4-(aminomethyl)octane-1,8-diamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Link to relevant study records

Reference

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:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material:
Manufacturer: Ascend Performance Materials Inc.
Lot no.: 1205635
- Purity, including information on contaminants, isomers, etc.:
100%



STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Ambient
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage:
stable
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis:
stable
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium:
highly soluble
- Reactivity of the test material with the incubation material used (e.g. plastic ware):
non-reactive

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding):
none
- Preliminary purification step (if any):
none

Target gene:

T/PE
his D 6610
his G 46
his D 3052
his G 46

Species / strain / cell type:

S. typhimurium TA 97a

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain / cell type:

S. typhimurium TA 100

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 1535

Metabolic activation:

with and without

Metabolic activation system:

An activation buffer containing 10% S9 obtained from the livers of Aroclor 1254-treated adult Sprague
Dawley® rats was prepared according to the manufacturer’s (Moltox) instructions. Each vial of Regensys B
(Moltox cat no. 60-201) was reconstituted with Regensys A (Moltox cat. no. 60-200). The solution was
transferred back into the Regensys A bottle. S9 (Moltox cat. no. 11-101) was then mixed with Regensys
A+B to yield a 10% S9 buffer stock solution. The stock was separated into aliquots in sterile 15 ml
conical tubes and refrigerated at 2-8°C until used.
The exogenous metabolic activation mixture was added to one set of all doses – each test article
concentration, vehicle control and positive control for each of the bacterial tester strains.

Test concentrations with justification for top dose:

Five concentrations (0.01, 0.05, 0.1, 0.5 and 1 μl/plate) of the test article were tested in each of five bacterial tester strains (Escherichia coli WP2 uvrA, and Salmonella typhimurium strains TA-97a, TA-1535, TA-98, and TA-100).

Vehicle / solvent:

tissue culture water

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

other:

Details on test system and experimental conditions:

Five concentrations (0.01, 0.05, 0.1, 0.5 and 1 μl/plate) of the test article were tested in each of five bacterial tester strains (Escherichia coli WP2 uvrA, and Salmonella typhimurium strains TA-97a, TA-1535, TA-98, and TA-100). Two sets of culture plates were dosed per concentration (+S9 and –S9). A vehicle
control and positive controls specific to each bacterial strain were treated in a similar manner as the test article concentrations. The plates were incubated at 37±2°C for 48 to 72 hours. Following incubation, the plates were stored at 2-8ºC, to stop bacterial growth, until the revertant colonies could be counted.

Due to failure of the positive and vehicle controls for tester strains WP2 uvrA and TA-1535 to meet the quality control acceptance criteria, the main assay was repeated with these tester strains. The controls for tester strain TA-1535 met the acceptance criteria, but those for tester strain WP2 uvrA did not. The assay was repeated a second time, with only tester strain WP2 uvrA, and all controls met the acceptance criteria. See Results and Discussion, Quality Checks (page 16). Only the results of valid assays were used as the basis for the study conclusion.

Statistics:

Plates were scored based on the number of revertant colony-forming units present per plate. The number of revertants of each test article plate were averaged and plotted versus concentration of the test article. The mean number of revertants of each dose was divided by the mean for the vehicle control value to obtain a ratio to vehicle. In evaluating the data, cytotoxicity of the test article as well as quality checks of the assay were taken into account5,6.

In general, a 2-fold increase with or without metabolic activation is considered a positive response. Doserelated increases approaching a 2-fold increase are deemed equivocal.

A negative result is determined by the absence of a dose-related increase in all five tester strains, again taking into account cytotoxicity of the test article as well as the quality checks of the assay.

Positive results from the bacterial reverse mutation test indicate that the material induces point mutations by base substitutions or frame shifts in the genome of either Salmonella typhimurium and/or Escherichia coli. Negative results indicate that under the test conditions, the test material is not mutagenic in the tested strains.

Species / strain:

S. typhimurium TA 97a

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Conclusions:

Under test conditions, test article Hexatran 100 did not have mutagenic potential in the Bacterial Reverse Mutation Assay.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)