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Toxicological information

Genetic toxicity: in vitro

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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

Constituent 1
Chemical structure
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:
N,N-bis[3-(dimethylamino)propyl]-N',N'-dimethylpropane-1,3-diamine
Test material form:
gas under pressure: refrigerated liquefied gas

Method

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.