A substance, defined by its quantitative composition, in which more than
one main constituent is present in a concentration ≥ 10% (w/w) and < 80% (w/w). A multi-constituent
substance is the result of a manufacturing process. The difference between mixture and multi-constituent
substance is that a mixture is obtained by blending of two or more substances without chemical reaction. A
multi-constituent substance is the result of a chemical reaction.
NC: Non corrosive.
OD: Optical Density.

PC: Positive Control, a replicate containing all components of a test system and treated with a substanceknown to induce a positive response. To ensure that variability in the positive control response across time
can be assessed, the magnitude of the positive response should not be excessive.
Performance standards (PS): Standards, based on a validated test method, that provide a basis forevaluating the comparability of a proposed test method that is mechanistically and functionally similar.
Included are; (i) essential test method components; (ii) a minimum list of Reference Chemicals selectedfrom among the chemicals used to demonstrate the acceptable performance of the validated test method;

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and (iii) the similar levels of reliability and accuracy, based on what was obtained for the validated test
method, that the proposed test method should demonstrate when evaluated using the minimum list ofReference Chemicals.
Relevance: Description of relationship of the test method to the effect of interest and whether it ismeaningful and useful for a particular purpose. It is the extent to which the test method correctly measuresor predicts the biological effect of interest. Relevance incorporates consideration of the accuracy(concordance) of a test method (20).
Reliability: Measures of the extent that a test method can be performed reproducibly within and between
laboratories over time, when performed using the same protocol. It is assessed by calculating intra- andinter-laboratory reproducibility (20).
Sensitivity: The proportion of all positive/active chemicals that are correctly classified by the test method.
It is a measure of accuracy for a test method that produces categorical results, and is an importantconsideration in assessing the relevance of a test method (20).
Skin corrosion in vivo: The production of irreversible damage of the skin; namely, visible necrosis
through the epidermis and into the dermis, following the application of a test chemical for up to four hours.
Corrosive reactions are typified by ulcers, bleeding, bloody scabs, and, by the end of observation at
14 days, by discoloration due to blanching of the skin, complete areas of alopecia, and scars.
Histopathology should be considered to evaluate questionable lesions.
Specificity: The proportion of all negative/inactive chemicals that are correctly classified by the test
method. It is a measure of accuracy for a test method that produces categorical results and is an importantconsideration in assessing the relevance of a test method (20).
Substance: means chemical elements and their compounds in the natural state or obtained by any
production process, including any additive necessary to preserve the stability of the product and any
impurities deriving from the process used, but excluding any solvent which may be separated without
affecting the stability of the substance or changing its composition.
(Testing) run: A single test chemical concurrently tested in a minimum of three replicate skin discs.
Test chemical: means what is being tested.
Transcutaneous Electrical Resistance (TER): is a measure of the electrical impedance of the skin, as a
resistance value in kilo Ohms. A simple and robust method of assessing barrier function by recording thepassage of ions through the skin using a Wheatstone bridge apparatus.
UVCB: substances of unknown or variable composition, complex reaction products or biologicalmaterials.

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