2,4,6-trinitrobenzene-1,3,5-triamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

purity of test substance, no. of cells per culture, no. of replications, vehicle, signs of toxicity, evaluation and interpretation of results, individual plate counts not reported

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1976

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

purity of test substance, no. of cells per culture, no. of replications, vehicle, signs of toxicity, evaluation and interpretation of results, individual plate counts not reported

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

pre-GLP

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,4,6-trinitrobenzene-1,3,5-triamine

EC Number:

221-297-5

EC Name:

2,4,6-trinitrobenzene-1,3,5-triamine

Cas Number:

3058-38-6

Molecular formula:

C6H6N6O6

IUPAC Name:

trinitrobenzene-1,3,5-triamine

Test material form:

not specified

Details on test material:

- Name of test material as cited in the report: TATB ; 1,3,5-triamino-2,4,6-trinitrobenzene
- Lot/batch No.of test material: B-318

Method

Target gene:

Histidine and tryptophan for S. typhimurium and E. Coli, respectively.

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Details on mammalian cell type (if applicable):

not applicable

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2

Details on mammalian cell type (if applicable):

not applicable

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-stimulated, rat liver-homogenate metabolic activation system was used

Test concentrations with justification for top dose:

5, 10, 50, 100, 500, 1000, 2500 and 5000 μg/plate, with and without S9-mix in S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 & TA 100 and E. coli WP2

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

other: 6-propiolactone (50 μg/plate for TA 1535); 2-Anthramine (20 μg/plate for TA 98, TA 100 and WP2)

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Anthramine

Remarks:

with metabolic activation

Details on test system and experimental conditions:

SOURCE OF TEST SYSTEM: S. typhimurium strains were obtained from Dr. Bruce Ames of the University of California at Berkeley. E. coli WP2 was obtained from Dr. D. McCalla.

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: Plates were incubated at 37 °C for 2 days.

Evaluation criteria:

None

Statistics:

None

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Any other information on results incl. tables

Table 7.6.1/1: Ames test - results

Test substance	Dose levels (µg/plate)	Mean no. Of revertants per plate
		Histidine revertants					Tryptophan revertants
		TA 1535	TA 1537	TA 1538	TA 98	TA 100	E. coli WP2
Without metabolic activation
Negative control	-	23	10	14	29	160	76
TATB	5	41	13	15	28	151	-
	10	30	7	12	32	126	81
	50	32	7	13	25	148	93
	100	35	8	16	21	148	78
	500	40	8	9	25	146	95
	1000	25	10	14	25	125	69
	2500	31	8	9	28	152	84
	5000	33	8	14	30	125	87
Positive control	*	680	>2000	310	40	160	72
With metabolic activation
Negative control	-	17	11	24	57	156	81
TATB	5	23	7	19	34	128	-
	10	20	9	28	25	151	91
	50	24	10	22	42	138	108
	100	22	7	20	39	132	87
	500	20	13	21	34	145	88
	1000	18	6	17	35	139	88
	2500	20	8	16	27	143	88
	5000	20	6	15	33	173	94
Positive control	**	-	-	-	>2000	>2000	1170

*Without metabolic activation - 9-Aminoacridine (100 μg/plate for TA 1537); 2-Nitrofluorene (50 μg/plate for TA 1538); 6-propiolactone (50 μg/plate for TA 1535); 2-Anthramine (20 μg/plate for TA 98, TA 100 and WP2)

**With metabolic activation - 2-Anthramine (20 μg/plate for TA 98, TA 100 and E. coli WP2)

Applicant's summary and conclusion

Conclusions:

Under the test conditions, test substance is not considered as mutagenic in S. typhimurium (TA1535, TA1537, TA 1538, TA98 and TA100) and E. coli WP2 strains.

Executive summary:

In a reverse gene mutation assay in bacteria, performed similarly to the OECD Guideline 471, strains of Salmonella typhimurium (TA1535, TA1537, TA 1538, TA98 and TA100) and Escherichia coli WP2 were exposed to test substance at the following concentrations using the plate incorporation method: 

5, 10, 50, 100, 500, 1000, 2500 and 5000 μg/plate, with and without S9-mix

 

Negative and positive control groups were also included in mutagenicity tests.

 

The mean numbers of revertant colonies are fell within acceptable ranges for negative control treatments, and were elevated by positive control treatments.

 

No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, at any dose level either with or without metabolic activation. 

 

Under the test conditions, test substance is not considered as mutagenic in S. typhimurium (TA1535, TA1537, TA 1538, TA98 and TA100) and E. coli WP2 strains.