4-hydroxy-2,2,6,6-tetramethylpiperidine-1-ethanol

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:

August 18, 1992 - September 10, 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study report which meets basic scientific principles

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-hydroxy-2,2,6,6-tetramethylpiperidine-1-ethanol

EC Number:

258-132-1

EC Name:

4-hydroxy-2,2,6,6-tetramethylpiperidine-1-ethanol

Cas Number:

52722-86-8

Molecular formula:

C11H23NO2

IUPAC Name:

1-(2-hydroxyethyl)-2,2,6,6-tetramethylpiperidin-4-ol

Details on test material:

- Physical state: solid; white
- Storage condition of test material: room temperature

Method

Target gene:

his, trp

Species / strain

Species / strain / cell type:

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

Range in the cytotoxicity test: 20.6 - 5000 µg/plate
Range in the mutagenicity test: 185.2 - 5000 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: see below

Remarks:

with and without metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours

NUMBER OF REPLICATIONS: two plates per test substance concentration

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

POSITIVE CONTROLS
WITHOUT S9:
TA 100, TA 1535: sodium azide in bidist.water, 5.0 µg/plate
WP2 uvrA: 4-nitroquinoline-N-oxide in DMSO, 2.0 µg/plate
TA 98: 2-nitrofluorene in DMSO, 20.0 µg/plate
TA 1537: 9(5)-aminoacridine in DMSO, 150.0 µg/plate

WITH S9
TA 100, TA 98, TA 1537: 2-aminoanthracene in DMSO, 2.5 µg/plate
TA 1535: cyclophosphamide in bidist.water, 400.0 µg/plate
WP2 uvrA: 2-aminoanthracene in DMSO, 50.0 µg/plate

Evaluation criteria:

The test substance is considered to be mutagenic in this test system if:
- A positive effect is observed in one strain and the effect can be reproduced in a confirmatory experiment.
- A positive effect is observed in two or more strains.
A positive effect is defined as an increase of the mean number of revertants per plate for any concentration above that of the negative control by at least a factor of 2.0 with strains TA 98, TA 1535, TA 1537 and WP2 uvrA, or by a factor of at least 1.5 with strain TA 100. Generally a concentration-related effect should be demonstrable. If equivocal results are obtained, the final decision has to be based on scientific judgement.

Results and discussion

Test results

Species / strain:

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

RANGE-FINDING/SCREENING STUDIES:
Six concentrations ranging from 20.6 to 5000 µg/plate were tested with strains S. typhimurium TA 100 and E.coli WP2 uvrA to determine the highest concentration to be used in the mutagenicity assay. The experiments were performed with and without metabolic activation. Normal background growth was observed with both strains. The numbers of revertant colonies were not reduced. From the results obtained, the highest concentration suitable for the mutagenicity test was selected to be 5000 µg/plate without and with metabolic activation.

COMPARISON WITH HISTORICAL CONTROL DATA:
Negative Controls were within the historical control range and positive controls produced the expected reactions.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the mutagenicity test normal background growth was observed with all strains at all concentrations. The numbers of revertant colonies were not reduced. The test substance exerted no toxic effect on the growth of the bacteria.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Experiment with metabolic activation:

Treatment/Strain	TA100	TA1535	WP2 uvrA	TA98	TA1537
Negative control	100.0	10.0	15.5	41.5	7.0
185.2 µg/plate	117.5	17.0	20.5	40.5	7.0
555.6 µg/plate	115.0	9.5	15.5	41.5	5.5
1666.7 µg/plate	118.0	11.0	14.0	37.0	4.5
5000.0 µg/plate	89.0	12.0	16.5	44.0	7.0
2-aminoanthracene	841.5		573.5	1086.0	139.5
cyclophosphamide		431.5

Experiment without metabolic activation:

Treatment/Strain	TA100	TA1535	WP2 uvrA	TA98	TA1537
Negative control	85.5	13.0	14.0	17.0	6.0
185.2 µg/plate	72.0	16.0	17.0	18.0	5.0
555.6 µg/plate	84.0	12.0	17.5	19.5	3.5
1666.7 µg/plate	84.0	16.5	14.5	12.5	3.5
5000.0 µg/plate	67.5	17.5	13.5	12.5	4.5
sodium azide	1066.0	790.0
4-NQO			789.0
2-nitrofluorene				1230.5
9-aminoacridine					1549.5

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Based on the results of these experiments and on standard evaluation criteria, it is concluded that the test item and its metabolites did not induce gene mutations in the strains of S. typhimurium and E. coli used.