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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Remarks:
Guideline study without detailed documentation.

Data source

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

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
-reliability scoring based on 1997 guideline
Deviations:
yes
Remarks:
-incubation temperature was not reported and less than 200 metaphase cells were scored per concentration and controls
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): 1,4-butandiol-bis (2,3-epoxypropylether)
- Physical state: Clear liquid
- Analytical purity: 93.9%
- Expiration date of the lot/batch: Stable until December 31, 1992
- Stability under test conditions: Reported as "stable 4h"
- Storage condition of test material: Dark at approximately 20 °C

Method

Species / strain
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver homogenate (S-9 fraction)
Test concentrations with justification for top dose:
Preliminary experiment: 1.0, 5.0, 10.0, 20.0, 30.0, 40.0, 50.0, and 100.0 μg/mL (-S9) and 100.0, 125.0, 150.0, 175.0, 200.0, and 250.0 μg/mL (+S9)
Main study:
0 or 10.0 μg/mL (7 hr, -S9)
0 or 100.0 μg/mL (7 hr, +S9)
0, 1.0, 5.0, and 10.0 μg/mL (18 hr, -S9)
0, 10.0, 50.0, and 100.0 μg/mL (18 hr, +S9)
0 or 10.0 μg/mL (28 hr, -S9)
0 or 100.0 μg/mL (28 hr, +S9)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Test substance was dissolved as a solution in minimum essential medium at appropriate concentrations.
- Justification for choice of solvent/vehicle: Not reported.
Controls
Untreated negative controls:
yes
Remarks:
untreated cells only
Negative solvent / vehicle controls:
yes
Remarks:
minimum essential medium
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: ethylmethanesulfonate (-S9) or cyclophosphamide (+S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hrs at 37 °C
- Expression time (cells in growth medium): 0.5, 11.5, and 21.5 hrs at 37 °C
- Fixation time (start of exposure up to fixation or harvest of cells): 7, 18, or 28 hrs

SPINDLE INHIBITOR (cytogenetic assays): Colcemide
STAIN (for cytogenetic assays): Staining for 10 minutes in approx. 2% orcein solution

NUMBER OF REPLICATIONS: Reported as 2 independent cell cultures but interpreted as duplicates.

NUMBER OF CELLS EVALUATED: 50 to 100 metaphases per concentration per preparation time (for all treated and control groups)

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: Yes
- Determination of endoreplication: No

Evaluation criteria:
Evaluation of the results was performed as follows:
-The test substance is classified as mutagenic if it induces a significantly increased aberration rate as compared with the solvent controls with one of the concentrations tested. The significance is obvious either by an enhancement of the rate clearly exceeding the control range or it is proven by adequate biometry.
-The test substance is classified as mutagenic if there is a reproducible concentration related increase in the aberration rate.
-The test substance is classified as non-mutagenic when it tests negatively both with and without metabolic activation.
Statistics:
The biometry of the results was performed with a one-sided Fisher - Exact test.

Results and discussion

Test results
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at concentrations of ≥40 μg/mL (-S9) and ≥175 μg/mL (+S9)
Vehicle controls validity:
valid
Untreated negative controls validity:
other: results were not reported
Positive controls validity:
other: Yes, valid at 18 h, which was the only time point at which the positive control was tested
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Results of the duplicates were not consistent at every concentration tested (100 μg/mL at 7 h and 50 μg/mL at 18 h).

Applicant's summary and conclusion

Conclusions:
Interpretation of results: positive without metabolic activation; positive with metabolic activation