ABSTRACT

Benzyltrimethylammonium chloride was not mutagenic to Salmonella typhimurium, TA100, TA1535, TA98, TA1537 and Escherichia coli WP2 uvrA, with or without an exogeneous metabolic activation system.

Benzyltrimethylammonium chloride slightly increased incidences of structural chromosomal aberrations with an exogeneous metabolic activation system.

SUMMARIZED DATA FROM THE STUDIES

1. Repeat Dose Toxicity １）

Purity	:	98%
Test species/strain	:	Rats/Crj:CD (SD)
Test method	:	Guidelines for 28-day Repeated Dose Toxicity Testing of Chemicals (Japan)
Route	:	Oral
Doses	:	0 (vehicle), 30, 60, 120 mg/kg/day
Vehicle	:	Water
Number of animals/group	:	Males, 5; females, 5
Administration period	:	Males and females, 28 days
Terminal kill	:	Males and females, days 29 or 43
GLP	:	Yes

　 Test results:

One female given 120 mg/kg died during the 4 th week of the dosing period. Increased salivation was observed in males given 60 mg/kg or more and females of the 120 mg/kg group, increased lacrimation and soiled fur were noted in 120 mg/kg males and females, and increased piloerection was observed in females of the 120 mg/kg group. Suppression of body weight gain and decreased food consumption were noted in males given 120 mg/kg. Hematological examination revealed increase of HGB, MCV and MCH levels in males of the 120 mg/kg group. Histopathological examination of the dead animal revealed hepatocellular swelling and eosinophilic bodies. NOELs for repeat dose toxicity are considered to be 30 mg/kg/day for males and 60 mg/kg/day for females.

2. Genetic Toxicity

2-1. Bacterial test １）

Purity	:	> 99.0%
Test species/strains	:	S. typhimurium TA100, TA1535, TA98, TA1537, E. coli WP2 uvrA
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals (Japan)
Procedures	:	Pre-incubation method
Solvent	:	D.W.
Positive controls	:	-S9 mix, AF-2 (TA100, TA98 and WP2 uvrA), Sodium azide (TA1535), 9-Aminoacridine (TA1537) +S9 mix, 2-Aminoanthracene (all strains)
Doses	:	156, 313, 625, 1250, 2500, 5000 μg/plate
S9	:	Rat liver, induced with phenobarbital and 5,6-benzoflavone
Plates/test	:	3
Number of replicates	:	2
GLP	:	Yes

　Test results:

This chemical did not induce gene mutations in the S. typhimurium and E. coli strains.

Genetic effects:
S. typhimurium TA100, TA1535, TA98 and TA1537

	+	?	-
Without metabolic activation:	[ ]	[ ]	[*]
With metabolic activation:	[ ]	[ ]	[*]

E. coli WP2 uvrA

	+	?	-
Without metabolic activation:	[ ]	[ ]	[*]
With metabolic activation:	[ ]	[ ]	[*]

2-2. Non-bacterial in vitro test (chromosomal aberration test) １）

Purity	:	> 99.0%
Type of cell used	:	Chinese hamster lung (CHL) cells
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals (Japan)
Solvent	:	Physiological saline
Positive controls	:	long-term treatment; Mitomycin C (24 h and 48 h) short-term treatment; Cyclophosphamide (+S9 and -S9 mix)
Doses	:	long-term treatment; 475, 950, 1900 μg/ml (24 h and 48 h) short-term treatment; 475, 950, 1900 μg/ml (+S9 and -S9 mix) short-term treatment; 1000, 1300, 1600, 1900 μg/ml (+S9 mix; confirmative examination)
S9	:	Rat liver, induced with phenobarbital and 5,6-benzoflavone
Plates/test	:	2
GLP	:	Yes

　Test results:

This chemical slightly increased incidences of structural chromosomal aberrations with an exogeneous metabolic activation system. Clear reproducibility was obtained the confirmation study.

Genetic effects:

	clastogenicity			polyploidy
	+	?	-	+	?	-
Without metabolic activation:	[ ]	[ ]	[*]	[ ]	[ ]	[*]
With metabolic activation:	[ ]	[*]	[ ]	[ ]	[ ]	[*]

1)	The tests were performed by the Biosafety Research Center, Foods, Drugs and Pesticides (An-pyo Center), Japan, 582-2 Shioshinden Arahama, Fukude-cho, Iwata-gun, Shizuoka, 437-12, Japan. Tel +81-538-58-1266 Fax +81-538-58-1393