Registration Dossier

Toxicological information

Genetic toxicity: in vivo

Currently viewing:

Administrative data

Endpoint:
genetic toxicity in vivo
Remarks:
Type of genotoxicity: other: Tripathy et al. (1990), Foureman et al. (1994) & Mitchell & Gerdes (1973): gene mutation
Type of information:
experimental study
Adequacy of study:
disregarded due to major methodological deficiencies
Reliability:
other: not rateable
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The references contained in the summary entry unsuitable test systems represent studies in drosophila melanogaster (Fourman P (1994), Tripathy NK (1990), Mitchell (1973)). These references are of limited value for risk assessment purposes, since this test system is no longer recommended as part of regulatory testing by many agencies worldwide and there is no up-to-date OECD guideline for its conduct. The existing OECD test guideline 477 has been withdrawn on 2nd April 2014. Interpretation of the relevance of both positive and negative results from such tests is therefore unclear and was not used for the current assessment. The information contained therein was included for information purposes only.
Data waiving:
other justification
Justification for data waiving:
other:

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Genetic toxicity of six carcinogens and six non-carcinogens in the Drosophila wing spot test
Author:
Tripathy, N.K. et al.
Year:
1990
Bibliographic source:
Mutat. Res. 242, 169-180
Reference Type:
publication
Title:
Chemical Mutagenesis Testing in Drosophila. X. Results of 70 coded chemicals tested for the National Toxicology Program
Author:
Foureman, P. et al.
Year:
1994
Bibliographic source:
Environ. Mol. Mutagen. 23, 208-227
Reference Type:
publication
Title:
Mutagenic effects of sodium and stannous fluoride upon Drosophila melanogaster
Author:
Mitchell, B. & Gerdes, R.A.
Year:
1973
Bibliographic source:
Fluoride 6, 113-117

Materials and methods

Principles of method if other than guideline:
Tripathy et al. (1990): stannous chloride was tested for its genotoxic activity in the Drosophila melanogaster wing spot test. 72-hour-old larvae trans-heterozygous for the recessive wing cell markers 'multiple wing hairs' (mwh) and 'flare' (fir 3) were fed 50 and 100 mM of the test compounds for a period of 48 hours.

Foureman et al. (1994):
Stannous chloride was tested for the ability to induce sex-linked recessive lethal (SLRL) mutations in postmeiotic and meiotic germ cells of male Drosophila menalogaster. Adult feeding (6500 ppm) and adult injection (12200 ppm) were chosen as exposure routes.

Mitchell & Gerdes (1973): Drosophila melanogaster was used to test the mutagenicity of stannous fluoride. LD50 and the sex-linked recessive lethal frequencies were determined.
GLP compliance:
not specified
Type of assay:
other: Tripathy et al. (1990): somatic mutation assay in Drosophila; Foureman et al. (1994) & Mitchell & Gerdes (1973): Drosophila SLRL test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Tripathy et al. (1990), Foureman et al. (1994) & Mitchell & Gerdes (1973): no data
Details on test material:
Tripathy et al. (1990):
- Name of test material (as cited in study report): Stannous chloride (source: Fluka)

Foureman et al. (1994):
- Name of test material (as cited in study report): stannous chloride (source: M and T Chemicals)
- Molecular formula: SnCl2
- Analysed purity: 99%+
- Batch No.: MT8-27-75

Mitchell & Gerdes (1973):
- Name of test material (as cited in study report): stannous fluoride
- Molecular formula: SnF2

Test animals

Species:
other: Tripathy et al. (1990), Foureman et al. (1994) & Mitchell & Gerdes (1973): Drosophila melanogaster
Strain:
other: Tripathy et al. (1990): outcross of two stocks (mwh and flr); Foureman et al. (1994): Canton-S & Basc stocks; Mitchell & Gerdes (1973): Oregon-R strain
Details on test animals and environmental conditions:
Tripathy et al. (1990):
Two stocks were outcrossed: (1) 'mwh', homozygous for mwh (multiple wing hairs 3-0.0); (2) 'flr', this stock has the genotype flr3/TM3, Ser (flare 3, 3-39.0).

Foureman et al. (1994):
Separate Canton-S and Basc stocks were maintained. Males to be exposed were collected from the Canton-S stocks. The Basc stocks supplied the balancer X-chromosome. P1 females used in the SLRL test were collected as virgins from this stock.

Mitchell & Gerdes (1973): no data

Administration / exposure

Route of administration:
other: Tripathy et al. (1990) & Mitchell & Gerdes (1973): oral: feed; Foureman et al. (1994): oral: feed & injection
Vehicle:
Tripathy et al. (1990):
- Vehicle(s)/solvent(s) used: 0.03 N HCl

Foureman et al. (1994):
- Vehicle(s)/solvent(s) used:
Adult feeding: ethanol
Adult injection: rapeseed oil or NaCl solution (not clearly stated)

Mitchell & Gerdes (1973):
- Vehicle(s)/solvent(s) used: water
Details on exposure:
Tripathy et al. (1990):
- Diet preparation: 1.5 g instant medium rehydrated with known concentrations of the test compound dissolved in 5 mL of the solvent.

Foureman et al. (1994):
no data

Mitchell & Gerdes (1973):
The various concentrations of the test compound were dissolved in 7% glucose solutions; strips of filter paper were saturated with the various test solutions and individually placed into half pint bottles.
Duration of treatment / exposure:
Tripathy et al. (1990): 48 hours
Foureman et al. (1994): 72 hours
Mitchell & Gerdes (1973): 24 hours
Frequency of treatment:
Tripathy et al. (1990) & Foureman et al. (1994): daily
Mitchell & Gerdes (1973): one treatment
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
Tripathy et al. (1990): 50 and 100 mM
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
Foureman et al. (1994): 6500 ppm (feeding) & 12200 ppm (injection)
Basis:
other: nominal in diet & nominal concentration
Remarks:
Doses / Concentrations:
Mitchell & Gerdes (1973): 5, 10, 15, 20, 25, and 30% (LD50 determination); 10, 15, 20, and 25% (sex-linked recessive lethal frequency measurement)
Basis:
nominal in diet
No. of animals per sex per dose:
Tripathy et al. (1990): 50-100 larvae
Foureman et al. (1994): no data
Mitchell & Gerdes (1973): approximately 100 flies
Control animals:
other: Tripathy et al. (1990): yes, concurrent vehicle... (see attached file)
Positive control(s):
Tripathy et al. (1990), Foureman et al. (1994) & Mitchell & Gerdes (1973): no data

Examinations

Tissues and cell types examined:
Tripathy et al. (1990):
marked wing primordial cells

Foureman et al. (1994):
postmeiotic and meiotic germ cells
A minimum of ~5000 chromosomes was scored from each of the treated and concurrent control groups unless the mutant frequency exceeded 1%.

Mitchell & Gerdes (1973):
The evaluation of sex-linked recessive lethals involves only those mutations which occur in the X chromosome of Drosophila melanogaster. They are lethal in the hemizygouse or homozygous condition.
Details of tissue and slide preparation:
Tripathy et al. (1990):
Eggs were collected for 8 hours from optimally fertile parents of the outcross kept on standard Drosophila cornmeal-agar-yeast medium. After 72 hours, the larvae (age: 72 hours) were washed out with 20% NaCl solution from the food. 50-100 larvae were transferred into plastic vials containing 1.5 g instant medium rehydrated with known concentrations of the compound dissolved in 5 mL of the solvent.
The experiments were conducted near the LD50 or at concentrations below that. Two independent experiments were conducted.

Foureman et al. (1994):
Toxicity tests were run on a series of exposures and, if possible, an exposure level was chosen that resulted in ~30% mortality after 72 hours of feeding or 24 hours after injections or inhalation.
The compound was first tested by feeding exposure. Glass fibre filters were saturated with the compound carried in a 5% sucrose solution (or other control solution) at the bottom of a standard vial. Solutions were renewed at 24 hours and 48 hours. After 72 hours of exposure, surviving males were mated.
In a second test, Canon-S males were injected with a solution containing the test chemical. At 24 hours post injection, toxicity was noted and survivors were mated.
For the SLRL test, each male was mated individually to three Basc virgin females, then transferred to fresh Basc virgin females every 2 to 3 days to make a total of three broods. F2 cultures were scored as presumptive lethals if the number of wild-type males was 0, 1, or <5% of the number of Basc males (or Basc/+ females). All putative lethals were confirmed through an additional generation.

Mitchell & Gerdes (1973):
Stannous fluoride was fed to Drosophila melanogaster. The adult feeding methods of Lüer (1964)* and Pelecanos and Alderson (1963)* were modified in the test treatments. The various concentrations of the test compound were dissolved in the 7% glucose solutions; strips of filter paper were saturated with the various test solutions and individually placed into half pint bottles. Approximately 100 flies were introduced into the bottle for a 24 hour treatment period. After treatment the flies were removed and placed into new food bottles for subsequent testing.
Three replications were used at each treatment level.
Two parameters were measured:
1) LD50 was determined by treating the flies for a 24 hour period and comparing the survival at different treatment concentrations. LD50 would be the concentration at which 50% of the flies survived a particular fluoride concentration.
2) Sex-linked recessive lethal frequencies were determined by the Muller-5 technique (1954)*.

*References:
- Lüer, H.: Untersuchung über die Mutagenität des TEM an Drosophila melanogaster. Arch. Geschwulstforsch., 6:77, 1964.
- Pelecanos, M. and Alderson, T.: The Mutagenic Response to Adult Feeding of Diethyl Sulfate in Drosophila. D. I. S., 37:116, 1963.
- Muller, H. J., Herskowitz, I. H., Abrahamson, S. and Oster, I. I.: A Non-Linear Relation Between X-Ray Dose and Recovered Lethal Mutations in Drosophila. Genetics, 39: 741 - 749, 1954.
Evaluation criteria:
Tripathy et al. (1990) & Mitchell & Gerdes (1973): no data

Foureman et al. (1994):
For a compound to be considered mutagenic, the mutant frequency in the treated series (treated frequency) must exceed 0.15% with a P value of <0.05, or the treated frequency must exceed 0.10% with a P value of <0.01. If the treated frequency is between 0.10% and 0.15% and the P value is between 0.1 and 0.01, or if the treated frequency is higher than 0.15% and the P value is between 0.1 and 0.05, the result is considered equivocal. All other results are considered negative.
Statistics:
Tripathy et al. (1990):
The statistical analysis was performed according to Frei and Würgler (1988)*.

Foureman et al. (1994):
SLRL assay: if two or more lethals were recovered among the progeny of one male, a Poisson analysis (Owen, 1962)* was performed to determine if these were part of a "cluster". In those cases in which a male was determined to have produced a cluster, the lethal and nonlethal tests for that P1 male were removed from the data. The corrected treated and control data were compared using a normal approximation to the binomial distribution, as suggested by Margolin et al. (1983)*. In addition, the treated data were compared to the historical control as described by Mason et al. (1992)*.

Mitchell & Gerdes (1973):
Pearson and Hartley test (Goldstein, 1964)*

*Reference:
- Frei, H., and F.E. Würgler (1988) Statistical methods to decide whether mutagenicity test data from Drosophila assays indicate a positive, negative, or inconclusive result, Mutation Res., 203, 297-308.
- Goldstein, A.: Biostatistics: An Introductory Text. New York, The Mac-Millan Company, 1964, p. 117.
- Owen D (1962): "Handbook of Statistical Tables." London: Addison-Wesley.
- Margolin BH, Collings BJ, Mason JM (1983): Statistical analysis and sample-size determinations for mutagenicity experiments with binomialresponses. Environ Mutagen 7: 705 - 716.
- Mason JM, Valencia R, Zimmering S (1992): Chemical mutagenesis testing in Drosophila. VIII. Reexamination of equivocal results. Environ Mol Mutagen 19: 227 - 234.

Results and discussion

Test resultsopen allclose all
Sex:
not specified
Genotoxicity:
other: Tripathy et al. (1990): negative
Toxicity:
not specified
Vehicle controls validity:
not specified
Negative controls validity:
not specified
Positive controls validity:
not specified
Sex:
male
Genotoxicity:
other: Foureman et al. (1994): negative
Toxicity:
not specified
Vehicle controls validity:
not specified
Negative controls validity:
not specified
Positive controls validity:
not specified
Sex:
not specified
Genotoxicity:
other: Mitchell & Gerdes (1973): positive
Toxicity:
not specified
Vehicle controls validity:
not specified
Negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
Tripathy et al. (1990):
In the experiments stannous chloride was tested for its mutagenicity at 2 concentrations, 50 and 100 mM, and the results were negative with respect to the induction of small single spots and inconclusive with respect to the induction of large single and twin spots.

Foureman et al. (1994):
No evidence of mutagenicity was observed for stannous chloride.

Mitchell & Gerdes (1973):
- lethal dose: a usable LD50 for SnF2 was not obtained because of the low solubility of the compound. High concentrations were accompanied by precipitation and therefore were not as reliable as lower concentrations.
- sex-linked recessive lethal mutation frequency: increase in the frequency of sex-linked lethals was observed; at the higher levels the differences were significant (15, 20, and 25 % treatment levels)

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): other: Summary entry, see IUCLID entry for further details.
No conclusion can be drawn from the above publications due to lack of quality, reliability and adequacy of the experimental data for the fulfilment of data requirements under REACH.