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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Principles of method if other than guideline:
Preincubation method according to Ames, Mutat. Res. 31, 347 (1975), Maron, Mutat. Res. 113, 173 (1983)
The test compound dissolved in 0.05 or 0.1 ml of solvent was supplemented with 0.5 ml of 59mix (metabolie activation method) or O.IM phosphate buffer pH7.4 (direct method) and 0.1 ml of tester strains whieh had been cultured in nutrient broth. The mixture was incubated for 20 min. at 37°C, then rapidly mixed with 2 ml of molten top agar containing 0.05µmol/ml of L-histidine and biotin for the Salmonella test. In the E. coli test 0.05/1 mol/
ml of L-tryptophan was used instead of L-histidine and biotin. Then the top agar mixture was rapidly poured onto a 30 ml of Vogel-Bonner minimal agar plate. All plates were incubated for 48 hours at 37°C and the numbers of revertant colonies were scored.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
o-chlortoluene, purity: 98 %

Method

Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA100, TA102, TA104, TA1535, TA98, TA1537, TA1538, Escherichia coli WP2uvrA, Escherichia coli WP2uvrA/pKM101
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1) +/-S9: 0.0763, 0.305, 1.22, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate;
2) +/-S9: 2.44 (only S.typh strains -S9), 4.88 (only S.typh strains -S9), 9.77, 19.5, 39.1, 78.1, 156, 313, 625 µg/plate
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
other: without activation: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, sodium azide, 9-aminoacridine, 2-nitrofluorene, 4-nitroquinoline-N-oxide, bleomycin, pyruvic aldehyde; with activation: 2-amino anthracene
Details on test system and experimental conditions:
IUCLID4 Type: Ames test

Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA100, TA102, TA104, TA1535, TA98, TA1537, TA1538, Escherichia coli WP2uvrA, Escherichia coli WP2uvrA/pKM101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: >= 19.5 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: other: Salmonella typhimurium TA100, TA102, TA104, TA1535, TA98, TA1537, TA1538, Escherichia coli WP2uvrA, Escherichia coli WP2uvrA/pKM101
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The positive controls were functional

Applicant's summary and conclusion

Executive summary:

method: an Ames test was conducted using a preincubation protocol in the absence of exogenous metabolic activation, and in the presence of liver S-9.

result: o-chlorotoluene was negative in the test with and without metabolic activation