3-chloro-o-toluidine

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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (5th strain is missing, which has an AT base pair at the primary reversion site. 2-Aminoanthracene is used as the sole indicator of the efficacy of the S9-mix.)

Data source

Reference

Reference Type:

secondary source

Title:

Unnamed

Year:

1996

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-chloro-o-toluidine

EC Number:

201-756-6

EC Name:

3-chloro-o-toluidine

Cas Number:

87-60-5

Molecular formula:

C7H8ClN

IUPAC Name:

3-chloro-2-methylaniline

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 97, TA 98, TA 100 and TA 1535

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats and syrian hamster treated with Aroclor 1254.

Test concentrations with justification for top dose:

0, 3, 10, 33, 100, 333, 666, 1000 µg/plate for strains TA 97 and TA 1535
0, 3, 10, 33, 100, 333, 666, 1000, 1666 µg/plate for strains TA 98 and TA 100

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-nitrofluorene or 4-nitro-o-phenylenediamine (TA 98); sodium azide (TA 100 and TA 1535); 9-aminoacridine (TA 97)

Remarks:

without S9 mix

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (all strains)

Remarks:

with S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes at 37°C
- Exposure duration: 48 hours

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 97, TA 98, TA 100 and TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

in the highest concentrations tested (see table)

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

TA 100

Dose	No Activation		No Activation		10% HLI		30% HLI		10% RLI		30% RLI
	(Negative)		(Negative)		(Negative)		(Negative)		(Negative)		(Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	128	3.3	118	7.2	123	8	109	3	127	6.1	111	3.7
3			119	4
10	113	8.3	116	2.4	118	10.8			109	3.2
33	115	7.5	112	2.7	116	11.8	106	3.5	122	4.8	113	4.6
100	96	6.5	110	3.8	130	6.4	107	4.2	109	14.6	108	4.3
333	116	3.3	94	5	133	12.7	108	5.5	131	9.5	103	5.8
666	0x	0			100	1.2			113	3
1000							104	4.2			106	1.5
1666							t				t
Positive Control	837	14.4	945	32.2	926	38.5	559	21.4	528	7.5	437	10.6

 

TA 1535

Dose	No Activation		No Activation		10% HLI		30% HLI		10% RLI		30% RLI
	(Negative)		(Negative)		(Negative)		(Negative)		(Negative)		(Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	11	1.5	9	1.7	13	1,5	10	1.9	16	2.3	11	1
3	8	1.5	11	0.3
10	8	0.9	8	1.5	15	2.3	11	1	14	1.5	9	0.7
33	7	0.6	10	1.5	12	1.2	9	1.2	12	1.5	9	0.9
100	8	1.5	10	0.7	11	1.8	10	1.9	10	0.7	7	1
333	7	1	8	0.7	15	2.3	10	2.1	11	1.9	7	0.3
666					6	0.9			4	0.6
1000							7	0.9			4s	0.6
Positive Control	877	21.1	907	46.4	149	6.7	259	19.7	124	9.5	138	10.1

 

TA 97

Dose	No Activation		No Activation		10% HLI		30% HLI		10% RLI		30% RLI
	(Negative)		(Negative)		(Negative)		(Negative)		(Negative)		(Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	160	9.9	135	6.1	132	13.9	170	7.2	157	11	186	4.6
3	168	9.7	127	7
10	170	8.8	139	0.9	137	9.2	176	7	135	11	181	4.7
33	176	9.9	143	7.3	156	15.9	180	8.1	159	9.9	187	6
100	183	6.2	120	9.8	151	5	174	11.5	158	14.7	179	8.3
333	188	7	98	0.9	140	12.4	181	9	178	5.5	176	10.4
666					118	5.5			108	1,2
1000							179	4.7			56s	8.9
Positive Control	581	18	583	58.3	1008	27.5	508	6.1	647	20,7	447	15.7

 

TA 98

Dose	No Activation		No Activation		10% HLI		30% HLI		10% RLI		30% RLI
	(Negative)		(Negative)		(Negative)		(Negative)		(Negative)		(Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/Plate	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM	Mean	± SEM
0	17	2	22	3.5	21	2.9	21	2.3	24	3.2	18	2.8
3			24	3.2
10	18	0.3	26	1.2	16	1.9			26	3.3
33	20	2.9	20	0.7	18	1.7	22	3.3	22	1	18	2.7
100	18	1.5	24	3.5	24	3.2	20	2.9	27	1.7	22	2.1
333	18	2.1	25	5.5	22	3.2	19	2.6	24	2.1	16	1.5
666	x				18	4.3			22	6.2
1000							17	2.2			15	1.2
1666							t				t
Positive Control	336	11	387	10	760	26.9	534	13.2	438	25.5	514	6.7

 

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; c = Contamination

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative