4-aminomethyl-2-methoxyphenol hydrochloride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

July 21, 1997

Deviations:

yes

Remarks:

no pre-experiment for toxicity performed

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-aminomethyl-2-methoxyphenol hydrochloride

EC Number:

230-468-3

EC Name:

4-aminomethyl-2-methoxyphenol hydrochloride

Cas Number:

7149-10-2

Molecular formula:

C8H11NO2.ClH

IUPAC Name:

4-aminomethyl-2-methoxyphenol hydrochloride

Test material form:

other: solid

Details on test material:

- Physical state: white solid
- Analytical purity: 99.77 %(w/w)
- Lot/batch No.: 77

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

his D 3052; rfa-; uvrB-; R-factor frame shift mutations

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

his G46; rfa-; uvrB-; R-factor base paired subst.

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

31.6; 100; 316.2; 1000; 2500; 5000 µg/plate;
as no pre-experiment had been conducted, 5000µg was selected as maximum concentration

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoantracene, 2AA

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

4-nitro-o-phenylene-diamine, 4-NOPD

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

10µg/plate 10% in DMSO

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk
- Cell density at seeding (if applicable): approx. 10E9 cells/mL

DURATION
- Preincubation period: 48h
- Exposure duration:
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):
S9 liver microsomal fraction was prepared by BSL Bioservice GmbH. Male Wistar Rats were induced with Phenobarbital and beta Naphtoflavone.

SPINDLE INHIBITOR (cytogenetic assays):

STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS: 3 plates per strain, dose and controls

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:

NUMBER OF CELLS EVALUATED:

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells):

CRITERIA FOR MICRONUCLEUS IDENTIFICATION:

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:
- Any supplementary information relevant to cytotoxicity:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable):

Evaluation criteria:

A test item is considered as mutagenic if:
- a dose-related increase in the number of revertant occurs and/or
- a reproducible biologically relevant positive response for at least one of the test points occurs
in at least one strain with or without metabolic activation

A biologically relevant increase is described as follows:
- if in strain TA 100 the numbers of reversions is at least twice as high
- if in strain TA 98 the number of reversions is at least three times higher
as compared to the spontaneous rate.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Applicant's summary and conclusion

Conclusions:

Vanillylamin hydrochloride did not cause gene mutations by base pair changes or frameshifts in the genome test strains used.

Executive summary:

Vanillylamin-HCl was investigated for its potential to induce gene mutations according to the plate incorporation test (experiment 1) and the precubation test (experiment 2) using Salmonella thyphimurium strains TA 100 and TA 98. No toxic effects of the test item were observed in both experiments up to the highest concentrations tested with both strains with and without metabolic activation. Under the test conditions Vanillylamin hydrochloride is not muatgenic.