title: Assessment of cosmetic ingredients in the in vitro reconstructed human epidermis test method EpiSkin™ using HPLC/UPLC-spectrophotometry in the MTT-reduction assay.
abstract: Cosmetics Europe recently established HPLC/UPLC-spectrophotometry as a suitable alternative endpoint detection system for measurement of formazan in the MTT-reduction assay of reconstructed human tissue test methods irrespective of the test system involved. This addressed a known limitation for such test methods that use optical density for measurement of formazan and may be incompatible for evaluation of strong MTT reducer and/or coloured chemicals. To build on the original project, Cosmetics Europe has undertaken a second study that focuses on evaluation of chemicals with functionalities relevant to cosmetic products. Such chemicals were primarily identified from the Scientific Committee on Consumer Safety (SCCS) 2010 memorandum (addendum) on the in vitro test EpiSkin™ for skin irritation testing. Fifty test items were evaluated in which both standard photometry and HPLC/UPLC-spectrophotometry were used for endpoint detection. The results obtained in this study: 1) provide further support for Within Laboratory Reproducibility of HPLC-UPLC-spectrophotometry for measurement of formazan; 2) demonstrate, through use a case study with Basazol C Blue pr. 8056, that HPLC/UPLC-spectrophotometry enables determination of an in vitro classification even when this is not possible using standard photometry and 3) addresses the question raised by SCCS in their 2010 memorandum (addendum) to consider an endpoint detection system not involving optical density quantification in in vitro reconstructed human epidermis skin irritation test methods.
authors.affiliation: L'Oréal Research & Innovation, Aulnay Sous Bois, France. Electronic address: nalepee@rd.loreal.com.
authors.forename: N
authors.initials: N
authors.lastname: Alépée
authors.affiliation: Chanel Parfums Beauté, Neuilly sur Seine, France.
authors.forename: J
authors.initials: J
authors.lastname: Hibatallah
authors.affiliation: Cosmetics Europe, The Personal Care Association, Brussels, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Klaric
authors.affiliation: Henkel AG & Co. KGaA, Düsseldorf, Germany.
authors.forename: K R
authors.initials: KR
authors.lastname: Mewes
authors.affiliation: Beiersdorf AG, Hamburg, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Pfannenbecker
authors.affiliation: The Procter & Gamble Company, Egham, Surrey, United Kingdom.
authors.forename: P
authors.initials: P
authors.lastname: McNamee
mesh_terms: D001681:Biological Assay; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003358:Cosmetics / Q000633:toxicity*; D004817:Epidermis / Q000187:drug effects*; D006801:Humans; D066298:In Vitro Techniques; D010084:Oxidation-Reduction; D023422:Skin Irritancy Tests; D013053:Spectrophotometry; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
chemical_list: D003358:Cosmetics; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue
keywords: Cosmetic ingredients; EpiSkin™; HPLC/UPLC-spectrophotometry; In vitro RhE test methods; MTT-reduction assay; skin irritation