Hexahydrocyclopenta[c]pyrrol-2(1H)-amine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

20 to 22nd August 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Test material form:

liquid

Specific details on test material used for the study:

batch 0048M-97F28

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Test concentrations with justification for top dose:

Each strain was exposed to five dose levels of the test substance (3 plates/dose-level) :
First experiment :
250, 500, 1000, 2000 and 4000 µg/plate : for the TA 1535, TA 1537, TA 98 and TA 100 strains without S9 mix
312.5, 625, 1250, 2500 and 5000 µg/plate : for the TA 102 strain without S9 mix and for all tester strains with S9 mix
Second experiment :
125, 250, 500, 1000 and 2000 µg/plate : for the TA 1535, TA 1537, TA 98 and TA 100 strains without S9 mix
312.5, 625, 1250, 2500 and 5000 µg/plate : for the TA 102 strain without S9 mix and for all tester strains with S9 mix

Vehicle / solvent:

Distilled water

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-Anthramine

Details on test system and experimental conditions:

see attached study report

Rationale for test conditions:

The experiments were performed according to the direct plate incorporation method (preliminary toxicity test and both mutagenicity experiments) : test substance solution (0.05 to 0.2ml), S9 mix (0.5ml) when riquired and bacterial suspension (0.1ml) were mixed with 2ml of overlay agar (containing traces of th relevant aminoacid and biotin and maintained at 45°C). Afetr rapid homogenization, the mixture was overlaid onto a Petri plate containing minimum medium.
After 48 to 72hours of incubation at 37°C, revertants were scored with an automatic counter.

Evaluation criteria:

Treatment of the results :
In each experiment, for each strain and for each experimental point, the number of revertants per plate was scored. The individual results and the mean number of revertants, with the corresponding standard deviation and ratio (mutants obtained in the presence of the test substance/mutants obtained in the presence of the vehicle), are presented in a tale.
Acceptance criteria :
This study would be considered valid since the following criteria ar fully met :
- the number of revertants in the vehicle controls is consistent with our historical data)
- the number of revertants in the positive controls is higher than that of vehicle controls and is consistent with our historical data.

Evaluation criteria :
A reproductible two-fold increase in the number of revertants compared with the vehicle controls, in any strain at any dose-level and/or evidence of a dose-relationship was considered as a positive result. Reference to historical data, or other considerations of biological relevance may also be taken into account in the evaluation of the data obtained

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

positive

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

positive

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

positive

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Under experimental conditions, the test substance 852 AMINAZA PURIFIEE showed mutagenic activity in this bacterial reverse mutation test on salmonella typhimurium