Phosphonium, trihexyltetradecyl-, chloride

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro mutagenicity in bacteria

A single reliable with restrictions (Klimisch 2) key study is available (Micheal S. Mecchi, 2003), performed similar to EPA OTS 798.5265 and conform with GLP requirements. In this study, trihexyltetradecylphosphonium chloride did not show mutagenic activity in the applied bacterium tester strains in the absence or presence of metabolic activation under the conditions of the test system

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-04-02 - 2003-11-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

GLP, guideline study; four bacterial strains used instead of five. Triplicate plating used only for vehicle controls

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

not specified

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Trade name: CYPHOS IL 101 Phosphonium Salt
Physical state: liquid
Composition of test material, percentage of components: Trihexyl(tetradecyl)phosphonium chloride (258864-54-9), > 95%
Lot/batch No.: WE01-001
Expiration date of the lot/batch: >1 year when stored at room temperature and protected from direct contact with water (hydrophobic)

Target gene:

Histidine locus

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from Aroclor-induced rat liver (from Molecular Toxicology, Inc)

Test concentrations with justification for top dose:

1.00, 3.33, 10.0, 33.3, 100, 333, 1000, 3330, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: The test article was observed to form a transparent, colorless, non-viscous solution in ethanol at a concentration of approximately 100 mg per mL.

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene, ICR-191

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Preincubation period:
- Exposure duration: 72 hours at 37°C

NUMBER OF REPLICATIONS: All doses of test article and positive controls were plated at one plate per dose.Vehicle controls were plated in triplicate.

DETERMINATION OF CYTOTOXICITY
- Method: The condition of the background bacterial lawn is evaluated both macroscopically and microscopically (using a dissecting microscope) for indications of cytotoxicity.

Evaluation criteria:

A test material is considered mutagenic if there is a reproducibly increasing dose-response curve of induced revertant colonies for at least 3 test concentrations. The minimal criteria for a positive response are a 2- to 3-fold increase in the number of revertants (at least 15 colonies) over the spontaneous number for the TA1535, TA1537, TA1538 and TA98 strains. In addition, a positive response must not be observed only at concentrations near toxic dose levels.

Species / strain:

S. typhimurium, other: TA1537, TA1535, TA100, TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Table 1: Mutagenicity assay Results - Individual Plate Counts

Concentration (µg test solution)	TA98						TA100						TA1535						TA1537
	-S9			+S9			-S9			+S9			-S9			+S9			-S9			+S9
0 (vehicle control)	11	11	9	27	26	22	59	66	63	90	93	93	14	7	8	15	14	20	10	7	5	6	5	6
	10±1			25±3			63±4			92±2			10±4			16±3			7±3			6±1
1.00	14			20			74			86			6			9			7			7
3.33	12			17			74			106			7			10			10			1
10.0	4			20			54			79			10			17			9			6
33.3	3			20			21			105			3			23			1			5
100	0			14			0			66			0			6			0			4
333	0			0			0			0			0			0			0			0
1000	0			0			0			0			0			0			0			0
3330	0			0			0			0			0			0			0			0
5000	0			0			0			0			0			0			0			0
Positive control
2-Nitrofluorene	243			-			-			-			-			-			-			-
benzo[a]pyrene	-			219			-			-			-			-			-			-
Sodium azide	-			-			1160			-			893			-			-			-
2-Aminoanthracene	-			-			-			477						58			-			80
ICR-191	-			-			-			-			-			-			942			-

Conclusions:

negative with and without metabolic activation

Trihexyltetradecylphosphonium chloride was not mutagenic in a bacterial reverse mutation assay up to cytotoxic concentrations.

Executive summary:

Trihexyltetradecylphosphonium chloride was examined for mutagenicity activity in the Salmonella/Mammalian-Microsome Reverse Mutation Screening Assay (Ames Test). This assay evaluated the test article for its ability to induce reverse mutations at the histidine locus in the genome of specific Salmonella typhimurium tester strains TA-98, TA-100, TA-1535 and TA-1537 both in the presence and absence of an exogenous metabolic activation system.

Positive controls (sodium azide, 2-Nitrofluorene, benzo[a]pyrene, ICR-191 and 2-aminoanthracene) were tested under the same experimental conditions.

No significant increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to the test material at levels from 1.00 to 5000 µg per plate, either in the presence or absence of the S-9 mix.

Under the conditions of this study, the test material Trihexyltetradecylphosphonium chloride was devoid of mutagenic activity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Trihexyltetradecylphosphonium chloride was examined for mutagenicity activity in the Salmonella/Mammalian-Microsome Reverse Mutation Screening Assay (Ames Test). This assay evaluated the test article for its ability to induce reverse mutations at the histidine locus in the genome of specific Salmonella typhimurium tester strains TA-98, TA-100, TA-1535 and TA-1537 both in the presence and absence of an exogenous metabolic activation system.

Positive controls (sodium azide, 2-Nitrofluorene, benzo[a]pyrene, ICR-191 and 2-aminoanthracene) were tested under the same experimental conditions.

No significant increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to the test material at levels from 1.00 to 5000 µg per plate, either in the presence or absence of the S-9 mix.

Under the conditions of this study, the test material Trihexyltetradecylphosphonium chloride was devoid of mutagenic activity.

Justification for classification or non-classification

Based on the result of the available in vitro study, trihexyltetradecylphosphonium chloride is considered not to be mutagenic and is not classified according to the CLP regulation.