ABSTRACT

The single dose oral toxicity test revealed an LD50 of more than 2000 mg/kg for both sexes.

In the repeat dose study, the test substance caused salivation in males dosed with 40 mg/kg or more and in females dosed with 200 mg/kg or more, lower values of food consumption in females dosed with 200 mg/kg or more and in males of the 1000 mg/kg group, and higher water intake in females of the 1000 mg/kg group. The test substance also caused slight anemia and higher values for liver weights in both sexes of the groups dosed with 200 mg/kg or more, as well as increased urinary volume and serum total cholesterol, and lowered urinary specific gravity in females of the 1000 mg/kg group. All of these changes were reversible and disappeared within a 14-day recovery period. NOELs are considered to be 8 mg/kg/day for males and 40 mg/kg/day for females in terms of general toxicological effects.

Tris(p-cumenyl) phosphate was not mutagenic in Salmonella typhimurium TA100, TA98, TA1535, TA1537 and Escherichia coli WP2 uvrA. Neither structural nor numerical chromosomal aberrations were induced in CHL/IU cells up to the concentration going 50% cell growth inhibition or the concentration of 10 mM, in the absence or presence of an exogenous metabolic activation system.

SUMMARIZED DATA FROM THE STUDIES

1. Single Dose Oral Toxicity 1)

Purity	:	99.7%
Test species/strain	:	Rat/Crj:CD (SD)
Test method	:	OECD Test Guideline 401
Dosage	:	500, 1000, 2000 mg/kg
Number of animals	:	Male, 5 ; Female, 5/group
Vehicle	:	Corn oil
GLP	:	Yes

　Test results:

No deaths were observed. Diarrhea was found in males dosed with 500 mg/kg or more and in females dosed with 100 mg/kg or more. Significant suppression in body weight gain was found in both sexes dosed with 1000 mg/kg or more one day after administration. No remarkable changes were found in any of the animals necropsied.

LD50: Male > 2000 mg/kg, Female > 2000 mg/kg

2. Repeat Dose Toxicity 1)

Purity	:	99.7%
Test species/strain	:	Rat/Crj:CD (SD)
Test method	:	Guidelines for 28-Day Repeat Dose Toxicity Testing of Chemicals (Japan)
Route	:	Oral (gavage)
Dosage	:	8, 40, 200, 1000 mg/kg/day
Number of animals	:	Male, 10 or 15 (0, 1000 mg/kg) Female, 10 or 15 (0, 1000 mg/kg)
Vehicle	:	Corn oil
Administration period	:	Male, Female ; 28 days
Terminal kill	:	Male, Female ; Day 29 or 43 (0, 1000 mg/kg)
GLP	:	Yes

　Test results:

Tris(p-cumenyl) phosphate caused increased salivation in males of the groups treated with 40 mg/kg or more and in females of the groups treated with 200 mg/kg or more, lower values of food consumption in females of the groups treated with 200 mg/kg or more and in males of the 1000 mg/kg group and higher water intake in females of the 1000 mg/kg group. Moreover, the test substance had effects on hematology resulting in a slight anemia and on liver weights in both sexes of the groups treated with 200 mg/kg or more. It also caused increased urinary volume and serum total cholesterol, and lowered urinary specific gravity in females of the 1000 mg/kg group. All of these changes were reversible and disappeared within a 14-day recovery period.

NOELs are considered to be 8 mg/kg/day for males and 40 mg/kg for females in terms of general toxicological effects under the conditions of the present study.

3. Genetic Toxicity

3-1 Bacterial test 2)

Purity	:	> 99%
Test species/strains	:	S.typhimurium TA100, TA1535, TA98, TA1537 E. coli WP2 uvrA
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals (Japan)
Procedures	:	Plate incorporation method
Solvent	:	DMSO
Positive controls	:	-S9, AF-2 (TA100, WP2, TA98), sodium azide (TA1535) and 9-aminoacridine (TA1537) +S9, 2-aminoanthracene (all strains)
Dosage	:	0, 312.5, 625, 1250, 2500, 5000μg/plate
S-9	:	Rat liver, induced with phenobarbital and 5,6-benzoflavone
Plates/test	:	3
Number of replicates	:	2
GLP	:	Yes

　Test results:

Minimum concentration of test substance at which toxicity was observed:

No toxicity was observed up to a concentration of 5000 μg/plate with or withoutmetabolic activation.

Genetic effects:
S. typhimurium TA100, TA1535, TA 98, TA1537

	+	?	-
with metabolic activation	[ ]	[ ]	[*]
without metabolic activation	[ ]	[ ]	[*]

E. coli WP2 uvrA

with metabolic activation	[ ]	[ ]	[*]
without metabolic activation	[ ]	[ ]	[*]

3-2 Non-bacterial in vitro test (chromosomal aberration test) 2)

Purity	:	99%
Type of cell used	:	Chinese hamster CHL/IU cells
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals (Japan)
Solvent	:	DMSO
Positive controls	:	-S9, Mitomycin C +S9, Cyclophosphamide
Dosage	:	-S9 (continuous treatment): 0, 0.04, 0.07, 0.14 mg/ml -S9 (short-term treatment): 0, 1.1, 2.3, 4.5 mg/ml +S9 (short-term treatment): 0, 1.1, 2.3, 4.5 mg/ml
S-9	:	Rat liver, induced with phenobarbital and 5,6-benzoflavone
Plates/test	:	2
GLP	:	Yes

　Test results:

Lowest concentration producing cytogenetic effects in vitro:

without metabolic activation (continuous treatment ): > 0.14 mg/ml

without metabolic activation (short-term treatment): > 4.5 mg/ml

with metabolic activation (short-term treatment): > 4.5 mg/ml

Genotoxic effects:

	clastogenicity			polyploidy
	+	?	-	+	?	-
without metabolic activation:	[ ]	[ ]	[*]	[ ]	[ ]	[*]
with metabolic activation:	[ ]	[ ]	[*]	[ ]	[ ]	[*]

1)	The tests were performed by Nihon Bioresearch Inc. Hashima Laboratory, 6-104 Majima, Fukuju-cho, Hashima, 501-62, Japan. Tel +81-58-392-6222 Fax +81-58-392-1284
2)	The tests were performed by the Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa 257, Japan. Tel +81-463-82-4751 Fax +81-463-82-9627