DEVELOPMENT OF AN IN VITRO COMBINED ASSAY METHOD, EVALUATING HEMOGLOBIN DENATURATION AND HEMOLYTIC ACTIVITY IN RAT RED BLOOD CELLS, FOR THE PREDICTION OF EYE IRRITANCY POTENCIAL

S. SUGAI1, K. MURATA1 AND M. WATANABE2
1Research and Development Department, Kumiai Chemical Co. Ltd., 1-4-26, Ikenohata, Taito-ku, Tokyo 110, Japan; 2Faculty of Pharmaceutical Sciences, Nagasaki University 1-14, Bunkyo-machi, Nagasaki 852, Japan

Regular article :AATEX 2(1):7-18

Abstract
Rat red blood cells were used in an in vitro method of evaluating the eye irritancy potential of chemicals in rabbits. Two assays were performed to investigate the effects of chemicals on proteins and lipids in the cell membrane; these effects were evaluated by determining hemoglobin denaturation and hemolytic activity, respectively. The relationship between the in vitro and in vivo eye irritancy potential (classified into 3 classes) of 117 chemicals of various chemical classes, including acids, alkalis, detergents, and solvents, was examined. Multivariate estimation with data from the two in vitro assays above was 82.9% predictive of the in vivo classification. Superior accuracy of prediction of eye irritancy potential was gained with analysis combining the data for two in vitro assays rather than by using each in vitro data set independently. These results suggest that such combined studies to determine the effects of chemicals on proteins and lipids in the cell membrane may be useful for the prediction of eye irritancy potential.

Key words: eye irritation, rabbit, in vitro, red blood cell, rat, hemoglobin denaturation, hemolysis, protein, lipid.


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