Irritation – skin (in vitro) Notified chemical. OECD TG 439 In vitro Skin Irritation: Reconstructed Human Epidermis Test Method. Vehicle Used as supplied. Remarks - Method EPISKINTM Reconstructed Human Epidermis Model. No significant protocol deviations. GLP Compliance. Negative and positive controls were used in parallel with the test substance. Test material Mean OD562 of triplicate tissues Relative mean Viability (%) SD of relative mean viability (%) Negative control 0.725 Set at 100 5.0 Test substance 0.863 119.1 9.6 Positive control 0.119 16.5 6.0 OD = optical density; SD = standard deviation Remarks - Results The MTT solution containing the test item did not turn blue, indicating that the test item did not directly reduce MTT. The controls gave responses within the expected range, confirming the validity of the test system. The study authors considered it unnecessary to perform IL-1α analysis as they deemed these results to be unequivocal. The notified chemical was non-irritating to the skin under the conditions of the test. Harlan (2013i) Irritation – skin Notified chemical. OECD TG 404 Acute Dermal Irritation/Corrosion. Species/Strain Rabbit/New Zealand White. Number of Animals 3 Males. Vehicle Test substance administered as supplied. Observation Period 14 days. Type of Dressing Semi-Occlusive. Remarks - Method No significant protocol deviations. GLP Compliance. Skin Reaction Mean Score* Animal No. Maximum Value Maximum Duration of Any Effect Maximum Value at End of Observation Period 1 2 3 Erythema/Eschar 0 0 0 1 < 24 hours 0 Oedema 0 0 0 0 0 0 *Calculated on the basis of the scores at 24, 48, and 72 hours for EACH animal.
Remarks - Results Very slight erythema was noted for 2 of the test animals immediately after patch removal, with symptoms persisting in 1 of these animals at the 1 hour observations. All treated skin sites appeared normal at the 24 hour observation. The notified chemical is slightly irritating to the skin. Harlan (2013j) Irritation – eye (in vitro) Notified chemical (20% w/v dilution). OECD TG 437 Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants. Controls Negative/Vehicle: 0.9% sodium chloride. Positive: 20% w/v Imidazole. Remarks - Method No significant protocol deviations. GLP Compliance. Negative and positive control items were tested concurrently. Test material Mean opacities of triplicate tissues (SD) Mean permeabilities of triplicate tissues (SD) IVIS (SD) Vehicle control 2.7 0.037 3.2 Test substance* 1.6 0.009 1.7 Positive control* 52.0 1.44 73.6 SD = Standard deviation; IVIS = in vitro irritancy score *Corrected for background values Remarks - Results The corneas treated with the test item and negative control were clear whereas the corneas treated with the positive control were cloudy. The controls gave satisfactory responses confirming the validity of the test system. The notified chemical was not corrosive or a severe eye irritant under the conditions of the test. Harlan (2013k) Irritation – eye Notified chemical. OECD TG 405 Acute Eye Irritation/Corrosion. Species/Strain Rabbit/New Zealand White. Number of Animals 3 Males. Observation Period 3 days. Remarks - Method No significant protocol deviations. GLP Compliance. Lesion Mean Score* Animal No. Maximum Value Maximum Duration of Any Effect Maximum Value at End of Observation Period
Test material Concentration (%) Mean OD570 of duplicate tissues Relative mean Absorbance (%) Without Irradiation Negative control 100 1.286 100 Test substance Test 1 0.316 1.436 111.7 Test 2 1.0 1.093 85.0 Test 3 3.16 1.072 83.3 Test 4 10 0.912 70.9 Positive control 0.05 0.567 44.1 With Irradiation Negative control 100 1.206 100 Test substance Test 1 0.316 1.192 98.9 Test 2 1.0 1.168 96.8 Test 3 3.16 0.918 76.1 Test 4 10 0.771 63.9 Positive control 0.05 0.066 5.5 SD= Standard Deviation Remarks - Results Cytotoxic effects were not observed after exposure of the skin equivalents to the test item suspension, neither in the presence or the absence of irradiation with artificial sunlight. The controls gave responses within the expected range. The notified chemical did not exhibit any phototoxic effects on the skin equivalents under the conditions of the test. Harlan (2013)