2,4-diaminobenzenesulphonic acid

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:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

SALMONELLA MUTAGENICITY TESTS: IV. RESULTS FROM THE TESTING OF 300 CHEMICALS;

Author:

ZEIGER,E, ANDERSON,B, HAWORTH,S, LAWLOR,T AND MORTELMANS,K;

Year:

1988

Bibliographic source:

Environmental and Molecular Mutagenesis, 11(SUPPL.12):1-158, 1988

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other:

Principles of method if other than guideline:

Gene mutation toxicity study was performed to evaluate the mutagenic nature of the test compound 2-aminobenzenesulphonic acid

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-aminobenzenesulphonic acid

EC Number:

201-810-9

EC Name:

2-aminobenzenesulphonic acid

Cas Number:

88-21-1

IUPAC Name:

2-aminobenzenesulfonic acid

Details on test material:

- Name of test material: 2-aminobenzenesulphonic acid- Molecular formula: C6H7NO3S- Molecular weight: 173.1913 g/mol- Substance type: Organic- Physical state: Solid- Purity: 95%

Method

Target gene:

Histidine auxotrophs

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100, TA1535

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

HAMSTER, LIVER, S-9, AROCLOR 1254 (10 OR 30%)

Test concentrations with justification for top dose:

0, 10, 33, 100, 333, 1000, 3333, 3334, 6667 µg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

For strains tested with S9

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

For strains TA100 and TA1535 tested in the absence of S9

Controls 3

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

For strains TA98 tested in the absence of S9

Controls 4

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

For strains TA97 tested in the absence of S9

Details on test system and experimental conditions:

Details on test system and conditionsMETHOD OF APPLICATION: preincubationDURATION- Preincubation period: 20 mins- Exposure duration: 48 hrs- Expression time (cells in growth medium): 48 hrs- Selection time (if incubation with a selection agent): No data- Fixation time (start of exposure up to fixation or harvest of cells): No dataSELECTION AGENT (mutation assays): No dataSPINDLE INHIBITOR (cytogenetic assays): No dataSTAIN (for cytogenetic assays): No dataNUMBER OF REPLICATIONS: At least five doses of each chemical were tested in triplicateNUMBER OF CELLS EVALUATED: No dataDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No dataOTHER EXAMINATIONS:- Determination of polyploidy: No data- Determination of endoreplication: No data- Other: No dataOTHER: No data

Evaluation criteria:

Evaluations were made at both the individual trial and overall chemical levels. Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of “ +W,” if only a single dose was elevated over the control, or if the increase seen was not dose related. The distinctions between a questionable mutagenic response and a nonmutagenic or weak mutagenic response, and between a weak mutagenic response and mutagenic response are highly subjective. It was not necessary for a response to reach twofold over background for a chemical to be judged mutagenic.A chemical was judged mutagenic (+) or weakly mutagenic (+ W) if it produced a reproducible dose-related reponse over the solvent control in replicate trials. A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a “+W” response, or if only single doses produced increases in his+ revertants in repeat trials. Chemicals were judged non-mutagenic (-) if they did not meet the criteria for a mutagenic or questionable response. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or non-mutagenicity.

Statistics:

No data available

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100, TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

Additional information on resultsTEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data- Effects of osmolality: No data- Evaporation from medium: No data- Water solubility: No data- Precipitation: No data - Other confounding effects: No dataRANGE-FINDING/SCREENING STUDIES: All chemicals were tested initially in a toxicity assay to determine the appropriate dose range for the mutagenicity assay. The toxicity assay was performed using TA100 or the system developed by Waleh et al. Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both.COMPARISON WITH HISTORICAL CONTROL DATA: No dataADDITIONAL INFORMATION ON CYTOTOXICITY: No data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Strain TA100

Dose	No Activation  (Negative)		No Activation  (Negative)		10% HLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		10% RLI  (Negative)		30% RLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	90	2	102	4.6	77	7.8	130	2.4	98	2.7	87	4.7	121	9
33	81	4.2	99	2	91	2.1	112	11.1	72	11.1	73	5.8	121	2.2
100	96	7.8	101	5.7	110	11	115	2	101	12.8	72	3.6	126	12.1
333	91	0.7	95	2	86	2.7	120	2.5	89	6.4	76	3.1	121	9.1
1000	80	7.3	91	7	91	2.7	112	5	84	7.3	78	2.1	116	2.3
3333	85	2.5	93	4.6	84	5.4	115	4	90	5.9	75	0.3	127	5
Positive Control	290	8.1	236	12.8	333	18.8	357	9.7	198	23.4	399	16.7

Strain TA1535

Dose	No Activation  (Negative)		No Activation  (Negative)		10% HLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		30% RLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	18	3.2	8	0.7	8	0.3	5	0	11	2	9	1.3
33	16	1.3	7	0.3	9	0.9	8	2	9	0.9	12	1.7
100	10	0.3	6	1.5	11	1.5	5	1.7	6	1	7	1.8
333	15	1.8	7	1.8	12	1.5	8	1.2	9	1.8	12	2.1
1000	16	1.2	8	1.5	11	0.9	7	0.9	8	1.8	6	1.5
3333	17	1.2	6	0.7	6	0.9	7	1	5	0.7	9	0.6
Positive Control	186	9.6	56	7.9	52c	2.5	40	4.2	39	2.9	58	3.9

Strain TA97

Dose	No Activation  (Not a Valid Test)		No Activation  (Negative)		No Activation  (Negative)		10% HLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		30% RLI  (Negative)		30% RLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	82	9.7	103c	2.5	90	1.7	107	4.9	184c	4.5	104	4.1	226	10.4	155	5.8
10															158	7.3
33	80	5	93	0.3	77	8.5	107	3.8	209	13.2	104	4.7	197	10.2	162	4.3
100	73	5.8	107	15.5	77	7.8	115	9.7	206	13.5	95	4.9	221	13.5	154	12.8
333	79	4.4	85	8	66	6.8	89	5.7	209	5.8	99	4.3	192	5.9	159	3.5
1000	79	2.7	94	14.3	58	2	100	9.6	227	10	105	2.7	196	12.5	171	36.9
3333	90	3.8	109c	0	78	3.5	103	9.2	203	12.8	99	2.9	233	22.8
Positive Control	114	13.3	237	11.1	331	27.1	475	22	715	12.2	239	2	338	8.4	435	3.1

Strain 98

Dose	No Activation  (Equivocal)		No Activation  (Equivocal)		No Activation  (Weak Positive)		10% HLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		30% RLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	19	4.2	14	0.9	16	2.7	32	2	24	0.6	30	0.9	20	0.6
33	15	2.1	14	2.8			28	5.2	22	2.7	24	1.2	19	2.8
100	15	0.9	19	4	20	4.4	24	4	23	4.1	24	2.4	20	2
333	19	1.2	16	2.2	18	3.2	33	1.2	17	1.3	24	2.1	18	2.6
1000	19	1.3	17	2.6	17	2.6	32	5.5	24	3	30	3.4	19	3.5
3333	33	1.9	24	2.3			31	2.6	18	1.9	35	5.8	23	3.8
3334					27	0.6
6667					39	3.8
Positivecontrol	164	12.5	143	18.1	148	7.2	154	9.6	90	6.4	72	5.8	87	5.7

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):negative with and withoutThe test compound 2-aminobenzenesulphonic acid failed to induce gene mutation in the Salmonella typhimurium strains TA100,TA1535,TA98 and TA97 with and without addition of S9 liver fractions from Aroclor induced hamsters and hence is not likely to classify for gene mutation in vitro..

Executive summary:

Gene mutation toxicity study was performed for the test chemical 2-aminobenzenesulphonic acid to evaluate its mutagenic nature. Preincubation protocol was followed at dose levels of 0, 10, 33, 100, 333, 1000, 3333, 3334, 6667 µg/plate using Salmonella typhimurium strains TA100,TA1535,TA98 and TA97 with and without addition of S9 liver fractions from Aroclor induced hamsters. The test compound was tested in triplicate in the mutagenic study. The test compound 2-aminobenzenesulphonic acid failed to induce gene mutation in the Salmonella typhimurium strains TA100,TA1535,TA98 and TA97 with and without addition of S9 liver fractions from Aroclor induced hamsters and hence is not likely to classify for gene mutation in vitro..