Sodium O,O-diisobutyl dithiophosphate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Sodium O,O-diisobutyl dithiophosphate

EC Number:

258-508-5

EC Name:

Sodium O,O-diisobutyl dithiophosphate

Cas Number:

53378-51-1

Molecular formula:

C8H18NaO2PS2

IUPAC Name:

sodium O,O-diisobutyl dithiophosphate

Test material form:

other: 51 % w/w solution in water, stabilized with X % NaOH

Method

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.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:

Under the present test conditions, 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 genetic toxicity HPRT test was performed according to OECD Guideline 476 under GLP compliance. Chinese hamster lung fibroblasts were used for strain. both with and without metabolic activation. No genotoxicity was detected. No cytotoxcity nor precipitates were detected.