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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:
weight of evidence
Study period:
NA to 1989-03-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non GLP study according to OECD Guideline 471 however only four strains of S.typhimurium (TA 1535, TA 100, TA 1537, and TA 98) were used.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report date:
1989

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
only four strains of S.typhimurium (TA 1535, TA 100, TA 1537, and TA 98) were used.
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Nitric acid
EC Number:
231-714-2
EC Name:
Nitric acid
Cas Number:
7697-37-2
Molecular formula:
HNO3
IUPAC Name:
nitric acid
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Nitric acid
- Analytical purity: 60% aqueous solution
- Other: test substance no.: 88/972

Method

Target gene:
histidine locus
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: see below in any other information on materials and methods
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S9
Test concentrations with justification for top dose:
The standard plate test and the preincubation test (with and without metabolic activation for all test strains): 20, 100, 500, 2500, and 5000 ug/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s): aqua dest.
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua dest. (vehicle of test substance)
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
with S9: for TA 100, TA 98, TA 1537 and TA 1535 at 10 ug/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua dest. (vehicle of test substance)
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without S9: for TA 1537 at 100 ug/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua dest. (vehicle of test substance)
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylendiamine
Remarks:
without S9: for TA 98 at 10 ug/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua dest. (vehicle of test substance)
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitroso-guanidine (MNNG)
Remarks:
without S9: for TA 100 and TA 1535
Details on test system and experimental conditions:
METHOD OF APPLICATION: experiment 1: in agar (plate incorporation); experiment 2: preincubation


DURATION
- Preincubation period: 20 minutes (experiment 2)
- Exposure duration: 48 hours (experiment 1 and 2)
- Selection time (if incubation with a selection agent): 48 hours (experiment 1 and 2)



SELECTION AGENT (mutation assays):
- histidine (S. typhimurium strains)



NUMBER OF REPLICATIONS:
- 3 (experiment 1 and 2)


DETERMINATION OF CYTOTOXICITY
- Method: The toxic effects of the test substance were detected by a substantial reduction in mean revertant colony counts or by the sparce or absent background bacterial lawn.

-Other: The positive controls were dissolved in DMSO

Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results
Statistics:
A statistical analysis of the data was not required.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
for TA 1537 at 5000 ug/plate for both experiment 1 and 2
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY: A weakly bacteriotoxic effect (reduced his - background growth, decrease in the number of his revertant) observed only using TA 1537 at 5000 ug/plate without S9 in the standard plate test and with S9 in the preincubation test.
Remarks on result:
other: all strains/cell types tested

Any other information on results incl. tables

Mutagenicity: An increase in the number of his + revertants was not observed both in the standard plate test and in the pre-incubation test either without S9 mix or after the addition of a metabolizing system.

Applicant's summary and conclusion

Conclusions:
Interpretation of results:
other: negative with and without metabolic activation

Under conditions of the study, the test substance was not mutagenic in the Ames test under the experimental conditions chosen in this study.