2-(4,6-diphenyl-1,3,5-triazin-2-yl)-5-((hexyl)oxy)phenol

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:

1992 - 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Reference 2

Reference Type:

other: amendment to CIBA-GEIGY924089 ["test dates"]

Title:

Unnamed

Year:

1994

Report date:

1994

Reference 3

Reference Type:

other: amendment to CIBA-GEIGY924089 ["Purity" and "Expiration date"]

Title:

Unnamed

Year:

1994

Report date:

1994

Reference 4

Reference Type:

other: amendment to CIBA-GEIGY924089 ["Appearance"]

Title:

Unnamed

Year:

1995

Report date:

1995

Reference 5

Reference Type:

other: synopsis of CIBA-GEIGY924089

Title:

Unnamed

Year:

1992

Report date:

1992

Reference 6

Reference Type:

other: expert statement to CIBA-GEIGY924089

Title:

Unnamed

Year:

1993

Report date:

1993

Reference 7

Reference Type:

other company data

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-(4,6-diphenyl-1,3,5-triazin-2-yl)-5-((hexyl)oxy)phenol

EC Number:

411-380-6

EC Name:

2-(4,6-diphenyl-1,3,5-triazin-2-yl)-5-((hexyl)oxy)phenol

Cas Number:

147315-50-2

Molecular formula:

C27 H27 N3 O2

IUPAC Name:

2-(4,6-diphenyl-1,3,5-triazin-2-yl)-5-(hexyloxy)phenol

Method

Target gene:

In the Salmonella typhimurium strains (TA 1535, TA 100, TA 1537, TA 98) the amino acid histidine locus is the target gene, in which induced back mutations will transform the histidine auxotrophy (his-) to histidine prototrophy (his+). The Salmonella typhimurium histidine (his) reversion system measures his- => his+ reversions. The Salmonella typhimurium strains are constructed to differentiate between base pair (TA 1535, TA 100) and frameshift (TA 1537, TA 98) mutations. The strain Escherichia coli (WP2 uvrA) is a tryptophan-auxotrophic strain (base-pair substitution).

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:

S9 mix (from liver homogenate of Aroclor 1254 treated rats)

Test concentrations with justification for top dose:

0, 312.5, 625, 1250, 2500, and 5000 µg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent used: DMSO (50 mg/mL, suspension)
- Justification for choice of solvent/vehicle: DMSO was chosen because of its solubility properties and its relative nontoxicity
- On the day of the experiment (immediately before the experiment), the test item was dissolved in DMSO

Controls

Untreated negative controls:

other: = vehicle control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

other: sodiuim azide, 4-NQO, 2-nitrofluorene (strains except TA 1537), 9-aminoacridine (TA 1537) - Without metabolic activation; 2-aminoanthracene (all strains), cyclophosphamide (all strains except TA 1537) - With metabolic activation

Remarks:

The solvent alone was used as the negative control.

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate method, no preincubation

Evaluation criteria:

The generally accepted conditions for the evaluation of the results are:
- mean colony counts of the control values of all strains are within the acceptable ranges
- results of the positive controls meet the criteria for a positive response

The test item is considered as mutagen in case of:
- reproducible doubling of the mean number of revertants per plate above that of the negative control at any concentration; Strains: S. typhimurium TA 98, TA 1535, TA 1537 and E. coli WP2 uvrA
- reproducible increase of the mean number of revertants per plate for any concentration above that of the negative control by at least a factor of 1.5 for strain S. typhimurium TA 100
- a concentration-related effect should be demonstrable.

Statistics:

A statistical analysis was not performed.

Results and discussion

Test results

Test results 1

Key result

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:

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

Vehicle controls validity:

valid

Untreated negative controls validity:

other: = vehicle control

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

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

Vehicle controls validity:

valid

Untreated negative controls validity:

other: = vehicle control

Positive controls validity:

valid

Additional information on results:

Concentration of test substance resulting in precipitation: approximately 600 - 5000.0 µg/plate.

Any other information on results incl. tables

Test item EC 411-380-6: Salmonella typhimurium/E.coli reverse mutation assay
Plate incorporation without S-9 mix: experiment I/II
Dose (µg/plate)	TA 100	TA 1535	WP2 uvrA	TA 98	TA1537
Negative control (solvent alone)	131.7 / 139.3	12.0 / 10.3	17.0 / 14.7	21.7 / 13.0	6.0 / 7.0
312.5	112.7 / 129.3	12.0 / 8.7	17.0 / 16.7	21.7 / 17.3	4.3 /
625	111.0 / 135.0	11.3 / 9.0	16 / 15.3	21.7 / 12.7	6.3 /
1250	112.0 / 112.0	11.3 / 7.0	17 / 14.0	18.0 / 12.3	4.3 /
2500	116.7 / 126.0	8.3 / 7.3	15 / 14.3	19.0 / 17.0	5.0 /
5000	109.3 / 111.7	10.3 / 7.3	19 / 13.3	21.7 / 14.0	3.7 /
Positive controls
- Sodum azide	961.7 / 1092.3	797.3 / 802.0
- 4-NQO			758.0 / 935.7
2-nitrofluorene				1209.7 / 1650.7
9-aminoacridine					1436.3 / 1456.0
Plate incorporation with S-9 mix: experiment I/II
Dose (µg/plate)	TA 100	TA 1535	WP2 uvrA	TA 98	TA1537
Negative control (solvent alone)	140.3 / 140.0	12.0 / 10.3	18.3 / 19.0	46.7 / 30.0	8.7 / 6.7
312.5	131.7/ 133.7	12.0 / 9.3	22.3 / 15.7	37.3 / 27.3	7.7 / 8.7
625	118.7 / 129.7	11.7 / 14.0	22.0 / 14.3	36.0 / 28.3	7.3 / 7.0
1250	120.0 / 112.0	10.7 / 12.7	21.3 / 13.3	38.0 / 22.3	7.3 / 6.7
2500	118.7 / 124.7	13.0 / 10.3	16.7 / 14.0	43.0 / 24.7	8.0 / 7.0
5000	129.0 / 96.7	10.7 / 9.7	16.7 / 20.3	38.3 / 30.0	5.0 / 4.3
Positive control
- 2-Aminoanthracene	1650.7 / 1432.3		1084.3 / 733.3	1917.7 / 1785.7	249.3 / 185.7
- Cyclophosphamide		374.0 / 362.3
Revertants/plate (I / II): mean from three plates

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative

Executive summary:

In a reverse gene mutation assay in bacteria, strains TA 1535, TA 1537, TA 98 and TA 100 of S. typhimurium and Escherichia coli strain WP2 uvrA were exposed to the test item (as described in section 1.2) dissolved in DMSO at concentrations of 312.5, 625, 1250, and 5000.0 µg/plate in the presence and absence of mammalian metabolic activation without pre-incubation. Concurrent positive and solvent controls were included. All concentrations were tested in triplicates. Concentrations of approximately 600 - 5000.0 µg/plate resulted in precipitation of the test item; no cytoxicity was noted up to the limit concentration. The positive controls induced the appropriate responses. No evidence of a mutagenic potential associated with the test substance was observed. This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.