3-({[(4-methylphenyl)sulfonyl]carbamoyl}amino)phenyl 4-methylbenzenesulfonate;N-(p-toluenesulfonyl)-N'-(3-(p-toluenesulfonyloxy)phenyl)urea

Genetic toxicity: in vitro

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD-guideline and GLP-compliant

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

432-520-2

EC Name:

-

Cas Number:

232938-43-1

Molecular formula:

C21 H20 N2 O6 S2

IUPAC Name:

3-{[(4-methylbenzenesulfonyl)carbamoyl]amino}phenyl 4-methylbenzene-1-sulfonate

Method

Target gene:

his trp

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

33, 100, 333, 1000, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide, 4-nitro-o-phenylene-diamine, methyl methane sulfonate, 2-aminoanthracene

Details on test system and experimental conditions:

For each strain and dose level including the controls, three plates were used.
The following materials were mixed in a test tube and poured onto the minimal agar plates (Experiment I):
100 microl test solution at each dose level, solvent (negative control) or reference mutagen solution (positive control),
500 microl S9 mix (for test with metabolic activation) or S9 mix substitution buffer (for test without metabolic activation),
100 microl Bacteria suspension (cf test system, pre-culture of the strains),
2000 microl Overlay agar
In the pre-incubation assay (Experiment II) 100 µl test solution, 500 µl S9 mix / S9 mix substitution buffer and 100 µl bacterial suspension were mixed in a test tube and shaken at 37° C for 60 minutes. After pre-incubation 2.0 ml overlay agar (45° C) was added to each tube. The mixture was poured on minimal agar plates.
After solidification the plates were incubated upside down for at least 48 hours at 37° C in the dark

Evaluation criteria:

A test item is considered positive if either a dose related and reproducible increase in the number of revertants or a biologically relevant and reproducible increase for at least one test concentration is induced. A test item producing neither a reproducible and dose related increase in the number of revertants, nor a biologically relevant and reproducibly positive response at any one of the test points is considered non-mutagenic in this system. A,mutagenic response is described as follows: A test item is considered mutagenic if in the strains TA 98, TA 100, and WP2 uvrA the number of reversions will be at least twice as high and in the strains TA 1535 and TA 1537 at least three times higher as compared to the spontaneous reversion rate.
Also, a dose-dependent and reproducible increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test item regardless whether the highest dose induced the above described enhancement factors or not.

Statistics:

No statistical evaluation of the data is required.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1537

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

Test results 2

Species / strain:

S. typhimurium TA 1535

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

Test results 3

Species / strain:

S. typhimurium TA 100

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

Test results 4

Species / strain:

S. typhimurium TA 98

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

Test results 5

Species / strain:

E. coli WP2 uvr A

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

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Revertants/plate (mean of three plates) in the Ames test with and without metabolic activation

	Revertants/plate, without S9 mix (mean of three plates)
Strain	TA 1535		TA1537		TA 98		TA 100		WP2 uvra
Experiment	I	II	I	II	I	II	I	II	I	II
Negative control	24	26	8	14	21	19	86	105	54	42
Solvent control	21	22	10	16	19	17	85	78	48	43
Positive control	611	1498	106	52	410	509	1025	1478	802	433
33	20	25	12	10	16	25	89	86	40	37
100	19	25	9	18	15	19	80	75	38	45
333	13	33	9	15	13	14	77	86	42	42
1000	21	23	10	14	16	17	69	73	42	37
2500	18	20	8	8	18	13	86	64	39	30
5000	19	15	4	8	12	13	79	43	29	34
	Revertants/plate, with S9 mix (mean of three plates)
Negative control	13	11	15	22	23	32	80	100	52	55
Solvent control	10	15	22	13	28	27	85	88	48	50
Positive control	132	109	140	48	314	312	503	510	275	228
33	10	12	13	25	24	29	93	94	48	51
100	10	13	10	15	18	29	85	92	48	45
333	8	12	11	15	25	29	76	89	49	46
1000	8	11	13	15	32	32	78	74	50	41
2500	9	10	12	10	30	23	81	76	45	38
5000	5	11	9	11	20	20	96	71	40	54
Experiment I: plate incorporation method, Experiment II: preincubation method

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

The test substance was not genotoxic when tested in the Ames test with different strains of Salmonella Typhimurium and the Escherichia coli strain WP2 uvrA according to the OECD 471 (1997).