Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
Acetamide was found to be not mutagenic in a study according to OECD Guideline 471.
Link to relevant study records
Reference
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:
2014-08-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Data obtained from a guideline study according to OECD Guideline 471 and therefore considered reliable without restrictions.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 97
Details on mammalian cell type (if applicable):
Mutation Details:
hisD6610, frame shift
uvrB, deletion
rfa, deletion
pKM101, plasmide
Additional strain / cell type characteristics:
other: histidine deficiency, UV sensitivity, biotine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistence
Species / strain / cell type:
S. typhimurium TA 98
Details on mammalian cell type (if applicable):
Mutation Details:
hisD3052, frame shift
uvrB, deletion
rfa, deletion
pKM101, plasmide
Additional strain / cell type characteristics:
other: histidine deficiency, UV sensitivity, biotine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistence
Species / strain / cell type:
S. typhimurium TA 100
Details on mammalian cell type (if applicable):
Mutation Details:
hisG46, base pair substitution
uvrB, deletion
rfa, deletion
pKM101, plasmide
Additional strain / cell type characteristics:
other: histidine deficiency, UV sensitivity, biotine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistence
Species / strain / cell type:
S. typhimurium TA 102
Details on mammalian cell type (if applicable):
Mutation Details:
hisG428, base pair substitution
rfa, deletion
pKM101, plasmide
pAQ1, plasmide
Additional strain / cell type characteristics:
other: histidine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistence, tetracyclin re-sistance
Species / strain / cell type:
S. typhimurium TA 1535
Details on mammalian cell type (if applicable):
Mutation Details:
hisG46, base pair substitution
uvrB, deletion
rfa, deletion
Additional strain / cell type characteristics:
other: histidine deficiency, UV sensitivity, biotine deficiency, lipopolysaccharide side chain deficiency
Metabolic activation:
with and without
Metabolic activation system:
S9-Mix
Test concentrations with justification for top dose:
First Experiment: 5000 / 1500 / 500 / 150 / 50 µg/plate
Second Experiment: 5000 / 2500 / 1250 / 625 / 313 µg/plate
Vehicle / solvent:
Demineralized water
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 4-Nitro-1,2-phenylene Diamine, 2-Amino-Anthracene
Species / strain:
S. typhimurium TA 98
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
Species / strain:
S. typhimurium TA 97
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
Species / strain:
S. typhimurium TA 100
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
Species / strain:
S. typhimurium TA 102
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
Species / strain:
S. typhimurium TA 1535
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

First Experiment:

Mean Revertants First Experiment

Strain

TA97a

TA98

TA100

TA102

TA1535

Induction

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

H2O

Mean

109

113

15

13

96

95

364

328

13

16

sd

1.2

5.6

2.1

4.6

4.7

1.7

18.3

38.6

3.2

2.3

DMSO

Mean

112

112

14

16

101

102

340

359

14

14

sd

5.5

1.7

3.6

1.2

2.6

8.2

40.6

26.0

3.2

2.6

Positive
Controls*

Mean

589

529

428

97

456

641

841

764

167

116

sd

50.6

89.9

56.0

6.1

85.1

57.5

72.6

238.3

12.1

13.3

f(I)

5.26

4.72

30.57

6.06

4.75

6.28

2.47

2.13

12.85

8.29

5000 µg/pl.

Mean

118

112

11

12

98

101

219

235

11

11

sd

9.7

1.5

3.1

4.0

1.5

7.1

21.6

16.7

2.3

1.2

f(I)

1.08

0.99

0.73

0.92

1.02

1.06

0.60

0.72

0.85

0.69

1500 µg/pl.

Mean

113

111

8

10

103

111

220

273

10

10

sd

2.5

1.5

1.5

0.6

11.5

28.0

13.9

47.4

1.5

0.0

f(I)

1.04

0.98

0.53

0.77

1.07

1.17

0.60

0.83

0.77

0.63

500 µg/pl.

Mean

114

110

10

10

109

90

245

231

15

15

sd

2.6

1.5

3.0

1.5

20.5

2.6

8.3

25.4

2.1

1.5

f(I)

1.05

0.97

0.67

0.77

1.14

0.95

0.67

0.70

1.15

0.94

150 µg/pl.

Mean

112

111

15

11

110

98

272

276

12

10

sd

1.5

1.0

1.5

1.2

14.2

9.1

50.0

20.0

1.5

0.0

f(I)

1.03

0.98

1.00

0.85

1.15

1.03

0.75

0.84

0.92

0.63

50 µg/pl.

Mean

114

109

13

9

103

99

228

260

11

12

sd

4.6

1.0

0.6

2.9

4.5

6.1

62.4

41.8

0.6

2.1

f(I)

1.05

0.96

0.87

0.69

1.07

1.04

0.63

0.79

0.85

0.75

Second Experiment:

Mean Revertants Second Experiment

Strain

TA97a

TA98

TA100

TA102

TA1535

Induction

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

H2O

Mean

109

116

13

17

98

106

303

307

11

12

sd

5.7

2.1

1.7

3.1

7.0

13.3

56.8

40.1

0.6

1.0

DMSO

Mean

113

114

14

19

106

105

329

284

10

14

sd

1.5

2.6

2.0

2.6

11.8

11.0

30.3

85.1

3.2

2.3

Positive
Controls*

Mean

489

579

439

94

608

649

884

873

108

92

sd

30.3

43.9

22.7

12.5

68.2

32.3

106.7

30.0

8.3

12.4

f(I)

4.33

5.08

31.36

4.95

6.20

6.18

2.69

3.07

9.82

6.57

5000 µg/pl.

Mean

111

117

12

14

115

112

264

244

14

14

sd

1.5

3.2

2.5

4.0

4.9

11.2

25.0

28.0

1.7

2.5

f(I)

1.02

1.01

0.92

0.82

1.17

1.06

0.87

0.79

1.27

1.17

2500 µg/pl.

Mean

114

114

15

17

118

123

263

291

14

11

sd

4.7

5.2

3.5

3.8

23.7

10.8

39.5

47.7

4.0

2.6

f(I)

1.05

0.98

1.15

1.00

1.20

1.16

0.87

0.95

1.27

0.92

1250 µg/pl.

Mean

111

127

21

18

95

101

315

273

10

13

sd

5.7

7.6

3.1

4.6

4.9

4.9

66.0

8.3

2.5

2.6

f(I)

1.02

1.09

1.62

1.06

0.97

0.95

1.04

0.89

0.91

1.08

625 µg/pl.

Mean

117

123

17

18

110

93

339

316

11

12

sd

6.8

2.6

1.7

2.1

5.8

1.2

29.5

52.9

4.0

1.5

f(I)

1.07

1.06

1.31

1.06

1.12

0.88

1.12

1.03

1.00

1.00

313 µg/pl.

Mean

119

128

16

15

101

97

267

283

12

12

sd

3.2

9.0

2.6

0.6

6.4

8.6

22.0

45.0

3.1

1.5

f(I)

1.09

1.10

1.23

0.88

1.03

0.92

0.88

0.92

1.09

1.00

Conclusions:
Interpretation of results (migrated information):
negative

The mutagenic potential of Acetamid was determined with the Bacterial Reverse Mutation Test following OECD 471 and EU B.13/14. The test item did not show mutagenic effects in both experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants (solvent only).
Cytotoxicity of the test item was not detected. The bacterial background lawn was visible and the number of revertants was not significantly decreased.
Therefore it can be stated, that under the test conditions, the test item Acetamid is not mu-tagenic in the Bacterial Reverse Mutation Test using Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535.
Executive summary:

The mutagenic potential of Acetamid was determined with the Bacterial Reverse Mutation Test following OECD 471 and EU B.13/14. Two valid experiments were performed.

 

First Experiment:

Five concentrations of the test item, dissolved in demineralized water (ranging from 50 to 5000 µg/plate) were used. Five genetically changed strains ofSalmonella typhimurium(TA97a, TA98, TA100, TA102 (genetically manipulated) and TA1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9-mix, rat liver S9 induced by Aroclor 1254) for 48 hours, using the plate incorporation method.

None of the concentrations caused a significant increase in the number of revertant colonies in the tested strains. The test item did not show any mutagenic effects in the first experiment.

The test item showed no precipitates on the plates in all tested concentrations.

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation.

 

Second Experiment:

To verify the results of the first experiment, a second experiment was performed, using five concentrations of the test item (ranging from 313 to 5000 µg/plate) and a modification in study performance (pre-incubation method).

The test item did not show mutagenic effects in the second experiment, either.

The test item showed no precipitates on the plates in all tested concentrations.

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation.

Under the conditions of the test, the test item did not show mutagenic effects towardsSalmonella typhimurium,strainsTA97a, TA98, TA100, TA102 and TA1535.

Therefore, no concentration-effect relationship could be determined.

 

The test itemAcetamidis considered as “not mutagenic under the conditions of the test”.

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

Additional information

Additional information from genetic toxicity in vitro:

The mutagenic potential of Acetamid was determined with the Bacterial Reverse Mutation Test following OECD 471 and EU B.13/14. The test item did not show mutagenic effects in both experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants (solvent only). Cytotoxicity of the test item was not detected. The bacterial background lawn was visible and the number of revertants was not significantly decreased. Therefore it can be stated, that under the test conditions, the test item Acetamid is not mu-tagenic in the Bacterial Reverse Mutation Test using Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535.


Justification for selection of genetic toxicity endpoint
Data obtained from a GLP compliant study according to OECD Guideline 471.

Justification for classification or non-classification

Based on the available in-vitro data, no classification is needed.