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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 1998 - April 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report date:
1999

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1H-imidazole-1-propylamine
EC Number:
225-730-9
EC Name:
1H-imidazole-1-propylamine
Cas Number:
5036-48-6
Molecular formula:
C6H11N3
IUPAC Name:
3-(1H-imidazol-1-yl)propan-1-amine
Details on test material:
- Name of test material (as cited in study report): N-(3-Aminopropyl)-imidazol
- Physical state: colorless liquid
- Analytical purity: 98.2 %
- Lot/batch No.: Abl. Nr. 33-0531

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S-9-Mix prepared from sprangue Dawlay rat livers after Arolor 1254 activation.
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500, 5000 µg/plate (Standard plate test with and without S-9-mix)
0, 20, 100, 500, 2500, 5000 µg/plate (Preincubation test with and without S-9-mix)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: see details
Details on test system and experimental conditions:
Standard plate test
Salmonella typhimurium
Test tubes containing 2-ml portions of soft agar (overlay agar), which consists of 100 ml agar (0.6% agar + 0.6% NaCI) and 10 ml amino acid solution (minimal amino acid solution for the determination of mutants: 0.5 mM histidine + 0.5 mM biotin) are kept in a water bath at 45°C, and the remaining components are added in the following order:
0.1 ml test solution or vehicle
0.1 ml fresh bacterial culture
0.5 ml S-9 mix (in tests with metabolic activation)
or
0.5 ml phosphate buffer (in tests without metabolic activation)
After mixing, the samples are poured onto Vogel-Bonner agar plates (minimal glucoseagar plates) within approx. 30 seconds.

Composition of the minimal glucose agar:
980 ml aqua dest.
20 ml Vogel-Bonner E medium
15 g Difco bacto agar
20 g D-glucose, monohydrate
After incubation at 3700 for 48 -72 hours in the dark, the bacterial colonies(his+ revertants) are counted.

Escherichia coli
Test tubes containing 2-ml portions of soft agar (overlay agar), which consists of 100 ml agar (0.6% agar + 0.6% NaCI) and 10 ml amino acid solution (minimal amino acid solution for the determination of mutants: 0.5 mM tryptophan) are kept in a water bath at 45°C, and the remaining components are added in the following order:
0.1 ml test solution or vehiele
0.1 ml fresh bacterial culture
0.5 ml S-9 mix (in tests with metabolic activation)
or
0.5 ml phosphate bufter (in tests without metabolic activation)
After mixing, the samples are poured onto minimal agar plates within approx.30 seconds.

Preincubation Test
0.1 ml test solution or vehicle, 0.1 ml bacterial suspension and 0.5 ml S-9 mix are incubated at 37°C for the duration of about 20 minutes using a shaker. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the agar plates within approx. 30 seconds.After incubation at 37°C for 48 -72 hours in the dark, the bacterial colonies are counted.

With S-9 mix
2-aminoanthracene (2-AA)
2.5 µg/plate, dissolved in DMS0 (strains: TA 1535, TA 100, TA 1537, TA 98)
60 µg/plate, dissolved in DMS0 (strain: Escherichia coli WP2 uvrA)

Without S-9 mix
N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
5 µg/plate, dissolved in DMSO (strains: TA 1535, TA 100)
4-nitro-o-phenylendiamine (NOPD)
10 µg/plate, dissolved in DMSO (strain: TA 98)
9-aminoacridine (AAC)
100 pg/plate, dissolved in DMS0 (strain: TA 1537)
4-nitroquinoline-N-oxide (4-NQO)
5 pg/plate, dissolved in DMSO (strain: E. coli WP2 uvrA)

The titer is generally determined only in the experimental parts with S-9 mix both for the negative controls (vehicle only) and for the two highest doses in all experiments.
Evaluation criteria:
Positive results:
A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least onr tester strain either without S-9 mix or after adding a metabolizing system.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: see additional information below
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: see additional information below
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Solubility: No test substance precipitation was found.
Toxicity: A weak bacteriotoxic effect (slight decrease in the number of revertans, slight reduction in the titer) was occasionally observed in the standard plate test depending on the strain and the conidtions from about 2500 µg/plate onward. In the preincubation assay bacteriotoxicity (reduced background growth, decrease in the number of revertants, reduction in the titer) was observed depending on the strain and the test conditions from about 500 µg - 2500 µg/plate onward.
Mutagenicity: An increase in the number of his+ or trp+ revertants was not observed in the standardplate test or in the preincubation test either without S-9 mix orafter the addition of a metabolizing system.
Remarks on result:
other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:
Interpretation of results: negative
According to the results of the present study, the test substance N-(3-Aminopropyl)-inidazol is not nutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay under the experimental conditions chosen here.
Executive summary:

N-(3-Aminopropyl)-imidazole was tested in the bacterial reverse mutation assay (OECD471) with S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA in concentrations between 20 -5000 µg/plate (BASF SE, 1999). An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either with or without S-9 mix. A weak bacteriotoxic effect (slight decrease in the number of revertans, slight reduction in the titer) was occasionally observed in the standard plate test depending on the strain and the conidtions from about 2500 µg/plate onward. In the preincubation assay bacteriotoxicity (reduced background growth, decrease in the number of revertants, reduction in the titer) was observed depending on the strain and the test conditions from about 500 µg - 2500 µg/plate onward. In conclusion, the test substance N-(3-Aminopropyl)-inidazol is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay under the experimental conditions chosen here.