Registration Dossier

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report date:
1992

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial gene mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Trisodium bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]-7-nitronaphthalene-1-sulphonato(3-)]chromate(3-)
EC Number:
260-906-9
EC Name:
Trisodium bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]-7-nitronaphthalene-1-sulphonato(3-)]chromate(3-)
Cas Number:
57693-14-8
Molecular formula:
C40H20CrN6O14S2.3Na
IUPAC Name:
trisodium bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]-7-nitronaphthalene-1-sulphonato(3-)]chromate(3-)
Test material form:
solid: particulate/powder

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9 mix
Test concentrations with justification for top dose:
0-8-40-20-1000-5000 µg/plate + 5 different positive control
Repeat: 0-125-250-500-1000-2000-4000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
other: nitro furatoin (TA 100); 4-nitro-1,2-phenylenediamine(TA 1537, TA98); 2- aminoanthracene
Details on test system and experimental conditions:
DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 4

DETERMINATION OF CYTOTOXICITY
- Method: background lawn density
Evaluation criteria:
Reproducible and dose-related increase of mutant count in at least one strain is considered a positive result.

Results and discussion

Test resultsopen allclose all
Species / strain:
E. coli WP2
Remarks:
not tested according to the OECD guideline 471 in force at the time the test was run.
Metabolic activation:
with and without
Genotoxicity:
not determined
Cytotoxicity / choice of top concentrations:
not determined
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
ambiguous
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
ambiguous
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

Summary of mean values without S9 mix

 dose (µg/plate)

 TA1535

TA100

 TA1537

TA 98

0

8

40

200

1000

5000

Na N3

NF

4 -NPDA

15

15

13

14

13

P

537

110 

117

118

112

125

P

243

12 

11

14

17

32

P

45

28 

34

38

50

68

P

65

 0

125

250

500

1000

2000

4000

Na N3

NF

4 -NPDA

12

10

10

14

7

11

P

747

102

92

98

108

115

109

P

460

 

9

19

17

19

30

23

P

74 

32

37

37

38

46

43

P

129 

 0

400

600

800

1000

1200

1400

1600

4 -NPDA

 

 

6

22

20

24

34

32

31

21

83 

33

52

56

56

66

65

59

62

291 

Summary of mean values with S9-mix

 dose (µg/plate)  TA 1535 TA100  TA 1537  TA 98 

 30 % S9mix

0

8

40

200

1000

5000

2 -AA

22

29

25

23

18

P

152 

159

165

182

172

192

P

663 

15

11

14

15

23

P

75 

34

46

38

57

85

P

464 

 10 % S9mix

0

125

250

500

1000

2000

4000

2 -AA

 

15

17

14

15

15

11

P

253

126

145

153

124

109

107

P

934 

13

14

13

18

23

22

P

443 

36

54

55

53

50

42

P

1304 

 30 % S9mix

0

400

600

800

1000

1200

1400

1600

2 -AA

   

13

12

14

15

26

22

18

24

116 

52

71

83

83

87

83

82

65

586 

Applicant's summary and conclusion

Conclusions:
Test substance showed equivocal mutagenic effects in 2 of 4 strains in Salmonella typhimurim.
Executive summary:

Method

The test material was examined in four strains of Salmonella typhimurium for gene mutations in bacteria (Ames test) in the presence and absence of a metabolic activation system (rat liver S9 mix). The strains employed were TA 98, TA 100, TA 1535, and TA 1537. All concentrations were tested in four plates each. The dose range was 0 (vehicle), 8 to 5000 µg/plate and several positive controls.

Results

Positive as well as negative controls gave the expected results. Plates treated with test item showed precipitation at concentrations of 4000 µg/plate and above. Cytotoxicity was noted at doses above 800 µg/plate. Equivocal mutagenic effects were observed in TA 1537 and TA 98. There were weak increases of revertant colonies, less than three times, mostly less than twice, the negative controls. However, no clear dose response relations could be determined, even when the dose was increased significantly.