Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation toxicity study was performed by Shimizu et al (1986) to determine the mutagenic nature of m- nitroacetophenone (CAS no 121 -89 -1; EC name: 3'-nitroacetophenone). The study was performed using Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system at dose levels of 0, 0.1, 0.5, 1, 5 or 10 mg/plate by the preincubation assay. All tests were performed in duplicate and repeated at least 3 times separately. Concurrent solvent and postitive controls were included in the study. The plates were inverted and incubated at 37°C in dark for 70 h. First the tests were carried out without metabolic activation and were terminated if the mutagenicity was positive. In case of negative results, tests with metabolic activation were carried out in addition. Colonies of his+ revertants were counted after incubation, and chemicals inducing more than twice the number of revertant colonies on the control plate were considered as mutagenic. m- nitroacetophenone failed to induce doubling of mutation in Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Link to relevant study records
Reference
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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
AMES Assay was performed to determine the mutagenic nature of m-Nitroacetophenone
GLP compliance:
no
Type of assay:
bacterial gene mutation assay
Specific details on test material used for the study:
- Name of the test material: m-Nitroacetophenone
- EC name: 3'-nitroacetophenone
- Molecular formula: C8H7NO3
- Molecular Weight: 165.147 g/mol
- Substance type: Organic
- Purity: 99%
-Impurity: 1%
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium, other: TA98, TA1538, TA1537, TA100, TA1535
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction was obtained from liver homogenate of male Sprague Dawley rats
Test concentrations with justification for top dose:
0, 0.1, 0.5, 1, 5 or 10 mg/plate
Vehicle / solvent:
- Solvent used: Sterilized DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation
- Cell density at seeding (if applicable): No data

DURATION
- Preincubation period: 15 mins
- Exposure duration: 70 hrs
- Expression time (cells in growth medium): 70 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data

SPINDLE INHIBITOR (cytogenetic assays): No data

STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: All tests were performed in duplicate and repeated at least 3 times separately

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: No data

NUMBER OF CELLS EVALUATED: No data

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): No data

CRITERIA FOR MICRONUCLEUS IDENTIFICATION: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data
- Any supplementary information relevant to cytotoxicity: The strains were observed for toxicity upon chemical treatment

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable): No data

- OTHER: No data
Rationale for test conditions:
No data
Evaluation criteria:
Colonies of his+ revertants were counted after incubation, and chemicals inducing more than twice the number of revertant colonies on the control plate were considered as mutagenic.
Statistics:
The numbers of revertant colonies indicate mean ± S.D., all these numbers revertant colonies were counted after 68-72 h incubation
Species / strain:
S. typhimurium, other: TA98, TA1538 and TA1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 10mg/plate
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
First the test were carried out without metabolic activation and was terminated if the mutagenicity was positive. In case of negative results, test with metabolic activation was carried out in addition.
Conclusions:
m- nitroacetophenone failed to induce mutation in Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of m- nitroacetophenone (EC name: 3'-nitroacetophenone). The study was performed using Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system at dose levels of 0, 0.1, 0.5, 1, 5 or 10 mg/plate by the preincubation assay. All tests were performed in duplicate and repeated at least 3 times separately. Concurrent solvent and postitive controls were included in the study. The plates were inverted and incubated at 37°C in dark for 70 h. First the tests were carried out without metabolic activation and were terminated if the mutagenicity was positive. In case of negative results, tests with metabolic activation were carried out in addition. Colonies of his+ revertants were counted after incubation, and chemicals inducing more than twice the number of revertant colonies on the control plate were considered as mutagenic. m- nitroacetophenone failed to induce doubling of mutation in Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Gene toxicity in vitro:

Various studies were reviewed to determine the mutagenic nature of m- nitroacetophenone (EC name: 3'-nitroacetophenone) . The studies on Salmonella typhimurium bacterial strains are as mentioned below:

Gene mutation toxicity study was performed by Shimizu et al ( Mutation Research, 1986) to determine the mutagenic nature of m- nitroacetophenone (CAS no 121 -89 -1; EC name: 3'-nitroacetophenone). The study was performed using Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system at dose levels of 0, 0.1, 0.5, 1, 5 or 10 mg/plate by the preincubation assay. All tests were performed in duplicate and repeated at least 3 times separately. Concurrent solvent and postitive controls were included in the study. The plates were inverted and incubated at 37°C in dark for 70 h. First the tests were carried out without metabolic activation and were terminated if the mutagenicity was positive. In case of negative results, tests with metabolic activation were carried out in addition. Colonies of his+ revertants were counted after incubation, and chemicals inducing more than twice the number of revertant colonies on the control plate were considered as mutagenic. m- nitroacetophenone failed to induce doubling of mutation in Salmonella typhimurium strain TA98, TA1538, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Another gene mutation toxicity study was performed by Zeiger et al (Environmental and molecular mutagenesis, 1987) to determine the mutagenic nature of m-Nitroacetophenone (CAS no 121 -89 -1; EC name: 3'-nitroacetophenone). The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and  TA97 with and without S9 metabolic activation system. The preincubation assay was performed. The test chemical, Salmonella culture, and S-9 mix or buffer were incubated at 37°C, without shaking, for 20 min. The top agar was added, and the contents of the tubes were mixed and poured onto the surface of petri dishes that contained Vogel- Bonner medium. The histidine-revertant (his') colonies arising on these plates were counted following 2 days incubation at 37°C. The plates were hand-counted when a precipitate was present; otherwise automatic colony counters were used. The chemical was tested initially in a toxicity assay to determine the appropriate dose range. At least five doses of the chemical were tested in triplicate. Experiments were repeated at least 1 wk following the initial trial. Each chemical was tested initially at half-log doses up to a dose that elicited toxicity; subsequent trials occasionally used narrower dose increments. Concurrent solvent and positive controls were run with each trial. The positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA 1537), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation was 2-aminoanthracene for all strains. m-Nitroacetophenone failed to induce mutation in Salmonella typhimurium strains TA100, TA1537 and TA1535 in the presence and absence of S9 metabolic activation system and TA97 and TA98 with S9. It however induced mutation in strains TA97 and TA98 in the absence of S9 activation system. m-Nitroacetophenone is however considered to be non-mutagenic in nature.

Based on the data available, m- nitroacetophenone (CAS no 121 -89 -1, EC name: 3'-nitroacetophenone) is not likely to classify as a gene mutant in vitro. Although mutagenic nature was observed for m-Nitroacetophenone in the absence of S9 metabolic activation system for the Salmonella typhimurium strains TA97 and TA98 (Zeiger et al, 1987), but the mutagenic nature of human concern is in the presence of S9 metabolic activation system. Since no mutagenic activity was observed in TA97 and TA98 in the presence of S9 metabolic activation system, hence it is considered to be non-mutagenic in nature.

Justification for classification or non-classification

Based on the data available, m- nitroacetophenone (CAS no 121 -89 -1) is not likely to classify as a gene mutant in vitro.