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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / micronucleus study
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 2009 - October 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: draft OECD guideline 487
Deviations:
no
GLP compliance:
yes
Type of assay:
in vitro mammalian cell micronucleus test

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium hexahydroxoantimonate
EC Number:
251-735-0
EC Name:
Sodium hexahydroxoantimonate
Cas Number:
33908-66-6
Molecular formula:
NaSb(OH)6
IUPAC Name:
sodium hexahydroxoantimonate
Constituent 2
Reference substance name:
sodiumhexahydroxoantimonate
IUPAC Name:
sodiumhexahydroxoantimonate
Details on test material:
- Name of test material (as cited in study report): Sodium hexahydroxoantimonate
- Substance type: inorganic
- Physical state: solid
- Analytical purity: 99.5%
- Purity test date: 2009-01-21
- Lot/batch No.: 61049-070901J100000
- Expiration date of the lot/batch: September 2010
- Stability under test conditions: stable
- Storage condition of test material: Keep container tightly closed.

Method

Target gene:
not applicable
Species / strain
Species / strain / cell type:
lymphocytes: human
Details on mammalian cell type (if applicable):
Range-Finder: Sex: Male; Age: 25, 27; Donor Identity: 9402, 9683
Micronucleus Experiment: Sex: Male; Age: 23, 27; Donor Identity: 9023, 9683
No volunteer was suspected of any virus infection or exposed to high levels of radiation or hazardous chemicals. The measured cell cycle time of the donors falls within the range 13 +/- 1.5 hours.
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Range Finder: 1.959-540 µg/mL with and without S9
Experiment 1 (3+21h): 10.68-540 µg/mL with and without S9
Experiment 2 (24+0h): 10.57-534.7 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: cell culture medium
- Justification for choice of solvent/vehicle: to achieve the highest possible treatment concentration without precipitation, the media replacement technique was chosen.
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
and cyclophosphamide and vinblastine
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 (culture stimulation with PHA)
- Exposure duration: 3 and 24 hours
- Expression time (cells in growth medium): 0 and 21 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 24 hours

SPINDLE INHIBITOR (cytogenetic assays): Cytochalasin B
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: RI:mono-, bi- and multinucleate cells to a minimum of 500 cells per culture, MNBN: 1000 binucleate cells from each culture (2000 per concentration) were analysed for micronuclei

DETERMINATION OF CYTOTOXICITY
- Method: replication index
Evaluation criteria:
For valid data, the test article was considered to induce clastogenic and/or aneugenic events if:
1. A statistically significant increase in the frequency of MNBN cells at one or more concentrations was observed
2. An incidence of MNBN cells at such a concentration that exceeded the normal range in both replicates was observed
3. A concentration-related increase in the proportion of MNBN cells was observed.

The test article was considered as positive in this assay if all of the above criteria were met.

The test article was considered as negative in this assay if none of the above criteria were met.

Results which only partially satisfied the above criteria were dealt with on a case-by-case basis. Evidence of a concentration-related effect was considered useful but not essential in the evaluation of a positive result. Biological relevance was taken into account, for example consistency of response within and between concentrations, or effects occurring only at high or very toxic concentrations.
Statistics:
Binomial Dispersion Test

Results and discussion

Test results
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none

- Precipitation: observed

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Caspase measurements on post treatment cell suspensions (taken during the Range-Finder Experiment), indicated no evidence of increased Caspase activity with increasing Sodium hexahydroxoantimonate concentration. Therefore, there was no indication of any Apoptotic activity as a result of test article treatment.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

It is concluded that Sodium hexahydroxoantimonate did not induce micronuclei in cultured human peripheral blood lymphocytes following treatments in the absence and presence of an Aroclor induced rat liver metabolic activation system (S-9). Concentrations were tested and analysed up to and in excess of the solubility limit in culture medium.