᪂つヨ㦂ἲᥦ᱌᭩             ᖹᡂ 29 ᖺ 10 ᭶ 1 ᪥                                 No. 2017-02



In vitro⭷ࣂࣜ࢔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 In vitro ⭷ࣂࣜ࢔ヨ㦂ἲ࡜ࡋ࡚ၟᴗⓗ࡟㈍኎ࡉࢀ࡚࠸ࡿヨ㦂ἲ࡜ࡋ࡚ Corrositex®ἲࡀ࠶ࡿࠋᮏヨ㦂ἲෆࡢ໬Ꮫ≀㉁᳨▱ࢩࢫࢸ࣒㸦Chemical Detection System㸸CDS㸧ࡢ㐺⏝ྍ⬟≀㉁࡟ไ⣙ࡀ࠶ࡾࠊഇ㝜ᛶࡀከ࠸ࡇ࡜࠿ࡽࠊከᵝ࡞≀㉁ࢆࢫࢡ࣮ࣜࢽࣥࢢホ౯ࡍࡿ┠ⓗ࡟ࡣ㐺ࡋ࡚࠸࡞࠸࡜⪃࠼ࡽࢀࡿࠋࡓࡔࡋࠊᮏヨ㦂ἲ࡟ࡼࡗ࡚㝧ᛶ࡜ุᐃࡉࢀࡓࡶ
ࡢࢆ㝧ᛶ≀㉁࡜ࡋ࡚㆑ูࡍࡿࡇ࡜࡟㝈ᐃࡍࢀࡤࠊ฼⏝ྍ⬟࡜ุ᩿ࡍࡿࠋCorrositex®௨እࡢ in vitro ⭷ࣂࣜ࢔ヨ㦂ἲ࡟ࡘ࠸࡚ࡣ▱ࡽࢀ࡚࠸࡞࠸ࡀࠊ㛤Ⓨࡉࢀࡓሙྜ࡟ࡣ༑ศ࡞ࣂ
ࣜࢹ࣮ࢩࣙࣥ⤖ᯝ࡟ᇶ࡙࠸࡚ࠊࡑࡢ⾜ᨻⓗ฼⏝ᛶࡢホ౯ࢆ⾜࠺࡭ࡁ࡛࠶ࡿࠋ
ࡇࡢᥦ᱌᭩ࡣࠊOrganisation for Economic Co-operation and Development (OECD) Test Guideline(TG) 435, In Vitro Membrane Barrier Test Method for Skin Corrosionࢆࡶ࡜࡟ࠊ⓶⭵⭉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 In vitro ⭷ࣂࣜ࢔ヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢ౑⏝ࢆᥦ᱌ࡍࡿࡶࡢ࡛࠶ࡿࠋ
 

኱㔝Ὀ㞝
㔝Ὀ㞝                      すᕝ⛅ె
JaCVAM ホ౯఍㆟ ㆟㛗               JaCVAM 㐠Ⴀጤဨ఍ ጤဨ㛗

i

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 Membrane Barrier Test Method for Skin Corrosion
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: Corrositex® is a commercially available in vitro membrane barrier test method.
We do not consider this test method to be suitable for predicting the skin corrosion
potential of test chemicals due to a significant number of false negative test results
and the limited applicability domain of the Chemical Detection System (CDS).
Since a positive result from this test method can, however, be consideredsufficient for predicting a test chemical to cause skin corrosion, this test could be
useful when used within that limitation. We do not know of any other in vitro
membrane barrier model, and if a new model is developed, its suitability for use ina regulatory context should be evaluated based on the results of a validation study.
This statement was prepared following a review of the Organisation for Economic
Co-operation and Development (OECD) Test Guideline (TG) 435 In Vitro Membrane Barrier
test method for skin corrosion 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 limited usefulness of thisassay.
Based on the above, we propose the In Vitro Membrane Barrier test method as a useful means
for assessing skin corrosion potential during safety assessments by regulatory agencies.

Yasuo
asuo Ohno
Chairperson
h i
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

vii

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In vitro⭷ࣂࣜ࢔ヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲ┠  ḟ
ホ౯఍㆟ሗ࿌᭩ ------------------------------------------------------------------------------------ 1
ホ౯ሗ࿌᭩ ------------------------------------------------------------------------------------------ 7
OECD GUIDELINE FOR THE TESTING OF CHEMICALS 435,
In Vitro Membrane Barrier Test Method for Skin Corrosion-----------------------------------21ix

xホ౯఍㆟ሗ࿌᭩

In vitro⭷ࣂࣜ࢔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

In vitro ⭷ࣂࣜ࢔ヨ㦂ࡣࠊ࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ࡢ௦᭰࡜ࡋ࡚㛤Ⓨࡉࢀࡓヨ㦂ἲ࡛࠶ࡿࠋᮏヨ㦂ἲ࡛ࡣࠊ⭉㣗ᛶ≀㉁࡟ࡼࡾᘬࡁ㉳ࡇࡉࢀࡿ⭷ࣂࣜ࢔㞀ᐖࢆᣦᶆ࡜ࡋ࡚⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࠋᮏヨ㦂ἲ࡟ࡘ࠸࡚ࡣࠊḢ⡿࡛ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࠊḢᕞ࡛ࡣ໬Ꮫ≀㉁ࡢ⓶⭵⭉㣗ᛶホ౯ࢆ┠ⓗ࡜ࡋ࡚ᢎㄆࡉࢀࠊ໬Ꮫ≀㉁ࡢ᭷ᐖᛶࡢศ㢮࣭⾲♧➼ࡢࡓࡵࡢヨ㦂ἲ࡜ࡋ࡚᥇⏝ࡉࢀ࡚࠸ࡿࠋࡲࡓࠊᮏヨ㦂ἲࡣ OECD㸦Organisation for Economic Co-operation and Development㸸⤒῭༠ຊ㛤Ⓨᶵᵓ㸧࡟࡚ࢸࢫࢺ࢞࢖
ࢻࣛ࢖ࣥ㸦TG㸧435 ࡜ࡋ࡚ᢎㄆࡉࢀ࡚࠸ࡿ 1)ࠋ
JaCVAM ホ౯఍㆟ࡣࠊ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾసᡂࡉࢀࡓࠕIn vitro ⭷ࣂࣜ࢔ヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩ࠖ2)ࢆ⏝࠸࡚ࠊᮏヨ㦂ἲࡢጇᙜᛶ࡟ࡘ࠸᳨࡚ウࡋࡓࠋ㸯㸬ヨ㦂ἲࡢᐃ⩏ྡ⛠㸸 In vitro ⭷ࣂࣜ࢔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ἲ࡛ࡣࠊ࢘ࢧࢠ⓶⭵ࡢ௦ࢃࡾ࡟ྜᡂࡢ㧗ศᏊ࡛ᵓᡂࡉࢀࡿேᕤࡢ⭷ࣂࣜ࢔㸦invitro ⭷ࣂࣜ࢔㸧ࢆ⏝࠸ࠊ໬Ꮫ≀㉁࡟ࡼࡿ⓶⭵⭉㣗ᛶⓎ⌧ᶵᵓࡢ㔜せ࡞ࢫࢸࢵࣉ࡛࠶ࡿ
⓶⭵ࡢࣂࣜ࢔ᶵ⬟ࡢ㞀ᐖࢆࠊ໬Ꮫ≀㉁᳨▱ࢩࢫࢸ࣒㸦Chemical Detection System㸸CDS㸧࡟ࡼࡗ᳨࡚ฟࡍࡿࡇ࡜࡟ࡼࡾࠊ⿕㦂≀㉁ࡢ⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿࠋ⿕㦂≀㉁ࢆ in vitro⭷ࣂࣜ࢔ࡢ⾲㠃࡟㐺⏝ࡍࡿ࡜ࠊ⭉㣗ᛶࡢ⿕㦂≀㉁ࡀ⓶⭵࡟స⏝ࡍࡿᶵᗎ࡜ྠᵝ࡟ࡋ࡚ᘬࡁ㉳ࡇࡍ⭷ࣂࣜ࢔ࡢᦆയ࡟ࡼࡾࠊ⿕㦂≀㉁ࡣ⭷ࣂࣜ࢔ࢆᾐ㏱࣭㏻㐣ࡋ࡚ CDS ഃ࡟฿㐩ࡋࠊpH ࢖ࣥࢪࢣ࣮ࢱ࣮ࡢⰍࡢኚ໬ࡲࡓࡣ࢖ࣥࢪࢣ࣮ࢱ࣮ᾮࡢ௚ࡢ≉ᛶࡢኚ໬ࢆ
ᘬࡁ㉳ࡇࡍ㸦໬Ꮫ཯ᛂࠊ㟁Ẽ໬Ꮫ཯ᛂ࡞࡝㸧ࠋ⭉㣗ᛶࡢ⛬ᗘࡣࠊ᭚㟢᫬㛫࡜ほᐹᮇ㛫࡟࠾ࡅࡿ CDS ࡢኚ໬ࡢ⛬ᗘ࠿ࡽุᐃࡍࡿࠋᮏヨ㦂ἲࡢࢩࢫࢸ࣒ࡣၟᴗⓗ࡟ࡣCorrositex®࡜ࡋ࡚㈍኎ࡉࢀ࡚࠸ࡿࠋ㸰㸬ホ౯࡟⏝࠸ࡓ㈨ᩱཬࡧホ౯ෆᐜࡢ⛉Ꮫⓗጇᙜᛶᮏヨ㦂ἲࡣࠊ⭉㣗ᛶࡢ⿕㦂≀㉁ࡀ⏕ࡁࡓ⓶⭵࡟స⏝ࡍࡿ㝿࡟ᘬࡁ㉳ࡇࡉࢀࡿ⭷ࣂࣜ࢔ᦆയࢆࠊேᕤࡢ
⭷ࢆ⏝࠸࡚ホ౯ࡍࡿࡶࡢ࡛࠶ࡾࠊ⓶⭵⭉㣗ᛶⓎ⌧ᶵᗎࡀཎ⌮ⓗ࡟⪃៖ࡉࢀ࡚࠸ࡿࠋ
Corrositex®ἲࡣࠊECVAM㸦European Centre for the Validation of Alternative Methods㸸Ḣᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮㸧࡟ࡼࡿࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀ࡞ࡉࢀ 3,4,5)ࠊESAC㸦ECVAM Scientific Advisory Committee㸸ECVAM⛉Ꮫㅎၥጤဨ఍㸧࡟ࡼࡾಙ㢗ᛶ࡜෌⌧ᛶࡀ㧗࠸࡜ホ౯ࡉࢀ࡚࠸ࡿ 6)ࠋࡇࡢ⤖ㄽࡣࠊ⡿ᅜࡢ௦᭰ἲ࡟㛵ࡍࡿ┬ᗇ㛫㐃⤡఍㆟㸦ICCVAM㸸Interagency Coordinating Committee on the Validation of Alternative Methods)࡟࠾࠸࡚ࡶ☜ㄆࡉࢀ࡚࠸ࡿ 7,8)ࠋࡇࢀࡽࡢ⤖ᯝࡣබ⾲ࡉࢀ࡚࠾ࡾࠊ㏱࡛᫂⊂❧࡞⛉Ꮫⓗホ౯ࡀ⾜ࢃࢀ࡚࠸ࡿ࡜⪃࠼ࡿࠋ᭦࡟ࠊᮏヨ㦂ἲࡣࠊOECD ࡟࠾࠸࡚ࡶホ౯ࡉࢀࠊTG435 ࡜ࡋ࡚ᢎㄆࡉࢀ࡚࠸ࡿ 1)ࠋᮏ㑥࡟

33࠾࠸࡚ࡣࠊJaCVAM ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍ࡀࠊࡇࢀࡽࡢ㈨ᩱࢆ⏝࠸࡚ᮏヨ㦂ἲࢆホ౯ࡋ࡚࠸ࡿ
2)ࠋ
௨ୖࡢⅬ࠿ࡽࠊCorrositex®ἲࡣ⓶⭵⭉㣗ᛶࢆホ౯ࡍࡿ᪉ἲ࡜ࡋ࡚ཎ⌮ⓗጇᙜᛶࡀ࠶ࡾࠊࡲࡓࠊࡑࡢホ౯࡟⏝࠸ࡓ㈨ᩱ࡞ࡽࡧ࡟ホ౯ෆᐜ࡟ࡘ࠸࡚ࡣࠊ⛉Ꮫⓗጇᙜᛶࡀ࠶ࡿ࡜⪃࠼ࡿࠋ㸱㸬ᮏヨ㦂ἲࡢ᭷⏝ᛶ࡜㐺⏝㝈⏺
ᮏヨ㦂ἲࡣࠊື≀ࢆ౑⏝ࡋ࡚࠾ࡽࡎࠊື≀⚟♴㠃࠿ࡽ௦᭰ἲ࡜ࡋ࡚ጇᙜ࡛࠶ࡿࠋ
ᮏヨ㦂ἲࡢࢩࢫࢸ࣒ࡣၟᴗⓗ࡟ࡣ Corrositex®࡜ࡋ࡚㈍኎ࡉࢀ࡚࠾ࡾࠊ⭷ࣂࣜ࢔ヨᩱࡢᛶ⬟ࡢᆒ୍ᛶࢆ
ಖࡘࡇ࡜ࡣẚ㍑ⓗᐜ᫆࡜ᛮࢃࢀࠊ≉ู࡞ᡭᢏࡶᚲせ࡞࠸࡜ᛮࢃࢀࡿࡇ࡜࠿ࡽࠊᢏ⾡⛣㌿ᛶࡣ㧗࠸࡜ุ᩿࡛ࡁࡿࠋࡲࡓࠊ⩦⇍ᗘ☜ㄆ≀㉁ࢆ⏝࠸࡚ᑓ㛛ᢏ⾡ࡢ⩦⇍࡟ࡘ࠸࡚☜ㄆࡍࡿࡇ࡜ࡀ࡛ࡁࠊከࡃࡢ⩦⇍ᗘ☜ㄆ≀㉁ࢆ୍ᐃࡢ᮲௳࡛ホ౯ࡍࡿࡇ࡜࡟ࡼࡾࠊCDS ࡢኚ໬ࡢ⛬ᗘ࡟ࡼࡿ⭉㣗ᛶศ㢮ࡢ⢭ᗘࢆ㧗ࡵࡿࡇ࡜ࡀ
࡛ࡁࡿ࡜᥎

ࡉࢀࡿࠋ

Corrositex®ἲࡢணᛶ࡟ࡘ࠸࡚ࠊICCVAM ࡟ࡼࡿ㞟ィ⤖ᯝ࡜ Botham ࠾ࡼࡧ Fentem ࡢㄽᩥ࡟グ㍕ࡉ

ࢀࡓឤᗘࠊṇ☜ᗘࠊ≉␗ᗘ࡛♧ࡍ࡜ࠊṇ☜ᗘ 73-80㸣ࠊឤᗘ 71-89㸣ࠊ≉␗ᗘ 65-76㸣࡛࠶ࡗࡓ 3,5,7)ࠋࡓ
ࡔࡋࠊࡇࢀࡽࡢ⤖ᯝࡣヨ㦂㛫࡛ᕪࡀ኱ࡁࡃࠊ⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡛ࡣࠊホ౯≀㉁㑅ᢥࡢᐈほᛶ࡞࡝ࡶ⪃៖ࡋ࡚ࠊFentem ࡢ⤖ᯝࢆ᥇⏝ࡍࡿࡇ࡜ࡀጇᙜ࡜ุ᩿ࡋ࡚࠸ࡿࠋࡇࡢ Fentem ࡢணᛶࡢ⤖ᯝࢆ⾲ 1 ࡟♧ࡋࡓ㸦ṇ☜ᗘ 73㸣ࠊឤᗘ 71㸣ࠊ≉␗ᗘ 76㸣࡛࠶ࡾࠊഇ㝜ᛶ⋡ 29㸣ࠊഇ㝧ᛶ⋡ 24%࡜࡞ࡿ㸧ࠋࡋࡓࡀࡗ࡚ࠊᮏヨ㦂ἲࡣഇ㝜ᛶ࡜࡞ࡿሙྜࡀከ࠸࡜ุ᩿ࡉࢀࡿࠋ
⓶⭵⭉㣗ᛶヨ㦂㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾసᡂࡉࢀࡓホ౯ሗ࿌᭩ 2)࡟ࡼࢀࡤࠊCorrositex®ἲࡢࡶࡗ࡜ࡶ኱
ࡁ࡞ၥ㢟Ⅼࡣࠊࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡛㑅ᢥࡉࢀࡓ≀㉁ࡢ୰࡛ࠊヨ㦂࡟㐺⏝࡛ࡁࡿ≀㉁ࡀ㝈ᐃࡉࢀ࡚࠸ࡿ
ࡇ࡜࡛࠶ࡿ㸦㐺ྜᛶヨ㦂࡟ࡼࡾ Botham ࡛ࡣ 70-76%ࠊFentem ࡛ࡣ 63%ࡋ࠿ヨ㦂ࢆᐇ᪋࡛ࡁ࡞࠿ࡗࡓ㸧ࠋ
Corrositex®ἲࡣཎ⌮ⓗ࡟ CDS ࡢⰍࡲࡓࡣ௚ࡢ≉ᛶࡢኚ໬ࢆ⏕ࡌ࡞࠸≀㉁࡟ࡣ㐺⏝࡛ࡁ࡞࠸ࠋ౛࠼ࡤࠊpHࡀ 4.5~8.5 ࡢ⠊ᅖࡢỈ⁐ᛶ≀㉁ࡣࠊከࡃࡢሙྜホ౯࡛ࡁ࡞࠸ࠋࡇࡢࡓࡵࠊ⭷ࣂࣜ࢔ࢆ⏝࠸ࡓヨ㦂ࢆ㛤ጞࡍࡿ๓࡟ࠊ㐺ྜᛶヨ㦂࡜ࡋ࡚ࠊ⿕㦂≀㉁ࡀ CDS ࡟ࡼࡗ᳨࡚ฟྍ⬟࠿࡝࠺࠿ࢆ☜ㄆࡋ࡚࠾ࡃᚲせࡀ࠶ࡿࠋࡇࡢࡼ࠺࡟㐺⏝࡛ࡁࡿ≀㉁࡟ࡘ࠸࡚ὀពࡍࡿᚲせࡀ࠶ࡿࡶࡢࡢࠊᮏヨ㦂ἲࡣᅛయ㸦Ỉ⁐ᛶ/㠀Ỉ⁐ᛶ㸧ࠊᾮయ㸦Ỉ⁐ᾮ/㠀Ỉ⁐ᾮ㸧࠾ࡼࡧங๣࡟㐺⏝࡛ࡁࡿࠋ⾲ 1. Corrositex®ࡢணᛶFentem et al.㸦1998㸧5)

ṇ☜ᗘ

73%

ឤᗘ

71%

≉␗ᗘ

76%

ホ౯≀㉁ᩘ

38/60

4
4

OECD ࡢ in vitro ⭉㣗ᛶヨ㦂࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ࠊ௚࡟ࠕTG431 ࣄࢺ⾲⓶ࣔࢹࣝヨ㦂ࠖ࠾ࡼࡧࠕTG430

⤒⓶㟁Ẽ᢬ᢠヨ㦂㸦TER㸧ࠖࡀ࠶ࡾࠊICCVAM ࡛ࡣᮏヨ㦂ἲ࡜ࡑࢀࡽࡢヨ㦂ἲ (EpiSkinτ, EpiDermτ࠾ࡼࡧ TER)࡟ࡘ࠸࡚≉␗ᛶࠊឤᗘࠊṇ☜ᛶ➼ࡀẚ㍑ࡉࢀ࡚࠸ࡿ 2,7,8)ࠋヨ㦂≀㉁ࡢᩘ㔞ࡸ✀㢮ࡀ␗࡞ࡗ࡚࠸
ࡿࡓࡵࠊ⤖ᯝࡢุᐃࡔࡅࢆࡶࡗ࡚ࠊ༢⣧࡟ᙜヱヨ㦂ἲࡢඃ㉺ᛶࢆホ౯ࡍࡿࡇ࡜ࡣᅔ㞴࡛࠶ࡿࡶࡢࡢࠊᮏ
ヨ㦂ἲࡣࠊ௚ࡢヨ㦂ἲ࡜ẚ࡭࡚ឤᗘࡀప࠸࡜ุ᩿ࡉࢀࡿ㸦EpiSkin™࡜ TER ࡢឤᗘࡣࡑࢀࡒࢀ 98.5%࡜
94%㸧2)ࠋ
௨ୖࡢⅬ࠿ࡽࠊCorrositex®ἲࡣ୍㒊ࡢ⿕㦂≀㉁࡜ΰྜ≀࡟࠾࠸࡚⭉㣗ᛶࡢ᭷↓ࢆホ౯࡛ࡁࡿ࡜⪃࠼ࡽࢀࡿࡶࡢࡢࠊࡑࡢ㐺⏝⠊ᅖࡣ⊃ࡃࠊࡲࡓࠊ㐺⏝ྍ⬟≀㉁ࡢ⠊ᅖࡀ୙᫂☜࡛࠶ࡿࠋࡋࡓࡀࡗ࡚ࠊᮏヨ㦂ἲࡣከᩘࡢ≀㉁ࢆࢫࢡ࣮ࣜࢽࣥࢢホ౯ࡍࡿ┠ⓗ࡟ࡣ㐺ࡋ࡚࠸࡞࠸࡜⪃࠼ࡽࢀࡿࠋࡲࡓࠊ௚ࡢヨ㦂ἲ࡜ẚ࡭࡚ഇ㝜ᛶࡀከ࠸ࡇ࡜࡟␃ពࡍࡿᚲせࡀ࠶ࡿࠋ࡞࠾ࠊin vitro ⭷ࣂࣜ࢔ヨ㦂ἲ࡟࠾ࡅࡿ᳨ฟ᪉ἲ࡜ࡋ࡚ࡣࠊⰍࡢኚ໬ࡢ௚࡟ࠊ໬Ꮫ཯ᛂࠊ㟁Ẽ໬Ꮫ཯ᛂࡶཎ⌮ⓗ࡟ࡣ㑅ᢥྍ⬟࡛࠶ࡾ 1)ࠊ㐺⏝⠊ᅖࢆᣑᙇ࡛ࡁࡿྍ⬟ᛶࡣ࠶ࡿࡀࠊࡑࡢࡼ࠺࡞᳨ฟ᪉ἲࢆྵࡵ
ࡓࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡣࡇࢀࡲ࡛࡟⾜ࢃࢀ࡚࠸࡞࠸࡜ᛮࢃࢀࡿࠋ

㸲㸬┠ⓗ࡜ࡍࡿ≀㉁ཪࡣ〇ရࡢẘᛶࢆホ౯ࡍࡿヨ㦂ἲ࡜ࡋ࡚ࡢࠊ♫఍ⓗཷࡅධࢀᛶཬࡧ⾜ᨻୖࡢ฼⏝ࡢྍ⬟ᛶ ♫఍ⓗཷࡅධࢀᛶ㸸ᮏヨ㦂ἲࡣࠊ≉ู࡞ᡭᢏࡀᚲせ࡛࡞࠸ヨ㦂ἲ࡛࠶ࡾࠊᮏヨ㦂ἲࡢࢩࢫࢸ࣒ࡣᕷ㈍ࡉࢀ࡚࠸ࡿࠋࡲࡓࠊ
⏕ࡁࡓື≀ࢆ⏝࠸࡞࠸࡜࠸࠺Ⅼ࡛ࠊ3Rs ࡢ⢭⚄࡟ྜ⮴ࡋ࡚࠾ࡾࠊ♫఍ⓗཷࡅධࢀᛶࡣ㧗࠸ࠋ ⾜ᨻୖࡢ฼⏝ᛶ㸸
Corrositex®ἲࡣ㐺⏝ྍ⬟≀㉁࡟ CDS ౑⏝ୖࡢไ⣙ࡀ࠶ࡾࠊഇ㝜ᛶࡀከ࠸ࡇ࡜࠿ࡽࠊከᵝ࡞≀㉁ࢆࢫࢡ
࣮ࣜࢽࣥࢢホ౯ࡍࡿ┠ⓗ࡟ࡣ㐺ࡋ࡚࠸࡞࠸࡜⪃࠼ࡽࢀࡿࠋࡓࡔࡋࠊᮏヨ㦂ἲ࡟ࡼࡗ࡚㝧ᛶ࡜ุᐃࡉࢀࡓ
ࡶࡢࢆ㝧ᛶ≀㉁࡜ࡋ࡚㆑ูࡍࡿࡇ࡜࡟㝈ᐃࡍࢀࡤࠊ฼⏝ྍ⬟࡜ุ᩿ࡍࡿࠋ
Corrositex®௨እࡢ in vitro ⭷ࣂࣜ࢔ヨ㦂ἲ࡟ࡘ࠸࡚ࡣ▱ࡽࢀ࡚࠸࡞࠸ࡀࠊ㛤Ⓨࡉࢀࡓሙྜ࡟ࡣ༑ศ࡞ࣂ
ࣜࢹ࣮ࢩࣙࣥ⤖ᯝ࡟ᇶ࡙࠸࡚ࠊࡑࡢ⾜ᨻⓗ฼⏝ᛶࡢホ౯ࢆ⾜࠺࡭ࡁ࡛࠶ࡿࠋ

ཧ⪃ᩥ⊩
1)

OECD (2015), Guidelines for the Testing of Chemicals, Test No. 435: In Vitro membrane barrier test method

2)

JaCVAM ⓶⭵⭉㣗ᛶ㈨ᩱ⦅⧩ጤဨ఍㸸⓶⭵⭉㣗ᛶヨ㦂ホ౯ሗ࿌᭩ In vitro ⭷ࣂࣜ࢔ヨ㦂ࢆ⏝࠸ࡓ⓶⭵⭉㣗ᛶヨ㦂௦᭰ἲࡢホ౯ሗ࿌᭩㸦2017 ᖺ 2 ᭶ 24 ᪥㸧㸬

3)Botham, P.A. et al. (1995), A prevalidation study on in vitro skin corrosivity testing: The report andrecommendations of ECVAM workshop 6. ATLA, 23, 219-255

4)

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

5
5

5)

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

6)

ECVAM. (2001), Statement on the application of the CORROSITEX® Assay for Skin Corrosivity Testing.
15th Meeting of ECVAM Scientific Advisory Committee (ESAC), Ispra, Italy. ATLA 29, 96-97.

7)

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

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 for assessing dermal
corrosivity potential of chemicals.

66ホ౯ሗ࿌᭩

In vitro ⭷ࣂࣜ࢔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. 435: In Vitro Membrane Barrier Test Method
for Skin Corrosion

2
8

せ᪨
࢘ࢧࢠࢆ⏝࠸ࡿ⓶⭵⭉㣗ᛶヨ㦂ࡢື≀ᐇ㦂௦᭰ἲ㸦௦᭰ἲ㸧࡜ࡋ࡚⤒῭༠ຊ㛤Ⓨᶵᵓ㸦OECD㸸

Organisation for Economic Co-operation and Development㸧࡟࡚ヨ㦂࢞࢖ࢻࣛ࢖ࣥ㸦TG: Test
Guideline㸧435 ࡜ࡋ࡚ᢎㄆࡉࢀࡓ in vitro ⭷ࣂࣜ࢔ヨ㦂ࡢ᭷⏝ᛶࢆホ౯ࡋࡓࠋಙ㢗ᛶ࡜ጇᙜᛶ࡜
࠸࠺どⅬ࡟࠾࠸࡚ࠊin vitro ⭷ࣂࣜ࢔ヨ㦂࡜ࡋ࡚ Corrositex ࢆホ౯ࡋࡓ⤖ᯝࠊᮏヨ㦂ἲࡣ୍㒊ࡢ≀㉁࡟࠾࠸࡚⭉㣗ᛶࡢ᭷↓ࢆホ౯࡛ࡁࡿ࡜⪃࠼ࡽࢀࡓࡀࠊࡑࡢ㐺⏝⠊ᅖࡣ⊃ࡃࠊࡲࡓࠊ㐺⏝ྍ⬟≀㉁ࡢศ㢮ࡀ୙᫂☜࡛࠶ࡿࡓࡵࠊ⭉㣗ᛶࡢホ౯࡟⏝࠸ࡿࡇ࡜ࡣᅔ㞴࡛࠶ࡿ࡜ุ᩿ࡋࡓࠋ

39

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㸧ࠋࡇࡢ᪉ἲࡣ⿕㦂≀㉁ࡢ่⃭ᛶࡸ⭉㣗ᛶࢆ᳨ฟࡍ
ࡿヨ㦂ἲ࡜ࡋ࡚㛗ࡃ౑⏝ࡉࢀ࡚ࡁࡓࡶࡢࡢࠊุᐃࢆ⫗║࡛⾜࠺ࡓࡵᐈほᛶ࡟ஈࡋࡃᐇ㦂㛫ࡸ᪋タ㛫࡛ࡢ෌⌧ᛶࡀஈࡋ࠸ࠊ᭦࡟ື≀࡟⃭ࡋ࠸③ࡳ࡜ࢫࢺࣞࢫࢆ୚࠼ࡿࡇ࡜ࡀ♫఍ⓗ࡟ၥ㢟࡜࡞ࡾࠊ௨๓ࡼࡾື≀ࢆ౑⏝ࡋ࡞࠸ື≀ᐇ㦂௦᭰ἲ㸦௨ୗࠊ௦᭰ἲ࡜グࡍ㸧ࡢ㛤Ⓨࡀษᮃࡉࢀ࡚࠸ࡓࠋ
ࡇࡢ௦᭰ἲ࡜ࡋ࡚ࠊ⤒῭༠ຊ㛤Ⓨᶵᵓ㸦OECD㸸Organisation for Economic Co-operation andDevelopment㸧࡟࡚ヨ㦂࢞࢖ࢻࣛ࢖ࣥ㸦TG: Test Guideline㸧435ࠊin vitro ⭷ࣂࣜ࢔ヨ㦂࡜ࡋ࡚ᢎㄆࡉࢀ࡚࠸ࡿ 2㸧ࠋࡇࡢヨ㦂ἲࡣࠊ㐺⏝ࡉࢀࡓ⭉㣗ᛶ≀㉁࡟ࡼࡾᘬࡁ㉳ࡇࡉࢀࡿ⭷ࣂࣜ࢔㞀ᐖࢆ᳨ฟࡍࡿࡇ࡜ࢆᣦᶆ࡟⓶⭵⭉㣗ᛶࢆホ౯ࡋ࡚࠸ࡿࠋᮏヨ㦂ἲࡣࣂࣜࢹ࣮ࢩࣙࣥヨ㦂ࡀᐇ᪋ࡉࢀࠊḢᕞ࡛ࡣ໬Ꮫ≀㉁ࡢ⓶⭵⭉㣗ᛶホ౯ࢆ┠ⓗ࡜ࡋ࡚ᢎㄆࡉࢀࠊ໬Ꮫ≀㉁ࡢࣜࢫࢡ⾲♧㆑ู➼࡟฼⏝ࡉࢀ࡚࠸ࡿࠋ≉࡟᫖௒࡛ࡣᅜ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ศ㢮࡟ᚑࡗ࡚ホ౯ࡉࢀࡿࢣ࣮ࢫࡀቑ࠼࡚࠸ࡿࠋ
ᡃࡀᅜ࡛᪤Ꮡࡢ໬Ꮫ≀㉁ࢆホ౯ࡍࡿሙྜࠊOECD ࡛ᢎㄆࡉࢀࡓヨ㦂᪉ἲ࡟ࡼࡿ⤖ᯝࡣࠊ୍⯡ⓗ࡟⾜ᨻⓗ࡟ཷࡅධࢀࡽࢀࡿࡀࠊ⌧ᅾࡲ࡛௦᭰ἲ࡛ࡢ⤖ᯝࢆࡶ࡜࡟⾜ᨻⓗ࡟ホ౯ࡉࢀࡓ౛ࡣከࡃ࡞
࠸ࠋᏳ඲ᛶホ౯࡟࠾ࡅࡿ௦᭰ἲࡢᬑཬࡀษᮃࡉࢀ࡚࠸ࡿ⌧≧࡟࠾࠸࡚ࠊᡃࡀᅜ࡛ࡶ⛉Ꮫⓗ࡟ጇᙜ࡜ࡉࢀࡓࡶࡢࡣ✚ᴟⓗ࡟ཷࡅධࢀࡿࡇ࡜ࡀᚲせ࡜࡞ࡗ࡚࠸ࡿࠋ
ࡇࢀࡽࡢ≧ἣ࡟㚷ࡳࠊᮏホ౯᭩࡛ࡣࠊ
OECD TG435 ࡟ᢎㄆࡉࢀࡓ in vitro ⭷ࣂࣜ࢔ヨ㦂࡜ࡋ࡚ࠊ
Corrositex ࡢ⭉㣗ᛶホ౯࡟࠾ࡅࡿ᭷⏝ᛶࢆホ౯ࡋࡓ 2㸧ࠋ
2. ヨ㦂ࣉࣟࢺࢥࣝᵓᡂࡢጇᙜᛶ
OECD ࢞࢖ࢻࣛ࢖ࣥ 435ࠕin vitro ⭷ࣂࣜ࢔ヨ㦂ࠖ2㸧ࡢཎ⌮࠾ࡼࡧ᪉ἲ࡟ࡘ࠸࡚㏙࡭ࡿࠋ㸯㸧ཎ⌮  㧗ศᏊ࡛ᵓᡂࡉࢀࡿ⭷ࣂࣜ࢔࠾ࡼࡧ໬Ꮫ≀㉁᳨▱ࢩࢫࢸ࣒㸦Chemical Detection
System㸸CDS㸧࠿ࡽ࡞ࡿࠋࡇࡢ᪉ἲࡢཎ⌮ࡣࠊ໬Ꮫ≀㉁࡟ࡼࡿ⓶⭵⭉㣗ᛶⓎ⌧ᶵᵓࡢ㔜せ࡞ࢫࢸࢵࣉ࡛࠶ࡿ⓶⭵ࡢࣂࣜ࢔ᶵ⬟ࡢ㞀ᐖࢆ᳨ฟࡍࡿࡶࡢ࡛࠶ࡿࠋၟᴗⓗ࡟ࡣᅗ㸯࡟♧ࡍࡼ࠺
࡟Corrositex࡜ࡋ࡚㈍኎ࡉࢀ࡚࠸ࡿ2)ࠋ
2㸧᪉ἲ

 ⿕㦂≀㉁ࡢ㐺⏝࡟ࡼࡿ⭷ࣂࣜ࢔ࡢୗࡢ pH ࡢኚ໬ࡲࡓࡣ࢖ࣥࢪࢣ࣮ࢱ࣮ᾮࡢ≉ᛶࢆྵࡴከᩘࡢⰍࡢኚ໬ࢆ᳨ฟࡋࠊ᭚㟢᫬㛫࡜ほᐹᮇ㛫࡜ࡢ㛵ಀ࡟࠾࠸࡚ㄪ࡭ࡿࡇ࡜࡟ࡼࡾࠊ⭉㣗ᛶࡢ⛬ᗘࢆุᐃࡍࡿࡶࡢ࡛࠶ࡿࠋ ⭷ࣂࣜ࢔ࢆ⏝࠸ࡿᮏヨ㦂ࡣࠊ౑⏝┠ⓗ࡜᝿ᐃࡋ࡚࠸ࡿ⿕㦂≀㉁࡟㐺ྜࡋ࡚࠸ࡿࡇ࡜ࢆணࡵ☜ㄆࡋ࡚࠾࠿ࡡࡤ࡞ࡽ࡞࠸ࠋ⭷ࣂࣜ࢔ヨ㦂ࢆ⾜࠺๓࡟㐺ྜᛶヨ㦂ࢆᐇ᪋ࡋ࡚ࠊ⿕㦂≀㉁ࢆCDS࡟ࡼࡗ᳨࡚ฟྍ⬟࠿ุ᩿ࡍࡿࠋCDSࡀ⿕㦂≀㉁ࢆ᳨ฟ࡛ࡁ࡞࠸ሙྜࠊ⭷ࣂࣜ࢔ヨ㦂ἲࡣࡑࡢ⿕㦂≀㉁ࡢ
⭉㣗ᛶࡢ᭷↓ࢆホ౯ࡍࡿ࡟ࡩࡉࢃࡋࡃ࡞࠸ࡢ࡛ࠊ␗࡞ࡿヨ㦂ἲࢆ⏝࠸ࡿࠋ410ᅗ㸯㸬TER ᴫせᅗ                  ᅗ㸯㸬Corrositex2㸧             
3. 㛤Ⓨ࠾ࡼࡧホ౯࡟౑ࢃࢀࡓ≀㉁ࡢศ㢮ࠊ㑅ᢥ⌮⏤ࡢጇᙜᛶ
Corrositex ࡢ෌⌧ᛶࡣ Botham et al ࡢㄽᩥ࡟ࡼࢀࡤ 3)ࠊࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ࡟࡚ᵝࠎ࡞࢝ࢸࢦࣜ
࣮ࡢ໬Ꮫ≀㉁ࢆྵࡴ 50 ࡢ⿕㦂≀㉁࡟ࡼࡾ᪋タ㛫෌⌧ᛶ࠾ࡼࡧண

ᛶࡀㄪ࡭ࡽࢀ࡚࠸ࡿࠋࡑࢀࡽ

ࡢ≀㉁ࣜࢫࢺࢆ ANNEX 1 ࡟♧ࡍࠋࡲࡓࠊFentem et al ࡢࣂࣜࢹ࣮ࢩࣙࣥ࡟࡚ 60 ≀㉁ࢆ⏝࠸࡚ࠊ෌⌧ᛶ࠾ࡼࡧண

ᛶࡀㄪ࡭ࡽࢀ࡚࠸ࡿ 4㸧ࠋࡑࡢࣜࢫࢺࢆ ANNEX 2 ࡟♧ࡍࠋࡇࢀࡽࡢ୰࡛ࠊ㐺ྜᛶヨ㦂࡛ホ౯ྍ⬟࡜ุ᩿ࡉࢀࠊホ౯࡟౑ࢃࢀࡓ≀㉁ࡣ 45 ≀㉁࡛࠶ࡿࠋ⡿ᅜࡢ௦᭰ἲ࡟㛵ࡍࡿ┬ᗇ㛫㐃⤡఍㆟㸦ICCVAM㸸Interagency Coordinating Committee on the Validation of Alternative Methods)ࡢ➨୕⪅ホ౯࡛ࡣࠊࡇࡢ 45 ≀㉁࡟ຍ࠼ࠊ〇㐀ඖࡢ IVI ♫࠿ࡽᥦ౪ࡉࢀ 118 ≀㉁ࠊྜィ 163 ≀㉁ࡢࢹ࣮
ࢱࡀ౑⏝ࡉࢀࡓ 5㸧ࠋ
4. ヨ㦂ἲࡢṇ☜ᛶࢆホ౯ࡍࡿࡓࡵ࡟⏝࠸ࡽࢀࡓ≀㉁ࡢ in vitro ࠾ࡼࡧཧ↷ࢹ࣮ࢱࡢ᭷↓ࣂࣜࢹ࣮ࢩ࡛ࣙࣥ⏝࠸ࡽࢀࡓ⿕㦂≀㉁ࡢከࡃࡣࠊḢᕞ௦᭰ἲホ౯ࢭࣥࢱ࣮(ECVAM㸸European
Centre for the Validation of Alternative Methods㸧ࡢ⓶⭵⭉㣗ᛶヨ㦂ࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑⏝ࡉࢀࡓ≀㉁࡛࠶ࡾࠊࡑࢀࡽࡢཧ↷ࢹ࣮ࢱࡣධᡭ࡛ࡁࡿࠋ୍᪉ࠊIVI ♫࠿ࡽᥦ౪ࡉࢀࡓ 118 ≀㉁ࡢࢹ࣮ࢱࡢᐈ
ほᛶࡣ☜ㄆ࡛ࡁ࡞࠿ࡗࡓࠋ
5. ヨ㦂ἲࡢṇ☜ᛶ㸦෌⌧ᛶ㸧 Botham et al (1995)ࡢࣂࣜࢹ࣮ࢩࣙࣥ࡟࠾ࡅࡿ᪋タ㛫෌⌧ᛶ࡛ࡣࠊ2 ᪋タࡀ 50 ࡢཧ↷≀㉁㸦25⭉㣗ᛶ≀㉁࠾ࡼࡧ 25 㠀⭉㣗ᛶ≀㉁㸧ࢆ⏝࠸࡚ホ౯ࡋࡓ 3)ࠋLab. A ࡛ホ౯࡛ࡁࡓ≀㉁ࡀ 38ࠊࡑࡢ511࠺ࡕṇ☜ᗘࡣ 74% (28/38)ࠊLab. B ࡛ホ౯࡛ࡁࡓ≀㉁ࡀ 35ࠊࡑࡢ࠺ࡕṇ☜ᗘࡣ 80% (28/35)࡛࠶ࡿ
࡜ࡉࢀ࡚࠾ࡾࠊ᪋タ㛫ࡢ෌⌧ᛶࡣ㧗࠸࡜ุ᩿ࡉࢀ࡚࠸ࡿࠋ
Fentem et al (1998)ࡢࣂࣜࢹ࣮ࢩࣙࣥ࡟࠾ࡅࡿ 60 ≀㉁ࡢ࠺ࡕࠊ20 ≀㉁࡛᪋タෆ࠾ࡼࡧ᪋タ㛫ࡢኚືࡀホ౯ࡉࢀࠊࡑࢀࡽࡢ㛫࡟᭷ព࡞ᕪࡣ࡞࠸࡜ุ᩿ࡉࢀࡓ 4)ࠋ࠸ࡃࡘ࠿ࡢ≀㉁ࡢ⤖ᯝࡀ᪋タ㛫࡛␗࡞ࡗ࡚࠸ࡓࡀࠊECVAM ࡣᮏヨ㦂ἲࡢಙ㢗ᛶ࡜෌⌧ᛶࡣ㧗࠸࡜ุ᩿ࡋࡓࠋࡇࡢ⤖ㄽࡣࠊECVAM⛉Ꮫㅎၥጤဨ఍(ESAC: ECVAM Scientific Advisory Committee, 2001㸧࡛ࡢホ౯ᚋ 6)ࠊICCVAM ࡟࠾࠸࡚ࡶ☜ㄆࡉࢀࡓ 7㸧ࠋ
6. ヨ㦂ἲࡢಙ㢗ᛶ Botham et al (1995)ࠊFentem et al (1998)࠾ࡼࡧICCVAM (1999)ࡢ㞟ィ⤖ᯝࢆ⾲1࡟♧ࡍ3,4,5㸧ࠋࡑࢀࡒࢀࡢሗ࿌࡟グ㍕ࡉࢀࡓṇ☜ᗘࠊឤᗘ࠾ࡼࡧ≉␗ᗘࡣࠊ73-80㸣ࠊ⣙71-89㸣ࠊ65-76㸣࡛࠶ࡾࠊヨ㦂㛫࡛ᕪࡀ኱ࡁ࠿ࡗࡓࠋࡶࡗ࡜ࡶ኱ࡁ࡞ၥ㢟Ⅼࡣࠊࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡛㑅ᢥࡉࢀࡓ≀㉁ࡢ୰࡛ࠊ
CDSࢆ⏝࠸ࡿ㐺ྜᛶヨ㦂࡟ࡼࡾBotham et al (1995)࡛ࡣ70-76%ࠊFentem et al (1998)࡛ࡣ64%ࡢ≀㉁࡛ࡋ࠿ヨ㦂ࢆᐇ᪋࡛ࡁ࡞࠿ࡗࡓ࡜࠸࠺⤖ᯝ࡛࠶ࡿࠋ౛࠼ࡤࠊpH ࡀ 4.5~8.5 ࡢ⠊ᅖࡢỈ⁐ᛶ≀㉁ࡣከࡃࡢሙྜࠊヨ㦂࡟㐺⏝࡛ࡁ࡞࠿ࡗࡓ㸦ࡇࢀࡲ᳨࡛ᰝࡉࢀࡓࡇࡢ pH ⠊ᅖࡢ໬Ꮫ≀㉁ࡢ 85%
ࡣࠊື≀ヨ㦂࡟࡚㠀⭉㣗ᛶ࡛࠶ࡗࡓ㸧ࠋᅛయ(Ỉ⁐ᛶ/㠀Ỉ⁐ᛶ)ࠊᾮయ(Ỉ⁐ᾮ/㠀Ỉ⁐ᾮ)࠾ࡼࡧங
๣࡟㐺⏝࡛ࡁࡿ࡜ࡉࢀ࡚࠸ࡿࡀࠊࣂࣜࢹ࣮ࢩࣙࣥヨ㦂࡟࠾ࡅࡿ㐺ྜヨ㦂࡟࠾࠸࡚ࠊ᳨ฟྍ⬟࡞ኚ໬ࡣ⏕ࡌ࡞࠿ࡗࡓ(ࡍ࡞ࢃࡕࠊࣂࣜࢹ࣮ࢩࣙࣥヨ㦂ἲࡢCDS࡛Ⰽࡢኚ໬ࢆ♧ࡉ࡞࠿ࡗࡓሙྜࡣ㐺⏝࡜ࡳ࡞ࡉࢀ࡞࠸)ࠋࡇࡢ⤖ᯝ࠿ࡽࠊᮏヨ㦂ἲࡣ୍㒊ࡢ⿕㦂≀㉁࡜ΰྜ≀ࡢ⭉㣗ᛶホ౯࡟ࡋ࠿㐺⏝࡛ࡁࡎࠊ௚ࡢ᪉ἲ࡜ྜࢃࡏ࡚฼⏝ࡉࢀࡡࡤ࡞ࡽ࡞࠸ࠋࡲࡓࠊࣂࣜࢹ࣮ࢩࣙࣥ⤖ᯝ࡜࣓࣮࣮࢝ࡢ⤖ᯝࢆྵࡴICCVAMࡢ㞟ィ⤖ᯝࡣࠊ㐺ྜྍ⬟࡞163≀㉁ࡢヲ⣽ࡸ㑅ᐃ⌮⏤ࡀ㈨ᩱ࡟᫂グࡉࢀ࡚࠾ࡽࡎࠊ㐺ษ࡞ホ౯ࡀ୙ྍ⬟࡛࠶ࡗࡓࠋࡼࡗ࡚ࠊᮏጤဨ఍ࡣࠊ⾲1ࡢ୰࡛ࠊFentem et al (1998)ࡢ⤖ᯝࡀࣂࣜࢹ࣮ࢩࣙࣥ⤖ᯝ࡜ࡋ࡚ጇᙜ࡞⤖ᯝ࡜ุ᩿ࡋࠊࡑࡢឤᗘࡀ71%࡛࠶ࡿࡇ࡜࠿ࡽᮏヨ㦂ἲࡢணᛶࡣ㧗ࡃ࡞࠸࡜ุ᩿ࡋࡓࠋホ౯࡛ࡁࡿ≀㉁ࡢ⠊ᅖࡣ᭷ᶵ㓟ࠊ↓ᶵ㓟ࠊ㓟↓Ỉศゎ≀㸦໬Ꮫ≀㉁࠿ࡽ┤᥋ࡲࡓࡣಟ㣭ࡶࡋࡃࡣ㒊ศⓗ࡞⨨᥮࡟ࡼࡗ࡚సࡾฟࡉࢀࡿ㓟࡜ᖜᗈࡃᐃ⩏ࡉࢀࡿࠋࡇࡢࢡࣛࢫ࡟
ࡣࠊ↓Ỉ≀ࠊࣁࣟ㓟ࠊሷ㢮ࡀྵࡲࢀࡿ㸧ࠊ࠾ࡼࡧ ሷᇶ࡞࡝࡟㝈ᐃࡉࢀࡿ࡜グ㍕ࡉࢀ࡚࠸ࡿࠋࣉ
ࣟࢺࢥࣝ࡟ࡼࢀࡤࠊ㝈ᐃࡉࢀࡓ≀㉁ศ㢮ࡢ⠊ᅖ࡛ࠊGHS⣽ศ㢮ࡶྍ⬟࡛࠶ࡿࡇ࡜ࢆᛕ㢌࡟⨨࠸࡚౑⏝ࡍ࡭ࡁ࡛࠶ࡿ࡜ࡉࢀ࡚࠸ࡿࡀࠊ⤖ᯝ࠿ࡽࡑࡢጇᙜᛶࢆㄞࡳྲྀࢀ࡞࠿ࡗࡓࠋ

⾲ 1. Corrositex ࡢணᛶ 3,4,5,7)

ICCVAM ࡢ⥲ィ

Botham et al. (1995)
Lab.A

Lab.B

Fentem et al.㸦1998㸧ṇ☜ᗘ

79% (128/163)

74% (28/38)

80% (28/35)

73%

ឤᗘ

85% (76/89)

81% (17/21)

89% (16/18)

71%

≉␗ᗘ

70% (52/74)

65% (11/17)

71% (12/17)

76%

ホ౯ᩘ

163

35/50

38/50

38/60

7. ௚ࡢ⛉Ꮫⓗ࡞ሗ࿌࡜ࡢẚ㍑ࡢ᭷↓12

 OECD ࡢ⭉㣗ᛶヨ㦂௦᭰ἲ࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ࠊ௚࡟ࠕTG430 ⤒⓶㟁Ẽ᢬ᢠヨ㦂ࠖ8㸧࠾ࡼࡧ
6

ࠕTG431 ࣄࢺ⾲⓶ࣔࢹࣝヨ㦂ࠖ9㸧ࡀᢎㄆࡉࢀ࡚࠸ࡿࠋࡇࢀࡽࡣ࠸ࡎࢀࡶࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࠊICCVAM ࡣࡇࢀࡽࡢヨ㦂ἲ(Rat Skin TER, EpiSkin™࠾ࡼࡧ TER)ࡢṇ☜ᗘࠊឤᗘ࠾ࡼࡧ
≉␗ᗘ࡟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㞴࡛࠶ࡿࡀࠊ㐺⏝ྍ⬟≀㉁࡛࠶ࢀࡤࠊᮏࣔࢹࣝࡣ௚ࡢヨ㦂ἲ࡜ྠ➼ࡢணᛶࢆ᭷ࡍࡿ࡜⪃࠼ࡽࢀࡿࠋ ホ౯ྍ⬟࡞໬Ꮫ≀㉁ࡣ௚ࡢヨ㦂ἲ࡜ẚ࡭࡚㝈ᐃⓗ࡛࠶ࡿࠋ௚ࡢヨ㦂ἲ࡜ẚ㍑ࡋ࡚ឤᗘࡀపࡃࠊ
ഇ㝜ᛶࡀከ࠸ࠋ

⾲ 2㸬ヨ㦂ἲࡢẚ㍑⤖ᯝ
EpiSkinTM(OECD,TER(ICCVAM,

10㸧

5㸧

CORROSITEX(Fentem,

2015)

1999)

1998)4㸧

ṇ☜ᗘ

89.6%

81%

73%

ឤᗘ

98.5%

94%

71%

≉␗ᗘ

79.3%

71%

76%

8. 㸱Rs ཎ๎࡬ࡢ㛵୚㸦ື≀⚟♴㠃࠿ࡽࡢጇᙜᛶ㸧 ື≀ࢆ౑⏝ࡋ࡚࠾ࡽࡎࠊື≀⚟♴㠃࠿ࡽ௦᭰ἲ࡜ࡋ࡚ጇᙜ࡛࠶ࡿࠋ
9. ヨ㦂ἲࡢ᭷⏝ᛶ࡜㝈⏺
 6㸬ヨ㦂ἲࡢಙ㢗ᛶ࡟ࡶグࡋࡓ㐺⏝㝈⏺࠿ࡽࠊከࡃࡢ໬Ꮫ≀㉁࠾ࡼࡧΰྜ≀࡟㐺⏝࡛ࡁ࡞࠸ྍ⬟ᛶࡀ࠶ࡿࠋุᐃࡶ⭉㣗ᛶࡢ᭷↓࡟㝈ࡽࢀ࡚࠾ࡾࠊࡑࡢ㝈⏺ࢆ༑ศ࡟ᢕᥱࡋ࡚౑⏝ࡍ࡭ࡁ࡛࠶ࡿࠋ
⾲ 3 ࡟♧ࡍ⩦⇍ᗘ☜ㄆ≀㉁ࢆṇࡋࡃศ㢮ࡍࡿࡇ࡜࡟ࡼࡾᑓ㛛ᢏ⾡ࡢ⩦⇍࡟ࡘ࠸࡚☜ㄆࡍࡿࡇ࡜
ࡀ࡛ࡁࡿ࡜ TG435 ࡟ࡣグ㍕ࡀ࠶ࡿࠋ

713⾲ 3 ⩦⇍ᗘ☜ㄆ୍ぴ⾲ 2㸧
1,2

UN GHS ༊ศ 3

VRM ༊ศ3

CASRN໬Ꮫ≀㉁ࢡࣛࢫ

In vivo ヨ㦂⤖ᯝ࡟ᇶ࡙ࡃIn vitro ヨ㦂⤖ᯝ࡟ᇶ࡙ࡃ࡯࠺⣲ࣇࣝ࢜ࣜࢻ஧Ỉ࿴≀

13319-75-0

↓ᶵ㓟

1A

1A

◪㓟

7697-37-2

↓ᶵ㓟

1A

1A

஬ሷ໬ࣜࣥ

10026-13-8 ↓ᶵ㓟ࡢ๓㥑≀㉁

1A

1A

ࣂࣞࣜࣝ㓟

638-29-9㓟ࢡࣟࣛ࢖ࢻ

1B

1B

Ỉ㓟໬ࢼࢺ࣒ࣜ࢘

1310-73-2

↓ᶵሷᇶ

1B

1B

1-(2-࢔࣑ࣀ࢚ࢳࣝ)ࣆ࣌ࣛࢪࣥ

140-31-8⬡⫫᪘࢔࣑ࣥ

1B

1B࣋ࣥࢮࣥࢫࣝ࣍ࣥ㓟ࢡࣟࣛ࢖ࢻ

98-09-9㓟ࢡࣟࣛ࢖ࢻ

1C

1C

N,N-ࢪ࣓ࢳࣝ࣋ࣥࢪࣝ࢔࣑ࣥ

103-83-3

࢔ࢽࣜࣥ

1C

1C

ࢸࢺ࢚ࣛࢳࣞࣥ࣌ࣥࢱ࣑ࣥ

112-57-2⬡⫫᪘࢔࣑ࣥ

1C

1C

࢜࢖ࢤࣀ࣮ࣝ

97-53-0ࣇ࢙ࣀ࣮ࣝ㢮

㠀⭉㣗ᛶ㠀⭉㣗ᛶ࢔ࢡࣜࣝ㓟ࣀࢽࣝ

2664-55-3

࢔ࢡࣜࣝ㓟㸭
࣓ࢱࢡࣜࣝ㓟㠀⭉㣗ᛶ㠀⭉㣗ᛶⅣ㓟Ỉ⣲ࢼࢺ࣒ࣜ࢘

144-55-8

↓ᶵሷᇶ

㠀⭉㣗ᛶ

㠀⭉㣗ᛶ

  ໬Ꮫ≀㉁

1㸸ୖ࡟グ㍕ࡋࡓ12ࡢ໬Ꮫ≀㉁ࡣࠊ⓶⭵⭉㣗ᛶ≀㉁࡟㛵ࡍࡿGHS ⣽༊ศࡢࡑࢀࡒࢀ࠿ࡽ3≀㉁ࡎ
ࡘ࡜㠀⭉㣗ᛶࡢ3≀㉁࡛࠶ࡿࠋࡇࢀࡽࡣ᳨ド῭ࡳཧ↷ヨ㦂ἲ࡜ᵓ㐀ⓗ࡟ࡶᶵ⬟ⓗ࡟ࡶ㢮ఝࡢヨ㦂ἲ࡟ࡘ࠸࡚ࠊࡑࡢ⢭ᗘ࡜ಙ㢗ᛶࢆ♧ࡍ┠ⓗ࡟౑⏝ࡉࢀࡿࡼ࠺≉ᐃࡉࢀ࡚࠸ࡿࠊ໬Ꮫ≀㉁᭱ᑠ
ࣜࢫࢺࡢ୰࡛ཧ↷໬Ꮫ≀㉁࡜ࡋ࡚ิᣲࡉࢀ࡚࠸ࡿ40ࡢ୰࠿ࡽ㑅ᢥࡋࡓ5,11)ࠋࡇࢀࡽࡢ໬Ꮫ≀㉁ࡣ
ᕷ㈍ᴗ⪅࠿ࡽᐜ᫆࡟㉎ධ࡛ࡁࠊUN GHS ⣽༊ศࡣ㧗ရ㉁ࡢin vivo ヨ㦂ࡢ⤖ᯝ࡟ᇶ࡙࠸࡚࠸ࡿ11)ࠋ
2㸸⣧ᗘࡣ90㸣௨ୖ
3㸸UN GHS ⣽༊ศ1A, 1B ࠾ࡼࡧ1C ࡟ࡣUN ໟ⿦➼⣭IࠊII ࠾ࡼࡧIII ࡀࡑࢀࡒࢀᑐᛂࡍࡿࠋ
NC; 㠀⭉㣗ᛶ
10. ࡑࡢ௚㸦≉チࡢ᭷↓࡞࡝㸧 ≉チ࡟ࡘ࠸࡚ࡣ♧ࡉࢀ࡚࠸࡞࠸ࠋ
11. ⤖ㄽ
ಙ㢗ᛶ࡜ጇᙜᛶ࡜࠸࠺どⅬ࡟࠾࠸࡚ࠊin vitro ⭷ࣂࣜ࢔ヨ㦂࡛࠶ࡿ Corrositex ࢆホ౯ࡋࡓ⤖ᯝࠊᙜヱヨ㦂ἲࡣ௚ࡢヨ㦂ἲ࡜ẚ࡭࡚ࠊホ౯ྍ⬟࡞໬ྜ≀ࡀ㝈ᐃࡉࢀࠊഇ㝜ᛶࡀከࡃ☜ㄆࡉࢀࡓࠋᙜ
ヱヨ㦂ἲ୍㒊ࡢ≀㉁࡟࠾࠸࡚⭉㣗ᛶࡢ᭷↓ࢆホ౯࡛ࡁࡿ࡜⪃࠼ࡽࢀࡓࡀࠊࡑࡢ㐺⏝⠊ᅖࡣ⊃ࡃࠊ814ࡲࡓࠊ㐺⏝ྍ⬟≀㉁ࡢࢡࣛࢫࡀ୙᫂☜࡛࠶ࡿࡓࡵࠊᮏጤဨ఍ࡣࠊᙜヱヨ㦂ἲࢆ⾜ᨻ┠ⓗࡢࡓࡵࡢ⭉㣗ᛶホ౯࡟⏝࠸ࡿࡇ࡜ࡣᅔ㞴࡛࠶ࡿ࡜ุ᩿ࡋࡓࠋ

12. ᩥ⊩
1) OECD (2015) Guideline for the testing of chemicals. 404, Acute Dermal Irritation/Corrosion
2) OECD (2006) Guidelines for the Testing of Chemicals, Test No. 435: In Vitro membrane barrier test
method3) Botham, P.A. et al. (1995) A prevalidation study on in vitro skin corrosivity testing: The report andrecommendations of ECVAM workshop 6. ATLA, 23, 219-255
4) 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
5) ICCVAM (1999) NIH Publication No.99-4495. Corrositex: An in vitro test method for assessing dermal
corrosivity potential of chemicals.
6) ECVAM (2001) Statement on the Application of the CORROSITEX Assay for Skin Corrosivity Testing.
15th Meeting of ECVAM Scientific Advisory Committee (ESAC), Ispra, Italy. ATLA 29, 96-97.
7) 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.
8) OECD (2015) Guideline for the testing of chemicals. 430, in vitro Skin Corrosion: Transcutaneouselectrical resistance test (TER).
9) OECD (2016) Guideline for the testing of chemicals. 431, in vitro Skin Corrosion: Human skin model
test.
10) OECD (2015) Series on Testing & Assessment No. 219, Performance Standards for the assessment ofproposed similar or modified in vitro reconstructed human epidermis (RhE) test methods for skincorrosion testing as described in TG 431
11) ICCVAM (2004) ICCVAM Recommended Performance Standards for In Vitro Test Methods for Skin
Corrosion. NIEHS, NIH Publication No. 04-4510. Available at:[http://www.ntp.niehs.nih.gov/iccvam/docs/dermal_docs/ps/ps044510.pdf].

915

ANNEX 1: Botham 1995 ࡢࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑ࢃࢀࡓ 50 ≀㉁

10

16

11
17

12
18

ANNEX 2: Fentem 1998 ࡢࣂࣜࢹ࣮ࢩ࡛ࣙࣥ౑ࢃࢀࡓ 60 ≀㉁

13

19

20

OECD/OCDE

435
Adopted:28 July 2015

OECD GUIDELINE FOR THE TESTING OF CHEMICALSIn Vitro Membrane Barrier Test Method for Skin Corrosion

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 435 provides an in vitro membrane barrier test method that can beused to identify corrosive chemicals. The test method utilizes an artificial membrane designed to respondto corrosive chemicals in a manner similar to animal skin in situ.
2.
Skin corrosivity has traditionally been assessed by applying the test chemical to the skin of living
animals and assessing the extent of tissue damage after a fixed period of time (2). Besides the present Test
Guideline, a number of other in vitro test methods have been adopted as alternatives (3) (4) to the standard
in vivo rabbit skin procedure (OECD TG 404) used to identify corrosive chemicals (2). The UN GHS
tiered testing and evaluation strategy for the assessment and classification of skin corrosivity and theOECD Guidance document on Integrated Approaches to Testing and Assessment (IATA) for Skin
Irritation/Corrosion recommend the use of validated and accepted in vitro test methods under modules 3
and 4 (1) (5). The IATA describes several modules which group information sources and analysis tools
and provides guidance on (i) how to integrate and use existing the test and non-test data for the assessmentof the skin irritation and skin corrosion potentials of chemicals and (ii) proposes an approach when further
testing is needed, including when negative results are found (5). In this modular approach, positive resultsfrom in vitro test methods can be used to classify a chemical as corrosive without the need for animal
testing, thus reducing and refining the use of animals in and avoiding the pain and distress that might occur
if animals were used for this purpose.
3.
Validation studies have been completed for the in vitro membrane barrier test method
commercially available as Corrositex® (6)(7)(8), showing an overall accuracy to predict skin corrosivity of
79% (128/163), a sensitivity of 85% (76/89), and a specificity of 70% (52/74) for a database of 163
substances and mixtures (7). Based on its acknowledged validity, this validated reference test method(VRM) has been recommended for use as part of a tiered testing strategy for assessing the dermal corrosion
hazard potential of chemicals (5) (7). Before an in vitro membrane barrier test method for skin corrosion
can be used for regulatory purposes, its reliability, relevance (accuracy), and limitations for its proposed
use should be determined to ensure that it is similar to that of the VRM (9), in accordance with the predefined performance standards (PS) (10). 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 andincluded in this Test Guideline. Currently, only one in vitro test method is covered by this Test Guideline,
the commercially available Corrositex® test method. .
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.

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4.
Other test methods for skin corrosivity testing are based on the use of reconstituted human skin
(OECD TG 431) (3) and isolated rat skin (OECD TG 430) (4). This Test Guideline also provides for
subcategorisation of corrosive chemicals into the three UN GHS Sub-categories of corrosivity and thethree UN Transport Packing Groups for corrosivity hazard. This Test Guideline was originally adopted in2006 and updated in 2015 to refer to the IATA guidance document and update the list of proficiency
substances.
DEFINITIONS
5.

Definitions used are provided in Annex 1.

INITIAL CONSIDERATIONS AND LIMITATIONS
6.
The test described in this Guideline allows the identification of corrosive test chemicals andallows the sub-categorisation of corrosive test chemicals according to the UN GHS (Table 1) (1). In
addition, such a test method may be used to make decisions on the corrosivity and non-corrosivity of
specific classes of chemicals, e.g., organic and inorganic acids, acid derivatives1, and bases for certaintransport testing purposes (7)(11)(12). This Test Guideline describes a generic procedure similar to thevalidated reference test method (7). While this Test Guideline does not provide adequate information onskin irritation, it should be noted that OECD TG 439 specifically addresses the health effect skin irritationin vitro (13). For a full evaluation of local skin effects after a single dermal exposure, the GuidanceDocument No. 203 on Integrated Approaches for Testing Assessment should be consulted (5).
Table 1. The UN GHS Skin Corrosive Category and Subcategories (1)Corrosive Category(category 1)(applies to authorities notusing subcategories)
Corrosive

Potential Corrosive
Subcategories(only applies to someauthorities)

Corrosive in >1 of 3 animals
Exposure

Observation

Corrosive subcategory 1A<3 minutes<1 hourCorrosive subcategory 1B>3 minutes / <1 hour

<14 days

Corrosive subcategory 1C>1 hour / <4 hours<14 days

7.
A limitation of the validated reference test method (7) is that many non-corrosive chemicals andsome corrosive chemicals may not qualify for testing, based on the results of the initial compatibility test(see paragraph 13). Aqueous chemicals with a pH in the range of 4.5 to 8.5 often do not qualify for testing;however, 85% of chemicals tested in this pH range were non-corrosive in animal tests (7). The in vitro
membrane barrier test methods may be used to test solids (soluble or insoluble in water), liquids (aqueous
or non-aqueous), and emulsions. However, test chemicals not causing a detectable change in thecompatibility test ( i.e., colour change in the Chemical Detection System (CDS) of the validated reference
test method) cannot be tested with the membrane barrier test method and should be tested using other test
methods.

1“Acid derivative” is a non-specific class designation and is broadly defined as an acid produced from a chemical
either directly or by modification or partial substitution. This class includes anhydrides, halo acids, salts, and othertypes of chemicals.

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PRINCIPLE OF THE TEST8.
The test system comprises two components: a synthetic macromolecular bio-barrier and achemical detection system (CDS); this test method detects via the CDS membrane barrier damage causedby corrosive test chemicals after the application of the test chemical to the surface of the synthetic
macromolecular membrane barrier (7), presumably by the same mechanism(s) of corrosion that operate on
living skin.
9.
Penetration of the membrane barrier (or breakthrough) might be measured by a number ofprocedures or CDS, including a change in the colour of a pH indicator dye or in some other property of theindicator solution below the barrier.
10.
The membrane barrier should be determined to be valid, i.e., relevant and reliable, for its
intended use. This includes ensuring that different preparations are consistent in regard to barrier
properties, e.g., capable of maintaining a barrier to non-corrosive chemicals, able to categorize thecorrosive properties of chemicals across the various UN GHS Sub-categories of corrosivity (1). The
classification assigned is based on the time it takes a chemical to penetrate through the membrane barrier tothe indicator solution.
DEMONSTRATION OF PROFICIENCY
11.
Prior to routine use of the in vitro membrane barrier test method, adhering to this Test Guideline,
laboratories should demonstrate technical proficiency by correctly classifying the twelve Proficiency
Substances recommended in Table 2. 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 (10)) provided that the same selection criteria as describedin Table 1 is applied.

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OECD/OCDE
Table 2: Proficiency Substances

1

Substance2

CASRN

Chemical ClassIn Vivo UN
GHS Subcategory3

In Vitro UN
GHS Subcategory3

Boron trifluoride dihydrate

13319-75-0

Inorganic acids

1A

1A

Nitric acid7697-37-2

Inorganic acids

1A

1A

Phosphorus pentachloride

10026-13-8

Precursors of inorganic
acids

1A

1A

Valeryl chloride

638-29-9Acid chlorides

1B

1B

Sodium Hydroxide

1310-73-2

Inorganic bases

1B

1B

1-(2-Aminoethyl) piperazine

140-31-8

Aliphatic amines

1B

1B

Benzenesulfonyl chloride

98-09-9

Acid chlorides

1C

1C

N,N-Dimethyl benzylamine

103-83-3

Anilines

1C

1C

Tetraethylenepentamine

112-57-2

Aliphatic amines

1C

1C

Eugenol

97-53-0

PhenolsNC

NC

Nonyl acrylate

2664-55-3

Acrylates/methacrylatesNC

NC

Sodium bicarbonate

144-55-8

Inorganic salts

NC

NC

1

The twelve substances listed above contain three substances from each of the three UN GHS subcategories forcorrosive substances and three non-corrosive substances, are readily available from commercial suppliers, and the UN
GHS subcategory is based on the results of high-quality in vivo testing. These substances are taken from the list of 40
reference substances that are included in the minimum list of chemicals identified for demonstrating the accuracy andreliability of test methods that are structurally and functionally similar to the validated reference test method, andwere selected from the 163 reference chemicals that were originally used to validate the reference test method(Corrositex®) (7) (10) (14). The goal of this selection process was to include, to the extent possible, chemicals that:were representative of the range of corrosivity responses (e.g., non-corrosives; UN Packing Groups I, II, and III
corrosives) that the validated reference test method is capable of measuring or predicting; were representative of thechemical classes used during the validation process; have chemical structures that were well-defined; induced
reproducible results in the validated reference test method; induced definitive results in the in vivo reference test;
were commercially available; and were not associated with prohibitive disposal costs (14).
2
Substances tested neat or with purity t 90%
3
The corresponding UN Packing groups are I, II and III, respectively, for the UN GHS Sub-categories 1A, 1B and 1C.
NC; Non-corrosive.

PROCEDURE
12.
The following paragraphs describe the components and procedures of an artificial membrane
barrier test method for corrosivity assessment (7) (15), based on the current VRM, i.e., the commerciallyavailable Corrositex®. The membrane barrier and the compatibility/indicator and categorisation solutions
can be constructed, prepared or obtained commercially such as in the case of the VRM Corrositex®. A
sample test method protocol for the validated reference test method is available (7). Testing should be
performed at ambient temperature (17-25ºC) and the components should comply with the following
conditions.
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Test Chemical Compatibility Test
13.
Prior to performing the membrane barrier test, a compatibility test is performed to determine if
the test chemical is detectable by the CDS. If the CDS does not detect the test chemical, the membrane
barrier test method is not suitable for evaluating the potential corrosivity of that particular test chemical
and a different test method should be used. The CDS and the exposure conditions used for thecompatibility test should reflect the exposure in the subsequent membrane barrier test.
Test Chemical Timescale Category Test
14.
If appropriate for the test method, a test chemical that has been qualified by the compatibility test
should be subjected to a timescale category test, i.e., a screening test to distinguish between weak andstrong acids or bases. For example, in the validated reference test method a timescale categorization test is
used to indicate which of two timescales should be used based on whether significant acid or alkaline
reserve is detected. Two different breakthrough timescales should be used for determining corrosivity and
UN GHS skin corrosivity Sub-category, based on the acid or alkali reserve of the test chemical.
Membrane Barrier Test Method Components
Membrane Barrier
15.
The membrane barrier consists of two components: a proteinaceous macromolecular aqueous gel
and a permeable supporting membrane. The proteinaceous gel should be impervious to liquids and solids
but can be corroded and made permeable. The fully constructed membrane barrier should be stored under
pre-determined conditions shown to preclude deterioration of the gel, e.g., drying, microbial growth,shifting, cracking, which would degrade its performance. The acceptable storage period should bedetermined and membrane barrier preparations not used after that period.
16.
The permeable supporting membrane provides mechanical support to the proteinaceous gel
during the gelling process and exposure to the test chemical. The supporting membrane should prevent
sagging or shifting of the gel and be readily permeable to all test chemicals.
17.
The proteinaceous gel, composed of protein, e.g., keratin, collagen, or mixtures of proteins,forming a gel matrix, serves as the target for the test chemical. The proteinaceous material is placed on thesurface of the supporting membrane and allowed to gel prior to placing the membrane barrier over theindicator solution. The proteinaceous gel should be of equal thickness and density throughout, and with noair bubbles or defects that could affect its functional integrity.
Chemical Detection System (CDS)18.
The indicator solution, which is the same solution used for the compatibility test, should respondto the presence of a test chemical. A pH indicator dye or combination of dyes, e.g., cresol red and methyl
orange that will show a colour change, in response to the presence of the test chemical, should be used.
The measurement system can be visual or electronic.
19.
Detection systems that are developed for detecting the passage of the test chemical through thebarrier membrane should be assessed for their relevance and reliability in order to demonstrate the range ofchemicals that can be detected and the quantitative limits of detection.

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OECD/OCDETEST PERFORMANCE
Assembly of the Test Method Components
20.
The membrane barrier is positioned in a vial (or tube) containing the indicator solution so that the
supporting membrane is in full contact with the indicator solution and with no air bubbles present. Careshould be taken to ensure that barrier integrity is maintained.
Application of the Test Chemical
21.
A suitable amount of the test chemical, e.g., 500 PL of a liquid or 500 mg of a finely powderedsolid (7), is carefully layered onto the upper surface of the membrane barrier and evenly distributed. An
appropriate number of replicates, e.g., four (7), is prepared for each test chemical and its corresponding
controls (see paragraphs 23 to 25). The time of applying the test chemical to the membrane barrier isrecorded. To ensure that short corrosion times are accurately recorded, the application times of the test
chemical to the replicate vials are staggered.
Measurement of Membrane Barrier Penetrations
22.
Each vial is appropriately monitored and the time of the first change in the indicator solution, i.e.,barrier penetration, is recorded, and the elapsed time between application and penetration of the membrane
barrier determined.
Controls
23.
In tests that involve the use of a vehicle or solvent with the test chemical, the vehicle or solvent
should be compatible with the membrane barrier system, i.e., not alter the integrity of the membrane
barrier system, and should not alter the corrosivity of the test chemical. When applicable, solvent (orvehicle) control should be tested concurrently with the test chemical to demonstrate the compatibility ofthe solvent with the membrane barrier system.
24.
A positive (corrosive) control with intermediate corrosivity activity, e.g., 110 ± 15 mg sodiumhydroxide (UN GHS Corrosive Sub-category 1B) (7), should be tested concurrently with the test chemical
to assess if the test system is performing in an acceptable manner. A second positive control that is of thesame chemical class as the test chemical may be useful for evaluating the relative corrosivity potential of acorrosive test chemical. Positive control(s) should be selected that are intermediate in their corrosivity(e.g., UN GHS Sub-category 1B) in order to detect changes in the penetration time that may beunacceptably longer or shorter than the established reference value, thereby indicating that the test system
is not functioning properly. For this purpose, extremely corrosive (UN GHS Sub-category 1A) or noncorrosive chemicals are of limited utility. A corrosive UN GHS Sub-category 1B chemical would allow
detection of a too rapid or too slow breakthrough time. A weakly corrosive (UN GHS Sub-category 1C)
might be employed as a positive control to measure the ability of the test method to consistently distinguishbetween weakly corrosive and non-corrosive chemicals. Regardless of the approach used, an acceptable
positive control response range should be developed based on the historical range of breakthrough times
for the positive control(s) employed, such as the mean ± 2-3 standard deviations. In each study, the exact
breakthrough time should be determined for the positive control so that deviations outside the acceptable
range can be detected.
25.
A negative (non-corrosive) control, e.g., 10% citric acid, 6% propionic acid (7), should also be
tested concurrently with the test chemical as another quality control measure to demonstrate the functional
integrity of the membrane barrier.
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Study Acceptability Criteria
26.
According to the established time parameters for each of the UN GHS corrosivity Sub-categories,
the time (in minutes) elapsed between application of a test chemical to the membrane barrier and barrier
penetration is used to predict the corrosivity of the test chemical. For a study to be considered acceptable,
the concurrent positive control should give the expected penetration response time (e.g. 8-16 min
breakthrough time for sodium hydroxide if used as a positive control), the concurrent negative control
should not be corrosive, and, when included, the concurrent solvent control should neither be corrosive nor
should it alter the corrosivity potential of the test chemical. Prior to routine use of a test method thatadheres to this Test Guideline, laboratories should demonstrate technical proficiency, using the twelve
substances recommended in Table 2. For new “me-too” test methods developed under this Test Guidelinethat are structurally and functionally similar to the validated reference test method (14) the pre-defined
performance standards should be used to demonstrate the reliability and accuracy of the new test methodprior to its use for regulatory testing (10).

Interpretation of Results and Corrosivity Classification of Test Chemicals
27.
The time (in minutes) elapsed between application of the test chemical to the membrane barrier
and barrier penetration is used to classify the test chemical in terms of UN GHS corrosive Sub-categories(1) and, if applicable, UN Packing Group (16). Cut-off time values for each of the three corrosive
subcategories are established for each proposed test method. Final decisions on cut-off times should
consider the need to minimize under-classification of corrosive hazard ( i.e., false negatives). In the present
Test Guideline, the cut-off times of Corrositex® as described in table 3 should be used as it represents the
only test method currently falling within the test guideline (7).
Table 3. Corrositex® prediction model
Mean breakthrough time (min.)UN GHS prediction3

Category 1 test chemicals1(determined by the method’s
categorization test)

Category 2 test chemicals2(determined by the method’s
categorization test)

0-3 min.

0-3 min.

Corrosive
optional Sub-category 1A> 3 to 60 min.

> 3 to 30 min.

Corrosive
optional Sub-category 1B> 60 to 240 min.

> 30 to 60 min.

Corrosive
optional Sub-category 1C> 240 min.

> 60 min.

Non-corrosive

1

Test chemicals with high acid/alkaline reserve (6)2Test chemicals with low acid/alkaline reserve (6)3UN GHS Subcategories 1A, 1B and 1C correspond to UN packing groups I, II and III respectively

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OECD/OCDEDATA AND REPORTING
Data
28.
The time (in minutes) elapsed between application and barrier penetration for the test chemical
and the positive control(s) should be reported in tabular form as individual replicate data, as well as means± the standard deviation for each trial.
Test Report
29.

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
by chemical identity (see above), quantitative occurrence and relevant
physicochemical properties of the constituents;
 Physical appearance, water solubility, and additional relevant physicochemical
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.
Vehicle:- Identification, concentration (where appropriate), volume used;- Justification for choice of vehicle.
In vitro membrane barrier model and protocol used, including demonstrated accuracy andreliability
Test Conditions:– Description of the apparatus and preparation procedures used;– Source and composition of the in vitro membrane barrier used;– Composition and properties of the indicator solution;– Method of detection;– Test chemical and control substance amounts;– Number of replicates;– Description and justification for the timescale categorisation test;– Method of application;– Observation times.
– Description of the evaluation and classification criteria applied;– Demonstration of proficiency in performing the test method before routine use by
testing of the proficiency chemicals.
Results:
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435– Tabulation of individual raw data from individual test and control samples for each
replicate;– Descriptions of other 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 ofChemicals (GHS), First 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,
Corrosion. 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. 430.): In Vitro Skin Corrosion:
Transcutaneous Electrical Resistance (TER). Organisation for Economic Cooperation and Development,
Paris.
(5)
Guidance Document on Integrated Approaches to Testing and Assessment of Skin
Irritation/Corrosion.Environment, Health and Safety Publications, Series on Testing and Assessment,(No. 203.). Organisation for Economic Cooperation and Development, Paris.
(6)
Fentem, J.H., Archer, G.E.B., Balls, M., Botham, P.A., Curren, R.D., Earl, L.K., Esdaile, D.J.,
Holzhutter, 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. Toxicology In Vitro 12, 483524.
(7)
ICCVAM. (1999). Corrositex®. An In Vitro Test Method for Assessing Dermal Corrosivity
Potential of Chemicals. The Results of an Independent Peer Review Evaluation Coordinated by
ICCVAM, NTP and NICEATM. NIEHS, NIH Publication (No. 99-4495.)(8)
Gordon V.C., Harvell J.D. and Maibach H.I. (1994). Dermal Corrosion, the Corrositex® System:
A DOT Accepted Method to Predict Corrosivity Potential of Test Materials. In vitro Skin ToxicologyIrritation, Phototoxicity, Sensitization. Alternative Methods in Toxicology 10, 37-45.
(9)OECD. (2005). Guidance Document on the Validation and International Acceptance of New or
Updated Test Methods for Hazard Assessment. Environmental, Health and Safety Publications. Series on
testing and Assesment (No. 34.).
(10) Performance Standards for the Assessment of Proposed Similar or Modified In Vitro Membrane
Barrier Test Method for Skin Corrosion in Relation to TG 435. Organisation for Economic Cooperation
and Development, Paris.
(11) ECVAM. (2001). Statement on the Application of the CORROSITEX£ Assay for Skin
Corrosivity Testing. 15th Meeting of ECVAM Scientific Advisory Committee (ESAC), Ispra, Italy.
ATLA 29, 96-97.
(12) U.S. DOT. (2002). Exemption DOT-E-10904 (Fifth Revision). (September 20, 2002).
Washington, D.C., U.S. DOT. Available at: [http://www.hazmat.dot.gov/exemptions/E10904.pdf].
(13) OECD. (2015). OECD Guideline for the Testing of Chemicals (No. 439.). In Vitro Skin
Irritation: Reconstructed Human Epidermis Test Method. . Organisation for Economic Cooperation andDevelopment, Paris.
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435(14) ICCVAM. (2004). ICCVAM Recommended Performance Standards for In Vitro Test Methodsfor
Skin
Corrosion.
NIEHS,
NIH
Publication
No.
04-4510.
Availableat:[http://www.ntp.niehs.nih.gov/iccvam/docs/dermal_docs/ps/ps044510.pdf].
(15) U.S.
EPA.
(1996).
Method1120,
Dermal[http://www.epa.gov/osw/hazard/testmethods/sw846/pdfs/1120.pdf].

Corrosion.

Available

at:(16) United Nations (UN). (2013). UN Recommendations on the Transport of Dangerous Goods,
Model Regulations, 18th Revised Edition (Part, Chapter 2.8), UN, 2013. Available at:[http://www.unece.org/fileadmin/DAM/trans/danger/publi/unrec/rev18/English/Rev18_Volume1_Part2.p
df]..

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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 (9).
Chemical: means a substance or a mixture.
Chemical Detection System (CDS): A visual or electronic measurement system with an indicatorsolution that responds to the presence of a test chemical, e.g., by a change in a pH indicator dye, or
combination of dyes, that will show a colour change in response to the presence of the test chemical or byother types of chemical or electrochemical reactions.
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 (9).
GHS (Globally Harmonized System of Classification and Labelling of Chemicals): a system proposing
the classification of chemicals (substances and mixtures) according to standardized types and levels ofphysical, health and environmental hazards, and addressing corresponding communication elements, suchas pictograms, signal words, hazard statements, precautionary statements and safety data sheets, so that toconvey 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 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.
Performance standards: Standards, based on a validated test method, that provide a basis for evaluating
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 selected from among thechemicals used to demonstrate the acceptable performance of the validated test method; and (iii) thesimilar levels of reliability and accuracy , based on what was obtained for the validated test method, thatthe proposed test method should demonstrate when evaluated using the minimum list of Reference
Chemicals (9).
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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 (9).
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 (9).
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 (9).
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 (9).
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.
Test chemical: means what is being tested.
UVCB: substances of unknown or variable composition, complex reaction products or biologicalmaterials.

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