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Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: Guidelines for 28 day repeated dose toxicity testing of chemicals Japan
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Reference substance name:
Tris (2-ethylhexyl) 1,2,4-benzenetricarboxylate
Cas Number:
3319-31-1
IUPAC Name:
Tris (2-ethylhexyl) 1,2,4-benzenetricarboxylate
Details on test material:
- Name of test material (as cited in study report): Tris (2-ethylhexyl) 1,2,4-benzenetricarboxylate
- Physical state: liquid
- Analytical purity:>99%
- Lot/batch No.: N-60601
- Stability under test conditions:
- Storage condition of test material: refrigerated when stable for 1W
- Other: soluble in oil; MW- 546.87; Mol formula C33H54O6

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan Kanagawa
- Age at study initiation: 6 weeks
- Weight at study initiation: males: 130-151g; females: 110-121g
- Fasting period before study: No
- Housing: 5/sex/group
- Diet (e.g. ad libitum): yes, Oriental yeast Co. Ltd sterilised by radiation
- Water (e.g. ad libitum): yes, tap water
- Acclimation period: 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23+/-2 degrees C
- Humidity (%): 55+/- 10%
- Air changes (per hr): 20
- Photoperiod (hrs dark / hrs light):12.00:12.00

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
A known weight of tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate was dissolved in corn oil

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil; tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate is poorly soluble in water
- Amount of vehicle (if gavage): 5 ml/kg
-Other: Supplied by Nakaraitesuku
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
No data except that concentration of preparations for 4 weeks were analysed and the results were in the range 98-102% confirming the accuracy of the formulation used.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
vehicel control (corn oil)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5 male and 5 female rats per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on 2 week preliminary study using doses of 0, 200, 600 and 1800 mg/kg.
- Rationale for animal assignment (if not random): randomised using stratified weight ranges
- Rationale for selecting satellite groups: no satellite groups
- Post-exposure recovery period: 5 male and 5 female control and 1000 tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate groups for 14 days.
Positive control:
No

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily for signs of ill health and moribund condition
- Cage side observations were included.: N/A no clinical signs and no deaths

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily for signs of ill health and moribund condition

BODY WEIGHT: Yes
- Time schedule for examinations: Immediately prior to the first exposure and again for each animal every week.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No - Not applicable as animals dosed by gavage

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No - Not applicable - Not a dietary study

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No - Not applicable as animals dosed by gavage:
-Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: terminal blood samples taken from the abdominal aorta at necropsy on completion of treatment and recovery periods
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes - 16h
- How many animals: all animals in 28 day and recovery groups
- Parameters were examined. yes, data reported for 28 days study only

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: terminal blood samples taken from the abdominal aorta at necropsy on completion of treatment and recovery periods
- Animals fasted: Yes - 16h
- How many animals: all animals in 28 days and recovery groups
- Parameters were examined: yes, data reported for 28 day study only

URINALYSIS: Yes
- Time schedule for collection of urine: 24h collection prior to terminal kills after 28 days treatment and recovery periods
- Parameters were examined: yes reported for 28 day treatment groups only

Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Organ weights: brain, liver, kidneys, spleen, adrenals, testes (male) and ovaries (females) for each animal.

HISTOPATHOLOGY: Yes
heart, liver, kidneys, spleen, adrenals and femoral bone marrow from rats of the control and high dosed groups, and kidneys from all dosage male.
Other examinations:
No
Statistics:
Bartlett’s test, Dunnett’s test or Kruskal-Wallis test depending on whether or not the data were nonhomogeneous or homogeneous. Fisher’s test for the pathological result. Jonckheere’s test for the correlation of dosage.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
No abnormality was detected during the study.
Mortality:
no mortality observed
Description (incidence):
No abnormality was detected during the study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The mean body weight of treatment groups of rats for males and females had no significant differences from the controls during the course of the study.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There was no significant difference between control and treatment groups throughout treatment and recovery periods for both sexes.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
at the end of dosing
Males and females : No dose-related significant changes were observed. In the examination of blood coagulating system, prothrombin time for males was slightly prolonged, but they were considered within the physiological fluctuation. For females, no significant changes in all test items.

after recovering period
Males : Haemoglobin was slightly increased for males at 1000 mg/kg group, but they were considered within the physiological fluctuation. In the examination of blood coagulation system, no significant changes were observed in all test items.
Females : No significant change in all tests.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
at the end of dosing
Males : No dose-related significant adverse treatment-related effect in clinical chemistry.
Females : At 300, and 1,000 mg/kg dosing, chlorine contents were low.

after recovering period
Males : At 1,000 mg/kg dosing, potassium contents were slightly high.
Females : At 1,000 mg/kg dosing, GOT were slightly high. But both changes were considered to be no meaning, because at the end of treatment these changes were not recognised.
Urinalysis findings:
no effects observed
Description (incidence and severity):
at the end of dosing
Males and Female: At 1,000 mg/kg dosing, some of rats (both sexes), amounts of urinary increased, but the mean urinary specific gravity values in the 1,000 mg/kg dosing group was not significant change from control group.

after recovering period
Males and Females: No dose-related significant change in all tests.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Organ weight changes:
at the end of dosing
Male : No dose-related change in all tested organs.
Female : Relative liver weight were slightly increased at 100 mg/kg dosing, but no doserelated change. Other organs, no significant change.

after recovering period:
Males : At 1,000 mg/kg dosing, relative kidney weight were slightly low.
Female : At 1,000 mg/kg dosing, absolute and relative adrenal weight were slightly high. But both changes were considered no related to dosing and recovering of this chemical.
Gross pathological findings:
no effects observed
Description (incidence and severity):
at the end of dosing
Males : Coloured patch/zone of lungs were observed 1 of 100 mg/kg, 2 of 300 mg/kg and 3 animals of 1,000 mg/kg dosing group. Also hypertrophy of the kidney,
hypertrophy of parathyroid, and etc. were observed. Amounts of eosinophilic body in the kidney were slightly increased in dosing group. But all these changes
were considered no related the dosing and recovering of this chemical, because the degree and rate of changes were same of all the group included control.
Females : Red patch/zone of thymus dilated lumen of the uterus and etc. were observed. But all these changes were considered no related the dosing and recovering of this chemical, because the degree and rate of changes were same of all the group included control.

after recovering period:
Males and Females: No dose-related significant change in all tests.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Any changes seen can be considered to be spontaneous and not related to treatment with tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate.
Histopathological findings: neoplastic:
not examined
Details on results:

HAEMATOLOGY: See attached Table

CLINICAL CHEMISTRY: See attached Table

URINALYSIS: See attached Table

ORGAN WEIGHTS: See attached Table

GROSS PATHOLOGY: See attached Table:

HISTOPATHOLOGY: NON-NEOPLASTIC: See attached Table

Effect levels

Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: The absence of effects in animals given 100, 300 and 1000 mg/kg/day tris(2-ethylhexyl) benzene-1,2,4-tricarboxylate.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Tables attached (Repeated Dose 28 -day Oral Toxicity Study_relevant tables.pdf)

Applicant's summary and conclusion

Conclusions:
No test substance related changes were noted in terms of clinical signs, body weight, food consumption, and haematology, blood chemical examination, urinalysis, and pathological findings. The NOEL for repeated dose toxicity for 28 days is considered to be 1,000 mg/kg/day for both sexes.
Executive summary:

This 28 -day repeated dose oral toxicity study was conducted according to Guidelines for 28 day repeated dose toxicity testing of chemicals Japan, similar to OECD Guideline 407 and in compliance with GLP criteria. Five rats per sex per dose were exposed once daily via gavage to 0 (vehicle corn oil), 100, 300 and 1,000 mg/kg/day test substance for 28 days. The post observation period was 2 weeks for 0 and 1,000 mg/kg/day dose.

No test substance related changes were noted in terms of clinical signs, body weight, food consumption, and haematology, blood chemical examination, urinalysis, and pathological findings. The NOEL for repeated dose toxicity for 28 days is considered to be 1,000 mg/kg/day for both sexes.

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