4-amino-N-(4-aminophenyl)benzenesulphonamide

Genetic toxicity: in vitro

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

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

Test material information

Constituent 1

Reference substance name:

4-amino-N-(4-aminophenyl)benzenesulphonamide

EC Number:

240-834-4

EC Name:

4-amino-N-(4-aminophenyl)benzenesulphonamide

Cas Number:

16803-97-7

Molecular formula:

C12H13N3O2S

IUPAC Name:

4-amino-N-(4-aminophenyl)benzenesulphonamide

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 / cell type:

S. typhimurium, other: TA 1535, TA 100, TA 1537 , TA 1538 and TA 9 8

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type 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 / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle:Complete solubility of the test substance in DMSO .

Controls

Untreated negative controls:

yes

Negative solvent / vehicle 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 experimental 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:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and 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

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 .

Remarks on result:

other: No mutagenic effect were observed .

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.