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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro: Ames test (OECD 471): negative with S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 with and without metabolic activation

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:
10 Sep - 11 Oct 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Reference:
Composition 1
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted 21 July 1997
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Ministerium für Umwelt und Verkehr, Baden-Württemberg, Germany
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Target gene:
his operon
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
co-factor 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:
5; 15; 50; 150; 500; 1500 and 5000 µg/plate with and without metabolic activation

Second experiment:
1,5; 5; 15; 50; 150; 500; 1500 and 5000 µg/plate without metabolic activation
15; 50; 150; 500; 1500 and 5000 µg/plate with metabolic activation
Vehicle:
- Vehicle(s)/solvent(s) used: ethanol
Negative controls:
yes
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
other: 2-aminoanthracene
Remarks:
+S9: 2-AA (0.8µg/plate for TA98, TA100; 0.9µg/plate for TA102, TA1535; 1.7 µg/plate for TA1537); -S9: NaN3 (0.7 µg/plate for TA100, TA1535), 2-NF (2.5 µg/plate for TA98), 9-AA (50 µg/plate for TA1537), Mitomycin C (0.15 µg/plate for TA102)
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48–72 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn

Statistics:
Mean values and standard deviation were calculated.
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
-S9: at 500 µg/plate; +S9: at 5000 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
+S9: at 5000 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
-S9: at 5000 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
-S9: at 500 µg/plate; +S9: at 5000 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
-S9: at 500 µg/plate; +S9: at 1500 µg/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation of the test substance was observed on the plates.

Table 1: Test Results of Experiment 1

EXPERIMENT 1

S9-Mix

Without

 

Test item (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

NC

44 ± 4

110 ± 16

269 ± 14

11 ± 5

9 ± 4

Solvent control (ethanol)

34 ± 11

106 ± 7

264 ± 12

12 ± 3

7 ± 2

5

-

-

283 ± 5

-

6 ± 2

15

37 ± 3

97 ± 10

292 ± 29

-

6 ± 1

50

44 ± 8

114 ± 11

248 ± 15

14 ± 6

6 ± 2

150

25 ± 4

109 ± 4

190 ± 10

11 ± 5

7 ± 3

500

33 ± 5

94 ± 3

170 ± 10 (T)

12 ± 7

5 ± 2

1500

34 ± 4

66 ± 5 (T)

-

9 ± 1

-

5000

-

-

-

8 ± 3

-

NaN3 (0.7)

-

491 ± 29

-

411 ± 9

-

2-NF (2.5)

463 ± 107

-

-

-

-

9-AA (50)

-

-

-

-

454 ± 49

Mitomycin C (0.15)

-

-

788 ± 28

-

-

S9-Mix

With

 

Test item (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

 

 

 

 

 

 

NC

38 ± 5

130 ± 16

251 ± 17

12 ± 5

16 ± 2

Solvent control (ethanol)

47 ± 4

113 ±13

255 ± 13

12 ± 4

18 ± 4

5

-

-

262 ± 16

-

-

15

-

108 ± 9

264 ± 16

12 ± 1

16 ± 5

50

52 ± 7

118 ± 13

240 ± 22

11 ± 2

16 ± 2

150

49 ± 4

118 ± 13

241 ± 16

13 ± 12

24 ± 3

500

42 ± 4

97 ± 7

224 ± 8

12 ± 2

20 ± 3

1500

52 ± 4

76 ± 6

-

11 ± 2

12 ± 5

5000

33 ± 4

-

-

-

-

2-AA (0.8)

749 ± 81

613 ± 58

-

-

-

2-AA (0.9)

-

-

378 ± 32

132 ± 30

-

2-AA (1.7)

-

-

-

-

177 ± 8

NC = Negative Control

T = bacteriotoxic

2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene; NaN3: sodium azide

Table 2: Test Results of Experiment 2

EXPERIMENT 2

S9-Mix

Without

 

Test item (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

NC

52 ± 8

92 ± 8

214 ± 21

12 ± 3

13 ± 3

Solvent control (ethanol)

47 ± 7

82 ± 5

222 ± 13

10 ± 2

8 ± 2

1.5

-

-

237 ± 8

-

-

5

-

86 ± 14

242 ± 10

-

-

15

-

80 ± 11

253 ± 6

-

9 ± 2

50

52 ± 15

78 ± 8

238 ± 9

10 ± 2

6 ± 2

150

47 ± 5

70 ± 9

205 ± 3

14 ± 3

7 ± 4

500

54 ± 8

59 ± 6 (T)

-

10 ± 4

5 ± 3 (T)

1500

59 ± 6

-

-

9 ± 3

6 ± 2 (T)

5000

33 ± 3 (T)

-

-

11 ± 2

-

NaN3 (0.7)

-

409 ± 18

-

322 ± 16

-

2-NF (2.5)

431 ± 18

-

-

-

-

9-AA (50)

-

-

-

-

228 ± 22

Mitomycin C (0.15)

-

-

605 ± 65

-

228 ± 22

S9-Mix

With

 

Test item (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

 

 

 

 

 

 

NC

48 ± 11

88 ± 15

259 ± 11

19 ± 5

11 ± 2

Solvent control (ethanol)

48 ± 7

97 ± 4

263 ± 15

21 ± 2

15 ± 4

15

-

-

255 ± 21

-

-

50

41 ± 3

103 ± 7

237 ± 13

20 ± 5

14 ± 3

150

51 ± 7

91 ± 10

235 ± 31

17 ± 4

16 ± 4

500

48 ± 5

111 ± 2

214 ± 19

18 ± 3

12 ± 4

1500

56 ± 9

87 ± 1

160 ± 12 (T)

15 ± 3

11 ± 2

5000

33 ± 5

19 ± 3 (T)

-

9 ± 3 (T)

1 ± 2 (T)

2-AA (0.8)

707 ± 23

619 ± 43

-

-

-

2-AA (0.9)

-

-

377 ± 20

133 ± 21

-

2-AA (1.7)

-

-

-

-

194 ± 9

NC = Negative Control

T = bacteriotoxic

2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene; NaN3: sodium azide

Conclusions:
Under the experimental conditions described, the test substance was not mutagenic in the Ames test using the Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and TA102 with and without metabolic activation.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Justification for selection of genetic toxicity endpoint
There is only one study available. This study is sufficient to fulfil the standard information requirements set out in Annex VII, 8.4 of Regulation (EC) No 1907/2006.

Justification for classification or non-classification

Based on the available data, there is no indication that the substance induces genetic toxicity. Nevertheless, no final decision on classification for genetic toxicity according to Regulation (EC) 1272/2008 can be made, as no information on chromosomal aberration and mutagenicity in mammalian cells/in vivo is available.