4-hydroxy-6-(3-sulphoanilino)naphthalene-2-sulphonic 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:

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: V. Results from the Testing of 311 Chemicals

Author:

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

Year:

1992

Bibliographic source:

Environmental and Molecular Mutagenesis Volume 19, Supplement 21: 2-141 (1992)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Procedure as described by Haworth et al 1983 with slight modification

Principles of method if other than guideline:

Preincubation assay was performed to evaluate the mutagenic nature of the test compound 4-(2-Naphthylamino)phenol

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

p-(2-naphthylamino)phenol

EC Number:

202-248-7

EC Name:

p-(2-naphthylamino)phenol

Cas Number:

93-45-8

IUPAC Name:

4-(2-naphthylamino)phenol

Details on test material:

- Name of test material: 4-(2-Naphthylamino)phenol
- Molecular formula: C16H13NO
- Molecular weight: 235.285 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: appr. 88%
- Impurities (identity and concentrations): No data

Specific details on test material used for the study:

- Name of test material: 4-(2-Naphthylamino)phenol
- IUPAC name: 4-(2-naphthylamino)phenol
- Molecular formula: C16H13NO
- Molecular weight: 235.285 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: appr. 88%
- Impurities (identity and concentrations): 12%

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 100, TA1535, TA 1537, TA 97 and TA98

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

10% and 30% hamster liver S9 and 10% and 30% rat liver S9

Test concentrations with justification for top dose:

0, 0.3, 1, 3, 10, 33, 66, 100, 166, 333 or 666 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, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days
- 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: No data available

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:

The plates were observed for a dose dependent increase in the number of Histidine- independent (his+) colonies.

Evaluations were made at both the individual trial and chemical levels.

Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase in 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 a weak increase was not dose-related. The distinctions between a questionable response and a nonmutagenic or weakly 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 trial to be judged mutagenic.

A chemical was judged mutagenic (+) or weakly mutagenic (+W) if it produced a reproducible, dose-related response over the solvent control, under a single metabolic activation condition, 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.

Statistics:

Mean, SEM

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 100, TA1535, TA 1537, TA 97 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:

No data available

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Results for the test chemical 4-(2-Naphthylamino)phenol

Dose (µg/plate)	TA100
	NA		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	91	5.4	89	6.2	139	5.6	102	9.0	154	4.4
0.3	125	5.8
1	104	10.0
3	91	2.9	82	7.3			98	6.7
10	85	0.7	105	21.6	134	15.7	108	17.0	181	8.9
33	46s	15.5	134	3.5	159	13.9	97	6.1	156	5.1
66
100			113	7.4	124	14.7	90	5.1	149	7.9
166			83	3.0			94	6.8
333					96	3.5			73	2.5
666					21s	9.4			0s	0.0
Positive control	352	7.2	681	36.9	612	23.9	414	7.2	550	31.4

 

 

Dose (µg/plate)	TA1535
	NA		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	22	3.8	14	4.5	12	0.3	12	1.8	12	0.9
0.3	23	2.3
1.0	25	2.7
10	23	1.8	11	2.3	13	2.1	15	3.0	11	0.9
33	16	1.8	15	1.5	11	2.0	12	1.5	10	1.2
66
100			13	2.2	13	1.9	11	2.1	13	2.2
166			12	2.2			8	0.9
333					13	2.7			7	2.6
666
Positive control	301	18.6	269	22.6	400	31.3	165	9.2	122	17.9

 

Dose (µg/plate)	TA1537
	NA		10% HLI		30% HLI
	Mean	SEM	Mean	SEM	Mean	SEM
0	10	2.2	10	1.2	12	0.9
0.3	9	0.0
1	8	3.9
3	7	1.2	10	3.2	9	0.3
10	5	1.2	8	0.9	9	2.2
33	6	2.1	10	0.9	9	2.2
66
100			11	1.9	9	2.0
166
333			6	2.6	10	1.3
666
Positive control	397	27.1	46	4.0	61	0.6

 

Dose (µg/plate)	TA97
	NA		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	189	9.6	190	20.5	167	12.3	193	10.8	174	11.7
0.3	198	2.3
1	195	3.5
3	202	7.1	205	8.0	173	13.6	193	0.7	174	4.9
10	175	12.0	194	5.0	156	14.3	204	9.0	186	12.3
33	184	12.1	170	3.5	201	7.0	208	4.5	182	13.3
66
100			121	13.0	189	9.3	190	16.5	159	23.1
166			67s	7.4			79	6.6
333					12	2.6			14	2.3
666
Positive control	433	9.6	332	11.3	343	18.3	316	6.7	351	12.2

 

Dose (µg/plate)	TA98
	NA		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	14	2.6	27	2.2	23	3.0	34	3.1	24	1.3
0.3	17	3.5
1	18	3.8
3	16	2.5	37	3.4			28	1.5
10	14	0.6	39	0.3	31	5.5	21	2.7	24	1.2
33	13	1.5	31	2.5	25	3.0	15	2.3	22	1.5
66
100			22	1.7	21	1.7	19	2.5	17	1.0
166			21	3.5			18	1.5
333					12	1.8			12	1.3
666					9	0.9			6	1.2
Positive control	401	33.4	392	31.0	376	29.1	193	5.8	164	9.8

Applicant's summary and conclusion

Conclusions:

4-(2-Naphthylamino)phenol did not induce gene mutation in the Salmonella typhimurium tester strains TA 100, TA1535, TA 1537, TA 97 and TA98 in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.

Executive summary:

Preincubation assay was performed to evaluate the mutagenic nature of the 4-(2-Naphthylamino)phenol. The study was performed as described by Haworth et al 1983 with slight modification. The study involved the use of test chemical dose levels of 0, 0.3, 1, 3, 10, 33, 66, 100, 166, 333 or 666 µg/plate and Salmonella typhimurium strains TA 100, TA1535, TA 1537, TA 97 and TA98 in the presence and absence of S9 metabolic activation system. 4-(2-Naphthylamino)phenol did not induce gene mutation in the Salmonella typhimurium tester strains in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.