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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study in Japanese only, however, basic data are given in English.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
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): Methyl dodecanoate
- Physical state: colorless clear liquid
- Analytical purity: 98%
- Lot/batch No.: GB01
- Stability under test conditions: Verified for stability for 4 hours
- Storage condition of test material: at room temperature

Method

Target gene:
his operon
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):
- Type and identity of media: minimal agar
Additional strain / cell type characteristics:
other:
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone.
Test concentrations with justification for top dose:
First and second experiment:
Without metabolic activation: 1.56, 3.13, 6.25, 12.5, 25 and 50 µg/plate (TA100 and TA1537); 6.25, 12.5, 25, 50, 100 and 200 µg/plate (TA1535 and TA98); 313, 625, 1250, 2500 and 5000 µg/plate (WP2)
With metabolic activation: 12.5, 25, 50, 100, 200 and 400 µg/plate (TA1537); 25, 50,100, 200, 400 and 800 (TA100 and TA1535); 50, 100, 200, 400, 800 and 1600 (TA98); 313, 625, 1250, 2500 and 5000 µg/plate (WP2)

Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
acetone
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: +S9 mix: 2-Aminoanthracene (0.5 (TA98), 1 (TA100), 2 (TA1535/1537), 10 µg/plate (WP2uvrA)); -S9 mix: 0.01 µg/plate 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, WP2, TA98), 0.5 µg/plate sodium azide (TA1535) and 80 µg/plate 9-Aminoacridine (TA1537)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates in each of two independent experiments


Evaluation criteria:
Colonies were counted for 3 plates. If the mean value is at least twice as high as the vehicle control and repeatedability or dose responsibility is observed, the substance is considered as positive . Otherwise it is considered as negative.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 25 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
without
Genotoxicity:
negative
Remarks:
at 25 µg/plate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 50 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 50 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 150 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 400 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 800 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: strain/cell type: TA 100
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test result of experiment 1

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates ± Standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA98

TA1537

0

99 ± 10.4

11 ± 2.5

19 ± 4.4

7 ± 2.6

1.56

107 ± 7.6

ND

ND

9 ± 4.0

3.13

107 ± 18.7

ND

ND

8 ± 1.5

6.25

95 ± 0.6

8 ± 0.6

17 ± 2.5

5 ± 1.2

12.5

100 ± 3.5

12 ± 7.5

13 ± 2.1

7 ± 2.6

25

86 ± 4.0

10 ± 1.0

19 ± 2.5

7 ± 2.5

50

72 ± 9.2

13 ± 3.5

19 ± 2.9

6 ± 2.5

100

-

11 ± 4.0

22 ± 2.3

-

200

-

11 ± 1.2

21 ± 1.5

-

Positive controls, –S9

Name

AF2

SA

AF2

9AA

Concentrations

(μg/plate)

0.01

0.5

0.1

80

Mean No. of colonies/plate

(average of 3 ± SD)

503 ± 22.3

197 ± 7.5

781 ± 42.9

772 ± 50.3

+

0

119 ± 10.4

15 ± 5.1

28 ± 3.5

11 ± 2.1

+

12.5

ND

ND

ND

15 ± 2.3

+

25

128 ± 17.8

12 ± 0.6

ND

11 ± 3.5

+

50

136 ± 8.6

16 ± 3.8

32 ± 4.4

8 ± 2.0

+

100

122 ± 21.7

16 ± 2.6

29 ± 0.6

11 ± 4.0

+

200

95 ± 16.5

10 ± 4.5

26 ± 2.1

17 ± 2.1

+

400

120 ± 19.7

12 ± 1.0

25 ± 6.7

7 ± 2.9

+

800

74 ± 7.1

9 ± 0.0

27 ± 1.7

 

+

1600

 

 

18 ± 4.7

 

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1

2

0.5

2

Mean No. of colonies/plate

(average of 3 ± SD)

1269 ± 167.2

317 ± 11.7

427 ± 35.4

252 ± 9.0

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

Base-pair substitution type

WP2uvrA

0

21 ± 1.0

313

26 ± 2.5

625

29 ± 1.5

1250

25 ± 2.6

2500

28 ± 5.0

5000

25 ± 3.5

Positive controls, –S9

Name

AF2

Concentrations

(μg/plate)

0.01

Mean No. of colonies/plate

(average of 3 ± SD)

106 ± 5.6

+

0

25 ± 2.6

+

313

26 ± 5.3

+

625

32 ± 6.8

+

1250

37 ± 7.6

+

2500

26 ± 3.5

+

5000

19 ± 7.0

Positive controls, +S9

Name

2AA

Concentrations

(μg/plate)

10

Mean No. of colonies/plate

(average of 3 ± SD)

1343 ± 79.7

Table 2: Test result of experiment 2

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates ± Standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA98

TA1537

0

122 ± 14.6

12 ± 2.6

24 ± 4.0

8 ± 4.6

1.56

142 ± 8.0

ND

ND

10 ± 4.0

3.13

136 ± 23.7

ND

ND

11 ± 3.1

6.25

112 ± 5.0

17 ± 1.5

19 ± 2.5

10 ± 4.0

12.5

115 ± 12.5

16 ± 4.0

20 ± 3.8

7 ± 2.5

25

129 ± 11.2

11 ± 3.5

21 ± 4.2

8 ± 3.1

50

108 ± 14.6

11 ± 2.0

17 ± 2.6

7 ± 3.5

100

 

12 ± 1.5

18 ± 2.1

 

200

 

14 ± 3.5

20 ± 3.5

 

Positive controls, –S9

Name

AF2

SA

AF2

9AA

Concentrations

(μg/plate)

0.01

0.5

0.1

80

Mean No. of colonies/plate

(average of 3 ± SD)

667 ± 16.2

240 ± 6.8

940 ± 12.7

1058 ± 53.3

+

0

131 ± 6.1

11 ± 2.1

31 ± 9.0

18 ± 4.4

+

12.5

ND

ND

ND

20 ± 2.6

+

25

151 ± 11.3

15 ± 1.0

ND

15 ± 6.0

+

50

147 ± 14.5

16 ± 2.6

33 ± 5.5

23 ± 6.8

+

100

143 ± 4.2

14 ± 4.0

30 ± 3.0

17 ± 0.6

+

200

129 ± 20.1

12 ± 4.0

31 ± 6.7

15 ± 1.7

+

400

102 ± 6.0

12 ± 3.2

28 ± 1.5

9 ± 0.6

+

800

89 ± 4.2

13 ± 3.1

22 ± 3.6

 

+

1600

 

 

12 ± 1.0

 

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1

2

0.5

2

Mean No. of colonies/plate

(average of 3 ± SD)

1227 ± 103.5

286 ± 14.8

493 ± 63.5

296 ± 3.2

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

Base-pair substitution type

WP2uvrA

0

27 ± 2.1

313

22 ± 4.9

625

30 ± 2.1

1250

22 ± 3.6

2500

26 ± 5.2

5000

24 ± 4.0

Positive controls, –S9

Name

AF2

Concentrations

(μg/plate)

0.01

Mean No. of colonies/plate

(average of 3 ± SD)

150 ± 41.0

+

0

22 ± 5.1

+

313

25 ± 1.5

+

625

22 ± 4.4

+

1250

28 ± 5.5

+

2500

17 ± 4.9

+

5000

23 ± 3.0

Positive controls, +S9

Name

2AA

Concentrations

(μg/plate)

10

Mean No. of colonies/plate

(average of 3 ± SD)

1531 ± 18.7

AF2: 2 -(2-Furyl)-3 -(5-nitro-2-furyl)acrylamide

SA: Sodium azide

9AA: 9-Aminoacridine

2AA: 2-Aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative