3-nitro-p-anisic acid

Genetic toxicity: in vitro

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: well performed GLP and OECD guideline like study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Company guideline following the test method published by: B.N. Ames et al (1973), B.N. Ames et al (1976), M.H.L. Green et al (1976), A.P. Alvares et al (1973), J. McCann (1975) et al, which was similar to OECD 471.

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-nitro-p-anisic acid

EC Number:

201-906-0

EC Name:

3-nitro-p-anisic acid

Cas Number:

89-41-8

Molecular formula:

C8H7NO5

IUPAC Name:

4-methoxy-3-nitrobenzoic acid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 (TN) induced rat liver S9-mix

Test concentrations with justification for top dose:

First experiment: 0, 4, 20, 100, 500, 2500, 10000 µg/plate
Second experiment: 0, 0.16, 0.8, 4, 20, 100, 500, 2500, 5000 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:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: N-Methyl-N-nitro-N-nitrosoguanidine, 2-Aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation method

DURATION
- Incubation period: 48 to 72 hour at 37 °c in the dark

NUMBER OF REPLICATIONS: three plates per dose, two independent experiments


DETERMINATION OF CYTOTOXICITY
- Method: A reduced rate of spontaneously occuring colonies as well as visible thinning of the bacterial lawn were used as indicator

Evaluation criteria:

- dose dependent increase in the number of revertant colonies
- relevant increases in the number of revertant colonies

Statistics:

mean of three plates

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

Without S9-Mix: dose a dependent increase of revertant colonies in Salmonella strainsTA 98, TA 100, TA 1537 and E. coli WP2uvrA. With S9-Mix: relevant increases of revertant colonies in Salmonella strains TA 98, TA 100, TA 1537 and E. coli WP.

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 100 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at doses of 2500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

3-Nitro-4-methoxybenzoesäure is mutagenic with and without exogenous metabolic activation in this bacterial reverse mutation assay.

Executive summary:

3-Nitro-4-methoxybenzoesäure was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA. The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 8 different doses from 0.16 µg/plate to 5000 µg/plate was used. Control plates (solvent control) without test item showed that the number of spontaneous revertant colonies was similiar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies. Toxicity: The test compound proved to be very toxic to the bacterial strains at 100 µg/plate. 5000 microgram/plate was chosen as top dose level for the mutagenicity study. Mutagenicity: In the absence of the metabolic activation system, the test compound gave a dose dependent increase in the number of revertant colonies with the bacterial strains Salmonella TA 98, TA 100, TA 1537, and E. coli WP2uvrA. Also in the presence of metabolic activation, treatment of the cells with 3-Nitro-4-methoxybenzoesäure results in relevant increases in the number of revertant colonies with the Salmonella strains TA 98, TA 100, TA 1537, and E. coli WP2uvrA. Summarizing, it can be stated that 3-Nitro-4-methoxybenzoesäure is mutagenic with and without exogenous metabolic activation in this bacterial reverse mutation assay.