2-methoxy-5-nitroanilinium chloride

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:

Mutagenicity evaluation of nitroanilines and nitroaminophenols in Salmonella typhimurium

Author:

Majdi M. Shahin

Year:

1985

Bibliographic source:

International Journal of Cosmetic Science 7, 277-289 (1985)

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:

Mutagenicity toxicity testing was performed to determine the mutagenic nature of 2-nitro-4-aminophenol

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-methoxy-5-nitroanilinium chloride

EC Number:

267-234-5

EC Name:

2-methoxy-5-nitroanilinium chloride

Cas Number:

67827-72-9

Molecular formula:

C7H9ClN2O3

IUPAC Name:

2-methoxy-5-nitroanilinium chloride

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material: 2-methoxy-5-nitroanilinium chloride
- IUPAC name: 2-methoxy-5-nitroanilinium chloride
- Molecular formula: C7H9ClN2O3
- Molecular weight: 204.6121g/mole
- Smiles: Cl.COc1ccc(cc1N)[N+](=O)[O-]
- Inchl: 1S/C7H8N2O3.ClH/c1-12-7-3-2-5(9(10)11)4-6(7)8;/h2-4H,8H2,1H3;1H
- Substance type: Organic
- Physical state: Solid powder

Specific details on test material used for the study:

- Name of test material: 2-nitro-4-aminophenol
- IUPAC name: 4-Amino-2-nitrophenol
- Molecular formula: C6H6N2O3
- Molecular weight: 154.1244 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: > 98%
- Impurities (identity and concentrations): No data

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA100, TA1537, TA1538 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:

S9 metabolic activation system was prepared from Aroclor 1254 induced male Wistar rats.

Test concentrations with justification for top dose:

0, 5, 10, 20, 50, 100, 250, 500 or 1000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 1,2-diamino-4-nitrobenzene and 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation )

DURATION
- Preincubation period: No data
- Exposure duration: 3 days
- Expression time (cells in growth medium): 3 days
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

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

NUMBER OF REPLICATIONS: Three plates were used per dose and the experiment was repeated twice

NUMBER OF CELLS EVALUATED: No data

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

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

OTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

A dose-dependent two-fold increase in numbers of revertants per plate was used as the criterion for genetic activity of the compounds tested. A 2.5-fold increase over the spontaneous level, even without clear evidence of dose-dependence, was also considered to be an indication of genetic activity if found in repeated experiments.

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98

Metabolic activation:

not specified

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

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutagenicity test of 2-nitro-4-aminophenol in Salmonella typhimurium in the presence (+) and absence (-) of an Aroclor-induced rat-liver microsomal metabolic activation system

Dose (µg/plate)	Revertant colonies/plate
	TA1535		TA100		TA1537		TA1538		TA98
	-	+	-	+	-	+	-	+	-	+
0	17.0	17.0	156.5	150.5	26.0	39.0	31.0	47.5	49.0	67.0
5	15.0	17.0	143.5	143.5	28.5	39.5	32.5	47.5	46.0	59.5
10	15.5	14.5	152.5	141.5	30.0	44.0	32.5	47.0	46.5	59.0
20	15.5	18.0	137.5	136.5	28.0	42.0	28.5	13.5	44.0	60.5
50	14.0	16.0	140.0	146.5	31.0	41.0	20.5	17.5	13.0	59.0
100	15.5	15.0	138.5	147.5	35.0	41.5	33.0	46.5	49.0	61.0
250	15.5	17.5	146.0	154.5	31.5	45.0	32.5	43.5	46.5	60.0
500	16.5	16.5	142.5	133.0	27.0	44.0	30.5	42.5	47.5	55.0
1000	14.0	14.5	151.5	132.5	17.0	30.5	26.5	41.0	39.0	44.0

 

Table: Summary of positive and negative controls for Salmonella typhimuium tester strains in the presence (+) and absence (-) of an Aroclor-1254-induced rat-liver microsomal metabolic activation system

	Dose (µg/plate)	Revertant colonies/plate
		TA1535		TA100		TA1537		TA1538		TA98
		-	+	-	+	-	+	-	+	-	+
Control, no additions	00	18.5	17.5	166.0	159.5	25.0	34.0	30.0	45.0	47.5	61.5
Control, DMSO	100µL	17.5	18.0	146.0	147.5	28.5	36.0	31.0	45.5	47.0	57.0
Control, 1,2-diamino-4-nitrobenzene	20µg	40.5	34.5	639.0	400.0	164.00	67.0	1990.0	1794.0	3018.5	2311.5
Control, 2-aminoanthracene	5µg	21.0	22.0	168.0	1649.0	23.5	258.0	29.5	1133.0	40.0	2050.0

 

Applicant's summary and conclusion

Conclusions:

2-nitro-4-aminophenol did not induce gene mutation in Salmonella typhimurium TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant.

Executive summary:

Mutagenicity toxicity testing was performed to determine the mutagenic nature of 2-nitro-4-aminophenol. The study was performed using Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels of 0, 5, 10, 20, 50, 100, 250, 500 or 1000 µg/plate. Plates were incubated for three days at 37°C before counting the revertant colonies. Concurrent solvent and negative control chemicals were also included in the study. A dose-dependent two-fold increase in numbers of revertants per plate was used as the criterion for genetic activity of the compounds tested. A 2.5-fold increase over the spontaneous level, even without clear evidence of dose-dependence, was also considered to be an indication of genetic activity if found in repeated experiments. 2-nitro-4-aminophenol did not induce gene mutation in Salmonella typhimurium TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant.