2-amino-4-nitrophenol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames assay

Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA100, TA1535, TA98 and TA1537 by AMES test. The test result was considered to be positive in all strain in the presence and absence of metabolic activation S9.

In vitro chromosomal abbreviation study

Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.

In vitro Mammalian cell gene mutation assay

Test chemical was evaluated for its mutagenic potential in mouse lymphoma by in vitro mammalian cell gene mutation. The test result was considered to be positive in Mouse lymphoma.

In vitro DNA damage and/or repair study

Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells in vitro sister chromatid exchange assay. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.

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Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in Salmonella Typhimurium TA98,TA 100,TA 1535 and TA 1537 by AMES test.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA98,TA 100,TA 1535 and TA 1537

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

RLI = induced male Sprague Dawley rat liver S9 HLI = induced male Syrian hamster liver S9

Test concentrations with justification for top dose:

0,10,30,100, 333,1000 and 3333µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: 95% Ethanol
- Justification for choice of solvent/vehicle: The test substance is soluble in 95% Ethanol.

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

95% Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: -S9 mix;4 nitro-O phenylenediamine (TA 98), Sodium azide (TA1535and TA 100) and 9-Aminoacridine(TA1537) +S9 mix; 2-Aminoanthracene(all strains)

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: Pre incubation method
NUMBER OF REPLICATIONS: Duplicate

Evaluation criteria:

The plates were observed for revertents mutants colonies per plates

Statistics:

Yes, SD ± Mean was observed.

Key result

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: Mutagenic effect were observed

Strain: TA100
Dose	No Activation  (Negative)		No Activation  (Negative)		10% RLI  (Negative)		10% RLI  (Weakly Positive)		10% HLI  (Positive)		10% HLI  (Weakly Positive)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	139	7.8	107	4	142	4.3	112	7.9	129	6.9	163	8.8
10					121	5.4	146	3.2
33	135	9.5	114	4.7	133	4.9	162	7.7	164	10.3	156	8.4
100	145	8.4	117	11.8	132	14.2	169	10.2	210	8.7	223	11
333	142	6.2	101	7.4	134	10	191	6.4	268	2.9	247	9
1000	146	10.3	97	11.6	122	2.7	180	3.5	289	17.3	282	5.9
3333	86 s	49.7	t						0 s	0	0 s	0
Positive Control	434	3.7	407	18.7	690	15.4	551	3.5	1290	45.3	1796	23.7

 

Strain: TA1535
Dose	No Activation  (Negative)		10% RLI  (Negative)		10% HLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	27	.9	46	3.8	42	4.5
33	29	3.7	28	1.7	41	2.1
100	25	3.5	21	1.8	36	.3
333	27	3	16	1.2	37	2.2
1000	28	5.2	0 s	0	32	3.5
3333	9 s	3.1	t		0 s	0
Positive Control	427	7.1	277	7.1	536	12.8

 

Strain: TA1537
Dose	No Activation  (Negative)		10% RLI  (Negative)		10% HLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	5	.9	10	1	5	.6
33	6	1	7	1.2	12	2.8
100	6	1.7	7	.9	12	4
333	6	.3	11	1.2	15	1.3
1000	10	2	12	2.3	10	2
3333	5	.3	9	1.7	0 s	0
Positive Control	172	4.7	167	14.8	500	5.5

 

Strain: TA98
Dose	No Activation  (Negative)		No Activation  (Positive)		10% RLI  (Negative)		10% RLI  (Positive)		10% HLI  (Positive)		10% HLI  (Positive)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	21	2	29	2.2	40	1.7	37	3.5	37	3.7	37	4.3
10					27	1.5	44	1.3
33	26	3.1	52	7.2	38	4.2	53	.7	50	3	54	3.2
100	30	2.7	64	4.6	50	2.3	57	1.9	67	3.6	62	3.7
333	17	2.3	64	6.6	55	3.5	75	6.8	94	4.7	86	.3
1000	17	1	84	7.1	56	4.7	83	1.2	242	19.6	113	13.5
3333	t		0 s	0					t		0 s	0
Positive Control	724	34	724	7.1	425	15.3	382	27	1514	107.5	1027	53.1

 

Abbreviations:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9
s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; t = Toxic;

Conclusions:

Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA100, TA1535, TA98 and TA1537 by AMES test. The test result was considered to be positive in all strain in the presence and absence of metabolic activation S9.

Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test.

GLP compliance:

not specified

Type of assay:

other: in vitro mammalian chromosome aberration test

Target gene:

Not specified

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

induced male Sprague Dawley rat liver S9

Test concentrations with justification for top dose:

-S9;0,150,199,249and 300 µg/mL
+S9; 0,2460,2760,2990 and 3520 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide
- Justification for choice of solvent/vehicle: The test substance is soluble in DMSO.

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: -S9 mix; Mitomycin-C +S9 mix; Cyclophosphamide

Details on test system and experimental conditions:

METHOD OF APPLICATION: In medium
DURATION
- Fixation time (start of exposure up to fixation or harvest of cells): 20.8 hours
NUMBER OF CELLS EVALUATED: 100 cells

Evaluation criteria:

The mammalian cells were observed for chromosome aberration, Chromosome gaps and breaks.

Statistics:

Yes, SD ± Mean was observed.

Key result

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: Mutagenic effects were observed

Dose µg/mL	Total Cells Examined	Total Aberrations			Complex Aberrations			Simple Aberrations			Other Abs
		No.of Abs	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs
Vehicle Control:	Negative (Not Specified)	0	100	2	0.020	2.0	2	0.020	2.0	0	0.000	0.0	0	0.000	0.0
	Dimethyl Sulfoxide	0	100	2	0.020	2.0	2	0.020	2.0	0	0.000	0.0	0	0.000	0.0
Test Chemical:	2-Amino-4-nitrophenol	150	100	12	0.120	3.0	0	0.000	0.0	1	0.010	1.0	11	0.110	2.0
		199	100	7	0.070	6.0	4	0.040	4.0	2	0.020	2.0	1	0.010	1.0
		249	100	12	0.120	11.0	6	0.060	6.0	3	0.030	3.0	3	0.030	3.0
		300	0	0	0.000	0.0	0	0.000	0.0	0	0.000	0.0	0	0.000	0.0
Positive Control:	Mitomycin-C	0.0625	50	18	0.360	26.0	16	0.320	24.0	2	0.040	4.0	0	0.000	0.0
Trend:				2.934			2.134			1.838
Probability:				0.002			0.016			0.033

Trial #:1_S9   Activation: Induced Rat Liver S9    Date: 03/10/1983    Harvest Time: 20.8 hour(s)   Trial Call: Positive
		Dose µg/mL	Total Cells Examined	Total Aberrations			Complex Aberrations			Simple Aberrations			Other Abs
				No.of Abs	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs	No.of Abs.	Abs Per Cell	% Cells With Abs
Vehicle Control:	Negative (Not Specified)	0	100	2	0.020	2.0	1	0.010	1.0	0	0.000	0.0	1	0.010	1.0
	Dimethyl Sulfoxide	0	100	1	0.010	1.0	0	0.000	0.0	0	0.000	0.0	1	0.010	1.0
Test Chemical:	2-Amino-4-nitrophenol	2460	100	14	0.140	5.0	1	0.010	1.0	1	0.010	1.0	12	0.120	3.0
		2760	100	103	1.030	28.0	35	0.350	19.0	15	0.150	11.0	53	0.530	8.0
		2990	50	74	1.480	34.0	16	0.320	18.0	6	0.120	10.0	52	1.040	14.0
		3520	0	0	0.000	0.0	0	0.000	0.0	0	0.000	0.0	0	0.000	0.0
Positive Control:	Cyclophosphamide	10	50	10	0.200	18.0	6	0.120	12.0	4	0.080	8.0	0	0.000	0.0
Trend:				6.835			5.479			3.964
Probability:				0.000			0.000			0.000

 

Conclusions:

Test chemical was evaluated for its mutagenic potential in Chinese hamster ovary cells by in vitro mammalian chromosome aberration test. The test result was considered to be mutagenic both in the presence and absence of metabolic activation.

Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian chromosome aberration test was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,150,199,249and 300 µg/mL

+S9; 0,2460,2760,2990 and 3520 µg/mL

Chromosome aberration, Chromosome gaps and breaks were observed in the presence or absence of metabolic activation. Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro mammalian chromosome aberration test. Hence the substance cannot be classified as mutagenic in vitro.

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in mouse lymphoma by in vitro mammalian cell gene mutation.

GLP compliance:

not specified

Type of assay:

other: in vitro mammalian cell gene mutation test

Target gene:

Not specified

Species / strain / cell type:

other: mouse lymphoma cells

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

not specified

Test concentrations with justification for top dose:

Experiment I; 0,25,50,100,150,200 and 300 µg/mL
Experiment II: 50,100,150,200 and 300 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide
- Justification for choice of solvent/vehicle: The test substance is soluble in DMSO.

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Methyl Methane Sulfonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: In culture dish
DETERMINATION OF CYTOTOXICITY; relative total growth

Evaluation criteria:

The mammalian cells were observed for mutagenic frequency in cells.

Statistics:

Yes, SD ± Mean was observed.

Key result

Species / strain:

other: mouse lymphoma cells

Metabolic activation:

not specified

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: Mutagenic effects were observed

Nonactivation Trial: 1 Experiment Call: Positive
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	111	101	122	37	43
			90	103	144	53
			75	101	94	42
			110	94	126	38
Test Chemical	2-Amino-4-nitrophenol	25	70	53	198	94	98*
			61	53	209	114
			52	62	134	85
		50	72	53	233	109	108*
			64	54	193	100
			61	50	211	115
		100	65	27	349	179	194*
			54	20	320	198
			48	20	296	206
		150	63	4	501	266	271*
			47	4	422	299
			50	10	370	248
		200					273*
			46	4	366	267
			52	3	433	278
		300
Positive Control	Methyl Methane Sulfonate	5	65	60	508	260	258*
			77	26	598	258
			65	60	503	257

 

Nonactivation Trial: 2 Experiment Call: Positive
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	93	90	98	35	29
			121r	126	92	25
			109	117	84	26
			111	93	89	27
Test Chemical	2-Amino-4-nitrophenol	50	69	43	141	69	71*
			77	42	177	77
			93	51	192	69
		75	114	57	251	74	73*
			79	44	165	70
			73	40	164	75
		100	92	24	206	75	82*
			89	36	243	91
			87	32	208	80
		150	93	17	365	131	118*
			86	20	280	108
			67	20	230	115
		200	86	18	287	111	133*
			68	6	315	156
			91	10	360	133
		300	100	8	393	131	143*
			67	5	295	148
			76	5	344	152
Positive Control	Methyl Methane Sulfonate	5	57	46	605	357	335*
			58	34	670	382
			75	62	592	265

 

Footnotes: 
Asterisks(*) indicate significant responses.
r = rejected value due to quality control criteria 
# = reduced sample size because of the loss of one culture dish due to contamination 

Conclusions:

Test chemical was evaluated for its mutagenic potential in mouse lymphoma by in vitro mammalian cell gene mutation. The test result was considered to be positive in Mouse lymphoma.

Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian cell gene mutation was performed .The test material was exposed to mouse lymphoma cells.The concentration of test material used in two experiment were mention below

Experiment I; 0,25,50,100,150,200 and 300 µg/mL

 Experiment II: 50,100,150,200 and 300 µg/mL

 

Mutagenic frequency were observed in the mammalian cells. Therefore test chemical was considered to bemouse lymphoma byin vitro mammalian cell gene mutation. Hence the substance can be classified as mutagenic in vitro.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

Test substance was evaluated for its mutagenic potential in male Mouse and rat by In vivo Mammalian Somatic cell study. The test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes.

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in B6C3F1 male Mouse by In vivo Mammalian Somatic cell study.

GLP compliance:

not specified

Type of assay:

other: In vivo Mammalian Somatic cell study.

Species:

mouse

Strain:

B6C3F1

Sex:

male

Details on test animals or test system and environmental conditions:

Not specified

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: Corn oil

Duration of treatment / exposure:

72 hour

Frequency of treatment:

3 times in 72 hours

Remarks:

0,4.687,9.375,18.75,37.5,75,150 and 300 mg/Kg

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

yes, concurrent vehicle

Positive control(s):

Positive controls; Cyclophosphamide
- Route of administration: Intraperitoneal Injection
- Doses / concentrations:25 mg/kg

Tissues and cell types examined:

Bone Marrow cells were examined.

Evaluation criteria:

An increase in the frequency of micronucleated polychromatic erythrocytes in treated animals is an indication of induced chromosome damage.

Statistics:

Yes SD± Mean was observed.

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: No mutagenic effect were observed

Vehicle Control	Corn Oil	0	5	0.700 ± 0.250
Test Chemical	2-Amino-4-nitrophenol	4.687	5	0.900 ± 0.290	0.309
		9.375	4	1.250 ± 0.320	0.116
		18.75	5	2.100 ± 0.480	0.004
		37.5	5	1.700 ± 0.250	0.021
		75	4	1.750 ± 0.520	0.020
		150	5	1.100 ± 0.240	0.173
		300	2	1.000 ± 0.500	0.284
Positive Control	Cyclophosphamide	25	5	16.200 ± 0.800	Value less than 0.0001

 

 

		Dose (mg/kg)	Number of Animals Scored	Mean Percent PCE ± SEM	Pairwise P
Vehicle Control	Corn Oil	0	5	41.170 ± 2.800
Test Chemical	2-Amino-4-nitrophenol	4.687	5	40.130 ± 2.570
		9.375	4	44.000 ± 1.640
		18.75	5	42.830 ± 3.470
		37.5	5	42.120 ± 2.510
		75	4	44.330 ± 2.950
		150	5	40.200 ± 3.900
		300	2	32.700 ± 8.500
Positive Control	Cyclophosphamide	25	5	43.200 ± 2.660

 

		Dose (mg/kg)	Number of Animals Scored	Mean MN-PCE/1000 PCE ± SEM	Pairwise P
Vehicle Control	Corn Oil	0	5	1.100 ± 0.190
Test Chemical	2-Amino-4-nitrophenol	18.75	5	1.300 ± 0.250	0.342
		37.5	5	1.600 ± 0.190	0.168
		75	5	1.100 ± 0.240	0.500
		150	5	0.700 ± 0.250	0.827
Positive Control	Cyclophosphamide	25	5	13.100 ± 0.830	Value less than 0.0001

 

 

		Dose (mg/kg)	Number of Animals Scored	Mean Percent PCE ± SEM	Pairwise P
Vehicle Control	Corn Oil	0	5	46.200 ± 0.000
Test Chemical	2-Amino-4-nitrophenol	18.75	5	54.840 ± 1.200
		37.5	5	44.600 ± 0.510
		75	5	58.640 ± 1.920
		150	5	57.320 ± 1.330
Positive Control	Cyclophosphamide	25	5	46.960 ± 1.900

Conclusions:

Test substance was evaluated for its mutagenic potential in B6C3F1 male Mouse by In vivo Mammalian Somatic cell study. The test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes.

Executive summary:

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in B6C3F1 male Mouse. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 4.687, 9.375, 18.75, 37.5,75,150 and 300 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Data for the various publication was reviewed to determine the mutagenic nature of 2-amino-4-nitrophenol (99-57-0). The studies are as mentioned below:

In Vitro studies

Ames assay

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

Supported by other AMES test. Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0, 10, 30,100, 333, 1000 and 3333 µg/plate. Mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance can be classified as gene mutant in vitro.

 

In vitro chromosomal abbreviation study

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian chromosome aberration test was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,150,199,249and 300 µg/mL

+S9; 0,2460,2760,2990 and 3520 µg/mL

Chromosome aberration, Chromosome gaps and breaks were observed in the presence or absence of metabolic activation. Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro mammalian chromosome aberration test. Hence the substance cannot be classified as mutagenic in vitro.

In vitro Mammalian cell gene mutation assay

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro mammalian cell gene mutation was performed .The test material was exposed to mouse lymphoma cells.The concentration of test material used in two experiment were mention below

Experiment I; 0,25,50,100,150,200 and 300 µg/mL

 Experiment II: 50,100,150,200 and 300 µg/mL

 

Mutagenic frequency were observed in the mammalian cells. Therefore test chemical was considered to be mutagenic in mouse lymphoma by in vitro mammalian cell gene mutation. Hence the substance can be classified as mutagenic in vitro.

In vitro DNA damage and/or repair study

Genetic toxicity in vitro study was assessed for test chemical. For this purpose in vitro sister chromatid exchange assay was performed .The test material was exposed toChinese hamster ovary cells inthe presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were mention below

-S9;0,5,16.7 and 50 µg/mL

+S9; 0,1740,2180 and 2670 µg/mL

 

The mammalian cells were observed fornumber of scored exchanges correlates to the number of DNA breakage and reunion eventsin the presence or absence of metabolic activation. The test article induced sister chromatid exchange in the cultured cells.  Therefore test chemical was considered to be mutagenic inChinese hamster ovary cells byin vitro sister chromatid exchange assay. Hence the substance cannot be classified as mutagenic in vitro.

In Vivo studies

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in B6C3F1 male Mouse. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 4.687, 9.375, 18.75, 37.5,75,150 and 300 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

In the study test substance was assessed for its possible mutagenic potential. For this purpose wasIn vivo Mammalian Somatic cell study in Fischer 344 male rat. The test substance was administrated by Intraperitoneal Injection by using corn oil as vehicle. The test substance was exposed at the concentration of 0, 3.91, 7.815, 15.63, 31.25, 62.5 and 125 mg/Kg for 72 hours. The dose was administrated thrice in 72 hours .The bone marrow cells were stained and observed for chromosome damage. No significant increase in the frequency of micronucleated polychromatic erythrocytes in treated animals was observed. Therefore test result was consider to be negative as there was no significant chromosome damage in micronucleated polychromatic erythrocytes. Hence the substance cannot be classified as mutant In Vivo.

 

 

 

Based on the data summarized, 2-amino-4-nitrophenol (99-57-0)did  not induce gene mutation in vivo while it induce gene mutation in vitro.The test chemical induce mutation in AMES test as well as in OECD 473 test (In vitro chromosomal abbreviation study) . The test chemical induce gene mutation in In vitro Mammalian cell gene mutation assay too. But as the test chemical did not induce gene mutatiomn in IN vivo study conducted in male rat and male mouse.As per CLP criteria

The classification in Category 2 is based on:

— positive evidence obtained from experiments in mammals and/or in some cases from in vitro experiments, obtained from:

— somatic cell mutagenicity tests in vivo, in mammals; or

— other in vivo somatic cell genotoxicity tests which are supported by positive results from in vitro mutagenicity assays.

Hence based on CLP criteria the test substance can not be classified as mutagenic.

Justification for classification or non-classification

Based on the data summarized and CLP criteria , 2-amino-4-nitrophenol (99-57-0) not likly induce gene mutation in vivo while it induce gene mutation in vitro.The test chemical induce mutation in AMES test as well as in OECD 473 test (In vitro chromosomal abbreviation study) .

The test chemical induce gene mutation in In vitro Mammalian cell gene mutation assay too.

But as the test chemical did not induce gene mutatiomn in In vivo study conducted in male rat and male mouse.As per CLP criteria

The classification in Category 2 is based on:

— positive evidence obtained from experiments in mammals and/or in some cases from in vitro experiments, obtained from:

— somatic cell mutagenicity tests in vivo, in mammals; or

— other in vivo somatic cell genotoxicity tests which are supported by positive results from in vitro mutagenicity assays.

As no evidence of somatic cell mutagenicity tests in vivo, in mammalswas observed

Hence based on CLP criteria the test substance not likly to be be classified as mutagen.