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Toxicological information

Genetic toxicity: in vitro

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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:
other: Ames etal, Mutation Research vol 31: 347, 1975
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
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