Tripropyleneglycol was studied for oral toxicity in rats in a single dose toxicity test at doses of 500, 1000 and 2000 mg/kg and in an OECD combined repeat dose and reproductive/developmental toxicity screening test at doses of 8, 40, 200 and 1000 mg/kg/day. Genotoxicity of tripropylene-glycol was also studied by the reverse mutation assay in bacteria and the chromosomal aberration test in cultured Chinese hamster lung (CHL/IU) cells.

The single dose oral toxicity test revealed an LD50 of above 2000 mg/kg for both sexes.
With regard to repeat dose toxicity, tripropyleneglycol caused changes such as increased salivation, significantly elevated values of absolute and relative liver weights as well as relative kidney weights in males of the 1000 mg/kg group, and higher relative liver weight values in females of the 1000 mg/kg group. The NOEL is considered to be 200 mg/kg/day for both sexes in terms of general toxicological effects. The substance showed no effects on any parameters related to reproductive/developmental toxicity. NOELs for reproductive performance of males and females and for pups development are considered to be both 1000 mg/kg/day.

Tripropyleneglycol 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 limit concentration of 10 mM, in the absence or presence of an exogenous metabolic activation system.

1. Single Dose Oral Toxicity 1)

Purity	:	> 98%
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	:	Distilled water
GLP	:	Yes

　Test results:

Neither deaths nor abnormal symptoms were observed. Body weight changes of each treated group were virtually the same as those of the control group. No remarkable changes were found in any of the animals necropsied. LD50 : Male > 2000 mg/kg, Female > 2000 mg/kg

2. Repeat Dose and Reproductive/Developmental Toxicity 1)

Purity	:	> 98%
Test species/strain	:	Rat/Crj:CD (SD)
Test method	:	OECD Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test
Route	:	Oral (gavage)
Dosage	:	8, 40, 200, 1000 mg/kg/day
Number of animals	:	Male, 12 ; Female, 12/group
Vehicle	:	Distilled water
Administration period	:	Male, 49 days Female, from 14 days before mating to Day 3 of lactation
Terminal kill	:	Male, Day 50 Female, Day 4 of lactation
GLP	:	Yes

　Test results:

<Repeat dose toxicity>

Increased Salivation was observed in the 1000 mg/kg males. The 1000 mg/kg males showed significantly higher values for absolute and relative liver weights and relative kidney weight, and the 1000 mg/kg females showed higher values for relative liver weight. Body weight gains in all dosed groups in both sexes were almost the same as those of the controls. Tripropyleneglycol did not cause any changes in data for food consumption, hematology, and blood chemistry or alterations in necropsy and histological findings. The NOEL is considered to be 200 mg/kg/day for males and females.

<Reproductive and developmental toxicity> The test substance showed no effects in the reproductive and developmental toxicity test. NOELs for reproductive performance of males and females and for pups development are considered to be both 1000 mg/kg/day.

3. Genetic Toxicity

3-1 Bacterial test 2)

Purity	:	> 98%
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	:	Water
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, 5000 μg/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 without metabolic 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	:	> 98%
Type of cell used	:	Chinese hamster CHL/IU cells
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals(Japan)
Solvent	:	Water for injection
Positive controls	:	-S9, Mitomycin C +S9, Cyclophosphamide
Dosage	:	-S9 (continuous treatment): 0, 0.48, 0.95, 1.90 mg/ml -S9 (short-term treatment): 0, 0.48, 0.95, 1.90 mg/ml +S9 (short-term treatment): 0, 0.48, 0.95, 1.90 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 ): > 1.90 mg/ml

without metabolic activation (short-term treatment): > 1.90 mg/ml

with metabolic activation (short-term treatment): > 1.90 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, Gifu 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