α,α-diphenylpiperidine-4-methanol hydrochloride

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:

January 21 to March 06 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD, GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Azacyclonol

EC Number:

204-092-5

EC Name:

Azacyclonol

Cas Number:

115-46-8

IUPAC Name:

diphenyl(piperidin-4-yl)methanol

Test material form:

solid - liquid: suspension

Details on test material:

- Name of test material (as cited in study report): 4-Piperidine Diphenyl Methanol (Azacyclonol) Base
- Physical state: white grain powder
- Analytical purity: 98.3 % (w/w)
- Lot/batch No.: 86000298
- Date of expiry: September 10, 1998
- Storage condition of test material: darkness at approximately 5 °C in refrigerator, protected from light
- Soluble and stable in DMSO.

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: Strains are deficient in the structure of their lipopolysaccharide layer and in DNA excision repair system. TA 98 and TA 100 also possess a modified postreplication DNA repair system which frequently causes an increase in the rate of mutations.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver post-mitochondrial fraction (S-9)

Test concentrations with justification for top dose:

First mutation experiment:
With and without metabolic activation: 4, 20, 100, 500, 2500 and 5000 µg/plate
Second mutation experiment:
With and without metabolic activation: 0.8, 4, 20, 100, 500, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

Solvent

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA1535 and TA 100 without S-9

Controls 2

Untreated negative controls:

yes

Remarks:

Solvent

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without S-9

Controls 3

Untreated negative controls:

yes

Remarks:

Solvent

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 without S-9

Controls 4

Untreated negative controls:

yes

Remarks:

Solvent

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA1535, TA 1537, TA 100 and TA 98 with S-9

Details on test system and experimental conditions:

A toxicity test using histidine-enriched agar plates and a dilution of the tester strain TA 100 (designated TA 100 D) was performed in parallel with the second mutation experiment.

Evaluation criteria:

Criteria for a valid assay:
The assay is considered valid if the following criteria are met:
- the solvent control data are within the laboratory's normal control range for the spontaneous mutant frequency.
- the positive controls induced increases in the mutation frequency which were bath statistically significant and within the laboratory's normal range.

Criteria for a positive response:
A test compound is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn.
b) it induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.
If the test substance does not achieve either of the above criteria, it is considered to show no evidence of mutagenic activity in this system.
The test results must be reproducible.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Strain-dependent toxicity was observed in a dose range of 2500 to 5000 µg/plate.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

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

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Strain-dependent toxicity was observed in a dose range of 500 to 2500 µg/plate and above without metabolic activation.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The test compound proved to be toxic to the bacterial strain TA 100 D in the presence and in the absence of metabolic activation at concentration of 2500 µg/plate.
Only in the first experiment an increased number of revertants was found in the tester strain TA 1537 in the absence of S9 -mix, but this effect was caused by the law spontaneous mutation frequency of the corresponding solvent control and therefore considered to be without biological relevance.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Azacyclonol is not mutagenic in these bacterial test systems either in the absence or in the presence of an exogenous metabolizing system.