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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
In a reverse gene mutation assay in bacteria (Ames Test), no mutagenicity was observed with or without metabolic activation (BASF, 1996).
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:
1996-05-07 to 1996-05-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline compliant study.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced rat liver S9 mix
Test concentrations with justification for top dose:
Experiment 1 and 2: 20, 100, 500, 2500, 5000 µg/plate
Vehicle / solvent:
DMSO
- Vehicle used: DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
with S9 mix (TA100, TA98, TA1537, TA 1535, WP2 uvrA)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitrosoguanidine
Remarks:
without S9 mix (TA100, TA1535)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylendiamine
Remarks:
without S9 mix (TA98)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without S9 mix (TA1537)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitrosoguanidine
Remarks:
without S9 mix (WP2 uvrA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: other: reduced background lawn
Evaluation criteria:
In general, a substance to be characterized as positive in the bacterial tests has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
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:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No bacteriotoxic effect (reduced his or trp background growth) was observed.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Raw data

Experiment 1 (Standard Plate test)

Dose (µg/plate)

Mean number of revertant colonies/3 replicate plates with different strains of Salmonella typhimurium and Escherichia coli

 

TA1535

TA1537

TA98

TA100

WP2 uvrA

 

 

 

Results with S9

Spontaneous Reversion

20

7

35

120

49

Positive control

206

116

828

1064

176

20

16

7

38

112

44

100

18

7

29

110

46

500

20

7

27

93

47

2500

12

6

26

97

42

5000

13

6

30

79

41

 

 

 

 

 

 

 

 

 

Results without S9

Spontaneous Reversion

20

7

23

106

44

Positive control

1430

658

1150

986

795

20

16

7

26

114

41

100

16

5

20

108

45

500

18

5

19

107

38

2500

19

7

20

118

44

5000

16

6

25

116

40

 

 

 

 

 

 

 

 

 

 

 

 

Experiment 2 (Pre-incubation test)

Dose (µg/plate)

Mean number of revertant colonies/3 replicate plates with different strains of Salmonella typhimurium and Escherichia coli

 

TA1535

TA1537

TA98

TA100

WP2 uvrA

 

 

 

Results with S9

Spontaneous Reversion

22

12

45

121

41

Positive control

137

111

675

782

208

20

20

13

48

113

35

100

19

9

45

124

39

500

20

11

41

118

32

2500

20

8

37

99

32

5000

19

7

39

107

35

 

 

 

 

 

 

 

 

 

Results without S9

Spontaneous Reversion

21

6

32

116

44

Positive control

868

632

1097

1119

791

20

20

6

33

114

45

100

16

4

33

123

38

500

20

4

38

115

41

2500

16

4

33

117

40

5000

19

4

35

110

38

 

 

 

 

 

 

Conclusions:
Interpretation of results (migrated information):
negative
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

In a reverse mutation assay according to OECD guideline 471, strains (TA1535, TA1537, TA100, TA98 and WP2 uvr A) of S. typhimurium and Escherichia coli were exposed to the test substance at concentrations of 20, 100, 500, 2500 and 5000 µg/plate in the presence and absence of mammalian metabolic activation (BASF, 40M0133/964048, 1996). Each concentration was tested in triplicate.

The positive controls induced the appropriate responses in the corresponding strains. There was no evidence or a concentration related positive response of induced mutant colonies over background. The test substance was not mutagenic in the S. typhimurium strains TA1535, TA1537, TA100 and TA98 and the E. coli strain WP2 uvr A under the experimental conditions with or without metabolic activation.


Justification for selection of genetic toxicity endpoint
GLP and guideline compliant study.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes. As a result the substance does not need to be classified and labelled for mutagenic toxicity under Regulation (EC) No 1272/2008, as amended for the seventh time in Regulation (EU) No 2015/1221.