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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:
guideline study with acceptable restrictions
Remarks:
Original report is only available in Japanese. The translation is in progress. Therefore, the report is cited by a peer-reviewed OECD SIDS.

Data source

Referenceopen allclose all

Reference Type:
other: summary report
Title:
Unnamed
Year:
1998
Reference Type:
publication
Title:
MHW, Japan (1998) Ministry of Health and Welfare, Toxicity Testing Reports of Environmental ChgmicaIs 6, 539-568
Author:
Ministry of Health and Welfare, Japan
Year:
2003
Bibliographic source:
cited in: OECD SIDS, 2-Dimethylaminoethylmethacrylate, CAS No: 2867-47-2, 07/2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9: Rat liver, induced with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose:
Without S9 mix: 156, 313, 625, 1250, 2500, 5000 µg/plate
With S9 mix: 156, 313, 625, 1250, 2500, 5000 µg/plate
Confirmative test without S9 mix: 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 µg/plate
Vehicle / solvent:
Distilled water
Controls
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Remarks:
Without S9 mix: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, TA98, WP2 uvrA), Sodium azide (TA1535) and 9-Aminoacridine (TA1537). With S9 mix: 2-Aminoanthracene (all strains).
Details on test system and experimental conditions:
By the preliminary test to decide the highest concentration, toxicity was observed at 5000 ug/plate in the direct method without S9 mix for TA 98 and TA 1537. Then the highest concentration was set at 5000 µg/plate for all tests.

- Procedure: Pre-incubation method
- No. of replicates: 2
- No. of plates/test: 3

Two trial tests were done for all cells and a confirmation test was conducted for TA 98 and TA 1537 which showed positive results in the trial tests.
Evaluation criteria:
1) The revertant colony increase should be more than two times of the control.
2) The revertant colony increase should increase proportionally to the concentration of the test substance (concentration dependency).
3) The same revertant colony increase should be observed repeatedly by more than two tests.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
without
Genotoxicity:
positive
Remarks:
at 2500 and 3000 µg/plate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
> 3500 µg/plate
Additional information on results:
In the two tests, the test substance caused a revertant colony increase of 2-times as much as that of the control to S. typhimurium TA 1537 at 2500 µg/plate without S9. However, a concentration dependency was not clear. Additionally, a revertant colony increasing tendency was observed for S. typhimurium TA98 at 2500 µg/plate and 5000 µg/plate without S9.
Consequently, to confirm the concentration dependent increase of revertant colonies at 2500 - 5000 µg/plate, the confirmation test was conducted for S. typhimurium strains TA 1537 and TA 98 by the direct method without S9. In this test, toxic effects were observed at 3500 µg/plate and more to TA 98 and TA 1537 without S9 mix. For TA 1537 a revertant colony increase was observed by more than 2-times of the control at 2500 and 3000 µg/plate. Furthermore, a concentration dependency was observed. For TA 98, although a revertant colony increase was observed at 2500 and 3000 µg/plate, it was less than 2-times as much as that of the control.
According to this study, the test substance is considered to be positive in this bacterial reverse mutation test. The number of the induced revertant colonies per mg was calculated to be 3.6/mg.

Any other information on results incl. tables

Number of revertant colonies (mean ± SD):

 

Dose/plate

(µg)

Without S9 mix

With S9 mix

Mean

SD

Mean

SD

1st Experiment

Salmonella typhimurium TA 98

0 (Water)

20

4

35

9

625

 

 

36

3

1250

20

4

42

7

2500

35

8

37

5

5000

34*

14

48

9

AF-2 0.1 µg

389

11

 

 

2-AA

 

 

275

23

 

Salmonella typhimurium TA 1537

0 (Water)

7

1

12

4

625

 

 

14

1

1250

7

1

15

2

2500

15

4

18

3

5000

4*

2

17

3

9-AA 80 µg

946

132

 

 

2-AA

 

 

89

12

 

2nd Experiment

Salmonella typhimurium TA 98

0 (Water)

26

6

37

7

625

 

 

35

5

1250

23

7

34

10

2500

39

2

40

6

5000

38*

14

43

14

AF-2 0.1 µg

406

31

 

 

2-AA

 

 

372

20

 

Salmonella typhimurium TA 1537

0 (Water)

6

1

19

2

625

 

 

19

5

1250

8

3

16

2

2500

15

3

17

2

5000

2*

2

27

3

9-AA 80 µg

964

18

 

 

2-AA

 

 

90

9

 

Confirmation Test

Salmonella typhimurium TA 98

0 (Water)

25

2

 

 

1000

23

2

 

 

1500

26

6

 

 

2000

33

3

 

 

2500

44

7

 

 

3000

42

6

 

 

3500

34*

10

 

 

4000

18*

3

 

 

4500

11*

4

 

 

5000

14*

4

 

 

AF-2 0.1 µg

365

26

 

 

 

Salmonella typhimurium TA 1537

0 (Water)

5

1

 

 

1000

7

2

 

 

1500

9

2

 

 

2000

8

1

 

 

2500

13

3

 

 

3000

15

5

 

 

3500

7*

2

 

 

4000

4*

1

 

 

4500

3*

1

 

 

5000

5*

3

 

 

9-AA 80 µg

933

29

 

 

 

* = Toxic effect was observed.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
positive without metabolic activation

The test substance was mutagenic in Salmonella typhimurium TA1537 without an exogenous metabolic activation system.
Executive summary:

The study was performed according to OECD TG 471 & TG 472 under GLP conditions.

The test substance induced mutations only in S. typhimurium TA1537 without metabolic activation at 2500 and 3000 µg/plate. The number of the induced revertant colonies/mg was calculated as 3.6/mg. Toxicity was observed at 5000 µg/plate (TA98, TA1537) without S9 mix. In a confirmation test, toxicity was observed at more than 3500 µg/plate (TA98, TA1537) without S9 mix.

Conclusion: The test substance was mutagenic in Salmonella typhimurium TA1537 without an exogenous metabolic activation system.