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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guidelien study with acceptable deviations

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Salmonella Mutagenicity Test results for 250 Chemicals
Author:
Haworth S, Lawlor T, Mortelmans K, Speck W, and Zeiger E
Year:
1983
Bibliographic source:
Environmental Mutagenesis Supplement 1:3-142
Reference Type:
publication
Title:
National Toxicology Program, NTP Technical report on toxicity studies of o-, m-, and p-Nitrotoluenes, administered in dosed feed to F344/N rats and B6C3F1 mice
Author:
US Department of Health and Human Services
Year:
1992
Bibliographic source:
NTP technical report series 23, NIH Publ.-No. 96-3346
Reference Type:
secondary source
Title:
BUA-Stoffbericht Nr. 41: Nitrotoluole
Author:
BUA (Beratergremium umweltrelevanter Altstoffe) (BUA)
Year:
1989
Bibliographic source:
Gesellschaft Deutscher Chemiker (Hrsg Behret) Bd 41, VCH (Verlag Chemie), Weinheim

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
Only 4 strain tested instead of 5. Positive control: 2-aminoantracene was only indicator of efficiency of S9-mix. Individual plate reading not reported.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3-nitrotoluene
EC Number:
202-728-6
EC Name:
3-nitrotoluene
Cas Number:
99-08-1
Molecular formula:
C7H7NO2
IUPAC Name:
1-methyl-3-nitrobenzene
Details on test material:
- Name of test material (as cited in study report): m-nitrotoluene
- Analytical purity: > 99%
- Lot Number: A9A

Method

Target gene:
his
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
microsomal enzyme reaction mix (S9-mix)
Test concentrations with justification for top dose:
EGG (LABORATORY): 0; 3.3; 10.0; 33.0; 100.0; 333.0 µg/plate
Vehicle / solvent:
- Vehicle/solvent used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (2-AA): in all strains in presence of rat and hamster S-9. 4-Nitro-o-phenylenediamine (NOPD): on TA98, without S-9. sodium azide (SA) : on TA100 and TA 1535, without S-9. 9-aminoacridine (9-AAD) was tested on 1537, without S-9.
Remarks:
The actual concentration for each positive control chemical used for each strain and activation condition was selected by the individual laboratory based on dose-response curves generated at the beginning of the testing program (see table 1)
Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48h
- Selection time (if incubation with a selection agent): 48h
- Concentration of S9 mix: 10% for both Hamster S9 mix and Rat S9 mix

SELECTION AGENT (mutation assays): L- histidine

NUMBER OF REPLICATIONS: 2 trial per strain and 3 dishes per dose




DETERMINATION OF CYTOTOXICITY
- Method: other: viability on complete medium and reduced numbers of revertant colonies per plate and/or thinning or absence of the bacterial lawn








Evaluation criteria:
A positive response was indicated by a reproducible, dose-related increase, whether it be twofold over background or not.
An equivocal response was defined as an increase in revertants which was not dose-related, not reproducible, or was of insufficient magnitude to support a determination of mutagenicity. A negative response was obtained when no increase in revertant colonies is observed following chemical treatment.

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
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:


RANGE-FINDING/SCREENING STUDIES: to select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA 100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10 mg/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity. Occasionally, in the earlier tests, the high dose was greater than 10 mg/plate.


Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537

Any other information on results incl. tables

 

TA 100

TA 1535

TA 1537

TA 98

Dose

NA

RLI

HLI

NA

RLI

HLI

NA

RLI

HLI

NA

RLI

HLI

0.0

142±4.6

105±8.5

106±5.9

29±4.4

10±3.2

13±1.5

5±0.9

6±1.5

9±2.3

21±3.6

25±3.5

29±2.8

3.3

135±6.0

126±4.5

110±5.7

35±2.2

10±2.0

12±0.9

8±0.3

6±0.3

11±1.8

15±0.3

29±0.9

25±1.5

10.0

136±2.5

115±9.6

110±3.8

29±6.0

11±2.1

11±1.3

5±1.2

7±2.4

10±1.9

20±3.1

24±1.9

26±1.5

33.3

151±6.1

128±3.7

105±7.7

33±1.2

9±0.7

8±0.9

7±1.2

7±1.2

6±1.7

19±1.8

32±2.2

25±3.4

100.0

143±10.4

125±1.3

116±5.0

30±1.5

11±0.6

8±0.3

3±1.0

8±2.3

13±2.5

22±1.8

24±4.6

28±1.2

333.3

120±8.4s

133±12.3s

106±9.5s

26±0.7s

10±1.2

13±1.7

2±0.9s

6±2.3

9±0.3

13±1.7s

23±4.5

27±3.3s

POS

2080±58.4

994±38.4

2190±78.4

1426±28.3

58±2.2

167±9.0

469±107.7

60±6.2

262±12.5

1707±61.0

779±32.7

2059±84.9

 

Table 1.Mutagenic responses of Salmonella strains TA100, TA 1535, TA 1537, and TA 98 (mean ± SEM) to m-nitrotoluene.

 Abbreviations: NA, not activated; RLI, rat liver S-9, Aroclor1254 induced; HLI, hamster liver S-9, Aroclor1254 induced, t=complete clearing background

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation
Executive summary:

Haworth (1983)

 

3-nitrotoluene was tested in the Ames test similar to OECD guideline 471 with deviations (Only 4 bacterial strains were used. The highest dose tested was 10 mg/plate. 2-Aminoanthracene was tested as sole indicator of the efficacy of the S9-mix. 4-Nitro-o-phenylenediamine was tested on TA 98), using preincubation method, in strains TA98, TA100, TA1535 and TA1537 ofSalmonella typhimirium.

Liver S9 fractions were prepared from male Sprague-Dawley rats and male Syrian hamsters that were injected, i.p., with Aroclor 1254. The concentrations of 3-nitrotoluene used were 0.0, 3.3, 10.0, 33.0, 100.0 and 333.0 µg/plate. The results were negative, 333.0 µg/plate resulted toxic for the strains TA 100 (with and without S9 mix), TA 1535 (without S9 mix), TA 1537 (without S9 mix) and TA 98[with (hamster) and without S9 mix].