saline (PBS) and then incubated with MTT bromide) for 3 hours at 37 °C (5% carbon dioxide, 90% humidity). Following overnight extraction at room ≥ temperature, the optical densities were determined (570 nm). Positive control samples (0.3% Triton X-100) were applied to the tissues in triplicate and exposed for 5 min, 15 min and 1 hr. Six replicates of the negative control (corn oil) were applied to tissues and exposed for 1 hour only. Samples were then treated the same as the test substance. The test substance was considered by the study authors to be an irritant if the ET50 value (exposure time required to reduce cell viability by 50%) was < 2 hour. Quality Assurance statement included. Test material Exposure period Mean OD570 of Relative mean ET50 Value triplicate tissues Viability (%) 1 1.709 100 - Negative control 20 min 1.866 109 Test substance 1 hr 1.849 108 > 2 hr (20%) 3 hr 1.849 108 20 min 1.631 95 Test substance 1 hr 1.786 105 > 2 hr (2%) 3 hr 1.706 100 5 min 1.474 86 15 min 1.123 66 21 min Positive control 1 hr 0.263 15 OD = optical density Remarks - Results Tissue viability after exposure to the test substance at 20% or 2% concentration was not reduced to 50% in any of the exposure periods. ≤ The positive control performed as expected. The test substance was considered to be non-irritating to the eye under the conditions of the test. CPT (1998b) A.11. Irritation eye (1) – Analogue 1 Similar to OECD TG 405 Acute Eye Irritation/Corrosion. Species/Strain Rabbit/New White Number of Animals 6 (Males and Females) Observation Period 72 hours Remarks - Method Test substance (0.1 mL) was instilled in the right eye (left eye remained untreated and acted as a control) and the eye remained unwashed. Eyes were examined 24 hours after exposure. Scoring was based on the Draize method (1944). No GLP statement provided. Remarks - Results None of the animals exhibited an adverse effect to the test substance.
saline (PBS) and then incubated with MTT bromide) for 3 hours at 37 °C (5% carbon dioxide, ≥ 90% humidity). Following overnight extraction at room temperature, the optical densities were determined (570 nm). Positive control samples (0.3% Triton X-100) were applied to the tissues in triplicate and exposed for 5 min, 15 min and 1 hr. Six replicates of the negative control (corn oil) were applied to tissues and exposed for 1 hour only. Samples were then treated the same as the test substance. The test substance was considered by the study authors to be an irritant if the ET50 value (exposure time required to reduce cell viability by 50%) was < 2 hour. Quality Assurance statement included. Test material Exposure period Mean OD570 of triplicate tissues Relative mean Viability (%) ET50 Value Negative control 1 1.709 100 - Test substance (20%) 20 min 1.866 109 > 2 hr 1 hr 1.849 108 3 hr 1.849 108 Test substance (2%) 20 min 1.631 95 > 2 hr 1 hr 1.786 105 3 hr 1.706 100 Positive control 5 min 1.474 86 21 min 15 min 1.123 66 1 hr 0.263 15 OD = optical density Remarks - Results Tissue viability after exposure to the test substance at 20% or 2% concentration was not reduced to ≤ 50% in any of the exposure periods. The positive control performed as expected. The test substance was considered to be non-irritating to the eye under the conditions of the test. CPT (1998b) A.11. Irritation – eye (1) Analogue 1 Similar to OECD TG 405 Acute Eye Irritation/Corrosion. Species/Strain Rabbit/New White Number of Animals 6 (Males and Females) Observation Period 72 hours Remarks - Method Test substance (0.1 mL) was instilled in the right eye (left eye remained untreated and acted as a control) and the eye remained unwashed. Eyes were examined 24 hours after exposure. Scoring was based on the Draize method (1944). No GLP statement provided. Remarks - Results None of the animals exhibited an adverse effect to the test substance.