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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:
March 5, 1985 - March 18, 1985
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report date:
1985

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3,6-dioxaoctamethylenediamine
EC Number:
213-203-6
EC Name:
3,6-dioxaoctamethylenediamine
Cas Number:
929-59-9
Molecular formula:
C6H16N2O2
IUPAC Name:
3,6-dioxaoctamethylenediamine
Test material form:
liquid
Details on test material:
- Physical state: liquid
- Appearance: colourless liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 5601-49-1

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:Room temperature in container received
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Soluble and stable

Method

Target gene:
Histidine locus
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA1535, TA1537, TA1538, TA98, TA100
Metabolic activation:
with and without
Metabolic activation system:
S-9
Test concentrations with justification for top dose:
50, 166, 500, 1666, and 5000 µg/plate.
Vehicle / solvent:
Distilled deonized water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
Remarks:
Negative controls without activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration:48-72 h at 37 degrees C

NUMBER OF REPLICATIONS: 3

- OTHER: 2 ml aliquots of molten top agar, 0.5 ml biotin and histadine, 0.1 ml tester strain, 0.1 ml test compound concentration were vortexed and poured onto minimal glucose plates in a thin layer. Plates needing activation were suplimented with 0.5 ml S-9 rat liver homogenate. Revertant colonies were counted with an electronic colony counter interfaced with a computer.
Evaluation criteria:
A positive result is a dose-related significant increase in the number of histidine dependent colonies as determined by the program developed by Fenton and Moore. A negative result is the absence of reproducible increase in the number of histidine dependent colonies.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
The test material was not found to increase mutation frequencies in any strain of S. typhimurium, either with or without metabolic activation. The substance is considered to not be mutagenic.
Executive summary:

The test article was received as a clear liquid. The solvent used throughout the assay was distilled/deionized water and 5000 microgram/plate.

Dose levels in a preliminary toxicity screen were 50, 166, 500, 1666 and 5000 microgram/plate. Strains TA1538 and TA100 of Salmonella typhimurium exhibited no inhibition of bacterial lawn growth at any of the dose levels tested.

Therefore , the top dose selected for the plate incorporation mutation assay was 5000 microgram/plate.

The test article was evaluated in strains TA 1535, TA1537, TA 1538 and TA100 of Salmonelle typhimurium both with and without metabolic activation preparation at dose of 50, 166, 500, 1666 and 5000 microgram/plate.

There were 0.10 mL of S-9 supernatant (31.1 mg protein/ml) per 1.0 ml of S9 -mix in the rat liver metabolic activation preparation.

The test article was negative in strains TA 1535, TA 1537, TA 1538, TA98 and TA100 of Salmonella typhimurium with and without metabolic activation preparation at dose of 50, 1661 500, 166 and 5000 microgram/plate. All solvent and positive controls used in the evaluation of the test article were within the acceptable limits of mean historical data.