Cetalkonium chloride

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

 

Ames test:

 

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The test chemical was dissolved in distilled water or DMSO as the solvent and used at dose level of 0, 0.01, 0.05, 0.1, 0.5, 1, 5 or 10 µg/plate. The exposure duration was 48 hrs at 37˚C. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer reviewed publication.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA98 and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 metabolic activation system

Test concentrations with justification for top dose:

0, 0.01, 0.05, 0.1, 0.5, 1, 5 or 10 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Distilled water or DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO or distilled water

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Distilled water or DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 4-nitroquinoline-N-oxide benzo(a)pyrene other: N-Methyl-N’-nitro-N-nitrosoguanidine, 2-Acetylaminofluorene, N-Nitrosodimethylamine

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar

DURATION
- Preincubation period: No data
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: No data

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

Rationale for test conditions:

Not specified

Evaluation criteria:

The plates were observed or a dose dependent increase in the number of revertants/plate

Statistics:

Mean value was observed.

Species / strain:

S. typhimurium, other: TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY:The test compound was toxic at dose level of 5 and 10 µg/plate in the absence of S9 metabolilc activation system

Remarks on result:

other: No mutagenic potential

Table 1. Mutagenicity of the test compound and the respective control chemicals

Compound	Dose (µg/plate)	No. of His+ revertants/plate
		TA100		TA98
		-S9	+S9	-S9	+S9
Distilled water	-	145	137	19	32
DMSO	-	151	141	23	28
4-Nitroquinoline 1-oxide	0.5	1588	148	167	36
Benzo[a]pyrene	2	4152	128	27	30
2-Acetylaminofluorene	5	178	928	35	614
N-Nitrosodimethylamine	50	128	1080	18	2292
Cetyltrimethylammonium chloride	0.01	123	139	12	29
	0.05	133	108	13	22
	0.1	105	120	10	40
	0.5	100	115	17	18
	1	96	129	15	26
	5	0*	132	0*	28
	10	0*	91	0*	18

Conclusions:

The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of test substance. The test chemical was dissolved in distilled water or DMSO as the solvent and used at dose level of 0, 0.01, 0.05, 0.1, 0.5, 1, 5 or 10µg/plate. The exposure duration was 48 hrs at 37˚C. Concurrent solvent and negative control chemicals were also included in the study. Test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Gene mutation in vitro:

Data available for the test chemical was reviewed to determine the mutagenic nature of Benzyldimethylhexadecylammonium chloride (CAS no 122 -18 -9). The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The test chemical was dissolved in distilled water or DMSO as the solvent and used at dose level of 0, 0.01, 0.05, 0.1, 0.5, 1, 5 or 10 µg/plate. The exposure duration was 48 hrs at 37˚C. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In the same study, In vitro mammalian cell transformation assay was performed to determine the mutagenic nature of the test chemical. The test chemical was dissolved in DMSO as the solvent and used at dose level of 0, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 50, 100 or 300µg/mL. Pregnant Syrian golden hamsters were killed on days 13 and 14 of gestation for preparation of target cells and feeder-layer cells respectively. Embryos without heads and viscera were minced with scissors,a nd trypsinized with 0.25% trypsin. Inocula of 10000000 embryo cells per 75-cm’ flask were incubated at 37°C in a humidified atmosphere of 10% CO2 in air. When they became confluent primary cultures were trypsinized, dispensed in lots of 5000000 cells in glass ampules, and stored in liquid nitrogen for use as target and feeder-layer cells. The assay takes 15 days from start to finish. On Day 0, an ampule of cryopreserved primary cells prepared as feeder-layer cells was rapidly thawed and plated in a 75-cm2 flask containing 20 ml of the culture medium. The medium was changed every day. On day 3, an ampule of cryopreserved primary cells prepared as target cells was also rapidly thawed and plated in a 75-cm2 flask. On day 4, the feeder cells which were shifting from a stage of logarithmic growth to a stationary phase were irradiated with 5000 R from a linear accelerator, trypsinized, and then plated at 6 x lo4 tells/60-mm dish in 2 ml of complete medium. On day 5, the target cells which were approximately 80 -90% confluent were trypsinized, and a suspension of 500 target cells in 2 ml of complete medium was then added to each of the dishes plated the day before with irradiated feeder-layer cells. On day 6, an appropriate dose of the test chemical in a volume of 4 ml was added, giving a total volume of 8 ml of medium in each dish. Nine dishes were used for each dose level in all the experiments (in a few cases only eight or seven dishes were used for controls). On day 14, the cultures werefixed with absolute methanol for 10min and stained with Giemsa solution for 45 min or more. The stained dishes were examined with a stereoscopic dissection microscope to count normal and transformed colonies. The exposure duration was 8 days. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce cell transformation in the Pregnant Syrian golden hamsters embryo cell line used and hence it is not likely to classify as a gene mutant in vitro.

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The test chemical was dissolved in distilled water or DMSO as the solvent and used at dose level of 0, 0.05, 0.2, 1, 10 or 50µg/plate in experiment 1 and 2 and at dose level of 0.01, 0.05, 0.2, 1.0 or 5.0µg/plate in experiment 3. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of rat, hamster and guinea pig isolated S9 metabolic activation system and also in the presence and absence of norharman and hence it is not likely to classify as a gene mutant in vitro.

Based on the data available for the test chemical, Benzyldimethylhexadecylammonium chloride (CAS no 122 -18 -9) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical, Benzyldimethylhexadecylammonium chloride (CAS no 122 -18 -9) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.