Propanamide, 3-amino-2,2-dimethyl-

Genetic toxicity: in vitro

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology BASF AG

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): 3-Amino-2,2-dimethyl-propanamide
- Physical state: solid, white granules
- Test substance No .: 04/0586-1
- Batch No: 04-00812_M4
- Purity: 99.5 %
- Storage condition of test material: room temperature (N2 conditions)

Method

Target gene:

His - (S. typhimurium)
Trp - (E. coli)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9-mix

Test concentrations with justification for top dose:

20 µg - 5000 µg/plate (SPT and PIT)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: solubility

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Remarks:

with metabolic activation: all strains

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine

Remarks:

without metabolic activation: TA 1535, TA 100

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without metabolic activation: TA 1537

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

without metabolic activation: E.Coli WP2 uvrA

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine

Remarks:

without metabolic activation: TA98

Details on test system and experimental conditions:

Experiment 1: Standard plate test
Test tubes containing 2 ml soft agar kept in a water bath at 42-45°C, and remaining components added in the following order:
0.1 ml test solution or vehicle
0.1 ml bacterial suspension
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 (S. Typhimurium) or minimal agar plates (E.Coli).

Experiment 2: Preincubation assay
0.1 mL test solution or vehicle, 0.1 mL bacterial suspension and 0.5 mL S9 mix (with metabolic activation) or phosphate buffer (without metabolic activation) 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.

Experiment 1 & 2:
In each experiment 3 test plates per dose or per control used; after incubation at 37°C for 48-72 hours in the dark, the bacterial colonies (his+ revertants or Trp + revertants) are counted.

Positive control:
with metabolic activation: 2.5 μg/plate 2-aminoanthracene for each Salmonella strain and 60µg/plate for E. coli WP2 uvrA;
without metabolic activation: 5 μg/plate N-methyl-N'-nitro-N-nitrosoguanidine for TA 100 and TA 1535, 10 μg/plate 4-nitro-o-phenylendiamine for TA 98, 100 μg 9-aminoacridine chloride monohydrate for TA 1537 and 10 μg/plate methyl-N- nitro-N-nitrosoguanidine for E. Coli WP2 uvrA, all positive substances were dissolved in DMSO.

The titer was determined and in regularly measurements the strain characteristics were checked. Sterility control performed.

Evaluation criteria:

Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
• The number of revertant colonies in the negative controls was within the normal range of the historical control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control articles both with and without S-9 mix induced a significant increase in the number of revertant colonies within the range of the historical control data or above.
• The titer of viable bacteria was > 10^8/ml.

Assessment criteria
The test chemical is considered positive in this assay if the following criteria are met:
• A dose-related and reproducible increase in the number of revertant colonies, i .e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Statistics:

Mean and standard deviation. No further data.

Results and discussion

Test results

Test results 1

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No precipitation of the substance was found.
No bacteriotoxic effect was observed.
An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolising system.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test (20 - 5000 µg/plate)
Strain	Metabolic activation system	Replicates	maximum revertant factor	dose dependency	Assessment
TA 98	no	3	1.0	no	negative
	yes	3	1.0	no	negative
TA 100	no	3	1.0	no	negative
	yes	3	1.0	no	negative
TA 1535	no	3	1.0	no	negative
	yes	3	0.9	no	negative
TA 1537	no	3	0.7	no	negative
	yes	3	1.2	no	negative
WP2 uvr A	no	3	1.0	no	negative
	yes	3	1.0	no	negative
Preincubation test (20 - 5000 µg/plate)
Strain	Metabolic activation system	Replicates	maximum revertant factor	dose dependency	Assessment
TA 98	no	3	1.0	no	negative
	yes	3	1.0	no	negative
TA 100	no	3	1.0	no	negative
	yes	3	1.0	no	negative
TA 1535	no	3	1.0	no	negative
	yes	3	0.9	no	negative
TA 1537	no	3	1.0	no	negative
	yes	3	1.2	no	negative
WP2 uvr A	no	3	1.1	no	negative
	yes	3	0.9	no	negative

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative