3-Morpholinone, 4-(4-aminophenyl)-

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:

other information

Study period:

Feb 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well reported GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

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:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-(4-aminophenyl)morpholin-3-one

EC Number:

610-147-8

Cas Number:

438056-69-0

Molecular formula:

C10 H12 N2 O2

IUPAC Name:

4-(4-aminophenyl)morpholin-3-one

Details on test material:

Anilinomorpholinon, fine yellow powder, batch number BXR3AR1, 96.2% content

Method

Target gene:

histidin gene locus

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

Anilinomorpholinon: 50-5000 µg/plate
Sodium-azide: 10 µg/plate (TA 1535)
Nitrofurantoin: 0.2 µg/plate (TA 100)
4-Nitro-1,2-phenylene diamine: 10 µg/plate (TA 1537), 0.5 µg/plate (TA 98)
Mitomycin C: 0.2 µg/plate (TA 102)
2-Aminoanthracene: 3 µg/plate

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

No solvent control was used since sufficient evidence was available in the literature and from testing laboratory experience, indicating that the solvents used had no influence on the spontaneous mutant counts of the used strains.

Positive controls:

yes

Positive control substance:

other: Sodium-azide, Nitrofurantoin, 4-Nitro-1,2-phenylene diamine, Mitomycin C, 2-Aminoanthracene

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA 1535 and TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA 100 and TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

lowest effective dose for TA 100: 158 µg/plate; lowest effective dose for TA 98: 500 µg/plate

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 102

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

lowest effective dose: 500 µg/plate

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: TA 1535 and TA 1537

Any other information on results incl. tables

There was no indication of a bacteriotoxic effect of Anilinomorpholinon up to and including 5000 µg/plate. No inhibition of growth was noted as well.

Applicant's summary and conclusion

Executive summary:

Anilinomorpholinon was investigated using the Salmonella/microsome plate incorporation test for point-mutagenic effects in doses up to and including 5000 µg/plate on the five histidine-auxotrophic Salmonella typhimurium LT2 strains TA 1535, TA 100, TA 1537, TA 98 and TA 102.

Doses up to and including 5000 µg/plate did not cause any bacteriotoxic effects. No inhibition of growth was noted as well.

Evidence of mutagenic activity of Anilinomorpholinon was seen. On Salmonella typhimurium TA 98, 100 and TA 102, a biologically relevant increase was found in the mutant count compared to the corresponding negative control. Positive response was found with and without S9 mix for TA 98 and TA 100 (effect was stronger with S9 mix) and only with S9 mix for TA 102. The lowest effective dose was 158 µg/plate for S. typhimurium TA 100 and 500 µg/plate for TA 98 and TA 102. The Salmonella/microsome test thus showed Anilinomorpholinon to have a mutagenic effect.