2,2'-phenyliminodiethanol

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment

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:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-phenyliminodiethanol

EC Number:

204-368-5

EC Name:

2,2'-phenyliminodiethanol

Cas Number:

120-07-0

Molecular formula:

C10H15NO2

IUPAC Name:

2-[(2-hydroxyethyl)(phenyl)amino]ethan-1-ol

Details on test material:

- Name of test material (as cited in study report): N,N-Di-(hydroxyethyl)-anilin
- Batch No.: 59-7325
- Purity: 97.7%
- Appearance, consistency: brown, solid
- Storage: room temperature

Method

Target gene:

- S. typhimurium: His-locus
- E. coli: Trp-locus

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 fraction of Aroclor 1254 induced rats

Test concentrations with justification for top dose:

- 1st experiment (standard plate test, all strains): 20, 100, 500, 2500, 5000 µg/plate
- 2nd experiment (preincubation test, all stains): 4, 20, 100, 500, 1000 µg/plate
- 3rd experiment (standard plate test, TA 98 and TA 100): 20, 100, 500, 100, 2000 µg/plate
- 4th experiment (standard plate test, TA 98): 750, 1000, 1500, 2000, 2500 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:

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

other: 2-aminoanthracene (2AA); aniline/o-toluidine; N-methyl-N'-nitro-N-nitrosoguanidine (MNNG); 4-nitro-o-phenylendiamine (NOPD)

Remarks:

With metabolic activation: 2AA (all strains); aniline/o-toluidine (tests with norharman); Without metabolic activation: MNNG (TA 100, TA1535); NOPD (TA 98); 9-aminoacridine (TA 1537); N-ethyl-N'-nitro-N-nitrosoguanidin (E.Coli)

Details on test system and experimental conditions:

-> 1st EXPERIMENT:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION: exposure duration: 48-72 hours
STRAINS TESTED: TA 1535, TA 1537, TA 98, TA 100, E.coli WP2

-> 2nd EXPERIMENT:
METHOD OF APPLICATION: preincubation
DURATION:
- Preincubation period: 20 minutes
- Exposure duration: 48-72 hours
STRAINS TESTED: TA 1535, TA 1537, TA 98, TA 100, E.coli WP2

-> 3rd EXPERIMENT:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION: exposure duration: 48-72 hours
STRAINS TESTED: TA 98, TA 100
OTHER: test with S9 mix incl. norharman

-> 3rd EXPERIMENT:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION: exposure duration: 48-72 hours
STRAINS TESTED: TA 98
OTHER: test with S9 mix incl. norharman

NUMBER OF REPLICATIONS: 3 test plates per dose or per control

DETERMINATION OF CYTOTOXICITY: Reduced his- background growth, decrease in the number of his revertants)

Evaluation criteria:

In general, a substance to be characterized as positive in the bacterial tests has to fulfil the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results

Test results 1

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:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON GENOTOXICITY
An increase in the number of his+ or trp+ revertants was not observed both in the routine standard plate test and in the preincubation test either without S9 mix or after the addition of a metabolizing system. However, after the addition of norharman there was a slight but dose-dependent increase in the number of mutant colonies from about 750 - 1500 µg/plate onward up to 2000 - 2500 µg/plate (factor 2.1 - 2.8) using the strain TA 98.

TEST-SPECIFIC CONFOUNDING FACTORS
Test substance precipitation was found from about 2500 µg/plate onward

CYTOTOXICIY
A bacteriotoxic effect (reduced his- background growth, decrease in the number of his revertants) was observed in the standard plate test depending on the strain and test conditions at doses ≥ 500 µg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative