The First Phase Validation Study of Alternative Methods to The Eye Irritation Test 8)Evaluation of Cytotoxicity Tests on HeLa and CHL Cells

The cytotoxicity test of MTT assay on HeLa cells and Crystal violet staining assay on CHL cells were evaluated as alternative methods to Draize eye irritation test. We tested 9 surfactants and isotonic sodium chloride solution and controlled the test procedures under common SOP among laboratories. The concentrations of test chemicals that showed 50% absorbance reduction to control (EC50) were determined, and was compared in vito scores. 8 labolaotories on HeLa and 7 them on CHL assay participated in these validation and the inter-baloratory coefficients of variation of these assays were average 0.29 on both assays. The correlation coefficient between the in vivo maximam total scores and their EC50s of surfactants was -0.817 and -0.902 on HeLa and CHL assay, respectively. The cytotoxicity of different cells showed similar EC50 Values. We conclude that the cytotoxicty test on HeLa and CHL cells expect alternative methods to Draize eye irritation test for surfactants. These tests should be validated for a wider range of chemicals used as cosmetic ingredients. A part of this study was supported by Research Grant for Health Sciences, MHW.