Assessment of cosmetic ingredients in the in vitro reconstructed human epidermis test method EpiSkin™ using HPLC/UPLC-spectrophotometry in the MTT-reduction assay. (June 2016)

• Record Type: Journal Article
• Authors: Alépée, N.; Hibatallah, J.; Klaric, M.; Mewes, K.R.; Pfannenbecker, U.; McNamee, P.
• Is Part Of: Toxicology in vitro. Volume 33(2016)
• Page Start: 105
• Page End: 117
• Publication Date: 2016-06
• Subjects: CAS RN Chemical Abstract Service Registry Number -- R2 coefficient of determination -- Cat. 1 Category 1 -- Cat. 2 Category 2 -- EIT eye irritation test -- DG SANCO European Commission Directorate General for Health and Consumer Protection -- EURL ECVAM European Union Reference Laboratory for Alternatives to Animal Testing -- FDA Food and Drug Administration -- HPLC-UPLC-spectrophotometry high/ultra high performance liquid chromatography-spectrophotometry -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide tetrazolium salt -- MSDS material safety data Sheet -- NSCLiving non-specific colour in living tissues -- NSCkilled non-specific colour in killed tissues -- NSMTT non-specific MTT reduction -- NC not classified -- OD optical density -- OECD organisation for economic co-operation and development -- % percentage -- RhCE reconstructed human cornea-like epithelium -- RHE reconstructed human epidermis -- RhT reconstructed human tissue -- R&I Research & Innovation -- SCCS scientific committee on consumer safety -- SD standard deviation -- SOP standard operating procedure -- TG test guideline -- UN GHS united nations globally harmonized system of classification and labelling of chemicals -- WLR within laboratory reproducibility; Cosmetic ingredients -- MTT-reduction assay -- HPLC/UPLC-spectrophotometry -- In vitro RhE test methods -- EpiSkin™ -- skin irritation; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2016.02.006
• Languages: English
• ISSNs: 0887-2333

Abstract

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. Highlights: HPLC/UPLC-spectrophotometry for formazan measurement in in vitro reconstructed human tissue test methods. Evaluation of 50 test items in EpiSkin™ test system. Reproducibility and concordance between OD and HPLC-UPLC-spectrophotometry demonstrated. Applicable for highly coloured chemicals interfering with standard detection method. Applicability to evaluation of chemicals with functionalities relevant to cosmetic products demonstrated.