Positive controls: i) without S9: N-ethyl-N’-nitro-N-nitrosoguanidine (WP2uvrA, TA100, TA1535), 9-Aminoacridine (TA1537), (TA98); ii) with S9: 1-Aminoanthracene (TA100, TA1535, TA1537, WP2uvrA), Benzo(a)pyrene (TA98). The dose for Test 2 was determined by the results of Test 1. Metabolic Test Substance Concentration (µg/plate) Resulting in: Activation Cytotoxicity in Cytotoxicity in Main Precipitation Genotoxic Effect Preliminary Test* Test Absent Test 1 - 150 > 5000 Negative ≥ Test 2 150 > 500 Negative ≥ Present Test 1 - 500 > 5000 Negative ≥ Test 2 500 > 1500 Negative ≥ * Not conducted Remarks - Results Positive and negative controls performed as expected confirming the validity of the test system. No significant increases in the frequency of the revertant colonies were recorded following treatment with the notified chemical at any dose level, with or without metabolic activation. The notified chemical was not mutagenic to bacteria the conditions of the test. Harlan (2014j) Genotoxicity in vivo – Notified chemical OECD TG 474 Mammalian Erythrocyte Micronucleus Test. EC Directive 2000/32/EC B.12 Mutagenicity - Mammalian Erythrocyte Micronucleus Test. Species/Strain Hsd: ICR (CD-1®) Route of Administration Oral gavage – Vehicle Arachis oil Remarks - Method Range finding test found no marked difference in toxic response between the sexes. It was therefore considered acceptable to use males only for the main test. Group Number and Sex Dose Sacrifice Time of Animals mg/kg bw I (vehicle control) 7 M 0 24 II (low dose) 7 M 250 24 III (mid dose) 7 M 500 24 IV (high dose) 7 M 1000 24 V (high dose) 7 M 1000 48 VI (positive control, CP) 5 M 50 24 CP=cyclophosphamide.
Positive controls: i) without S9: N-ethyl-N’-nitro-N-nitrosoguanidine (WP2uvrA, TA100, TA1535), 9-Aminoacridine (TA1537), (TA98); ii) with S9: 1-Aminoanthracene (TA100, TA1535, TA1537, WP2uvrA), Benzo(a)pyrene (TA98). The dose for Test 2 was determined by the results of Test 1. Metabolic Activation Test Substance Concentration (µg/plate) Resulting in: Cytotoxicity in Preliminary Test* Cytotoxicity in Main Test Precipitation Genotoxic Effect Absent Test 1 - ≥ 150 > 5000 Negative Test 2 ≥ 150 > 500 Negative Present Test 1 - ≥ 500 > 5000 Negative Test 2 ≥ 500 > 1500 Negative * Not conducted Remarks - Results Positive and negative controls performed as expected confirming the validity of the test system. No significant increases in the frequency of the revertant colonies were recorded following treatment with the notified chemical at any dose level, with or without metabolic activation. The notified chemical was not mutagenic to bacteria the conditions of the test. Harlan (2014j) Genotoxicity – in vivo Notified chemical OECD TG 474 Mammalian Erythrocyte Micronucleus Test. EC Directive 2000/32/EC B.12 Mutagenicity - Mammalian Erythrocyte Micronucleus Test. Species/Strain Hsd: ICR (CD-1®) Route of Administration Oral – gavage Vehicle Arachis oil Remarks - Method Range finding test found no marked difference in toxic response between the sexes. It was therefore considered acceptable to use males only for the main test. Group Number and Sex of Animals Dose mg/kg bw Sacrifice Time I (vehicle control) 7 M 0 24 II (low dose) 7 M 250 24 III (mid dose) 7 M 500 24 IV (high dose) 7 M 1000 24 V (high dose) 7 M 1000 48 VI (positive control, CP) 5 M 50 24 CP=cyclophosphamide.
STD1555-FR-Final.DOCX