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Elevated temperatures accelerate chemical reactions, including hydrolysis and oxidation, which can break peptide bonds or alter their structure, diminishing their efficacy

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The current standard operating procedure by the European Center for the Validation of Alternative Methods (ECVAM) recommends the use of MTT reduction as parameter to make prediction on the skin irritancy of substances using EpiSkin TM test method, while IL-1 endpoint is regarded as a useful adjunct to MTT assay 48