Registration Dossier

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

in vitro gene mutation study in mammalian cells
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian cell gene mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium O,O-diisobutyl dithiophosphate
EC Number:
EC Name:
Sodium O,O-diisobutyl dithiophosphate
Cas Number:
Molecular formula:
Sodium O,O-diisobutyl phosphorodithioate
Test material form:
other: 51 % w/w solution in water, stabilized with X % NaOH
Details on test material:
- Substance type: Liquid
- Physical state: Light brown liquid
- Molecular formula (if other than submission substance): C8H18NaO2PS2
- Molecular weight (if other than submission substance): 264
- Smiles notation (if other than submission substance): CC(C)COP(=S)([S-])OCC(C)C.[Na+]
- InChl (if other than submission substance): InChI=1/C8H19O2PS2.Na/c1-7(2)5-9-11(12,13)10-6-8(3)4;/h7-8H,5-6H2,1-4H3,(H,12,13);/q;+1/p-1


Target gene:
HPRT locus. This gene is situated in man, hamster and other species at the X-chromosome. The gene product, hypoxanthine guanine phosphoribosyl transferase, an enzyme which is not vital for the cell, activates the purine analogues 8-azaguanine and 6-thioguanine (used in this assay) to toxic metabolites. Mutants which do not synthesize the active enzyme are resistant to high concentrations of 6-thioguanine or 8-azaguanine. Such mutants can be obtained by several types of mutations since a simple destruction of a gene ('forward mutation') is sufficient.
Species / strain
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: Dulbecco's modified Eagle-Medium supplemented with 10% fetal calf serum2, penicillin2 (100 U/mL) and streptomycin2 (100 µg/mL) called DMEM-FCS. Cultures are incubated at 37°C in a humidified atmosphere (90%) containing 10% CO2. For subculturing, a trypsin (0.05%)-EDTA (ethylene¬diamine¬tetra¬acetic acid, 0.02%) solution in modified Puck's salt solution A2 is used. Exposure to the test item in the presence of S9 mix is performed in Dulbecco's phosphate buffered saline (PBS)2 which additionally contained 20 mM HEPES (N'-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid)2 pH 7.4 (PBS-HEPES). The cells are periodically checked for the absence of mycoplasma contamination by using the HOECHST stain 33258. The spontaneous mutation rate is continuously monitored.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes/no
- Periodically "cleansed" against high spontaneous background: yes
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
a microsomal preparation derived from Aroclor 1254-induced rat liver.
Test concentrations with justification for top dose:
Preliminary study: 10, 25, 100, 250, 1000, 2500 and 5000 μg IBP1-Na/mL
Main study: (4h) 156.3, 312.5, 625, 1250, 2500 μg IBP1-Na/mL. (24h) 78.13, 156.3, 312.5, 625, 1250 μg IBP1-Na/mL.
Untreated negative controls:
Negative solvent / vehicle controls:
True negative controls:
Positive controls:
Positive control substance:

Results and discussion

Test results
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
Untreated negative controls validity:
Positive controls validity:
Remarks on result:
other: all strains/cell types tested
Migrated from field 'Test system'.

Applicant's summary and conclusion

Interpretation of results:
negative with metabolic activation
negative without metabolic activation

Under the present test conditions the substance, IBP1-Na, tested up to a cytotoxic concentration, caused no mutagenic effect in the Chinese hamster lung fibroblasts (V79) carried out without and with metabolic activation.
Remark: The test solution contains NaOH due to the production method.
Executive summary:

The test was performed according to the OECD guideline 476 under GLP compliance. Chinese hamster lung fibroblasts was used as strain. Metabolic activation was used both with and without. Blank, vehicle control and positive control was tested. No genotoxicity, no cytotoxicity was detected.