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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006-02-15 to 2006-03-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesamt für Umweltschutz und Gewerbeaufsicht
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Glycine, N-methyl-, N-coco acyl derivs., sodium salts
EC Number:
263-193-2
EC Name:
Glycine, N-methyl-, N-coco acyl derivs., sodium salts
Cas Number:
61791-59-1
Molecular formula:
Not applicable
IUPAC Name:
disodium 2-(N-methyldodecanamido)acetate 2-(N-methyltetradecanamido)acetate
Test material form:
not specified

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102 and TA 1535
Additional strain / cell type characteristics:
other: histidin-deficient
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
Test concentrations with justification for top dose:
First experiment:
- 1.5, 5, 15, 50, 149 µg/plate with and without metabolic activation

Second experiment:
- 37.5, 75, 150 µg/plate with and without metabolic activation
Vehicle / solvent:
water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other:
Remarks:
NPD: 80 μg/plate in water, -S9, TA 97a, TA 98, TA 102; Sodium azide: 6 μg/plate in water, -S9, TA100 and TA1535; 2-AA: 3 μg/plate in DMSO, +S9, TA 98; BaP: 40 μg/plate in DMSO, +S9, TA 97a, TA 100, TA 102, TA 1535
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 12 h
- Exposure duration: 48 h

NUMBER OF REPLICATIONS:
- dublicates each in two independent experiments.

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:
The test material may be considered positive in this test system if the following
criteria are met:
For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at one strain with or without S9 mixture or there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies with increasing dose levels
Statistics:
Mean values and standard deviation were calculated

Results and discussion

Test results
Key result
Species / strain:
other: S. typhimurium: TA 97a, TA 98, TA 100, TA 102 and TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
above 150 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
- The highest concentration used in the study was 150 µg/plate, based on the cytotoxicity at higher concentrations.

Any other information on results incl. tables

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 2 plates ± Standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA102

TA98

TA97a

0

114 ± 49

9 ± 2.4

9 ± 2.4

12 ± 0.5

130 ± 34.5

1.5

71 ± 15

6 ± 4

164 ± 25

8 ± 1

124 ± 36

5

97 ± 22

7 ± 2

125 ± 9

8 ± 1

97 ± 38

15

86 ± 12

6 ± 2

106 ± 19

6 ± 3

80 ± 7

50

90 ± 16

8 ± 1

101 ± 10

4 ± 2

83 ± 31

150*

113 ± 41

8 ± 3

139 ± 7

14 ± 6

120 ± 55

Positive controls, –S9

Name

NaN3

NaN3

NPD

NPD

NPD

Concentrations

(μg/plate)

6

6

80

80

80

Mean No. of colonies/plate

(average of 2 ± SD)

366 ± 101

1001 ± 0

657 ± 122

205 ± 98

1001 ± 0

+

0

138 ± 38

6 ± 2.6

158 ± 27.9

15 ± 2.4

101 ± 26.9

+

1.5

74 ± 13

5 ± 2

154 ± 44

12 ± 7

93 ± 42

+

5

127 ± 22

7 ± 1

156 ± 26

9 ± 6

115 ± 38

+

15

104 ± 14

8 ± 3

146 ± 9

8 ± 5

86 ± 31

+

50

103 ± 17

11 ± 3

159 ± 23

12 ± 7

101 ± 21

+

150*

78 ± 9

9 ± 1

115 ± 32

8 ± 5

102 ± 31

Positive controls, +S9

Name

2AA

2AA

2AA

BP

2AA

Concentrations

(μg/plate)

3

3

3

40

3

Mean No. of colonies/plate

(average of 2 ± SD)

1218 ± 251

130 ± 12

382 ± 42

200 ± 48

718 ± 231

Plate incorporation

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 2 plates ± Standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA102

TA98

TA97a

0

145 ± 17

12 ± 3.4

186 ± 3.6

8 ± 2.2

153 ± 27.3

37.5

158 ± 45

7 ± 3

136 ± 47

7 ± 3

122 ± 54

75

131 ± 19

8 ± 4

128 ± 42

13 ± 5

125 ± 20

150*

146 ± 7

6 ± 4

129 ± 28

11 ± 5

115 ± 37

Positive controls, –S9

Name

NaN3

NaN3

NPD

NPD

NPD

Concentrations

(μg/plate)

6

6

80

80

80

Mean No. of colonies/plate

(average of 2 ± SD)

1001 ± 0

1001 ± 0

1001 ± 0

1001 ± 0

1001 ± 0

+

0

153 ± 7

13 ± 1.7

137 ± 43.2

10 ± 3.4

141 ± 23.6

+

37.5

95 ± 16

8 ± 3

86 ± 25

10 ± 4

104 ± 31

+

75

104 ± 21

11 ± 3

117 ± 15

10 ± 4

156 ± 29

+

150*

120 ± 10

5 ± 2

134 ± 44

6 ± 2

89 ± 28

Positive controls, +S9

Name

2AA

2AA

2AA

BP

2AA

Concentrations

(μg/plate)

3

3

3

40

3

Mean No. of colonies/plate

(average of 2 ± SD)

1001 ± 0

1001 ± 0

1001 ± 0

105 ± 6

1001 ± 0

Preincubation

*Remark: 150 µg/plate was the highest concentration of test substance due to toxicity testing

NPD: 4-Nitro-1,2-phenylene diamine

NaN3: sodium azide

2AA: 2-Amino-anthracene

BaP: Benzo-a-pyrene

Applicant's summary and conclusion

Conclusions:
Interpretation of results: negative