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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
August 1998 to May 1999
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: well documented study performed according to OECD test guideline and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report Date:
1999

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Tri (hexyl, octyl, decyl) citrate
- Substance type: pure active substance
- Physical state: liquid
- Lot/batch No.: DK 4/141-68CT
- Expiration date of the lot/batch: December 1999
- Stability under test conditions: pure: at least 1 year, in vehicle: at least 2 hours
- Storage condition of test material: room tempeature in the closed container

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen
- Age at study initiation: 7-8 weeks
- Weight at study initiation: 150-180 g (females); 207-242 g (males)
- Fasting period before study: no data
- Housing: individually in Macrolon cages on Altromin saw fiber bedding
- Diet: Altromin 1324 maintanance diet for rats and mice, totally -pathogen-free-TPF (ad libitum)
- Water: tap water ad libitum
- Acclimation period: 12-14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 55 +/- 10
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: suspension of test article in carboxymethylcellulose (1% in aqua ad inject.)

VEHICLE
- Justification for use and choice of vehicle (if other than water): due to its non-toxic characteristics
- Amount of vehicle (if gavage): 5 ml/kg bw
- Lot/batch no. (if required): 36H0738
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
150 mg/kg bw/d
Basis:
actual ingested
Remarks:
Doses / Concentrations:
500 mg/kg bw/d
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: according to the results of the acute oral toxicity study and a preliminary study. The highest dose level was chosen with the aim to induce toxic effects but no death or severe suffering.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily, at least twice daily observation for mobidity and mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before th first exposure and once a week thereafter


BODY WEIGHT: Yes
- Time schedule for examinations: prior to first appliation (day 0) and once a week thereafter (days 7, 14, 21, 28 or day of sacrifice, respectively)


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes


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


OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: at necropsy by puncture of the abdominal aorta of the anaestesized animals
- Anaesthetic used for blood collection: Yes (identity)
- Animals fasted: Yes
- How many animals: all
- Parameters checked: Haematocrit )Hct), Haemoglobin (Hb), erythrocyte count (RBSC), total leucocyte count (WBC), platelet count, blood clotting time, differential laucocyte count


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at necropsy by puncture of the abdominal aorta of the anaestesized animals
- Animals fasted: Yes
- How many animals: all
- Parameters checked:nAspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase )(AP), cholesterol (Chol), total protein (TP), glucose (GLU), urea, creatinine (CREA), albumin (ALB), Na, K


URINALYSIS: Yes
- Time schedule for collection of urine: at necropsy by puncture of the urine bladder
- Metabolism cages used for collection of urine: No
- Animals fasted: Yes
- Parameters checked: pH, glucose, total protein


NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: once before the first exposure and in the fourth exposure week
- Dose groups that were examined: all
- Battery of functions tested: sensory reactivity, grasping reflex with grip strength, motor activity, flexion reflex, equilibrium, righting reflex and auditory startle

Other: rectal body temperature, counts of fecal boluss and pools of urine
Sacrifice and pathology:
GROSS PATHOLOGY: Yes: full, detailed gross necropsy including careful examination of the external surfce of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents

Organ weights: liver, kidneys, adrenals, testes, epididymides, thymus, spleen, brain and heart

Organ preparation: the following tissues were preserved from all animals of all groups: all gross lesions, brain, spinal cord, liver, kidneys, adrenals, stomach, small and large intestines, thumys, thyroid, spleen, lung and trachea, heart, gonads, accessory sex organs, urinary bladder, lymph nodes, peripheral nerve, bone marrow

HISTOPATHOLOGY: Yes: on the preserved organs and tissues of all animals in the control and high dose groups
Statistics:
One-way analysis of variance (ANOVA) was carried out, followed by Student's t-test to reveal any differences between control-and test groups. For parameters indicating no compound relatd alterations and/or showing ahigh degree of variation (both in control and test groups) no statistical evaluation was carried out. Therefore, biological relevance was discussed.
In the evaluation of laboratory parameters all values within a range of the mean values +/- the two fold standard deviation (x +/- 2s) are considered to be "normal" values within a "normal" population.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
All rats with the test item survived throughout the test period without showing clinical-toxic effects.

BODY WEIGHT AND WEIGHT GAIN
No significant differences in weight gain between test and control groups could be detected. A slightly diminished weight gain was observed in all dose groups compared to controls without reaching statistical significance and with no dose dependency.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
All animals showed normal food intake and no significant compound related reduction in food consumption was found.

HAEMATOLOGY
No significant differences are found for all parameters determined.

CLINICAL CHEMISTRY
No dose dependent and toxicologically relevant differences were found for all parameters determined.

URINALYSIS
No dose dependent and toxicologically relevant differences were found for all parameters determined.

NEUROBEHAVIOUR
No abnormalities or differences compared to the control group were observed.

ORGAN WEIGHTS
There were no significant results in relative and absolute organ weights for both sexes and any of the groups in liver, spleen, heart, brain and testes. In the low dose group in females the mean relative weights of kidneys were significantly higher and the mean absolute and relative weights of thymus were significantly lower than those in the corresponding control group. No dose dependency was found as values of the high dose groups corresponded to those of the control group. The mean absolute weight of adrenals in the male high dose group was slightly lower than in the corresponding control group, reaching statistical significance. The mean relative weight of the epididymides was significantly higher in all dose groups, the absolute weight was higher in the low dose group.

GROSS PATHOLOGY
No treatment-related effects observed.

HISTOPATHOLOGY: NON-NEOPLASTIC
No treatment-related differences in incidence or severity of morphological changes observed.

Effect levels

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
male/female

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

The mean weight gain in all groups was not as high as expected according to the standard growth curve for this strain. Explanation for this diminished weight gain in all anmals might be the daily treatment.

Applicant's summary and conclusion

Conclusions:
There was no indication that 2-Hydroxypropane-1,2,3-tricarboxylic acid, tri (hexyl, octyl, decyl) ester was toxic in male or female rats after repeated oral exposure up to 1000 mg/kg bw/d.
Executive summary:

There was no indication that 2-Hydroxypropane-1,2,3-tricarboxylic acid, tri (hexyl, octyl, decyl) ester was toxic in male or female rats after repeated oral exposure up to 1000 mg/kg bw/d.