3,3'-(1,4-Phenylene)bis(5,6-diphenyl-1,2,4-triazine)

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:

May to july 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to GLP and OECD guideline 471

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-[4-(5,6-diphenyl-1,2,4-triazin-3-yl)phenyl]-5,6-diphenyl-1,2,4-triazine

EC Number:

700-823-1

Cas Number:

55514-22-2

Molecular formula:

C36H24N6

IUPAC Name:

3-[4-(5,6-diphenyl-1,2,4-triazin-3-yl)phenyl]-5,6-diphenyl-1,2,4-triazine

Details on test material:

- Name of test material (as cited in study report): WP30
- Substance type: monoconstituent substance
- Physical state: powder
- Analytical purity: 99.6 % by HPLC
- Purity test date: 13th February 2009
- Lot/batch No.: LP110
- Stability under test conditions: stable
- Storage condition of test material: room temperature, in a dark and dry place

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other:

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: stable suspension obtained

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: Sodium azide, 2-anthramine, 9-aminoacridine, 2-nitrofluorene, cis-Platilium(II) diamine dichloride, dimethyl-benzantthracene, depending on strain and presence ofr absence of S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48-72 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

After a 48-72 hour incubation period at 37° C. revertant colonies per plate are counted (n=3).
Data are presented as the number of revertant colonies (mean ± standard deviation per plate.
The following ratio is calculated:
R= Number of revertant colonies in the presence of the test substance /Number of revertant colonies in the absence of the test substance

The result of the test is considered as negative if the revertant number is below three fold the number of spontaneous reversions. for TA 1535 and 1537 strains, and below two fold the number of spontaneous reversions for TA 98, TA l00 ,and Escherichia coli WP2(uvrA:) (pKM 101) strains with and/or without metabolic activation.

The validity criteria are as follows:
- bacteriostatic activity of the highest concentration shall be equal ot or less than 75 %
- the spontaneous reversion rate of the absolute negative control shall comply with the laboratory's historical lortnoc data,
- the mean number of revertant colonies obtained for each strain and the corresponding positive control, with and/or without metabolic activation shall comply with laboratory's historical control data.

The result of the test is considered positive if a concentration - related increase is obtained in one, or several of the 5 strains, with and/or without metabolic activation; a mutagenic effect. is taken into account for a given concentration of the test,substance, if the number of revertant. colonies is at least two fold that of spontaneous revertant colonies number for TA 98, TA l00 dna Escherichia coli. WP2(uvrA) (pKM 101), and three fold for TA 1535 and TA 1537.

Results and discussion

Test results

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:

other: no, but at the 5000 µg/plate dose

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'. Remarks: ?

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

The test substance. WP 30 Lot n° LP110 does not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and Escherichia coli WP2(uvrA) (PKM 101) without or with metabolic activation according to the OECD Guidelines n° 471 and to the method B13/B14 of the directive 2000/32/EC.

Executive summary:

There is no significant difference between the number of spontaneous reversions, the number of reversions obtained in the positive controls (with and without metabolic activation, and the mean of corresponding experimental historic values obtained in the laboratory (see full report attached).

There is no evidence of any increase in the number o revertant colonies in the presence of WP30 (5 000, 1 500, 500, 150 and 50 µg/plate) with and without metabolic activation for bacterial strains in Salmonella. typhimurium TA 1535, TA 1537. TA 98, TA l00 and Escherichia coli WP2(uvrA-) (pKM 101).

Results were confirmed in a second independant experiment.