Mean OD550 of Relative mean Mean OD550 of Relative mean duplicate tissues viability (%) duplicate tissues viability (%) 1.834 100 1.773 100 Negative control 1.634 89 1.726 97 Test substance 0.395 22 0.304 17 Positive control OD = optical density Remarks - Results The test substance showed no corrosive effects. The positive and negative controls gave satisfactory results, confirming the validities of the test systems. The notified chemical was non-corrosive to the skin under the conditions of the test. Bioservice (2012c) Irritation skin (in vitro) – Notified chemical OECD TG 439 In vitro Skin Irritation: Reconstructed Human Epidermis Test Method EPISKIN-SM™ Reconstructed 3D Human Epidermis Model Vehicle Distilled water Remarks - Method The test substance (10 mg) was applied to the tissues (pre-moistened with 5 µL distilled water) in triplicate. Following exposure periods of 15 minutes (room temperature) and 1 hour (room temperature; test 2), the tissues were rinsed, treated with MTT bromide] and then incubated at 37 °C for 3 hours. In a preliminary test the test substance was shown not to directly reduce MTT. Positive and negative controls were run in parallel with the test substance: Negative control (NC): phosphate buffered saline Positive control (PC): 5% sodium dodecyl sulphate in distilled water Test material Mean OD550 of triplicate Relative mean SD of relative mean tissues Viability (%) viability 1.233 100 4.8 Negative control 1.259 102.1 5 Test substance 0.096 7.8 0.4 Positive control OD = optical density; SD = standard deviation Remarks - Results The test substance showed no irritation effects. The positive and negative controls gave satisfactory results, confirming the validities of the test systems. The notified chemical was non-irritating to the skin under the conditions of the test. Bioservice (2012d) Irritation eye (in vitro) –
Mean OD550 of duplicate tissues Relative mean viability (%) Mean OD550 of duplicate tissues Relative mean viability (%) Negative control 1.834 100 1.773 100 Test substance 1.634 89 1.726 97 Positive control 0.395 22 0.304 17 OD = optical density Remarks - Results The test substance showed no corrosive effects. The positive and negative controls gave satisfactory results, confirming the validities of the test systems. The notified chemical was non-corrosive to the skin under the conditions of the test. Bioservice (2012c) Irritation – skin (in vitro) Notified chemical OECD TG 439 In vitro Skin Irritation: Reconstructed Human Epidermis Test Method EPISKIN-SM™ Reconstructed 3D Human Epidermis Model Vehicle Distilled water Remarks - Method The test substance (10 mg) was applied to the tissues (pre-moistened with 5 µL distilled water) in triplicate. Following exposure periods of 15 minutes (room temperature) and 1 hour (room temperature; test 2), the tissues were rinsed, treated with MTT bromide] and then incubated at 37 °C for 3 hours. In a preliminary test the test substance was shown not to directly reduce MTT. Positive and negative controls were run in parallel with the test substance: Negative control (NC): phosphate buffered saline Positive control (PC): 5% sodium dodecyl sulphate in distilled water Test material Mean OD550 of triplicate tissues Relative mean Viability (%) SD of relative mean viability Negative control 1.233 100 4.8 Test substance 1.259 102.1 5 Positive control 0.096 7.8 0.4 OD = optical density; SD = standard deviation Remarks - Results The test substance showed no irritation effects. The positive and negative controls gave satisfactory results, confirming the validities of the test systems. The notified chemical was non-irritating to the skin under the conditions of the test. Bioservice (2012d) Irritation – eye (in vitro)