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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from Study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
The substance "Sulfamid" was tested for mutagenicity in the Ames test by Standard plate metod.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Name of test material : 4-amino-N-(4-aminophenyl)benzenesulphonamide
- Molecular formula : C12H13N3O2S
- Molecular weight : 263.32 g/mol
- Smiles notation : O=S(=O)(c1ccc(cc1)Nc1ccc(cc1)N)N
- InChl : 1S/C12H13N3O2S/c13-9-1-3-10(4-2-9)15-11-5-7-12(8-6-11)18(14,16)17/h1-8,15H,13H2,(H2,14,16,17)
- Substance type : Organic
- Physical state : Solid
Specific details on test material used for the study:
- Name of test material : 4-amino-N-(4-aminophenyl)benzenesulphonamide
- Molecular formula : C12H13N3O2S
- Molecular weight : 263.32 g/mol
- Smiles notation : O=S(=O)(c1ccc(cc1)Nc1ccc(cc1)N)N
- InChl : 1S/C12H13N3O2S/c13-9-1-3-10(4-2-9)15-11-5-7-12(8-6-11)18(14,16)17/h1-8,15H,13H2,(H2,14,16,17)
- Substance type : Organic
- Physical state : Solid

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: J. 9045, V . 134
- Purity : 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material;+4C

Method

Target gene:
Histidine
Species / strain
Species / strain:
other: TA 1535, TA 100, TA 1537 , TA 1538 and TA 9 8
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
other: All strains have a defective excision repair system (uvrB) and reduced hydrophilic polysaccharide layer(rfa)
Cytokinesis block (if used):
Not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix is prepared by injecting male Sprague-Dawley rats liver with Aroclor 1254
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500 and 5000 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle:Complete solubility of the test substance in DMSO .
Controls
Negative controls:
yes
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: + S-9 mix ;2-aminoanthracene for TA 100, TA 98, TA 1538, TA 153T and T - S-9 mix ; N-methyl-N'-nitro-N-nitrosoguanidine for TA 100 and TA 153 5 4-nitro-o-phenylendiamine for TA 98 and TA 153 8 9-aminoacridine chloride monohydrate for TA 153T.
Details on test system and conditions:
METHOD OF APPLICATION: Standard plate test
DURATION
- Exposure duration:48 hours

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

- OTHER:3 test plates per dose or per control was observed

Rationale for test conditions:
Not specified .
Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationshi p
- reproducibility of the results .
Statistics:
Mean± Standard deviation was observed.

Results and discussion

Test results
Key result
Species / strain:
other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: No mutagenic effect were observed .
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY: A weakly bacteriotoxic effect (slight decrease in the number of his+ revertants) was observed depending on the strain and test conditions at doses, ≥2500 µg/plate .

Any other information on results incl. tables

Table 1

Strain : TA 1535

Dose

MCG/Pl

Revertants / plate

Titer Dil

Exp-6

Quotient

-S9

M

SD

+S9

M

SD

-S9

+S9*

Negative control DMSO

14

17

3

15

17

2

35

1.0

1.0

17

 

 

18

 

 

42

 

 

20

 

 

19

 

 

47

 

 

20

13

14

3

17

19

2

 

0.8

1.1

18

 

 

20

 

 

 

 

 

12

 

 

19

 

 

 

 

 

100

15

16

1

21

18

3

 

0.9

1.1

15

 

 

15

 

 

 

 

 

17

 

 

19

 

 

 

 

 

500

22

18

5

10

11

3

33

1.0

0.7

20

 

 

9

 

 

38

 

 

18

 

 

5

 

 

41

 

 

2500

23

18

5

10

11

3

3

1.0

0.7

14

 

 

9

 

 

 

 

 

16

 

 

15

 

 

 

 

 

5000

12

12

2

13

12

2

17

0.7

0.7

11

 

 

12

 

 

24

 

 

14

 

 

10

 

 

21

 

 

Positive control 2-AA 10

 

 

 

351

305

40

 

 

17.6

 

 

 

289

 

 

 

 

 

 

 

 

276

 

 

 

 

 

Positive control MNNG      5

1940

2030

82

 

 

 

 

119.4

 

2100

 

 

 

 

 

 

 

 

2050

 

 

 

 

 

 

 

 

*S-9 Fraction/Co-factor=3.7                EXP:EXP to 10

 

 

 

 

 

 

Table : 2                                Strain: TA100

Dose

MCG/Pl

Revertants / plate

Titer Dil

Exp-6

Quotient

-S9

M

SD

+S9

M

SD

-S9

+S9*

Negative control DMSO

118

115

10

107

112

6

63

1.0

1.0

124

 

 

119

 

 

67

 

 

104

 

 

110

 

 

52

 

 

20

115

113

10

124

121

7

 

1.0

1.1

105

 

 

127

 

 

 

 

 

120

 

 

113

 

 

 

 

 

100

109

108

3

109

113

8

 

0.9

1.0

105

 

 

108

 

 

 

 

 

110

 

 

122

 

 

 

 

 

500

113

118

6

131

124

7

 

1.0

1.1

117

 

 

117

 

 

 

 

 

124

 

 

123

 

 

 

 

 

2500

126

120

16

109

108

7

35

1.0

1.0

131

 

 

115

 

 

47

 

 

102

 

 

101

 

 

53

 

 

5000

87

79

7

87

72

16

23

0.7

0.6

73

 

 

56

 

 

29

 

 

76

 

 

74

 

 

41

 

 

Positive control 2-AA 10

 

 

 

2400

2283

126

 

 

20.4

 

 

 

2150

 

 

 

 

 

 

 

 

2300

 

 

 

 

 

Positive control MNNG      5

1850

1780

62

 

 

 

 

15.4

 

1730

 

 

 

 

 

 

 

 

1760

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table : 3                          Strain: TA1537

Dose

MCG/Pl

Revertants / plate

Titer Dil

Exp-6

Quotient

-S9

M

SD

+S9

M

SD

-S9

+S9*

Negative control DMSO

10

10

2

8

9

2

28

1.0

1.0

12

 

 

11

 

 

37

 

 

9

 

 

9

 

 

31

 

 

20

8

11

3

12

10

2

 

1.1

1.1

14

 

 

11

 

 

 

 

 

11

 

 

8

 

 

 

 

 

100

12

12

1

10

9

2

 

1.2

1.0

13

 

 

7

 

 

 

 

 

12

 

 

11

 

 

 

 

 

500

9

10

1

13

10

3

 

0.9

1.1

11

 

 

9

 

 

 

 

 

9

 

 

8

 

 

 

 

 

2500

10

9

1

10

8

2

44

0.8

0.9

8

 

 

8

 

 

32

 

 

8

 

 

7

 

 

36

 

 

5000

3

3

2

7

6

2

14

0.3

0.6

2

 

 

6

 

 

21

 

 

5

 

 

4

 

 

22

 

 

Positive control 2-AA 10

 

 

 

127

130

12

 

 

13.9

 

 

 

143

 

 

 

 

 

 

 

 

119

 

 

 

 

 

Positive control MNNG      100

568

674

93

 

 

 

 

65.3

 

713

 

 

 

 

 

 

 

 

742

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table No. 4                             Strain: TA 98

Dose

MCG/Pl

Revertants / plate

Titer Dil

Exp-6

Quotient

-S9

M

SD

+S9

M

SD

-S9

+S9*

Negative control DMSO

24

23

3

37

34

4

36

1.0

1.0

26

 

 

34

 

 

44

 

 

20

 

 

30

 

 

29

 

 

20

28

23

4

31

35

6

 

1.0

1.0

20

 

 

33

 

 

 

 

 

21

 

 

42

 

 

 

 

 

100

23

24

3

32

36

5

 

1.0

1.1

27

 

 

36

 

 

 

 

 

21

 

 

41

 

 

 

 

 

500

18

21

2

30

32

4

 

0.9

1.1

22

 

 

29

 

 

 

 

 

22

 

 

37

 

 

 

 

 

2500

21

21

3

32

37

4

21

0.9

1.1

19

 

 

38

 

 

23

 

 

24

 

 

40

 

 

41

 

 

5000

8

9

4

17

15

5

17

0.4

0.5

13

 

 

19

 

 

12

 

 

6

 

 

10

 

 

19

 

 

Positive control 2-AA 10

 

 

 

1250

1180

66

 

 

35.0

 

 

 

1120

 

 

 

 

 

 

 

 

1170

 

 

 

 

 

Positive control MNNG      5

833

903

65

 

 

 

 

38.7

 

915

 

 

 

 

 

 

 

 

961

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table No. 5                        Strain: TA 1538

Dose

MCG/Pl

Revertants / plate

Titer Dil

Exp-6

Quotient

-S9

M

SD

+S9

M

SD

-S9

+S9*

Negative control DMSO

14

14

1

30

28

2

50

1.0

1.0

14

 

 

27

 

 

48

 

 

15

 

 

26

 

 

39

 

 

20

15

17

2

21

26

5

 

1.2

0.8

17

 

 

26

 

 

 

 

 

18

 

 

30

 

 

 

 

 

100

10

13

3

28

29

6

 

0.9

0.1

13

 

 

35

 

 

 

 

 

16

 

 

24

 

 

 

 

 

500

13

12

2

23

23

2

 

0.8

0.8

10

 

 

24

 

 

 

 

 

12

 

 

21

 

 

 

 

 

2500

10

7

3

24

19

6

16

0.5

0.7

4

 

 

20

 

 

36

 

 

7

 

 

12

 

 

6

 

 

5000

4

4

1

14

12

2

0

0.3

0.4

4

 

 

11

 

 

0

 

 

3

 

 

12

 

 

0

 

 

Positive control 2-AA                10

 

 

 

800

989

241

 

 

35.8

 

 

 

908

 

 

 

 

 

 

 

 

1260

 

 

 

 

 

Positive control MNNG      5

760

592

147

 

 

 

 

41.3

 

531

 

 

 

 

 

 

 

 

486

 

 

 

 

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
The substance "Sulfamid" (16803-97-7)was tested for mutagenicity in the Ames test (standard plate test) both in the presence and in the absence of a metabolizing system obtained from rat liver (S-9 mix)using the strains TA 1535, TA 100, TA 153T, TA 1538 and TA 98. The test result was considered to be negative both in the presence and absence of metabolic activation.
Executive summary:

In Vitro genetic toxicity study "Sulfamid" (16803-97-7) was assessed for its possible mutagenic potential. For this purpose is Bacterial Reverse mutation assay was performed according to OECD guideline 471. The test substance was exposed to the Salmonella Typhimurium strains TA 1535, TA 100, TA 153T, TA 1538 and TA 98 at the concentration of 0, 20, 100, 500, 2500 and 5000 µg/plate. The test substance was tested by standard plate method both in the presence and in the absence of a metabolizing system obtained from rat liver (S-9 mix). Cytotoxicity was also observed. No mutagenic effects were observed in the test. Therefore "Sulfamid" was considered to be non mutagenic in the strains TA 1535, TA 100, TA 153T, TA 1538 and TA 98. Hence the substance cannot be classified as gene mutant in vitro.