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Toxicological information

Genetic toxicity: in vitro

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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
Study period:
1994-10-12 to 1994-10-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted 26 May 1983
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
29 December 1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): 1,4-Butanediol dimethacrylate

Method

Target gene:
his-locus
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Experiment 1 (plate incorporation test): 33.3, 100.0, 333.3, 1000.0, 2500.0, 5000.0 µg/plate
Experiment 2 (pre-incubation test): 10.0, 33.3, 100.0, 333.3, 1000.0, 2500.0 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility properties + relative nontoxicity to the bacteria
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
other: 4-nitro-o-phenylene-diamine
Remarks:
without metabolic activation
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and preincubation

DURATION
- Preincubation period: 60 minutes
- Exposure duration: at least 48 h

NUMBER OF EXPERIMENTS: 2 (one performed as pre-incubation assay, one as plate incorporation assay), 3 plates per concentration each

DETERMINATION OF CYTOTOXICITY
- Method: Toxicity of the test article may be evidenced by a reduction in the number of spontaneous revertants, a clearing of the bacterial background lawn, or by degree of survival of treated cultures.

OTHER:
automatic colony count with “AUTOCOUNT” (Artek systems, Biosys)
Evaluation criteria:
- corresponding background growth on negative control and test plates
- normal range of spontaneous revertants: TA1535 (10-29), TA 1537 (5-28), TA 1538 (12-37), TA 98 (15-57), TA 100 (77-189)
- test material is considered mutagenic if either a dose related and reproducible increase in revertant numbers or a significant and reproducible increase in revertant numbers for at least one test concentration in induced
- a significant increase in revertants means at least twice as high in TA 100 and a three times higher number in TA 1535, TA 1537, TA 1538, TA 89 compared to the negative control
Statistics:
no appropriate statistical method available

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
No substantial increases in revertant colony numbers of any of the five tester strains were observed following treatment with 1,4-Butanediol dimethacrylate at any dose level, either in the presence or absence of metabolie activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of significance.

TEST-SPECIFIC CONFOUNDING FACTORS
- no data

RANGE-FINDING/SCREENING STUDIES:
- performed with 3.3, 10.0, 33.3, 100.0, 333.3, 1000.0, 2500.0 and 5000.0 µg/plate in TA 98 and TA 100
- normal background growth was observed up to 5000 µg/plate

COMPARISON WITH HISTORICAL CONTROL DATA:
controls were within the range of historical control data

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- in the plate incorporation assay concentrations of 2500 µg/plate (without metabolic activation) and higher reduced the number of revertants in TA 1535, TA 1537 and TA 1538
- in the pre-incubation assay the number of revertants was reduced at concentrations of 1000 µg/plate (without metabolic activation) and higher in TA 1535 and TA 1537; at 2500 µg/plate (with metabolic activation) in TA 1537 and at 2500 µg/plate (without metabolic activation) in TA 1538

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

In this study 1,4-BDDMA did not induce mutant colonies over background and is therefore considered non-mutagenic.
Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, adopted 26 May 1983, strains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 of S. typhimurium were exposed to 1,4-BDDMA in DMSO at concentrations of 33.3, 100.0, 333.3, 1000.0, 2500.0, 5000.0 µg/plate in a plate incorporation assay and 10.0, 33.3, 100.0, 333.3, 1000.0, 2500.0 µg/plate in a pre-incubation assay in the presence and absence of mammalian metabolic activation (S9 mix).

1,4-BDDMA was tested up to cytotoxic concentrations. There was no evidence of induced mutant colonies over background.

The positive controls induced the appropriate responses in the corresponding strains.

This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.

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