3-amino-5-hydroxynaphthalene-2,7-disulphonic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

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:

Errol Zeiger, Beth Anderson, Steve Haworth, Timothy Lawlor, and Kristien Mortelmans

Year:

1988

Bibliographic source:

Environmental and Molecular Mutagenesis Volume 11, Supplement 12: 1-158 (1988)

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

The test chemical 6-Amino-4-nitro-1-phenol-2-sulfonic acid was studied to for its ability to induce mutations in strains of Salmonella typhimurium.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-amino-2-hydroxy-5-nitrobenzenesulfonic acid

Cas Number:

96-67-3

Molecular formula:

C6H6N2O6S

IUPAC Name:

3-amino-2-hydroxy-5-nitrobenzenesulfonic acid

Details on test material:

- Name of test material (as cited in study report): 6-Amino-4-nitro-1-phenol-2-sulfonic acid
- Molecular formula (if other than submission substance): C6H6N2O6S
- Molecular weight (if other than submission substance): 234.187 g/mol
- Substance type: Organic

Specific details on test material used for the study:

- Name of test material: 6-Amino-4-nitro-1-phenol-2-sulfonic acid
- IUPAC name: 3-amino-2-hydroxy-5-nitrobenzenesulfonic acid
- Molecular formula: C6H6N2O6S
- Molecular weight: 234.187 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data available

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

10% and 30% HLI and RLI S-9 (9,000 g supernatant) fractions were prepared from Aroclor 1254-induced, male Sprague- Dawley rat and male Syrian hamster livers

Test concentrations with justification for top dose:

0, 100, 333, 1000, 3333, 5000 or 10000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: No data available

Controls

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

sodium azide

other: 4-nitro-o-phenylenediamine (TA98; -S9) and 2-aminoanthracene (all strains; +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- 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 available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available

Rationale for test conditions:

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 two fold 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 nonmutagenic (-) 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 nonmutagenicity.

Statistics:

Mean ± SD

Results and discussion

Test results

Species / strain:

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

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:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-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. 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: Yes

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutagenicity of 6-Amino-4-nitro-1-phenol-2-sulfonic acid

Dose (µg/plate)	TA100
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	96	7.1	86	5.8	103	3.1	93	2.9	122	7.3
100	82	3.5	84	8.1	96	7.3	106	2.2	113	6.4
333	77	3.6	89	4.5	97	10.3	84	3.8	120	3.3
1000	86	1.5	93	5.9	104	17.0	88	2.4	130	3.6
3333	84	2.7	99	5.6	89	3.0	85	1.9	98	6.7
5000	81	7.6	119	7.5	107	10.7	84	0.6	126	8.8
10000
Positive control	240	25.2	524	21.2	313	15.4	594	14.2	361	23.9

 

Dose (µg/plate)	TA1535
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	21	1.5	11	1.3	8	1.2	9	1.2	12	1.5
100	22	2.0	7	0.3	10	1.2	10	0.6	14	2.4
333	20	3.8	10	1.5	12	3.0	12	1.0	11	1.5
1000	15	1.8	10	3.9	10	3.5	12	1.9	10	2.4
3333	25	3.6	9	1.2	11	1.3	12	0.6	10	1.5
5000	15	1.5	5	1.3	6	1.2	8	2.6	10	2.4
10000
Positive control	127	6.1	76	2.8	83	3.8	184	7.9	92	1.2

 

Dose (µg/plate)	TA97
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	83	6.4	145	6.1	179	33.9	173	2.9	172	8.0
100	83	8.6	136	2.7	205	14.3	250	5.5	160	3.6
333	103	7.1	118	8.0	218	9.3	217	11.5	183	9.0
1000	95	12.4	134	2.4	216	39.1	154	16.0	191	0.3
3333	97	2.3	122	2.3	212	16.9	120	2.0	126	7.1
5000
10000			119	7.2
Positive control	284	12.5	755	5.8	648	10.9	868	25.0	423	10.8

 

Dose (µg/plate)	TA98
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	20	2.9	21	4.0	35	3.7	26	2.8	21	2.2
100	16	2.2	28	2.6	31	3.8	20	3.5	25	2.4
333	15	0.0	25	0.9	31	1.0	20	1.7	22	3.0
1000	16	2.8	23	1.0	26	1.8	27	1.7	20	1.7
3333	20	2.7	20	2.3	27	2.4	26	4.7	21	5.0
5000	17	1.2	29	4.1	21	2.0	21	2.4	30	1.5
10000
Positive control	187	11.2	202	13.9	86	8.0	115	1.5	99	6.6

Applicant's summary and conclusion

Conclusions:

6-Amino-4-nitro-1-phenol-2-sulfonic acid is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system.

Executive summary:

6-Amino-4-nitro-1-phenol-2-sulfonic acid was studied for its ability to induce mutations in strains of Salmonella typhimurium.

 

The test compound was dissolved in DMSO and was tested at concentration of 0, 100, 333, 1000, 3333, 5000 or 10000 µg/plate using Salmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of 10 % and 30 % rat and hamster liver S9 metabolic activation system. Preincubation assay was performed with a preicubation for 20 mins. The plates were observed for histidine independence after 2 days incubation period. Concurrent solvent and positive controls were included in the study.

 

6-Amino-4-nitro-1-phenol-2-sulfonic acidis not mutagenic to theSalmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system.