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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:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study. Read across was performed with butyl methacrylate.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9: Rat liver, induced with phenobarbital and 5,6-benzoflavone 
Test concentrations with justification for top dose:
-S9 mix: 9.77, 19.5, 39.1, 78.1, 156, 313 and 625 µg/plate
+S9 mix: 9.77, 19.5, 39.1, 78.1, 156, 313, 625 and 1250 µg/plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9 mix; 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (TA100, TA98, WP2 uvrA), Sodium azide (TA1535), 9-Aminoacridine (TA1537)  +S9 mix; 2-Aminoanthracene(all strains) 
Details on test system and experimental conditions:
100 μl of the solvent used, the test substance solution and the positive control substance was placed in a capped tube, and then 500 μl of 0.1M sodium phosphate buffer (pH7.4) for the direct method, or 500 μl of the S9 mix for the metabolic activation method was added. Next, after adding 100 μl of the pre-incubated test bacteria strain suspension, this was subject to 20 minutes of shaking incubation (pre-incubation) at 37°C using an incubator shaker. After incubation was completed, 2 ml of top agar was added and the contents mixed. Then, the mixed solution was poured and evenly spread on the plate. After the layered top agar solidified, each plate was sealed with cellophane tape and incubated for 48 hours under conditions of 37°C using an incubator.
Three plates per dose were used. Also, to verify reproducibility, these tests were independently performed two times.
Evaluation criteria:
The number of reverse mutated colonies increased to nearly double that of the control solvent, and when the reproducibility and the test substance dose dependence was confirmed, it was deemed positive
Statistics:
A statistical analysis was not performed

Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
>= 156 µg/plate in the 5 strains without S9. >=313 µg/plate (TA100, Ta1535, TA98, TA1537) and >= 625 µg/plate (WP2 uvrA) with S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion