Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Some information in this page has been claimed confidential.

Administrative data

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:
06 May - 29 Aug 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
September 1995
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
September 1995
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Method

Target gene:
his operon (S. typhimurium strains)
trp operon ( E. coli strains)
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
co-factor supplemented post-mitochondrial fraction (S9-mix), prepared from livers of male rats treated with Aroclor 1254
Test concentrations with justification for top dose:
0, 10, 33, 100, 333 and 1000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
- S9: sodium azide (1 μg/plate, TA1535 and TA100); 9-aminoacridine (75 µg/plate, TA 1537); 2-nitrofluorene (1 µg/plate, TA98 and TA 1538); methylmethanesulfonate (1000 µg/plate, WP2 uvrA); +S9: 2-aminoanthracene (1 μg/pate, all strains)
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h
- Expression time (cells in growth medium): 48 to 72 h

DETERMINATION OF CYTOTOXICITY
- Method: inspection of the bacterial background lawn wit a dissecting microscope
Evaluation criteria:
Revertant colonies were counted and the mean and standard deviation were calculated and compared to the controls.
All Salmonella tester strains must demonstrate the presence of the deep rough mutation and the deletion of the uvrA gene. Cultures of the TA98 and TA100 strains must demonstrate the presence of the pKM101 plasmid R-factor. All WP2 uvrA cultures must demonstrate the deletion of the uvrA gene. All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls. Tester strain titers must be above 30.000.000 cells/ml. The mean of each positive control must be at least three-fold increased to the controlls. A minimum of three non-toxic dose levels are recquired to evaluate assay data.
Statistics:
Mean and standard deviation were calculated

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
other: Unacceptable vehicle control values for the tester strain TA1537 and TA 98 were repeated.
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
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, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: >100 µg/plate

RANGE-FINDING/SCREENING STUDIES: Yes

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test results of experiment 1:

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates)

Base-pair substitution type

Frameshift type

TA 1535

TA1537

TA98

TA100

 

TA 1538

WP2uvrA

-

Vehicle

12

4

17

111

5

21

 -

10

3

4

15

115

6

16

 -

33

5

5

19

113

6

11

-

100

9

4

19

98

7

12

-

333

7

5

11

116

4

10

-

1000

8

6

21

120

7

13

Positive

controls

- S9

Name

SA

9AA

2NF

SA

2NF

MMS

Concentrations

(μg/plate)

1.0

75

1.0

1.0

1.0

1000

Number of colonies/plate

241

40

105

377

179

143

+

Vehicle

14

5

18

133

7

11

 +

10

9

5

27

114

12

16

+

33

9

3

21

113

8

13

+

100

8

7

26

108

9

13

+

333

9

4

27

115

6

8

+

1000

10

5

17

111

9

13

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1.0

1.0

1.0

1.0

1.0

10

Number of colonies/plate

72

127

888

904

783

57

SA: sodium azide

9AA : 9-aminoacridine

MMS: methylmethanesulfonate

2-AA: 2-aminoanthracene

2NF: 2-nitrofluorene

 

 

Table 2: Test results of experiment 2/3:

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates)

Base-pair substitution type

Frameshift type

TA 1535

TA1537

TA98

TA100

 

TA 1538

WP2uvrA

-

Vehicle

14

10

23

126

11

27

 -

10

7

13

16

117

7

27

 -

33

9

15

23

124

8

19

-

100

5

13

22

120

6

18

-

333

12

9

17

110

11

16

-

1000

8

14

17

125

5

21

Positive

controls

- S9

Name

SA

9AA

2NF

SA

2NF

MMS

Concentrations

(μg/plate)

1.0

75

1.0

1.0

1.0

1000

Number of colonies/plate

429

757

125

601

221

195

+

Vehicle

8

5

19

147

14

24

 +

10

9

7

17

142

16

30

+

33

10

5

19

136

18

25

+

100

10

5

20

132

13

31

+

333

10

4

17

138

12

19

+

1000

10

7

19

125

12

19

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1.0

1.0

1.0

1.0

1.0

10

Number of colonies/plate

85

97

530

647

1041

88

SA: sodium azide

9AA : 9 -aminoacridine

MMS: methylmethanesulfonate

2-AA: 2 -aminoanthracene

2NF: 2 -nitrofluorene

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative