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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Four in vitro genotoxicity studies were negative for genotoxicity endpoints with and without metabolic activation. Positive control substances in these studies induced positive effects.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27/09/2001 - 08/02/2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test
Deviations:
no
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
Mycoplasma-free tk+/-
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Experiment 1 and 2:
0 (saline), 400, 800, 1200, 1600 and 2022 µg/mL
Both with and without metabolic activation.
See Table 1.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: saline
Untreated negative controls:
yes
Remarks:
saline diluted 10-fold in the treatment medium
Negative solvent / vehicle controls:
yes
Remarks:
as above
True negative controls:
not specified
Positive controls:
yes
Remarks:
4-nitroquinoline-1-oxide (without S9) benz(a)pyrene (with S9)
Positive control substance:
other: as above
Details on test system and experimental conditions:
Dinotefuran was dissolved in saline, under subdued lighting conditions, immediately prior to assay to give the required concentration. Stock solutions were filter-sterilised and further dilutions were made using saline. The dinotefuran solutions were protected from light and used within 2 hours of preparation. No change in osmolality occurred at the highest concentration tested.
Evaluation criteria:
The test article was considered to be mutagenic if all the following criteria were met:
• the assay was valid
• the mutant frequency at one or more doses was significantly greater than that of the negative control (p<0.05)
• there was a significant dose-relationship as indicated by the linear trend analysis (p<0.05).
Statistics:
Statistical significance of mutant frequencies (total wells with clones) was carried out according to the UKEMS guidelines. Thus the control log mutant frequency (LMF) was compared with the LMF from each treatment concentration based on Dunnett's test for multiple comparisons, and secondly the data was checked for a linear trend in mutant frequency with treatment concentration using weighted regression. The test for linear trend is one-tailed, therefore negative trend was not considered significant. These tests required the calculation of the heterogeneity factore to obtain a modified estimate of variance.
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Genotoxicity without metabolic activation:

Experiment 1:
No: relative survival at 2022µg/mL was 102.68 without activation.

Experiment 2:
No: relative survival at 2022µg/mL was 76.16 without metabolic activation.

See Table 3

Genotoxicity with metabolic activation:

Experiment 1:
No: relative survival at 2022µg/mL was 118.06% with metabolic activation.

Experiment 2:
No: relative survival at 2022µg/mL was 100.28% with metabolic activation.

See Table 3

Cytotoxicity:

No: no statistically significant increases in mutation frequency occurred at any dose level in the absence or presence of metabolic activation in either independent experiment. The proportion of small colony mutants for the solvent controls without and with metabolic activation ranged from 38 to 41% in experiment 1 and from 52 to 55% in experiment 2. Marked increases in the numbers of both small and large colony mutants occurred in response to both positive control materials.

Analysis of dinotefuran formulations demonstrated achieved concentrations within 100 ± 10% of nominal concentrations. The assay acceptance criteria were met and the study is considered valid.

See Table 2.

Remarks on result:
other: all strains/cell types tested

Table 1: Dose levels in two preliminary studies and main assay

Study

Dose levels ofdinotefuranused (µg/mL)

Cytotoxicity ± S9, 3h

62.5

125

250

500

1000

2022

 

 

 

Cytotoxicity-S9, 24h

7.81

15.63

31.25

62.5

125

250

500

1000

2022

Experiment 1 in main assay ± S9, 3h

400

800

1200

1600

2022

 

 

 

 

Experiment 2 inmain assay-S9, 24h; +S9, 3h

400

800

1200

1600

2022

 

 

 

 

 

Table 2: Summary of relative survival in the cytotoxicity range-finding study

Compound

Dose level

Relative survival (%):

 

(µg/mL)

3-hour exposure (-S9)

3-hour exposure (+S9)

24-hour exposure (-S9)

DMSO

0

= 100

= 100

= 100

Dinotefuran

7.81

-

-

150.17

 

15.63

-

-

122.63

 

31.25

-

-

116.22

 

62.5

47.60

58.55

101.05

 

125

103.20

72.40

111.20

 

250

97.81

82.08

109.29

 

500

93.43

94.94

133.42

 

1000

132.27

66.25

97.63

 

2022

87.00

70.00

109.58

- not assayed


Table 3: Relative survival and mutant frequency – main experiments

Concn.

Experiment 1

(mg/mL)

3-hr exposure (-S9)

3-hr exposure (+S9)

 

%RSa

RTGb

MFc

%RSa

RTGb

MFc

0

= 100

1.00

67.92

= 100

1.00

81.65

400

100.42

0.87

70.05

103.94

1.11

80.01

800

98.63

1.06

70.70

116.66

1.18

54.15

1200

112.29

1.12

60.56

105.90

1.12

64.32

1600

103.08

0.99

85.24

105.69

1.14

68.82

2022

102.68

0.91

81.84

118.06

1.03

61.05

NQO 0.05

0.01

0.02

0.04

 

102.70

94.13

-

-

 

0.85

0.54

-

-

 

149.24

337.64

-

-

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

BP

2.0

3.0

 

-

-

 

-

-

 

-

-

 

91.27

81.81

 

0.76

0.69

 

365.23

525.10

 

Experiment 2

 

24-hr exposure (-S9)

3-hr exposure (+S9)

 

%RSa

RTGb

MFc

%RSa

RTGb

MFc

0

= 100

1.00

72.69

= 100

 

101.87

400

96.10

1.04

61.05

105.89

 

78.81

800

88.13

0.99

62.01

112.71

 

82.82

1200

83.95

1.09

85.11

116.11

 

129.05

1600

107.55

1.01

56.23

101.87

 

105.00

2022

76.16

0.94

82.33

100.28

 

107.68

NQO 0.05

0.01

0.02

0.04

 

-

-

61.06

55.07

 

-

-

0.84

0.92

 

-

-

194.58

196.02

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

BP

2.0

3.0

 

-

-

 

-

-

 

-

-

 

102.04

62.54

 

0.58

0.41

 

423.87

606.07

a% relative survival adjusted by post-treatment cell count;

brelative total growth;

c5-TFT resistant mutants/106viable cells 2 days after treatment;

- not assayed

 

Conclusions:
Interpretation of results :negative

In both experiments relative survival at 2022µg/mL was at least 76.16 % with and without metabolic activation. No statistically significant increases in mutation frequency occurred at any dose level in the absence or presence of metabolic activation in either independent experiment. Marked increases in the numbers of both small and large colony mutants occurred in response to both positive control materials.

It was concluded that dinotefuran technical and/or metabolites does not induce mutation at the tk locus of L5178Y mouse lymphoma cells at concentrations up to 10mM, and is considered not mutagenic in this test system.
Endpoint:
in vitro DNA damage and/or repair study
Remarks:
Type of genotoxicity: DNA damage and/or repair
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12/06/1996 - 10/07/1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, Guideline study
Qualifier:
according to guideline
Guideline:
other: JMAFF 59 NohSan no. 4200 (1985)
Deviations:
no
GLP compliance:
yes
Type of assay:
other: Unscheduled DNA synthesis in mammalian cells in vitro
Species / strain / cell type:
bacteria, other: Bacillus subtilis: M45 Rec- and H17 Rec+
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
0, 62.5, 250, 1000, 4000 and 16000µg/disc dinotefuran with and without metabolic activation.
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Remarks:
Kanamycin sulfate (without S9); Streptomycin sulfate (with S9)
Negative solvent / vehicle controls:
yes
Remarks:
As above
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Mitomycin C (without S9); Trp-P-1 (with S9)
Details on test system and experimental conditions:
An 8mm paper disc (2 discs/dose) was treated with 20 µL solvent or test substance solution and placed on the spore-innoculated agar without metabolic activation. With metabolic activation, an 8mm paper disc (2 discs/dose level) was treated with 20 µL coenzyme solution and 20 µL solvent or test substance solution and placed on the spore-innoculated agar containing S9 mix.
Evaluation criteria:
Results are judged positive when the difference in diameter of growth inhibition zones between strains H17 and M45 is more than 5mm, and negative if less than 2.5mm. Where differences in inhibition zone diameter are between 2.5 and 5.0mm the results are evaluated by consideration of dose-response and reproducibility.
Statistics:
Not reported
Key result
Species / strain:
other: Bacillus subtilis: M45 Rec- and H17 Rec+
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Preliminary study with and without metabolic activation:
No growth inhibition occurred at any dose level in either strain with or without metabolic activation (Table 1).
Based on these results, dinotefuran in DMSO solvent was tested in H17 and M45 (approx. 2 x 107cells/mL) at concentrations of 0 (DMSO only), 1000, 2000, 4000, 8000 and 16000µg/disc with and without S9 metabolic activation.

Main study with and without metabolic activation:
DMSO solvent alone and all doses of DINOTEFURAN employed, up to and including the highest dose, 16000µg/disc, produced no growth inhibition of either strain of B. subtilis either with or without S9 metabolic activation (Table 2). The negative control substances, KM without S9 and SM with S9 produced differences in the inhibition zone diameters of 1.7 and 1.1mm, respectively, whereas the positive control substances, MMC without S9 and Trp-P-1 with S9, produced differences of 7.2 and 6.0mm, respectively.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Table 1:Summary of growth inhibition for the dose range-finding study

Compound

Dose level

± S9

Growth inhibition zone (mm):

 

(µg/disc)

 

M45 (Rec-)

H17 (Rec+)

Difference (M45 –H17)

DMSO

0

-

0

0

0

Dinotefuran

1000

-

0

0

0

 

2000

-

0

0

0

 

4000

-

0

0

0

 

8000

-

0

0

0

 

16000

-

0

0

0

KM

10

-

10.0

8.4

1.6

MMC

0.01

-

6.9

0

6.9

DMSO

0

+

0

0

0

Dinotefuran

1000

+

0

0

0

 

2000

+

0

0

0

 

4000

+

0

0

0

 

8000

+

0

0

0

 

16000

+

0

0

0

SM

100

+

3.3

2.7

0.6

Trp-P-1

3

+

5.6

0

5.6

 

Table 2:Summary of growth inhibition for the main assay

Compound

Dose level

± S9

Growth inhibition zone (mm):

 

(µg/disc)

 

M45 (Rec-)

H17 (Rec+)

Difference (M45 – H17)

DMSO

0

-

0

0

0

Dinotefuran

1000

-

0

0

0

 

2000

-

0

0

0

 

4000

-

0

0

0

 

8000

-

0

0

0

 

16000

-

0

0

0

KM

10

-

10.5

8.8

1.7

MMC

0.01

-

7.2

0

7.2

DMSO

0

+

0

0

0

Dinotefuran

1000

+

0

0

0

 

2000

+

0

0

0

 

4000

+

0

0

0

 

8000

+

0

0

0

 

16000

+

0

0

0

SM

100

+

4.0

2.9

1.1

Trp-P-1

3

+

6.0

0

6.0

 

Conclusions:
Interpretation of results :
negative

It was concluded that under the conditions of the study, dinotefuran and/or metabolites does not exhibit DNA-damaging activity in B. subtilis under the conditions of the study at doses up to 16000µg/disc
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14/06/1996 - 15/10/1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF 59 NohSan no. 4200 (1985)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMHW, Part 1 (1990); Japan Ministry of Labor, Appendix 1, Notification nos. 143 (1987)
Deviations:
no
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test
Species / strain / cell type:
other: Chinese hamster lung (CHL/IU)
Details on mammalian cell type (if applicable):
Mycoplasma-free
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
2000 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: saline for direct method and S9 method.
Untreated negative controls:
yes
Remarks:
saline
Negative solvent / vehicle controls:
yes
Remarks:
as above
True negative controls:
not specified
Positive controls:
yes
Remarks:
Mitomycin C (without S9) Cyclophosphamide (with S9)
Positive control substance:
other: as above
Details on test system and experimental conditions:
Dinotefuran was dissolved in saline. For preparation of the original solution and serial dilution of dinotefuran, the solution in 11-fold of each final concentration was prepared. In the cell growth inhibition test, 101.6 mg of dinotefuran dissolved in 4.6 mL of saline (final concentration is 2000 mg/mL which was equal to 10 mM) both with and without S9. In the main study, 146.3 mg of of dinotefuran dissolved in 6.65 mL of saline without S9 and 86.8 mg of dinotefuran dissolved in 3.9 mL of saline with S9 (final concentration is 2000 mg/mL which was equal to 10 mM).

Mitomycin C (MMC - 0.03µg/mL) was the positive control material for 24 and 48 hour exposures without activation and cyclophosphamide (CP - 12µg/mL) for 6 hour exposures with and without activation.
Evaluation criteria:
The test substance was to be considered as, negative (-) if the total incidence of structural aberrant cells (including gaps)or numberical aberration cells obtained from 2 dishes (total 200 metaphase) was less than 5%, suspected positive (±) if it was more than 5% and less than 10%, and positive (+) if it was more than 10%.
Statistics:
Not conducted
Species / strain:
other: Chinese hamster lung (CHL/IU)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Genotoxicity without metabolic activation: No.
Structural or numerical aberrations were not induced after 6 hours exposure to dinotefuran either in the absence of metabolic activation. Cell survival was unaffected at all dose levels after 6 hours exposure.

MMC in the absence of metabolic activation produced marked increases in the incidences of structural aberrations, notably chromatid breaks and exchanges.

See Table 1 and Table 2.

Genotoxicity with metabolic activation:

No. Structural or numerical aberrations were not induced after 6 hours exposure to dinotefuran either in the presence of metabolic activation. Cell survival was unaffected at all dose levels after 6 hours exposure.

Cyclophosphamide in the presence of metabolic activation produced marked increases in the incidences of structural aberrations, notably chromatid breaks and exchanges.

See Table 1 and Table 2.

Cytotoxicity:
No. In the main study, structural or numerical aberrations were not induced by dinotefuran either after 24 or 48 hours exposure to dose levels up to 2000 µg/mL. However, slight growth inhibition occurred at 2000 µg/mL, 28% after 24 hours and 38% after 48 hours exposure.

See Table 1 and Table 2.


Remarks on result:
other: all strains/cell types tested

Table 1: Cell growth rate (%) relative to solvent control

Dose level

Preliminary study exposed for:

Main study exposed for:

(µg/mL)

48hr (-S9)

6hr (+S9)

24hr (-S9)

48hr (-S9)

6hr (+S9)

6hr (-S9)

0

=100

=100

=100

=100

=100

=100

7.81

99

100

-

-

-

-

15.6

88

93

-

-

-

-

31.3

96

102

-

-

-

-

62.5

93

88

-

-

-

-

125

92

85

-

-

-

-

250

92

85

-

-

-

-

500

88

90

85

93

100

103

1000

82

88

84

95

99

104

2000

65

100

72

62

104

106

MMC 0.03

CP 12.0

-

-

-

-

70

-

74

-

-

86

-

99

 

Table 2: Summary of results from the main assay

Compound

Dose

Cell survival

No. cells

No. cells with structural aberrations:

Total less

Polyploidy

 

(µg/mL)

(%)

 

gap

ctb

cte

csb

cse

other

gaps (%)

(%)

 

 

6-hour exposure, without S9

Saline

0

100

200

0.5

0

0

0

0

0

0.5

0.5

Dinotefuran

500

103

200

0

0

0.5

0

0

0

0.5

1.0

 

1000

104

200

0.5

0

0

0

0

0

0.5

0

 

2000

106

200

0

0.5

0.5

0

0

0

1.0

1.0

CP

12.0

99

200

1.0

0

0

0.5

0

0

1.5

0.5

 

 

6-hour exposure with S9

Saline

0

100

200

0

0.5

0

0

0

0

0.5

1.0

Dinotefuran

500

100

200

0

0.5

0

0

1.0

0

1.5

1.0

 

1000

99

200

0

0.5

0

0

0

0

0.5

2.5

 

2000

104

200

0

0

0

0

1.0

0

1.0

1.0

CP

12.0

86

200

0

15.5

43.5

0

0.5

0

51.0

1.0

 

 

24-hour exposure, without S9

Saline

0

100

200

0.5

0.5

0

0

1.5

0

2.5

0

Dinotefuran

500

85

200

0

2.5

2.0

0

0

0

4.5

0

 

1000

84

200

0.5

1.0

1.0

0

0

0

2.5

0.5

 

2000

72

200

0.5

2.0

0.5

0.5

0

0

3.5

0

MMC

0.03

70

200

2.0

42.5

38.5

1.0

0.5

0

65.5

0.5

 

 

48-hour exposure, without S9

Saline

0

100

200

1.0

1.5

0

0.5

0

0

3.0

2.5

Dinotefuran

500

93

200

0

1.0

0.5

0

0.5

0

2.0

0.5

 

1000

95

200

1.0

0

0

0

1.0

0

2.0

0.5

 

2000

62

200

0

1.0

0

0

0

0

1.0

1.5

MMC

0.03

74

200

2.0

36.0

66.5

0

0.5

0

76.0

0.5

ctb-chromatid break;

cte-chromatid exchange;

csb-chromosome break;

cse-chromosome exchange including dicentric and ring chromosomes;

other-otheraberrations including fragmentation;

gap - includes both chromatid and chromosome gaps

Conclusions:
Interpretation of results: negative

In the main study, structural or numerical aberrations were not induced by dinotefuran either after 24 or 48 hours exposure to dose levels up to 2000µg/mL. However, slight growth inhibition occurred at 2000µg/mL, 28% after 24 hours and 38% after 48 hours exposure. Similarly, structural or numerical aberrations were not induced after 6 hours exposure to dinotefuran either in the presence or absence of metabolic activation. Cell survival was unaffected at all dose levels after 6 hours exposure.

It was concluded that under the conditions of this study dinotefuran and/or metabolites does not induce structural or numerical chromosomal aberrations in Chinese hamster lung cells at dose levels up to and including 2000µg/mL (equivalent to 10 mM).
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
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1994
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
1994
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EU Method B.14
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
Qualifier:
according to guideline
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
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
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
Key result
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'.

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

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

Genetic toxicity in vivo

Description of key information

2019 in vivo MN study additionally covered the endpoint aneugenicity and was negative.

Additional information

Justification for classification or non-classification