Tris{4-[(4-acetylphenyl)sulfanyl]phenyl}sulfonium hexafluorophosphate

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 guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

hexafluoro-λ⁵-phosphanuide; tris({4-[(4-acetylphenyl)sulfanyl]phenyl})sulfanium

EC Number:

700-373-6

Cas Number:

953084-13-4

Molecular formula:

C42 H33 O3 S4 . F6 P

IUPAC Name:

hexafluoro-λ⁵-phosphanuide; tris({4-[(4-acetylphenyl)sulfanyl]phenyl})sulfanium

Method

Target gene:

His (Salmonella typhimurium)
Trp (E. coli)

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA

Metabolic activation:

with and without

Metabolic activation system:

phenobarbital and β-naphthoflavone - induced rat liver S9 mix

Test concentrations with justification for top dose:

1st Experiment (SPT): 0; 22; 110; 550; 2750 and 5500 μg/plate (TA 1535, TA 100, TA 1537, TA 98, E. coli WP2 uvrA)
2nd Experiment (SPT): 0; 0.4; 2; 10; 50 and 100 μg/plate (TA 1535, TA 100, TA 1537); 0; 0.8; 4; 20; 100 and 200 μg/plate (TA 98). Reason: Strong bacteriotoxicity was observed in the 1st Experiment.
3rd Experiment (PIT): 0; 0.2; 1; 5; 25 and 50 μg/plate (TA 1535, TA 100, TA 1537); 0; 0.4; 2; 10; 50 and 100 μg/plate (TA 98); 0; 22; 110; 550; 2750 and 5500 μg/plate (E. coli)

Vehicle / solvent:

Dimethylsulfoxide (DMSO)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

other: 2-aminoanthracene

Details on test system and experimental conditions:

POSITIVE CONTROLS:
With S9 mix:
• 2-aminoanthracene (2-AA)
- 2.5 μg/plate, dissolved in DMSO
- strains: TA 1535, TA 100, TA 1537, TA 98
- 60 μg/plate, dissolved in DMSO
- strain: Escherichia coli WP2 uvrA
Without S9 mix:
• N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
- 5 μg/plate, dissolved in DMSO
- strains: TA 1535, TA 100
• 4-nitro-o-phenylenediamine (NOPD)
- 10 μg/plate, dissolved in DMSO
- strain: TA 98
• 9-aminoacridine (AAC)
- 100 μg/plate, dissolved in DMSO
- strain: TA 1537
• 4-nitroquinoline-N-oxide (4-NQO)
- 5 μg/plate, dissolved in DMSO
- strain: E. coli WP2 uvrA

METHOD OF APPLICATION: in agar

DURATION
- Exposure duration: 48 – 72 hours at 37 °C

NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY: by a decrease in the number of revertants, clearing or diminution of the background lawn (= reduced his- or trp- background growth), reduction in the titer

Evaluation criteria:

Generally, the experiment is considered valid if the following criteria are met:
• The number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control items both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
• The titer of viable bacteria was ≥ 10 e8/mL.

The test item is considered positive in this assay if the following criteria are met:
• A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9 mix or after adding a metabolizing system.
A test item is generally considered non-mutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in at least two experiments carried out independently of each other.

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

depending on the strain and test conditions

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY: A bacteriotoxic effect was occasionally observed depending on the strain and test conditions from about 10 μg/plate onward. No bacteriotoxicity was observed with E. coli WP2uvrA up to the highest applied dose of 5500 μg/plate in the absence and presence of metabolic activation. In the preincubation assay bacteriotoxicity was observed depending on the strain and test conditions from about 25 µg/plate onward using four salmonella tester strains. In the preincubation assay with E. coli WP2uvrA weak bacteriotoxicity was observed only in the presence of metabolic activation at 5500 μg/plate.

PRECIPITATION: Test item precipitation was found from 550 μg/plate onward with and without S9 mix.

MUTAGENICITY: The test item did not lead to a relevant increase in the number of revertant colonies either without S9 mix or after adding a metabolizing system in three experiments carried out independently of each other (standard plate test and preincubation assay). (see table 1)

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Number of revertants in the control or after treatment with the test substance in the a.) standard plate test and b.) preincubation test

a.) Standard plate test (0.4 - 100 µg/plate)
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 1535	no	20	0.9	no	negative
	yes	17	0.9	no	negative
TA 100	no	121	1.0	no	negative
	yes	132	1.1	no	negative
TA 1537	no	12	1.0	no	negative
	yes	14	0.9	no	negative
TA 98	no	26	1.1	no	negative
	yes	29	1.0	no	negative
b.) Standard plate test (22 - 5500 µg/plate)
E. coli WP2 uvrA	no	47	1.1	no	negative
	yes	50	1.1	no	negative
Preincubation test (0.2 - 50 µg/plate)
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 1535	no	14	1.0	no	negative
	yes	16	0.9	no	negative
TA 100	no	95	1.0	no	negative
	yes	97	1.0	no	negative
TA 1537	no	9	0.9	no	negative
	yes	9	0.9	no	negative
TA 98	no	32	1.1	no	negative
	yes	36	1.1	no	negative
E. coli WP2 uvrA	no	40	1.1	no	negative
	yes	38	1.2	no	negative

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative