N,N-bis[3-(dimethylamino)propyl]-N',N'-dimethylpropane-1,3-diamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: other: in vitro micronucleus assay

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECG 487 - In Vitro Mammalian Cell Micronucleus Test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian cell micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

N,N-bis[3-(dimethylamino)propyl]-N',N'-dimethylpropane-1,3-diamine

EC Number:

251-459-0

EC Name:

N,N-bis[3-(dimethylamino)propyl]-N',N'-dimethylpropane-1,3-diamine

Cas Number:

33329-35-0

Molecular formula:

C15H36N4

IUPAC Name:

(3-{bis[3-(dimethylamino)propyl]amino}propyl)dimethylamine

Test material form:

gas under pressure: refrigerated liquefied gas

Method

Metabolic activation:

with and without

Test concentrations with justification for top dose:

The test substance was suspended in culture medium (RPMI 1640) and the highest test concentration of the test substance was 5000 g/mL. The starting solution (5000 g/mL) was diluted until formation of concentration series (5000, 2500, 1000, 500 and 250 g/mL). Fresh solutions of test substance were prepared before each experiment.

In the experiment with metabolic activation the concentration 5000 μg/mL was highly cytotoxic. Then the concentration of 2500 μg/mL was used as the highest one for the analysis of genotoxic effect with metabolic activation.

In experiments without metabolic activation the concentration 5000 μg/mL was used as the highest test concentration for analysis of genotoxicity without metabolic activation.

Vehicle / solvent:

20 l of the appropriate concentration of test substance solution in medium was added to lymphocyte culture (2.5 mL growth medium RPMI-M + ca 150-210 l human peripheral blood) in the presence and absence of a metabolic activation system (18.5 l S9 post mitochondrial fraction + 18.5 l cofactors). Duplicate cultures were used for each concentration and control.
The cultures were treated by test substance for 48 hours after mitogenic stimulation,

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

other: Colchicine

Details on test system and experimental conditions:

Principle of test is the detection of binucleated cells with micronuclei, which are induced by the test substance in human peripheral blood lymphocytes. Lymphocytes are cultured in growth medium and test substance is added to them. Cell cycle is then stopped by cytochalasin B, cultures are sampled and microscopic preparations are prepared. Preparations are then analysed by microscope. Genotoxicity is indicated by increased incidence of binucleated cells with micronuclei.
Experiments with and without metabolic activation with short treatment (3 hours) are done at first. If both experiments of the short treatments are negative or equivocal, subsequently, extended exposure treatment without metabolic activation is performed.

The human peripheral blood lymphocytes used for testing were obtained from healthy non smoking donors.

Results and discussion

Test results

Species / strain:

lymphocytes:

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Concentration 5000 μg/mL was highly cytotoxic in experiment with metabolic activation.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

In the in vitro micronucleus test using human peripheral blood lymphocytes, the test substance POLYCAT 9 Catalyst was non mutagenic for the human peripheral blood lymphocytes in experiments both without and with metabolic activation.