Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1987
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Report date:
1987

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
Safepharm Standard Test Method M26 based on the following publications:
1. Ames, B.N., Durson, W.E., Yamasaki, E., and Lee, F . D. Proc. Nat. Acad.Sci. USA (1970), 70, 22B5
2. Ames, B.N. , Mccann, J. and Yamasake, E., Mutation Research (1975), 31, 347.
3. McCann, J., Coi, E., Yamasaki, E., and f'l!les, B.N. Proc. Nat. Acad. Sci. USA (1975) 75, 5135.
Deviations:
not specified
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
(ethyl-3-oxobutanoato-O'1,O'3)(2-dimethylaminoethanolato)(1-methoxypropan-2-olato)aluminium(III), dimerised
EC Number:
402-370-2
EC Name:
(ethyl-3-oxobutanoato-O'1,O'3)(2-dimethylaminoethanolato)(1-methoxypropan-2-olato)aluminium(III), dimerised
Cas Number:
149057-70-5
Molecular formula:
C14H28AlNO6
IUPAC Name:
octaaluminium(3+) octakis((2Z)-4-ethoxy-4-oxobut-2-en-2-olate) octakis(1-methoxypropan-2-olate) octakis(2-(dimethylamino)ethan-1-olate)
impurity 1
Chemical structure
Reference substance name:
1-methoxypropan-2-ol
EC Number:
203-539-1
EC Name:
1-methoxypropan-2-ol
Cas Number:
107-98-2
Molecular formula:
C4H10O2
IUPAC Name:
1-methoxypropan-2-ol
Test material form:
liquid
Details on test material:
purified with vacum distillation, analyzed by Gel permeation chromatography as stated in one of the study reports

Method

Target gene:
histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
derived from salmonella T. LT2, supplied by Pr. Ames
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction is obtained from animals pre-treated with Arochlor 1254 (a mixture of polychlorinated biphenyls) and was stated to be from protein level 37.l mg/ml.
Test concentrations with justification for top dose:
12500 / 2500 / 500 / 100 / 20 µg/plate
Vehicle / solvent:
acetone
Controlsopen allclose all
Untreated negative controls:
no
Remarks:
see solvent control
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
N-ethyl-N-nitro-N-nitrosoguanidine
other: 4 Nitro-0-phenyldiamine
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Evaluation criteria:
For a substance to be considered positive in this test system, it should have induced a dose-'related and statisically significant increase in mutation rate (of at least twice the spontaneous reversion rate) in one or more strains of bacteria in the presence and/or absence of the S-9 microsomal enzymes. To be considered negative the number of induced revertants compared to spontaneous revertants should be less than twofold at all dose levels employed, the intervals of which should be between
3 and 5 fold and extend to the limits imposed by toxicity or solubility.
Statistics:
not specified

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
solvent control is negative control

Any other information on results incl. tables

a) Preliminary Toxicity Study

The zones of inhibition observed for each tester strain and for each concentration of SA2/1/0A were scored as being+, + or - for bacteriostatic/ bactericidal activity. There was no evidence of toxicity at any of the concentrations of the test material and the main mutation studies were carried out using SA2/1/0A at concentrations in the range 20 ug/plate to 12,500 ug/plate.

b) Mutation Study

The results for the checks on viability and spontaneous reversion rate for each tester strain are shown in Table 2. The overnight culture of each strain was found to be in the required range of 108 to 109 bacteria per ml and the spontaneous reversion rate for each was found to be within the expected range. Individual plate counts together with the mean number of revertant colonies obtained for each tester strain following incubation with the test material, with and without metabolic activation.

No significant increase in the nunber of revertant colonies was recorded for any of the strains at any of the dose levels employed in this study, either with or without metabolic activation. All counts of revertant colonies were similar to those recorded for the negative control plates and were within the range expected for spontaneous reversion for each strain . The positive control substances all produced marked increases in the number of revertant colonies and the activity of the S-9 fraction was found to be satisfactory.

Applicant's summary and conclusion

Conclusions:
The test material , SA2/1/OA was found to exhibit no evidence of mutagenic activity under the conditions of this experiment
Executive summary:

In an ames test which protocol is similar to OECD 471 guidelines, the test material , SA2/l/OA was found to exhibit no evidence of mutagenic activity under the conditions of this experiment.