The quantitive chlorioallantoic membrane test using trypan blue stain to predict the eye irritancy of liquid...

The chorioallantoic membrane-trypan blue staining assay (CAM-TBS) is used to evaluate the potential ocular irritation caused by liquid scintillation cocktails constituted by complex mixtures, including surfactants and other potential irritants. The harmful effect of these substances is determined by the amount of trypan blue adsorbed by the CAM. In the membrane previously treated with the scintillation liquids this amount was concentration dependent only in certain cases, irrespective of the water solubility of the mixtures. In general, it showed a high correlation (r=0.986) with the scores obtained in the Draize eye irritation test. In the present study, only two cocktails presented values of trypan blue adsorption higher than those recorded for their eye irritation in vivo, irrespective of the solvent nature. Unlike the classical HET-CAM procedure, this assay is objective and allows the evaluation of opaque and coloured substances without interfering in the determination of irritancy. Despite these advantages, the method is not suitable for complex mixtures of products that induce ocular irritation in small quantities.