Administrative data
Type of genotoxicity: gene mutation
13 June 2005 - 9 August 2005
Attached justification
Cross-reference
Data source
Materials and methods
Test guideline
#1 - Test guideline
adopted 1997
#2 - Test guideline
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#3 - Test guideline
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#4 - Test guideline
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#5 - Test guideline
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Test material
Appearance: Clear and colourless liquid
Storage conditions: Room temperature (ca. 20ºC), in the dark
Batch number: 52834
Expiry date: December 2006
Purity: >98%
Method
thymidine
Species / strain
#1 - Species / strain
CELLS USED
- Suitability of cells: heterozygous at the thymidine kinase locus, TK +/-
MEDIA USED
- Type and identity of media:
R0: RPMI 1640, buffered with 2 mg/mL sodium bicarbonate, supplemented with 2.0 mM L-glutamine and 50 µg/mL gentamicin.
R10p: R0, supplemented with 0.1% v/v Synperonic F68, 1.0 mM sodium pyruvate and HiDHS at 10% v/v.
R30p: R0, supplemented with 0.02% v/v Synperonic F68, 1.0 mM sodium pyruvate and HiDHS at 30% v/v.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
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S9
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- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: the test item was found to be soluble
Controls
#1 - Controls
- methylmethanesulfonate
In the absence of S9 mix; solvent: DMSO
#2 - Controls
- 3-methylcholanthrene
In the presence of S9 mix; solvent: DMSO
Exposure period (with metabolic activation): 3 hours
Exposure period (without metabolic activation): 24 hours
Expression time:
2 days
Selection time:
10-14 days
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency
Data are presented for concentrations tested up to the maximum exposure of 1041 µg/mL (10mM) for freely soluble compounds, in accordance with current guidelines.
The test agent was regarded as negative if: The Induced mutation frequency (test concentration MF minus mean control MF) for any test concentration was less than the Global Evaluation Factor (126 x 10-6). If the IMF of any test concentration exceeded the GEF, a linear trend test was applied: If the linear trend test was negative, the result was regarded as negative. If the linear trend test was positive, this indicated a positive, biologically relevant response.
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Model and software
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Any other information on materials and methods incl. tables
Results and discussion
Test results
#1 - Test results
#2 - Test results
#3 - Test results
Preliminary toxicity test (Relative suspension growth):
Concentrations from 2 to 1041 µg/mL:
3 hours exposure :
- without S9 mix:127% to 44%
- with S9 mix: 98% to 89%
24 hours exposure :
- without S9 mix: 93% to 78%
Additional information about applicability domain and reliability of (Q)SAR predictions
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Similar substances with experimental data
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Any other information on results incl. tables
Overall remarks, attachments
Attachments
Applicant's summary and conclusion
MBD did not demonstrate mutagenic potential up to limit concentrations (1041 µg/mL; 10mM) in this in vitro mammalian cell gene mutation assay (OECD 476, GLP), under the experimental conditions described.
In a mammalian cell gene mutation assay (the thymidine kinase (tk) locus (TK+/-), OECD 476, GLP) L5178Y mouse lymphoma (3.7.2c) cells cultured in vitro were exposed to 3-Methyl-1,3-butandiol (MBD, purity >98%) dissolved in sterile water at concentrations of 32.53, 65.06, 130.13, 260.25, 520.5, 1041 µg/mL in the presence and absence of mammalian metabolic activation (S9 mix). The cells were exposed for either 3 hours or 24 hours in the absence of exogenous metabolic activation (S9 mix) or 3 hours in the presence of S9 mix.
3-Methyl-1,3-butandiol was tested up to the maximum recommended concentration specified in OECD TG 476 (10 mM). There were no increases in induced mutation frequency that exceeded the Global Evaluation Factor (GEF) or the mutant frequency of the concurrent solvent control at any of the concentrations tested within acceptable levels of cytotoxicity. The positive controls induced an acceptable increase in mutation frequency. The results for the solvent controls were within acceptable ranges of historical control data.
This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 476 for in vitro mutagenicity (mammalian forward gene mutation) data.