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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:
1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (S. typhimurium TA102 or E. choli strains not included; limited documentation of results)

Data source

Reference
Reference Type:
publication
Title:
Structural specificity of aromatic compounds with special reference to mutagenic activity in Salmonella typhimurium - a series of chloro- or fluoro-nitrobenzene derivates
Author:
Shimizu, M. et al.
Year:
1983
Bibliographic source:
Mutation Research 116: 217-238

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
S. typhimurium TA102 or E. choli strains not included; limited documentation of results
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 2,3-Dichloronitrobenzene
- Analytical purity: 99%
- Source: Tokyo Kasei Kogyo Co. Ltd.

Method

Target gene:
his-operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix prepared from livers of PCB-induced (500 mg/kg bw) male Sprague-Dawley rats.
Test concentrations with justification for top dose:
25.6, 51.2, 102.4, 204.8, 409.6, 819.2, 1638.4, 3276.8 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
0.05 ml DMSO per plate
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
Migrated to IUCLID6: 2 and 10 µg/plate without metabolic activation. The lower concentration was tested in strain TA100, the higher concentration in strain TA1535.
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
Migrated to IUCLID6: 2 and 5 µg/plate without metabolic activation. The lower concentration was tested in strain TA98, the higher concentration in strain TA1538.
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Migrated to IUCLID6: 100 µg/plate without metabolic activation; tested in strain TA1537.
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene at 5 µg/plate with metabolic activation; tested in all strains.
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation


DURATION
- Preincubation period: 15 min
- Exposure duration: 3 days
- Selection time (if incubation with a selection agent): 3 days


SELECTION AGENT (mutation assays): minimal agar


NUMBER OF REPLICATIONS: The test was performed in duplicate and repeated at least 3 times separately.


DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency
Evaluation criteria:
The test material was considered as mutagenic if it induced more than twice the number of revertant colonies on the control plate.

Results and discussion

Test resultsopen allclose all
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:
At 1638.4 µg/plate in most strains, at 3276.8 µg/plate in all strains both with and without metabolic activation.
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
At 819.2 µg/plate without metabolic activation and at higher concentrations with and without metabolic activation.

Any other information on results incl. tables

Table of results

Compound

Dose per plate

His+ revertants/plate

 

 

TA98

TA1538

TA1537

TA100

TA1535

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

 

DMSO (control)

0.05 ml

28±6

30±5

22±7

24±5

8±3

11±4

181±23

175±36

32±8

30±9

N-ethyl-N-nitro-N-nitrosoguanidine

2 µg

 

 

 

 

 

 

1994±377

 

 

 

10 µg

 

 

 

 

 

 

 

 

2489±287

 

2-nitrofluorene

2 µg

1798±258

 

 

 

 

 

 

 

 

 

5 µg

 

 

1659±228

 

 

 

 

 

 

 

9-aminoacridine

100 µg

 

 

 

 

1288±198

 

 

 

 

 

2-aminoanthracene

5 µg

 

1249±202

 

753±62

 

132±36

 

1549±269

 

174±23

2,3-Dichloronitrobenzene

25.6 µg

31±3

28±3

26±3

30±4

10±2

12±4

206±23

212±24

33±4

28±3

51.2 µg

36±7

32±4

20±2

27±3

12±3

10±3

176±18

201±19

34±5

32±3

102.4 µg

34±5

32±3

20±3

30±3

6±1

10±3

212±22

197±15

39±7

40±7

204.8 µg

28±3

28±2

26±4

32±5

9±2

11±3

242±32

223±33

41±8

29±2

409.6 µg

32±4

29±3

16±2

25±3

10±3

9±2

232±28

207±22

35±5

33±4

819.2 µg

29±3

26±3

3±8*

19±4

8±2

6±1

198±16

174±19

30±3

30±3

1638.4 µg

2±11*

15±6*

0*

0*

0*

8±2

110±32*

128±21*

16±3

21±4

3276.8 µg

0*

0*

0*

0*

0*

0*

0*

0*

0*

0*

* indicates toxic effects

0 indicates no revertants detectable

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative
Executive summary:

Shimizu (1983)

The mutagenic activity of a series of chloro- and fluoro-nitrobenzene derivates was investigated in a bacterial gene mutation assay conducted according to Ames' procedure (1973, Proc. Natl. Acad. Sci. 70:782 -786). The preincubation assay was performed with Salmonella typhimurium TA98, TA1538, TA1537, TA100 and TA1535 both in presence and absence of a metabolic activator (PCB-induced rat liver S9) at 25.6, 51.2, 102.4, 204.8, 409.6, 819.2, 1638.4, 3276.8 µg 1,2 -dichloro-3 -nitrobenzene per plate. Clear cytotoxic effects were observed at the two highest concentrations with and without metabolic activation.

1,2 -dichloro-3 -nitrobenzene did not induce mutations in the bacterial mutation assay in either the presence or absence of the metabolic activator in any of the strains tested, while the vehicle and positive controls included in the test showed the expected results.