2,2`-[6-(phenylamino)-heteromonocycle-2,4-diyl]diphenol

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:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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)

Version / remarks:

adopted July 21,1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Commission Directive 2000/32/EC, L1362000,Annex 40

Version / remarks:

May 19, 2000

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines: "Kanpoan No. 287 -- Environment Protection Agency" "Eisei No. 127 -- Ministry of Health & Welfare" "Heisei 09/10/31 Kikyoku No. 2 -- Ministry of International Trade & Industry"

Principles of method if other than guideline:

study performed as plate incorporation test (experiment I) and as pre-incubation test (experiment II);

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2`-[6-(phenylamino)-1,3,5-triazine-2,4-diyl]diphenol

Cas Number:

1248-66-4

Molecular formula:

C21H16N4O2

IUPAC Name:

2,2`-[6-(phenylamino)-1,3,5-triazine-2,4-diyl]diphenol

Method

Target gene:

TA 1537: his C 3076; rfa-; uvrB-
TA 98: his D 3052; rfa-; uvrB-; R-factor
TA 1535: his G 46, rfa-; uvrB-;
TA100: his G 46; rfa-; uvrB-, R-factor
WP2 uvrA: trp-. uvrA-

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

from rat liver S9

Test concentrations with justification for top dose:

Pre-experiment/ experiment I: 3, 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate;
experiment II: 33, 100, 333, 1000, 2500 and 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

with and without activation

Controls 2

Positive controls:

yes

Remarks:

TA 1535, TA 100

Positive control substance:

sodium azide

Remarks:

without activation

Controls 3

Positive controls:

yes

Remarks:

TA 1537, TA 98

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

without activation

Controls 4

Positive controls:

yes

Remarks:

WP2 uvrA

Positive control substance:

methylmethanesulfonate

Remarks:

without activation

Controls 5

Positive controls:

yes

Remarks:

TA 1535, TA 1537, TA 98, TA 100, WP2 uvrA

Positive control substance:

other: 2-aminoanthracene

Remarks:

with activation

Controls 6

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Remarks:

with and without activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation;
DURATION
- Preincubation period: 60 min at 37°C
- Exposure duration: 48 h at 37°C

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

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:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Under the conditions of the assay, the test item is non-mutagenic in the bacterial reverse mutation assay.

Executive summary:

The test item was assessed for its potential to induce gene mutations in the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, and the Escherichia coli strain WP2 uvrA.
The assay was performed in two independent experiments both with and without liver
microsomal activation. Each concentration and the controls were tested in triplicate. The
test item was tested at the following concentrations:
Pre-Experiment/Experiment I: 3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate
Experiment II: 33; 100; 333; 1000; 2500; and 5000 µg/plate
The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without metabolic activation in both independent experiments.
No toxic effects, evident as a reduction in the number of revertants, occurred in the test groups with and without metabolic activation in experiment I. In experiment II, toxic effects were observed in strain TA 1537 at 5000 µg/plate without metabolic activation and from 1000 up to 5000 µg/plate with metabolic activation.
No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with test item at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
Appropriate reference mutagens were used as positive controls. They showed a distinct increase of induced revertant colonies.
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.