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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 19-27, 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study generated according to internationally accepted testing guidelines.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Slags, ferronickel-manufg.
EC Number:
273-729-7
EC Name:
Slags, ferronickel-manufg.
Cas Number:
69012-29-9
Molecular formula:
This is a UVCB substance, consisting of oxides of aluminum, iron, magnesium and silicon. Due to the nature of the substance, specific molecular formula is not relevant.
Details on test material:
Slag from Ferronickel Converter containing Iron in the form of the element and its oxides, metalic oxides and Nickel.
Solid / dark brown to grey.
Chemical name: Slags, Ferronickel-manufg.

Method

Target gene:
HIS- to HIS+ reversions
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Additional strain / cell type characteristics:
other: histidine operon mutation
Metabolic activation:
with and without
Metabolic activation system:
S9 liver microsomal fraction
Test concentrations with justification for top dose:
The test item was extracted in a polar extraction medium [0.9% NaCl Lot No: Delta-Select 251002T-1 (physiological solution)] and in a non-polar extraction medium (DMSO Lot No: Applichem 9U009953) for 72+/-2h at 37(+/-1)C at a weight/volume ratio of 0.2mg/L according to ISO 10993-3, 2003 and ISO 10993-12, 2007. After extraction, the extracts were centrifuged at room temperature for 5 minutes at 1000g.
10, 20, 40, 60, 80 and 100% of test material.
Vehicle / solvent:
Solution 0,9% w/v NaCl or DMSO
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Negative controls (extraction medium and A.dest.) were treated in the same way as the treatment groups.
Positive controls:
yes
Remarks:
without metabolic activation on tester strains TA100, TA1535
Positive control substance:
other: NaN3, Sodium Azide
Remarks:
Dissolved in Aqua dest, Concentration: 10μg/plate
Positive controls:
yes
Remarks:
without metabolic activation on tester strains TA98, TA1537
Positive control substance:
other: 4-NOPD, 4-nitro-o-phenylene-diamine
Remarks:
Dissolved in DMSO, Concentrations: 10μm/plate for TA98, 40μg/plate for TA1537
Positive controls:
yes
Remarks:
without metabolic activation on tester strains TA102
Positive control substance:
methylmethanesulfonate
Remarks:
Dissolved in Aqua dest, Concentration: 1μL/plate
Positive controls:
yes
Remarks:
with metabolic activation on tester strains TA98, TA100, TA1535, TA1537 and TA102
Positive control substance:
other: 2-AA, 2-amino-anthracene
Remarks:
Dissolved in DMSO, Concentrations: 2.5μg/plate, 10μg/plate for TA102
Details on test system and experimental conditions:
S typhimurium strains TA 98, TA 100, TA1535, TA1537 and TA102 in nutrient broth at the late exponential or early statonary phase of growth
with or without metabolic activation (S9 substitution buffer/ S9 mix respectively)
All salmonella strains contain mutations in the histidine operon, thereby imposing a requirement for histidine in the growth medium.
The properties of the S.typhimurium strains with regard to membrane permeability, ampicillin- and tetracycline-resistance, as well as normal spontaneous mutation rates are checked regularly. In this way, it is ensured that the experimental conditions set up by Ames are fulfilled.
Evaluation criteria:
Cytotoxicity can be detected by a clearing or rather a diminution of the background lawn or a reduction in the number of revertants down to a mutation factor of approximately <0.5 in relation to the extract medium control.
The mutation factor is calculated by dividing the mean value of the revertant counts through the mean values of the extract medium control
a test item is considered mutagenic if
-a clear and dose related increase in the number of revertants occurs
-and/or a biologically relevant positive response for at least one of the dose groups occurs
Statistics:
According to OECD guidelines the biological relevance is the criterion for the interpretation of results and a statistical evaluation is not regarded necessary

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
not determined
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
No toxic effects of the test item extracts were observed in any of the five tester strains used up to the highest concentrations evaluated (with and without metabolic evaluation) in experiment I and II.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
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 with or without metabolic activation

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item extracts did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, extracts (polar and non-polar) of Slags,ferronickel manufg./Electric furnace slag/Converter slag are considered to be non-mutagenic in this bacterial reverse mutation assay.
Executive summary:

Ames test was performed on five S typhimurium strains (TA 98, TA 100, TA1535, TA1537 and TA102) with or without metabolic activation according to OECD 471. The test item was diluted either in 0.9% NaCl in water ir in DMSO to final concentrations of 10, 20, 40, 60, 80 and 100%. No toxic effects were noted in all tester strains at all concentrations in experiment I (with Aqua dest) and in experiment II (with DMSO). No biologically relevant increase in revertant colonies were observed following treatment with polar or non-polar extracts of the test item at any concentration level neither in the presence or absence of metabolic activation. The reference mutagens caused an distinct increase confirming the validity of the experiment. It can be deduced that the test item is not mutagenic for bacteria under the present experimental conditions.

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