reaction product of alumina and sulfuric acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 04.Feb.2016 to 18.Mar.2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

certified by Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Kaiser-Friedrich-Str. 7, D-55116 Mainz, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

reaction product of alumina and sulfuric acid

EC Number:

812-508-7

Cas Number:

1093112-19-6

Molecular formula:

unspecified, UVCB substance

IUPAC Name:

reaction product of alumina and sulfuric acid

Test material form:

solid: particulate/powder

Method

Target gene:

Salmonella typhimurium genes (histidine deficiency, UVsensitivity/biotine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistance, teracycline resistance)
hisD6610 (frame shift), hisD3052 (frame shift), hisG46 (base pair substitution), uvrB (deletion), rfa (deletion), pKM101 (plasmid), pAQ1 (plasmid)

Species / strain

Species / strain / cell type:

bacteria, other: TA97a, TA98, TA100, TA102, TA1535

Additional strain / cell type characteristics:

other: histidine deficiency, UV sensitivity/biotine deficiency, lipopolysaccharide side chain deficiency, ampicillin resistance, tertacycline resistance

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix

Test concentrations with justification for top dose:

1. experiment (plate incorporation method): 5000/1500/500/150/50 µg/plate
2. experiment (pre-incubation method): 5000/2500/1250/625/313/156 µg/plate
top dose according to guideline for soluble, non-cytotoxic substances

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Remarks:

water

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Remarks:

4-nitro-1,2-phenylene, sodium azide, 2-amino-anthracen, benz-o-pyrene

Details on test system and experimental conditions:

Genotype confirmation was performed before stock culture preparation. Bacteria were checked for growth before the experiments. Per strain and dose 3 plates with and 3 plates without S9-Mix were used. For the 1. experiments plates were prepared by the plate incorporation method. for the 2. experiment by the pre-incubation method, both described in the guideline.
All strains used require histidine, so they are cultured on a histidine containing agar. All strains except TA102 are UV sensitive, therefore they are streaked on a biotine and histidine-biotone plate and half of the palte was covered with an aluminium foil to protect them against light. during radiation with a UV-lamp. All strains have a lipopolysaccharide side chain deficiency, which can detected by crystal violet sensitivity (deep rough). All strains except TA1535 are ampicillin resistant. TA102 is resistant to tetracycline too. So each strain was streaked on an apicilline and on aa ampicilline-tetracycline agar plate. The non-resistant strains served as controls.
After preparing different plates with test substance or control in- or excluding metabolic activation the plates were incubated by 37+/-1°C in the dark for 24h. To identify spotaneous revertants 3 replicates without activation were incubated for 48h. The density (titre) of cells was determined after incubation for 48h in 2 replicants. As toxicity control served 2 replicants with the maximal dose and with and without metabolic activation, respectively, and an incubation tíme of 48h. A sterility control with and without activation and without bacteria was also incubated for 48h in 4 replicates.
The colonies were counted visually.

Rationale for test conditions:

according to guideline and coresponding SOP of the laboratory

Evaluation criteria:

A substance is considered to have mutagenic potential, if an increase of revertants colonies exceeding a factor of 2 in at least one strain.

Statistics:

Microsoft Excel (R) was used to calculate mean values, standard deviations of each treatment and controls

Results and discussion

Test results

Key result

Species / strain:

bacteria, other: salminella thyphimurium TA97a, TA98, TA100, TA102 and TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Remarks:

although the test substance showed small precipitations on the plates

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In both experiments no mutagenic potential was found under the conditions of this test.

Remarks on result:

not determinable

Remarks:

the test substance is not mutagenic under the conditions of this test

Applicant's summary and conclusion

Conclusions:

no muatgenic potential found under the test conditions of guideline 471