3-chloroaniline

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

3-chloroaniline was found to be non-genotoxic.

Link to relevant study records

Reference

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)

Qualifier:

according to guideline

Guideline:

other:

Principles of method if other than guideline:

In the standard protocol (preincubation) for conducting the Ames assay, a test tube containing a suspension of one strain of Salmonella typhimurium (or E. coli) plus S9 mix or plain buffer without S9, is incubated for 20 minutes at 37º C with the test chemical. Control cultures, with all the same ingredients except the test chemical, are also incubated. In addition, positive control cultures are prepared; these contain the particular bacterial tester strain under investigation, the various culture ingredients, and a known potent mutagen*. After 20 minutes, agar is added to the cultures and the contents of the tubes are thoroughly mixed and poured onto the surface of Petri dishes containing standard bacterial culture medium. The plates are incubated, and bacterial colonies that do not require an excess of supplemental histidine or tryptophan appear and grow. These colonies are comprised of bacteria that have undergone reverse mutation to restore function of the histidine- or tryptophan- manufacturing gene. The number of colonies is usually counted after 2 days.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

S. typhimurium TA 1535

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

S. typhimurium TA 1537

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

S. typhimurium TA 97

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

10% & 30% RLI = induced male Sprague Dawley rat liver S9 and 10% & 30% HLI = induced male Syrian hamster liver S9

Test concentrations with justification for top dose:

33 - 2000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide (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: 2-aminoanthracene

Remarks:

For strains tested with S9

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

For strains TA100 and TA1535 tested in the absence of S9

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

Remarks:

For strains TA97 and TA1537 tested in the absence of S9

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine

Remarks:

For strain TA98 tested in the absence of S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Strain: TA100

Dose	No Activation  (Negative)		No Activation  (Negative)		30% RLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		10% HLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	118	11.9	98	4.4	120	5.5	121	2.4	112	2.2	94	4.2
33	126	8.8	94	2.3	139	5.2	110	3.5	104	7.2	101	3.8
100	123	8.4	95	3.8	139	10	120	9.5	101	2.3	103	9.4
333	125	9.4	98	6.7	127	3.2	130	4.8	107	2.3	97	7.1
1000	111	4.8	61	7.9	116	7.2	122	9	103	3.8	89	3.3
1500			48	21.2							12	10
2000	64	3.5			74	9.7	100	9
Positive Control	476	8	310	6	560	58	728	9	570	13.7	409	10.7

Strain: TA1535

Dose	No Activation  (Negative)		No Activation  (Negative)		30% RLI  (Negative)		30% HLI  (Negative)		10% RLI  (Negative)		10% HLI  (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	26	4.2	20	3	13	.9	13	1.9	10	1	10	0
33	18	1.9	15	3.2	9	2	11	1.7	10	1.8	12	1.5
100	29	2.5	16	.9	13	1.8	9	1.5	10	1.8	8	2
333	27	3.2	14	2	8	.9	9	.7	8	.7	13	2.3
1000	23	4.7	12	1	7	1.7	13	.9	8	1.8	10	1.2
1500			15	5							4	1
2000							12	2.5
Positive Control	291	5.8	220	12	143	.3	228	53.1	145	9.8	59	2.8

Conclusions:

Interpretation of results (migrated information):
negative

In an Ames test , 3-chloroaniline, in dimethyl sulfoxide from doses 33 - 2000 µg/plate was not mutagenic in Salmonella typhimurium strains TA100 , TA 1535, TA1537 , TA 97 and TA 98 with and without addition of S9 liver microsome fractions from Aroclor induced rats.The same has been obtained for hamsters as well.

Executive summary:

In an Ames test , 3-chloroaniline, in dimethyl sulfoxide from doses 33 - 2000 µg/plate was not mutagenic in Salmonella typhimurium strains TA100 , TA 1535, TA1537 , TA 97 and TA 98 with and without addition of S9 liver microsome fractions from Aroclor induced rats.The same has been obtained for hamsters as well.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Various studies were reviewed for genetic toxicity from reliable sources having Klimish rating 2 and 4 for the target and the read across substance for CAS: 108-42-9 considering the weight of evidence approach.

The summary of the results are presented below

Sr.No	Endpoint	Interpretation of Results	Species/Strain	Conclusion	Sources
1	In vitro Genetic toxicity	Negative with and without metabolic activation	S. typhimurium TA 100, TA 1535, TA 1537,TA 97 and TA 98	In an Ames test , 3-chloroaniline, in dimethyl sulfoxide from doses 33 - 2000 µg/plate was not mutagenic in Salmonella typhimurium strains TA100 , TA 1535, TA1537 , TA 97 and TA 98 with and without addition of S9 liver microsome fractions from Aroclor induced rats.The same has been obtained for hamsters as well.	Publication data for CAS : 108-42-9
2	In vitro Genetic toxicity	Negative with and without metabolic activation	S. typhimurium TA 100, TA 1535, TA 1537 and TA 98	In an Ames test , 3-chloroaniline, in dimethyl sulfoxide from doses 1 - 3333 µg/plate was not mutagenic in Salmonella typhimurium strains TA100 , TA 1535, TA1537 and TA 98 with and without addition of S9 liver microsome fractions from Aroclor induced rats.The same has been obtained for hamsters as well.	Publication data for CAS : 108-42-9
3	In vitro Genetic toxicity	Negative with and without metabolic activation	Salmonella typhimurium strains TA92, TA94, TA98, TA100, TA1535, TA1537	In bacterial gene mutation assay on Salmonella typhimurium strains TA92, TA94, TA98, TA100, TA1535, TA1537 with and without metabolic activation it was found that 3 -Chloroaniline does not exhibit positive gene mutation effect.	Publication data for CAS : 108-42-9
4	Chromosome aberration	Negative without metabolic activation	Chinese hamster lung (CHL) cells	Based on the prediction for in-vitro mammalian chromosome aberration test on Chinese hamster Lung (CHL) without S9 metabolic activation it was estimated that 3-chloroaniline does not exhibit positive chromosomal abberation effect.	Predicted data for CAS : 108-42-9
5	Chromosome aberration	Negative without metabolic activation	CHO-LB CELLS	In mammalian chromosome aberration test , 4-chlorobenzene-1,3-diamine, in dimethyl sulfoxide from doses 148; 494; 14800 µg/ml was not mutagenic in CHO-LB CELLS with addition of S9 liver microsome fractions from Aroclor induced rats.	Publication data for RA CAS : 5131-60-2

 

Based on the above results it can be concluded that substance CAS: 108-42-9 is expected to show the similar toxicological effect based on the effects observed on the other category members. Thus it is concluded that the substance is non-genotoxic. Thus 3-chloroaniline is considered to be non-mutagenic as the per the CLP regulation.

Justification for selection of genetic toxicity endpoint

In an Ames test , 3-chloroaniline, in dimethyl sulfoxide from doses 33 - 2000 µg/plate was not mutagenic in Salmonella typhimurium strains TA100 , TA 1535, TA1537 , TA 97 and TA 98 with and without addition of S9 liver microsome fractions from Aroclor induced rats.The same has been obtained for hamsters as well.

Justification for classification or non-classification

3-chloroaniline is considered to be non-mutagenic as the per the CLP regulation.