m-phenylenediamine

Genetic toxicity: in vitro

Administrative data

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

No reference to OECD guideline. Deviation from OECD guideline with the use of 4-o-Tolylazo-o-toluidine as positive control. Study conducted in 1975.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Only 4 strains of bacteria were used. Strain TA1538 was used and Strain TA98 was not. Positive controls deviate from those recommended in guideline.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

m-phenylenediamine

EC Number:

203-584-7

EC Name:

m-phenylenediamine

Cas Number:

108-45-2

Molecular formula:

C6H8N2

IUPAC Name:

m-phenylenediamine

Details on test material:

-Purity: not reported
-Name of test material (as cited in study report): m-phenylenediamine

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1537

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbitol-stimulated rat liver homogenate

Test concentrations with justification for top dose:

1, 10, 50, 100, 250, 500, 750, 1000 micrograms/plate.

Vehicle / solvent:

no data

Controls

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 4-o-Tolylazo-o-toluidine (for TA1538 and metabolic activation) and N-methyl-N'-nitro-N-nitrosoguanidine

Details on test system and experimental conditions:

In the absence of metabolic activation, approximately 5E8 bacteria were added to 2 mL of top agar (0.6% Difco agar, 0.6% NaCl, 0.050 mM of L-histidine, 0.05 mM of biotin). After the test chemical was added to the top agar, the solution was mixed and poured onto the surface of a minimal agar plate with Vogel-Bonner E medium containing 1.5% agar and 2% glucose.

The metabolic activation system consisted of 0.15 mL of the 9000 x g supernatant of homogenized rat liver, MgCl2, KCl, glucose-6-phosphate, TPN, and a sodium phosphate buffer (pH 7.4). This mixture (0.25 mL) was added directly to the top agar immediately before it was poured over the minimal agar plate. The plates were incubated at 37 deg C for 40 hours. The number of histidine-positive (his+) revertant colonies on each plate then was counted. All tests were performed in duplicate.

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION: 40 hours

SELECTION AGENT (mutation assays): histidine

NUMBER OF CELLS EVALUATED: 5E8

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 5

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Any other information on results incl. tables

In Vitro Salmonella Typhimurium Assay With m-PDA.

 

Chemical	Metabolic Activation	Amount of Compound Added per Plate (µg)	Average Number of Histidine-Positive Revertants per Plate
			TA100	TA1535	TA1537	TA1538
Negative Control	-		72	11	5	9
	+		84	11	6	15
4-o-Tolylazo-o-toluidine (Positive control for TA1538 and metabolic activation)	-	25				10
	+	25				195
N-methyl-N’-nitro-N-nitrosoguanidine	-	2		600
m-phenylenediamine	-	1	65	9	4	4
	-	10	64	8	4	2
	-	50	78	9	5	9
	-	100	80	10	3	12
	-	250				9
	-	500	83	8	2	17
	-	750
	-	1000	74	11	5	21
	+	1	87	14	8	10
	+	10	78	11	7	17
	+	50	80	9	6	79
	+	100	85	14	6	177
	+	250				423
	+	500	84	14	11	570
	+	750				623
	+	1000	79	10	8	971

 

Applicant's summary and conclusion

Conclusions:

The test substance was mutagenic in strain TA 1538 with metabolic activation in the S. typhimurium assay, but negative when activation was not used. It was negative in all other strains, with or without activation.

Executive summary:

The test substance was mutagenic in strain TA 1538 with metabolic activation in the S. typhimurium assay, but negative when activation was not used. It was negative in all other strains, with or without activation.