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EC number: 468-710-7 | CAS number: 754-12-1
Table 1. Group mean tail intensity and percentage of ghost cells±SD in isolated male rat hepatocytes after exposure.
in air (ppm)
* Mann-Whitney p-value: 0.0079
Table 2. Group mean tail intensity and percentage of ghost cells±SD in isolated male rat lung cells after exposure.
Table 3. The group mean number of PE per 500 E and group mean number of MPE per 4000 PE observed in the micronucleus test in male rats after exposure.
PE per 500 E
MPE per 4000 PE
* Mann-Whitney p-value: 0.0088
** Mann-Whitney p-value: 0.0040
According to OECD guidelines 474 and 489 and in compliance with GLP, the test material was examined for its potential to cause damage to the chromosomes and/or the mitotic apparatus of erythroblasts by analysis of erythrocytes as sampled in bone marrow (micronucleus test), and to cause primary DNA damage (such as single and double strand DNA breaks, alkali labile sites and incomplete repair sites) in lung and liver cells (comet assay) of rats, after administration of the test material by inhalation. For both assays, rats were exposed to clean air (negative control; group 1) or three concentrations of the test material (i.e. 5000, 15000 or 50000 ppm; groups 2, 3 and 4, respectively) by inhalation on three successive days (i.e. 6 hours exposure on the first two days and 2 hours exposure on the third day). The interval between start of the first and second exposure was about 24 hours, and the interval between the start of the second and third exposure ranged from 19 to 25 hours. Rats were sacrificed within 3 to 6 hours after the start of the last exposure. The inhalation route was selected because humans may be exposed to the test material by inhalation. The rats of the positive control groups were treated once intraperitoneally (i.p., 10 mL/kg-bw) with Mitomycin C (MMC; 1.5 mg/kg-bw; group 5) ca. 24 hours prior to sacrifice (micronucleus test), once i.p. (10 mL/kg-bw) with Methyl methane sulfonate (MMS; 40 mg/kg-bw; group 6) 2 to 6 hours prior to sacrifice, or once orally (20 mL/kg-bw; group 7) by gavage with 2-acetylaminofluorene (2-AAF, 50 mg/kg-bw) 12 to 15 hours before sacrifice. Prior to each dosing the rats were weighed and the dosing volume was adjusted to the body weight. For the comet assay, the liver of rats of groups 1, 2, 3, 4 and 7 were perfused to obtain single cells, and immediately following liver perfusion (groups 1, 2, 3 and 4) or sacrifice (group 6), lung tissue was dissected and processed to obtain single cells for preparation of slides for comet analysis. For the micronucleus test, immediately following liver perfusion (groups 1, 2, 3 and 4) or sacrifice (group 5), the bone marrow cells of one of the femurs of each rat were collected and processed into smears for microscopic examination. The negative and positive controls of both the micronucleus test and comet assay showed the expected responses and therefore the study was considered valid. Under the conditions used in this study, it is concluded that test material did neither induce chromosomal damage and/or damage to the mitotic spindle apparatus in bone marrow cells, nor induced primary DNA damage in liver and lung cells, of rats exposed to the test material by inhalation on three successive days up to the maximum concentration tested of 50000 ppm. The TK study performed with the test substance (study no. 20567) demonstrated the presence of the test material in blood, indicating that the negative responses observed in this study are not due to lack of systemic availability of the test material or its metabolites.
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