3(or 4)-methylbenzene-1,2-diamine

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: Pre-GLP study, fully reported, essentially to guideline. No E coli or TA102 strains used.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

Duplicate plates used. Four test concentrations, up to limit.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3(or 4)-methylbenzene-1,2-diamine

EC Number:

248-145-0

EC Name:

3(or 4)-methylbenzene-1,2-diamine

Cas Number:

26966-75-6

Molecular formula:

C7H10N2

IUPAC Name:

3(or 4)-methylbenzene-1,2-diamine

Details on test material:

3,4-TDA, CAS 496-72-0. 97% purity

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Rat S-9, induced with Aroclor 1254.

Test concentrations with justification for top dose:

1000, 1710, 2924, 5000,

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

other:

Remarks:

Positive control S9 varied according to tester strain and use of S9.

Details on test system and experimental conditions:

Plate incopporation method. Duplicate plates.

Statistics:

None

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

3.4-TDA was marginally active in strains TA98 and TA1538, although no dose-response relationship was evident.

3,4-TDA was more toxic to cells in the absence of S9, and was particularly toxic to TA98 and TA 1538 strains. All doses tested were toxic.

Any other information on results incl. tables

Bacterial Mutagenicity of 3,4-TDA

Chemical	Amount per plate (ug)	S-9^b	Revertants per plate^a
			TA98	TA100	TA1535	TA1537	TA1538
Negative control	-	-	13	54	54	6	8
Negative control	-	+	17	55	52	7	11
Positive control^c	-	-	558	237	78	122	1209
Positive control^d	-	+	1178	242	0	379	1156
3,4-TDA	1000	-	11	60	46	4	6
3,4-TDA	1710	-	20	66	51	8	6
3,4-TDA	2924	-	7	42	15	7	6
3,4-TDA	5000	-	4	14	1	4	4
3,4-TDA	1000	+	48	68	70	12	56
3,4-TDA	1710	+	54	64	58	13	56
3,4-TDA	2924	+	59	62	62	9	48
3,4-TDA	5000	+	48	55	58	5	26

^a Average of two plates

^b Aroclor 1254 induced Fisher 344 rat liver.

^c The following positive controls were used without activating medium: dinitrotoluene (500 ug, TA1535 and TA100), quinacrine mustard (10ug, TA1537) and nitrofluorene (100 ug, TA1538 and TA98)

^d The following positive controls were used with activating medium, anthramine (100 ug, TA1535 and TA100), aminoquinoline (100 ug, TA1537) and acetamidofluorene (500 ug, TA1538 and TA98)

Toxicity of TDA Isomers for S. Typhimurium

Chemical	Amount per plate (ug)	S-9^b	Relative Survival^a
			TA98	TA100	TA1535	TA1537	TA1538
3,4-TDA	1000	-	0.107	0.463	0.803	0.794	0.341
3,4-TDA	1710	-	0.094	0.473	0.767	0.811	0.302
3,4-TDA	2924	-	0.068	0.471	0.598	0.712	0.185
3,4-TDA	5000	-	0.018	0.411	0.407	0.326	0.042
3,4-TDA	1000	+	0.224	0.471	0.901	0.872	0.320
3,4-TDA	1710	+	0.247	0.501	0.904	0.868	0.313
3,4-TDA	2924	+	0.240	0.422	0.816	0.861	0.309
3,4-TDA	5000	+	0.194	0.385	0.773	0.875	0.309

^a The amount of chemical indicated, + or - rat S-9, was added to a tube of top agar containing 0.1 ml of 1 10^-6 dilution of the relevant tester strain. The mixture was poured onto a nutrient agar plate, was allowed to harden, and the plates were incubated at 37°C for 24 hr.

Relative Survival = Number of colonies on plate + chemical

................................Number of colonies on control plate

^b Aroclor 1254 induced Fisher 344 rat liver.