Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
12/07/1996 - 02/10/1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1994
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
1994
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Method B.14
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
Deviations:
no
Principles of method if other than guideline:
Deviations: Yes, in addition to the required 4 S. typhimurium strains, one strain of E. coli was also included since this is a requirement for Japanese authorities. The deviation does not affect the validity of the study.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
S. typhimurium: histidine-auxotrophic strains
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
E. coli: tryptophan-auxotrophic strain
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Pre-incubation: 0 (solvent control), 313, 625, 1250, 2500 and 5000g/plate
Main assay: 0 (solvent control), 1.2, 4.9, 20, 78, 313, 1250 and 5000 µg/plate dinotefuran and the relevant positive controls both with and without metabolic activation.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
Same as negative control
True negative controls:
not specified
Positive controls:
yes
Remarks:
Without S9: 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2), 9-Aminoacridine and Sodium azide With S9: 2-Aminoanthracene. See Table 1.
Positive control substance:
other: See above positive controls
Details on test system and experimental conditions:
METHOD OF APPLICATION: dissolved in medium
DURATION
- Preincubation period: 20 minutes at 37°C
Evaluation criteria:
Evaluated for colonies by an automatic counter and for precipitation and growth inhibition of the background lawn by microscopy.

Dinotefuran was considered positive if at least one dose produced a mean reversion frequency in a given strains that was at least two times greater than that of the corresponding negative control plates, and the response was dose-dependent and reproducibility were observed.
Statistics:
Not reported

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:
no cytotoxicity
Remarks:
Dinotefuran did not influence the growth of any strain tested at dose levels of up to 5000µg/plate.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Genotoxicity without metabolic activation:

Dose range-finding study:
No, normal background growth occurred in all strains without metabolic activation. None of the strains showed an appreciable increase in the reversion frequency at any of the dose levels tested.

Main study:
No appreciable increase in the reversion frequencies occurred in any strain tested in the dose range 313 - 5000µg/plate.
See Table 2 and Table 3.

Genotoxicity with metabolic activation:

Dose range-finding study:
No, normal background growth occurred in all strains with metabolic activation. None of the strains showed an appreciable increase in the reversion frequency at any of the dose levels tested.

Main study:
No appreciable increase in the reversion frequencies occurred in any strain tested in the dose range 313 - 5000µg/plate.
See Table 2 and Table 3.

Cytotoxicity:

Dose range-finding study:
No appreciable cytotoxicity was observed at up to 5000µg/plate, the highest concentration evaluated.

Main study:
Dinotefuran did not influence the growth of any strain tested at dose levels of up to 5000µg/plate.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Strain specific positive control substances and dose levels employed

Strain

Without S9

With S9

Positive control

Dose (µg/plate)

Positive control

Dose (µg/plate)

S. typhimurium TA100

AF-2

0.01

2-AA

1.0

S. typhimurium TA98

AF-2

0.01

2-AA

2.0

S. typhimurium TA1535

NaN3

0.5

2-AA

2.0

S. typhimurium TA1537

9-AA

80.0

2-AA

1.5

E. coli WP2uvrA

AF-2

0.01

2-AA

10.0

AF-2:2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide;NaN3: sodium azide; 9-AA:9-Aminoacridine;

2-AA:-Aminoanthracene

 

Table 2: Summary of the incidences of revertant colonies for the dose range-finding assay

Treatment

Dose

Mean (n = 3) ± SD revertant colonies/plate for strain:

 

(µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

 

 

Without S9

DMSO

0

119

7

14

11

3

Dinotefuran

1.2

138

4

17

9

4

 

4.9

128

5

13

13

3

 

20

129

9

17

10

4

 

78

129

6

16

10

2

 

313

134

7

13

8

4

 

1250

122

6

15

5

4

 

5000

124

6

17

9

3

AF-2

0.01

829

-

-

-

-

NaN3

0.5

-

180

-

-

-

AF-2

0.01

-

-

104

-

-

AF-2

0.1

-

-

-

475

-

9-AA

80

-

-

-

-

376

 

 

With S9

DMSO

0

121

8

15

17

9

Dinotefuran

1.2

106

10

18

22

9

 

4.9

119

10

15

19

8

 

20

121

10

14

17

8

 

78

120

10

12

19

5

 

313

116

7

18

13

7

 

1250

118

10

13

14

6

 

5000

112

10

11

14

8

2-AA

1

1006

-

-

-

-

 

2

-

208

-

-

-

 

10

-

-

845

-

-

 

0.5

-

-

-

286

-

 

2

-

-

-

-

71

 


Table 3: Summary of the incidence of revertant colonies for the main assay

Treatment

Dose

Mean (n = 3) ± SD revertant colonies/plate for strain:

 

(µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

 

 

Without S9

DMSO

0

101

5

25

12

3

Dinotefuran

313

111

7

26

12

3

 

625

104

7

18

7

3

 

1250

121

9

22

11

2

 

2500

103

6

19

9

5

 

5000

103

9

18

16

5

AF-2

0.01

727

-

-

-

-

NaN3

0.5

-

244

-

-

-

AF-2

0.01

-

-

106

-

-

AF-2

0.1

-

-

-

460

-

9-AA

80

-

-

-

-

578

 

 

With S9

DMSO

0

104

9

24

18

12

Dinotefuran

313

102

8

24

21

8

 

625

107

10

25

16

8

 

1250

111

9

27

26

10

 

2500

112

7

28

25

13

 

5000

106

7

25

19

13

2-AA

1

665

-

-

-

-

 

2

-

210

-

-

-

 

10

-

-

799

-

-

 

0.5

-

-

-

209

-

 

2

-

-

-

-

81

Applicant's summary and conclusion

Conclusions:
Interpretation of results:
negative

No appreciable cytotoxicity was observed in all strains, with and without metabolic activation at up to 5000µg/plate, the highest concentration evaluated. None of the strains showed an appreciable increase in the reversion frequency at any of the dose levels tested.

In the main assay, dinotefuran did not influence the growth of any strain tested at dose levels of up to 5000µg/plate. No appreciable increase in the reversion frequencies occurred in any strain tested in the dose range 313 - 5000µg/plate. In contrast, the positive control substances produced marked increases in the number of revertant colonies in all strains tested.

Under the conditions of this study, dinotefuran and/or metabolites does not induce gene mutations in the strains of S. typhimurium and E. coli used in the study at doses up to 5000µg/plate.