Test No. 496: In vitro Macromolecular Test Method for Identifying Chemicals Inducing
Serious Eye Damage and Chemicals Not Requiring Classification for Eye Irritation or
Serious Eye Damage
The in vitro macromolecular test method is a biochemical in vitro test method that
can be used to identify chemicals (substances and mixtures) that have the potential
to induce serious eye damage as well as chemicals not requiring classification for
eye irritation or serious eye damage. The in vitro macromolecular test method contains
a macromolecular reagent composed of a mixture of proteins, glycoproteins, carbohydrates,
lipids and low molecular weight components, that when rehydrated forms a complex macromolecular
matrix which mimics the highly ordered structure of the transparent cornea. Corneal
opacity is described as the most important driver for classification of eye hazard.
Test chemicals producing protein denaturation, unfolding and changes in conformation
will lead to the disruption and disaggregation of the highly organised macromolecular
reagent matrix, and produce turbidity of the macromolecular reagent. Such phenomena
is quantified, by measuring the changes in light scattering (at a wavelength of 405
nm using a spectrometer), which is compared to the standard curve established in parallel
by measuring the increase in OD produced by a set of calibration substances.
Published on July 04, 2023Also available in: French
In series:OECD Guidelines for the Testing of Chemicals, Section 4: Health Effectsview more titles