2-Propenenitrile, reaction products with ethylenediamine, hydrogenated

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

and OECD Guideline 472

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-Propenenitrile, reaction products with ethylenediamine, hydrogenated

EC Number:

272-727-3

EC Name:

2-Propenenitrile, reaction products with ethylenediamine, hydrogenated

Cas Number:

68909-99-9

Molecular formula:

C3 H3 N .C2 H8 N2

IUPAC Name:

ethane-1,2-diamine; prop-2-enenitrile

Details on test material:

- Name of test material (as cited in study report): Mixture of N3/N4-amine

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S-9-Mix prepared from sprangue Dawlay rat livers after Aroclor 1254 activation.

Test concentrations with justification for top dose:

0, 20, 100, 500, 2500, 5000 µg/plate (standard plate test)
0, 4, 20, 100, 500, 2500 µg/plate (preincubation test)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Controls

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

other: see details

Details on test system and experimental conditions:

Standard plate test
Test tubes containing 2 ml portions of saft agar kept in a water bath at 45'C, and the remaining companents are added in the following order:
0.1 ml test solution or vehicle
0.1 ml bacterial suspension
0.5 ml S-9 mix (in tests with metabolic activatian) or
0.5 ml phosphate buffer (in tests without metabalic activatian)
After mixing, the samples are poured onto Vogel-Bonner agar plates. After incubation at 37'C for 48 -72 haurs in the dark, the bacterial calonies (his+ revertants) are counted.


Preincubation test
0.1 ml test solution or vehicle, 0.1 ml bacterial Suspension and 0.5 ml S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the agar plates within approx.30 seconds.


Both Tests
In each experiment 3 Test plates per dose per control used. After incubation at 37°C for 48 -72 haurs in the dark, the bacterial colonies (his+ revertants) are counted.
Posivite Control:
with S-9 mix: 2.5 µg 2-aminoanthracene (2-AA)(dissolved in DMSO) for the strains TA 100, TA 98, TA 1537and TA 1535
60 µg 2-aminoanthracene (2-AA)(dissolvecl in DMSO) for the strain E. coli WP2 uvrA
without S-9 mix: 5 µg N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) (dissolved in DMSO) for the strains TA 100 and TA 1535
10 µg 4-nitro-o-phenylendiamine (NOPD) (dissolved in DMSO) for the strain TA 98
100 µg 9-aminoacridine (AAC) chloride monohydrate (dissolved in DMSO) for the strain TA 1537
10 µg N-ethyl-N'-nitro-N-nitroso-guanidin (ENNG) (dissolved in DMSO) for the strain E. coli WP2 uvrA
The Titer was determined and in regularly measurements the strain characteristics were checked. Sterility control performed.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Additional information on results:

Solubility: No test substance precipitation was found.
Toxicity: A bacteriotoxic effect (reduced his- or trp- background gowth, decrease in the number of his+ or trp+ revertants, deduction in the titer) was observed at dose >= 2500 µ/plate
Mutagenicity: An increase in teh muber of his+ or trp+ revertants was not observed both in the standard plate test and in the preincubation test eitherwithout S-9 mix or after the addition of a metabolizing system.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

According to the results of the present study, the test substance Mixture of N3/N4-Amine is not mutagenic in the Ames test and in the Escherichia coli -reverse mutation assay under the experimental conditions chosen here.