QUANTITATIVE EVALUATION FOR THE PREDICTION OF EYE IRRITATION USING HEMOGLOBIN

T. HAYASHI1, H. ITAGAKI2, T. FUKUDA1, U. TAMURA1 and S. KATO2
1Shiseido Product Research Center, 2Shiseido Safety and Analytical Research Center, 1050 Nippa-cho, Kohoku-ku, Yokohama 723, Japan

Regular article :AATEX 2(1):25-31

Abstract
The toxicological effects of chemicals on eye tissues are usually assessed by animal using procedures such as the Draize eye irritation test. Various alternative techniques have proposed, e.g. the SIRC and HeLa cultivated cell method and the CAM method. Based on the hypothesis that irritation is due to damages of the cellular plasma membrane and cellular proteins, we selected hemoglobin as a model protein and developed new method for the prediction of eye irritation by measuring the denaturation of hemoglobin spectrophotometrically. Twelve kinds of surfactants were examined and the results were compared with previously reported Draize scores. Correlation analysis, factorial analysis and multi-regression analysis were applied and we found that:
correlations between the hemoglobin denaturation ratio (HDR%) and Draize scores were reasonably high; factorial analysis showed that two factors control hemoglobin denaturation, a low concentration factor (less than 0.25%) and a high concentration factor (more than 1.0%); a higher correlation was obtained by multi-regression analysis, namely, both corneal score and total eye irritation score were estimated with an accuracy of r=0.878 and r=0.861.


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