o-TOLUIDINE CAS N°: 95-53-4 - UNEP Chemicals

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OECD SIDS o-TOLUIDINE 5. TOXICITY ID: 95-53-4 DATE: 07.01.2005 168 The positive control was functional (2) o-toluidine hydrochloride was tested negative in TA 98 with and without metabolic activation systems; o-toluidine hydrochloride was tested negative in TA 100 in the absence of S9-mix and in the presence of 30 % S9-mix prepared from rat liver but yielded positive results in the presence of 30 % S9-mix from hamster liver. The positive controls were functional. Reliability : (2) valid with restrictions only 4 strains used, no cytotox. concentration given, no information on GLP Flag : Critical study for SIDS endpoint 11.08.2004 (171) Type : HGPRT assay System of testing : Chinese hamster V 79 cells Test concentration : Trial 1: see freetext TC Trial 2: see freetext TC Cycotoxic concentr. : with and without serum: preliminary cytotoxicity study, with different doses and exposure times with and without S9-mix Metabolic activation : with and without Result : negative Method : other: as described by McMillan and Fox (1979) Year : 1985 GLP : no data Test substance : other TS: o-toluidine, no data on purity Result : Trial 1: Little evidence of toxicity (except at very high concentrations) was observed Trial 2: o-Toluidine did not induce increases in mutant frequencies over control levels Positive controls were functional Test condition : METHOD: 1. INHIBITION OF COLONY FORMING ABILITY Cells were exposed to the test substance in the presence and in the absence of serum and in the presence and absence of S9-mix; exposure time: 1 hour doses: highest non-tox.dose: with and without serum, +/-S9-mix, 0.1mg/ml; conc. for 50 % kill: with Serum, +/-S9-mix: >3mg/ml, without Serum: -S9-mix, 3mg/ml, +S9mix, 2mg/ml 2. MUTATION ASSAY Thioguanine was used to select for HGPRT-deficient mutants. Exposure to the testsubstance was for 60 min. in the absence of serum and in the presence and absence of S9-mix. Doses tested were chosen to include at least one that induced measureable cytytotoxicity in colony forming assay and one higher dose. without serum, +/- S9-mix: 0, 0.25, 0.30, 0.50, 1.00, 2.00 mg/ml expression time: 5, 8, 12 days CONTROLS Positive control cyclophosphamide and ethylmethansulfonate in the presence and in the absence of S9 mix, respectively negative control Solvent: DMSO UNEP PUBLICATIONS
OECD SIDS o-TOLUIDINE 5. TOXICITY ID: 95-53-4 DATE: 07.01.2005 METABOLIC ACTIVATION SYSTEM: S9 mix liver homogenates from with phenobarbital and betanaphthoflavone induced rats Reliability : (2) valid with restrictions evaluation of assay systems,in accordance with the guidline Flag : Critical study for SIDS endpoint 21.05.2004 (172) (173) Type : Mammalian cell gene mutation assay System of testing : Chinese hamster V79 cells Test concentration : +S9-mix: 0, 1, 10, 50, 100, 500 µg/ml; -S9-mix: 0, 10, 50, 100, 500, 1000 µg/ml Cycotoxic concentr. : > 500 µg/ml Metabolic activation : with and without Result : positive Method : other: replating method Fox, M. (1975): Mutat. Res. 29, 449-466 Year : 1984 GLP : no data Test substance : other TS: o-toluidine, no data on purity Result : o-toluidine was positive without S9 mix Test condition : --cells from normal young adult male Chinese hamsters --Incubation with test substance in the presence and in the absence of a metabolic activation system for 3 hours at 38°C, then cells were rinsed twice with Hanks's solution and incubated in normal medium for a mutation expression time of 6 days --then treatment with trypsin and replated in medium containing 6TG for 14 days --cell colonies formed were fixed and stainde and the number of 6TGresistant colonies was scored --METABOLIC ACTIVATION: S9 mix liver homogenates with phenobarbital and 5,6-benzoflavone induced rats --the observed number of induced 6TG-resistant mutant colonies induced by o-toluidine was corrected for the decrease in the colony-forming activity of cells in normal medium due to the cytotoxic action of the chemical. The number of induced mutants was calculated by subtracting the number of colonies in untreated controll cultures (no further data). Reliability : (2) valid with restrictions assay system evaluation, well documented Flag : Critical study for SIDS endpoint 19.05.2004 (173) (174) (175) Type : Mouse lymphoma assay System of testing : L5178Y TK-/TK+ Test concentration : +S9-mix: 120-903 µg/ml in DMSO; -S9-mix: 80-1072 mg/ml in DMSO Cycotoxic concentr. : +S9-mix: ca. 350 µg/ml; -S9-mix: ca: 500 µg/ml Metabolic activation : with and without Result : negative Method : other: according Clive et al., 1972, 16, 77-87 and Mutation Res. 31, 17-29 (1975) Year : 1985 GLP : no data Test substance : other TS: o-toluidine, no data on purity Result : o-Toluidine was not considered mutagenic in either the presence or absence of liver S9 activation mix. Careful examination of the data does suggest, however, that this chemical was more cytotoxic in the presence of S9, along with a slight increase in mutant frequency. The positivecontrols were functional. UNEP PUBLICATIONS 169
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o-TOLUIDINE CAS N°: 95-53-4 - UNEP Chemicals

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