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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
end 2008-02-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, OECD 471 (1997 July 21st)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Ethyldiisopropylamine
EC Number:
230-392-0
EC Name:
Ethyldiisopropylamine
Cas Number:
7087-68-5
Molecular formula:
C8H19N
IUPAC Name:
ethylbis(propan-2-yl)amine
Details on test material:
- Name of test material (as cited in study report): Ethyldiisopropylamine
- Physical state: liquid
- Analytical purity: 99.89%
- Purity test date: 2006-11-29
- Lot/batch No.: 2811061
- Stability under test conditions: no data
- Storage condition of test material: protected from moisture and at temperature <50°C

Method

Target gene:
histidine operon
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
312.5, 625, 1250, 2500, and 5000µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol (Batch V61963106L, Carlo Erba, Val de Reuil, France)
- Justification for choice of solvent/vehicle: solubility
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: NaN3 1µg/plate (TA1535-TA100), 9-AA 50 µg/plate (TA1537), 2-NF 0.5µg/plate (TA98), MMC 0.5µg/plate (TA102) +S9: 2-anthramine 2µg/plate (TA 1535-TA1537-TA98), Benzo(a)pyrene5µg/plate (TA100), 2-anthramine 10µg/plate (TA102)
Details on test system and experimental conditions:
METHODS OF APPLICATION:
Plate incorporation (both experiments without S9 mix, and the first experiment with S9mix);
test item solution (0.025mL),
+/-S9mix or phosphate buffer pH7.4 (0.5mL)
bacterial suspension (0.1mL)
2mL of overlay agar

preincubation (the second experiment with S9mix)
DURATION : 60min at 37°C
test item solution (0.025mL),
S9mix(0.5mL)
bacterial suspension (0.1mL)
after preincubation: 2mL of overlay agar

READING after 48 to 72hours incubation at 37°C

DETERMINATION OF CYTOTOXICITY
- Method: number of revertants
Evaluation criteria:
A reproductible 2-fold increase (TA98, TA100, TA102) or 3-fold increase (TA1535, TA1537) in the number of revertants compared with the vehicle controls, in any strain at any dose-level and/or evidence of dose-relationship was considered as positive result.

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at doses > 2500µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. The study was therefore considered valid.
Since the test item was freely soluble and non-toxic, the highest dose-level was 5000 µg/plate, according to the criteria specified in the international guidelines.
The selected treatment-levels were 312.5, 625, 1250 2500 and 5000 µg/plate, for both mutagenicity experiments with and without S9 mix.
Without S9 mix, no toxicity was induced in any of the five strains tested, in either experiment.
With S9 mix, using the direct plate incorporation method, no toxicity was induced in any of the five strains tested.
In the preincubation method, moderate to strong toxicity was induced at dose-levels = 2500 µg/plate, in all tester strains tested.
The test item did not induce any noteworthy increase in the number of revertants, either with or without S9 mix, in any of the five strains.
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

Ethyldiisopropylamine did not show any mutagenic activity in the bacterial reverse mutation test with S. typhimurium.
Executive summary:

The genotoxic activity of ethyldiisopropylamine EDIPA is evaluated in the Ames test on Salmonella typhimurium according to the OECD 471 guideline (Haddouk, 2008). Doses of 312.5, 625, 1250, 2500, and 5000µg/plate are tested on five strains (TA 98, TA 100, TA 102, TA 1535 and TA 1537) according to two methods: direct plate incorporation (the first experiment and the second without S9mix) and the preincubation method (60 min at 37°C - second experiment with S9mix). The number of revertants colonies is evaluated 48 to 72 hours later. Cytotoxicity was observed for the highest doses (2500µg/plate and higher) with and without metabolic activation in any strain. No genotoxic activity was observed for all doses with and without metabolic activation on the 5 tested strains. Ethyldiisopropylamine did not induce genotoxicity in the Ames test on Salmonella typhimurium in the absence and presence of metabolic activation.