Tetramethylenediamine

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

Study period:

3 days

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Not according to OECD guidelines (study in 1977), but following Ames etal, Mutation Research vol 31: 347, 1975; very well documented tests

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1977

Report date:

1977

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Ames etal, Mutation Research vol 31: 347, 1975

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tetramethylenediamine

EC Number:

203-782-3

EC Name:

Tetramethylenediamine

Cas Number:

110-60-1

Molecular formula:

C4H12N2

IUPAC Name:

butane-1,4-diamine

Details on test material:

Test compound received on July 22, 1977: cloudy yellow liquid.

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 38, TA 98 and TA 100

Additional strain / cell type characteristics:

other: Saccharomyces cerevidiae, strain D4

Metabolic activation:

with and without

Metabolic activation system:

S-9 Homogenate (Sprague-Dawley adult male rat liver, induced by Aroclor 1254 5 days prior to kill)

Test concentrations with justification for top dose:

0.00100 ul, 0.01000 ul, 0.10000 ul, 1.00000 ul, 5.00000 ul

Vehicle / solvent:

TPN (4 umol), Glucose-6-phosphate (5 umol), Sodium phosphate (dibasic) (100 umol), MgCl2 (8 umol), KCl (33 umol), Homogenate fraction equivalent to 25 mg of wet tissue (0.1-0.15 ml 9.000 x 9 supernatant of rat liver).

Controls

Negative solvent / vehicle controls:

yes

Remarks:

demethylsulfoxide or deionized water

Positive controls:

yes

Remarks:

Non activation: Methylnitrosoguanidine (MNNG) in water or saline, 2-Nitrofluorene (NF) in dimethylsulfoxide, Quinacrine mustard (QM) in water or saline. Activation: 2-Anthramine (ANTH) in dimethylsulfoxide, 2-Acetylaminofluorene (AAF) in demethylsulfoxide

Details on test system and experimental conditions:

Approximately 10exp8 cells from an overnight culture of each indicator strain were added to separate test tubes containing 2.0 ml of molten agar supplemented with biotin and a trace of histidine. For non-activation tests, at least 4 dose levels of the test compound were added to the content of the appropriate tubes and poured over the surfaces of selective agar plates. In activation tests, a minimum of 4 different concentrations of the test chemical were added to the appropriate tubes with cells. Just Prior to pouring, an aliquot of reaction mixture (0.5 ml containing the 9,000 x g liver homogenate) was added to each of the activation overlay tubes, wich were then mixed, and the contents poured over the surface of a minimal agar plate and allowed to solidify. The plates were incubated for 48 hours at 37°C, and scored for the number of colonies growing on each plate. Positive and solvent controls using both directly active positive chemicals and those that require metabolic activation were run with each assay.

Evaluation criteria:

The numbers of colonies on each plate were counted and recored on printed forms. These raw data were ananlyzed in a computer program and reported in a print out. The results are presented as revertants per plate for each indicator strain employed in the assay. The positiev and solvent controls are provided as reference points.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 38, TA 98 and TA 100 and Saccharomyces cerevisiae, D4

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: no, except for TA-1538 (dose 5 ul)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

DAB was tested over a series of concentrations such that there was either quantitative or qualitative evidence of some chemically-induced physiological effects at the high dose level. The low dose in all cases was below a concentration that demonstrated any toxic effect. The dose range employed for the evaluation of this compound was from 0.001 ul to 5 ul per plate. DAB was toxic to the strain TA-1538 at 5 ul per plate.
The results of the tests conducted on DAB in the absence of a metabolic system were all negative.
The results of the tests conducted on DAB in the presence of the rat liver activation system were all negative.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Revertants per plate

Test	Species/tissue	TA-1535	TA-1537	TA-1538	TA-98	TA-100	D4+
Solvent control		17	13	18	35	85	108
Positive control		>1000	314	945	411	>1000	>1000
DAB 0.00100 ul		7	12	12	28	56	84
DAB 0.01000 ul		7	10	8	30	50	78
DAB 0.10000 ul		8	15	6	39	50	75
DAB 1.00000 ul		8	15	7	28	38	102
DAB 5.00000 ul		4	10	0	24	19	69
Solvent control	Rat/liver	22	17	18	56	125	24
Positive control	Rat/liver	316	228	324	558	895	50
DAB 0.00100 ul	Rat/liver	17	15	16	36	52	25
DAB 0.01000 ul	Rat/liver	23	27	15	35	49	9
DAB 0.10000 ul	Rat/liver	23	18	10	42	57	10
DAB 1.00000 ul	Rat/liver	30	23	22	45	46	33
DAB 5.00000 ul	Rat/liver	24	14	11	42	29	30

Applicant's summary and conclusion

Conclusions:

DAB did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered not mutagenic under these test conditions.

Executive summary:

1,4 Butaandamine did not demonstrate mutagenic activity in any of the assays with or without metabolic activation (S9) in Salmonella typhimurium TA-1535, TA-1537, TA-1538, TA-98, TA-100, and Saccharomyces cerevisiae D4, and was considered not mutagenic under these test conditions