Appendix B: Toxicological Investigations Irritation eye (in isolated chicken eye test) vitro – Notified chemical OECD TG 438 Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants Vehicle None Remarks - Method The purity of the test substance was reported as 95.2%. The test substance was directly administered to the isolated chicken eyes. The control eyes and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Corneal thickness and corneal opacity were measured at all time points. Fluorescein retention was measured on two occasions, at base line (t = 0) and approximately 30 minutes after the post-treatment rinse. Positive control: imidazole Negative control: saline (0.9% w/w sodium chloride) Test Substance Observation Value ICE Class 1% I Mean maximum corneal swelling at up to 75 min 1% I Mean maximum corneal swelling at up to 240 min 0.67 II Mean maximum corneal opacity 1.00 II Mean fluorescein retention 1 × I, 2 × II Overall ICE Class Positive Control Observation Value ICE Class 1% I Mean maximum corneal swelling at up to 75 min 6% II Mean maximum corneal swelling at up to 240 min 3.83 IV Mean maximum corneal opacity 2.67 IV Mean fluorescein retention 1 × II, 2 × IV Overall ICE Class Negative Control Observation Value ICE Class 0% I Mean maximum corneal swelling at up to 75 min 0% I Mean maximum corneal swelling at up to 240 min 0.00 I Mean maximum corneal opacity 0.00 I Mean fluorescein retention 3 × I Overall ICE Class Remarks - Results The test substance and the positive control (imidazole) were stuck on the surface of the cornea. Gentle rinsing with 20 mL saline was performed at each observation time point. The surface of the cornea was not cleared 240 minutes after the post-treatment rinse. The notified chemical was not corrosive or a severe eye irritant under the conditions of the test. The notified chemical also was not considered as a non-irritant and an study was required for classification. in vivo CiToxLAB (2014g) Irritation eye – Notified chemical
Appendix B: Toxicological Investigations Irritation – eye (in vitro isolated chicken eye test) Notified chemical OECD TG 438 Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants Vehicle None Remarks - Method The purity of the test substance was reported as 95.2%. The test substance was directly administered to the isolated chicken eyes. The control eyes and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Corneal thickness and corneal opacity were measured at all time points. Fluorescein retention was measured on two occasions, at base line (t = 0) and approximately 30 minutes after the post-treatment rinse. Positive control: imidazole Negative control: saline (0.9% w/w sodium chloride) Test Substance Observation Value ICE Class Mean maximum corneal swelling at up to 75 min 1% I Mean maximum corneal swelling at up to 240 min 1% I Mean maximum corneal opacity 0.67 II Mean fluorescein retention 1.00 II Overall ICE Class 1 × I, 2 × II Positive Control Observation Value ICE Class Mean maximum corneal swelling at up to 75 min 1% I Mean maximum corneal swelling at up to 240 min 6% II Mean maximum corneal opacity 3.83 IV Mean fluorescein retention 2.67 IV Overall ICE Class 1 × II, 2 × IV Negative Control Observation Value ICE Class Mean maximum corneal swelling at up to 75 min 0% I Mean maximum corneal swelling at up to 240 min 0% I Mean maximum corneal opacity 0.00 I Mean fluorescein retention 0.00 I Overall ICE Class 3 × I Remarks - Results The test substance and the positive control (imidazole) were stuck on the surface of the cornea. Gentle rinsing with 20 mL saline was performed at each observation time point. The surface of the cornea was not cleared 240 minutes after the post-treatment rinse. The notified chemical was not corrosive or a severe eye irritant under the conditions of the test. The notified chemical also was not considered as a non-irritant and an in vivo study was required for classification. CiToxLAB (2014g) Irritation – eye Notified chemical