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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
Study period:
1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study. However, the updated and currently applicable guideline states use of 5 strains instead of 4, which are used here.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1991

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
a fifth strain was not included (requested in the updated and currently applicable OECD guideline)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Test material form:
liquid: viscous
Details on test material:
- Substance type: Technical product.
- Storage condition of test material: No data.

Method

Target gene:
histidine gene
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Liver S9-mix prepared from Aroclor 1254-induced Wistar or Sprague Dawley rats
Test concentrations with justification for top dose:
100, 333, 1000, 3330 and 5000 µg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Sodium azide (TA1535 -S9), 9-aminoacridine (TA1537 -S9), daunomycine (TA98 -S9), methylmethanesulfonate (TA100 -S9), 2-aminoanthracene (TA1535, TA1537, TA100, TA98 +S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 48 hours.

NUMBER OF REPLICATIONS: two separate experiments were conducted with three replicates per dose level.
Evaluation criteria:
A test substance is considered positive in the Ames test if:
1) It induced at least a 2-fold, dose related increase in the number of revertants with respect to the number induced by the solvent control in any of the tester strains, either with or without metabolic activation.
2) The positive response should be reproducible in at least one independently repeated experiment.
Statistics:
The revertant colonies have been counted automatically with a colony counter or manually, if less than 40 colonies per plate were present.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium, other: TA98, TA1537
Metabolic activation:
without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA98, TA1537
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA100, TA1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
The strains TA1537 and TA98, in the presence of S9-mix, and the strains TA1535 and TA100, in the presence and absence of S9-mix showed
negative responses over the entire dose range of the test substance, in two independently repeated experiments. However, in the absence of S9-mix the test substance did induce a reproducible 3- to 21-fold, dose-related increase in the number of revertant colonies in tester strains TA1537 and
TA98.

Any other information on results incl. tables

Table 1: Summary of the results

Parameter

Strain

TA1535

TA1537

TA98

TA100

Metabolic activation

with

without

with

without

with

without

with

without

Genotoxicity

no

no

no

yes

no

yes

no

no

Cytotoxicity

no

no

no

no

no

no

no

no

Vehicle control valid

yes

yes

yes

yes

yes

yes

yes

yes

Positive control valid

yes

yes

yes

yes

yes

yes

yes

yes

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
positive

2-Amylanthraquinone was tested in the Ames test for its ability to induce mutations in five histidine dependent Salmonella typhimurium strains. Two independent mutation tests were performed, each in presence and absence of a metabolic activation system (S9-mix). The bacterial strains were exposed to 0-5000 μg/plate 2-amylanthraquinone. In the absence of S9-mix the test substance induced a reproducible 3- to 21-fold, dose-related increase in the number of revertant colonies in tester strains TA1537 and TA98. The test substance is considered to be mutagenic under the experimental conditions employed.