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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

-bacterial reverse mutation test (Ames): negative; study according to OECD TG 471, with and without metabolic activation, GLP, RL1

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1998-07-30 to 1998-09-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted 21 July 1997
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
adopted December 29, 1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine locus (Salmonella typhimurium strains) and tryptophan locus (E. coli strain)
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1st and 2nd test: 0, 62, 185, 556, 1667 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: DMSO was chosen as vehicle because the test substance decomposes in water.
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
N-ethyl-N-nitro-N-nitrosoguanidine
benzo(a)pyrene
other: N-ethyl-N-nitrosourea (without metabolic acitivation); 2-aminoanthracene (with metabolic activation)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: none
- Exposure duration: 3 days

NUMBER OF REPLICATIONS: Two experiments, all determinations in triplicate

DETERMINATION OF CYTOTOXICITY
- Cytotoxicity is defined as a reduction in the number of revertant colonies and/or a clearing of the background lawn of bacterial growth
Evaluation criteria:
The study is considered valid if the mean colony counts of the control values of the strains are within acceptable ranges and if the results of the positive controls meet the criteria for a positive response.
A response is considered to be positive if the mean number of revertant colonies on the test plates is increased two-fold or more compared to that on the vehicle control plates.
A test substance is considered to be mutagenic if a concentration-related increase or if a positive response reproducible in both assays is observed.
A test substance is considered to be not mutagenic in the bacterial gene mutation test if it produces neither a dose-related increase in the mean number of revertant colonies nor a reproducible positive response at any of the test points.
Statistics:
No statistical analysis is required.
Key result
Species / strain:
other: S. typhimurium TA 98, TA 100, TA 1535, Ta1537 and E.coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: A precipitate on the plates was observed in the first test only at 1667 µg/plate and higher.

COMPARISON WITH HISTORICAL CONTROL DATA:The mean number of his+ and trp+ revertant colonies of the negative controls were within the acceptable range, and the positive controls gave the expected increase in the mean number of revertant colonies

ADDITIONAL INFORMATION ON CYTOTOXICITY: The test substance Glycolide was not toxic to any strain, as was evidenced by the absence of decrease in the mean number of revertant colonies
Conclusions:
It is concluded that the results obtained with the test substance 1,4-Dioxane-2,5-dione (min 99.5%) in a reverse gene mutation assay in bacteria according to OECD guideline 471 (adopted July 21, 1997) and EU method B.13/14 (December 29, 1992) with the strains TA 1535, TA 1537, TA 98, TA 100 of S. typhimurium and E. coli  WP2 uvrA in the presence and absence of mammalian metabolic activation (S9 mix), indicate that 1,4-Dioxane-2,5-dione (Glycolide) was not mutagenic under the conditions employed in this study.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In a test according to the reverse gene mutation assay in bacteria according to OECD guideline 471 (adopted July 21, 1997) and EU method B.13/14 (December 29, 1992) with the S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli  WP2 uvrA  in the presence and absence of mammalian metabolic activation (S9 mix), 1,4-Dioxane-2,5-dione (Glycolide) was applied in the following concentrations: 62, 185, 556, 1667 and 5000 µg/plate. Precipitation was observed at 1665 µg/plate and higher.

In all strains and in both assays,1,4-Dioxane-2,5-dione (Glycolide) did not cause a two-fold or greater increase in the mean number of revertant colonies appearing in the test plates compared to the background spontaneous reversion rate observed with the negative control. Thus, 1,4-Dioxane-2,5-dione (Glycolide) was considered not mutagenic under the conditions of the test.

Justification for classification or non-classification

According to CLP Regulation EC No 1272/2008 and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) no classification and labelling for mutagenic toxicity is necessary.