1,4-Diazabicyclo[2.2.2]octane-2-methanol

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

Study period:

12 - 28 Dec 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Data source

Reference

Reference Type:

other: study report (translation)

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted Jul 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: “Standards for Mutagenicity Tests using Microorganisms” (Notification No. 77 and 120) and the “Amendment of the Reporting Form of the Results of the Mutagenicity Tests using Microorganisms” (Notification No. 653), Ministry of Labour, Japan

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

{1,4-diazabicyclo[2.2.2]octan-2-yl}methanol

EC Number:

692-731-2

Cas Number:

76950-43-1

Molecular formula:

C7H14N2O

IUPAC Name:

{1,4-diazabicyclo[2.2.2]octan-2-yl}methanol

Details on test material:

- Name of test material (as cited in study report): [trade name]
- Substance type: pale yellow solid
- Analytical purity: 90.7% (subsequent information by sponsor)
- Impurities (identity and concentrations): 1,5-diazabicyclo[3.2.2]nonane-3-ol 8.9% (subsequent information by sponsor)
- Storage condition of test material: at room temperature in the dark, under nitrogen substitution
- Stability: stable at room temperature and shading (not reactive to lights nor to air)

Method

Target gene:

his operon (for S. typhimurium strains)
trp operon (for E. coli strain)

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:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone

Test concentrations with justification for top dose:

preliminary study: 1.2, 4.9, 20, 78, 313, 1250 and 5000 μg/plate
main test 1 and 2: 313, 625, 1250, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle used: water
- Justification for choice of solvent/vehicle: Based on the information from the sponsor that the test substance was soluble at 5% and more in water.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: benzo[a]pyrene (B[a]P), 2-aminoanthracene (2AA), 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2), 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine x 2HCl (ICR-191), sodium azide (NaN3)

Remarks:

+S9: B[a]P (5 µg/plate, TA100, TA98, TA1537), 2AA (2 µg/plate, TA1535; 10 µg/plate, WP2 uvrA); -S9: AF-2 (0.01 µg/plate, TA100, WP2 uvrA; 0.1 µg/plate, TA98); ICR-191 (1 µg/plate, TA1537); NaN3 (0.5 µg/plate, TA1535)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition

Evaluation criteria:

If the number of revertant colonies on the test plates increased significantly in comparison with that on the control plates (based on twice as many as that of the negative control), and dose-response and reproducibility were also observed, the test substance was to be judged positive.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

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 nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

PRECIPITATION
No precipitation on the plates was observed.

RANGE-FINDING/SCREENING STUDIES:
No growth inhibition by the test substance was observed in any strain with and without metabolic activation. Therefore, as the highest dose level of the test substance in the main tests, the 5000 μg/plate dose was selected for all strains.

COMPARISON WITH HISTORICAL CONTROL DATA:
The revertant colonies of the positive controls and solvent controls were within the limits (mean±3SD) of historical control data.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative