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Diss Factsheets

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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
, E. coli or S. typhimurium strain TA 102 not tested
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
(diethylamino)acetonitrile
EC Number:
221-130-6
EC Name:
(diethylamino)acetonitrile
Cas Number:
3010-02-4
Molecular formula:
C6H12N2
IUPAC Name:
2-(diethylamino)acetonitrile
Details on test material:
- Name of the test substance used in the study report: Diethylaminoacetonitril
- Physical state: Colorless to yellow liquid
- Storage:+ 4 °C to + 6 °C

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
- The Salmonella strains are checked for the following characteristics at regular intervals: deep rough character; UV sensitivity; ampicillin resistance.
- Histidine auxotrophy is automatically checked in each experiment via the spontaneous rate.
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver S-9 mix
Test concentrations with justification for top dose:
Fist Experiment: 0, 20, 100, 500, 2500 and 5000 µg/plate
2nd Experiment: TA 1535; TA 100; TA 1537: 0, 15, 30, 60, 120 and 250 µg/plate; TA 98: 0, 3, 6, 12, 25 and 50 µg/plate
3rd Experiment: 0, 6, 12, 25, 50 and 100 µg/plate
Vehicle / solvent:
distilled water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene, N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitro-o-phenylendiamine, 9-aminoacridine chloride monohydrate
Remarks:
With S9: 2-aminoanthracene (all strains), Without S9: N-methyl-N-nitro-N-nitrosoguanidine (TA100, TA 1535), 4-nitro-o-phenylendiamine (TA 98), 9-aminoacridine chloride monohydrate (TA 1537)
Details on test system and experimental conditions:
METHOD OF APPLICATION: standard plate method and preincubation method
- 1st Experiment (standard plate test with and without S-9 mix)
- 2nd Experiment: (standard plate test with and without S-9 mix)
- 3rd Experiment: (preincubation test with and without S-9 mix)

DURATION
- Preincubation period: 20 min (preincubation test only)
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY: reduced his- background growth, decrease in the number of his+ revertants
Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.

Results and discussion

Test results
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
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
No increase in the number of his+ revertants was observed both in the standard plate test and in the preincubation test, either without S-9 mix or after the addition of a metabolizing system.

A bacteriotoxic effect (reduced his- background growth, decrease in the number of his+ revertants) was observed depending on the strain and testconditions from about 25 µg - 250 µg/plate onward.

No test substance precipitation was found.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
negative