[No public or meaningful name is available]

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26/07/1996 to 19/03/1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

Annex V (28 day study)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

liquid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Inc.
- Age at study initiation: Five weeks old.
- Weight at study initiation: Males: 146.8-167.0 g; Females: 118.4-142.6 g.
- Fasting period before study: None.
- Housing: Individually in hanging stainless steel cages with wire-mesh floor.
- Diet (e.g. ad libitum): Ad libitum.
- Water (e.g. ad libitum): Ad libitum.
- Acclimation period: Animals acclimatised but period not specified.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25.
- Humidity (%): 45-65.
- Air changes (per hr): 10-15.
- Photoperiod (hrs dark / hrs light): 12/12.

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Test substance was accurately weighed and dissolved with olive oil to make a 10.0 w/v% solution, and the solution was prepared more than once per week. Dilutions were made from the 10% solution.

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data.
- Concentration in vehicle: 0.08, 0.4, 2.0, 10%.
- Amount of vehicle (if gavage): Dependent on dose.
- Lot/batch no. (if required): No data.
- Purity: No data.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

28 days

Frequency of treatment:

Daily (7 days/week)

Doses / concentrations

Remarks:

Doses / Concentrations:
8, 40, 200 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

Six

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on results of a preliminary dose range-finding study conducted with 50, 250 and 1000 mg/kg bw/day.
- Post-exposure recovery period in satellite groups: 14 days recovery period with no dosing with test substance.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once per day.

DETAILED CLINICAL OBSERVATIONS: No data.

BODY WEIGHT: Yes.
- Time schedule for examinations: Two days before dosing started, Day 1 of dosing, then Days 3, 5, 8, 10, 12, 15, 17, 19, 22, 24, 26 and 28. During the recovery period animals were weighed on Recovery Day 1, 3, 5, 8, 10, 12 and 14.

FOOD CONSUMPTION: Measured once before dosing, then twice per week (no further details).

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.

WATER CONSUMPTION: No.

OPHTHALMOSCOPIC EXAMINATION: No.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the dosing and recovery periods.
- Anaesthetic used for blood collection: Yes (ether).
- Animals fasted: Yes, overnight.
- How many animals: All animals.
- Parameters checked in table No.1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the dosing and recovery periods.
- Animals fasted: Yes, overnight.
- How many animals: All animals.
- Parameters checked in table No.1 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: 16-hour urine samples collected at the end of the dosing and recovery periods.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, overnight.
- Parameters checked in table No.1 were examined.

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table 2)
HISTOPATHOLOGY: Yes (see table 2)

Statistics:

Data regarding body weights, food consumption, haematological examination, blood chemistry, urine volume and organ weights were analysed using Bartlett's test for homogeneity of variance. If the variances were homogeneous at a significance level of 5%, one way analysis of variance was performed. When there was a significant difference in this analysis, the difference between the vehicle control group and each of the treatment group analysed by Dunnett's test. If the variances were not homogeneous in the Bartlett's test, Kruskal-Wallis's test was used. When there was a significance in this test, the difference between the control group and each of the treatment groups was analysed by nonparametric Dunnett's test.

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):

not examined

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: There were no deaths. During the dosing period, unkempt hair was observed in males of the 1000 mg/kg bw/day group. Although salivation was observed in all groups including the vehicle control group and frequency of this sign was different between the vehicle control group and the 1000 mg/kg bw/day group, it was not considered to be an adverse toxicological effect because the sign was observed just after dosing and it had no related other findings.

BODY WEIGHT AND WEIGHT GAIN: There was no adverse effect on body weights of any group.

FOOD CONSUMPTION: No adverse effects.

HAEMATOLOGY: At the end of the dosing period there were no differences between treated and control animals. At the end of the recovery period the highest dose group had a decrease in mean corpuscular volume.

CLINICAL CHEMISTRY: At the end of the dosing period, an increase in serum calcium level and a decrease in serum chloride level were observed in males of the 1000 mg/kg bw/day group. Increases in serum GPT, gamma-GTP activities, total cholesterol, total protein levels and decreases in serum cholinesterase activity and A/G ratio were observed in females of the 1000 mg/kg bw/day groups. A decrease in serum GOT activity was observed in males of all treated animals. These changes were concluded to be non-adverse.

URINALYSIS: No adverse effects.

ORGAN WEIGHTS: At the end of the dosing period, an increase in relative liver weight was observed in both sexes of the groups dosed with 200 mg/kg bw/day and over. Increases in absolute liver weight were observed in both sexes of the 1000 mg/kg bw/day group. An increase in relative kidney weight was observed in males of the 1000 mg/kg bw/day group. At the end of the recovery period there was a increase in relative kidney weights in males of the 1000 mg/kg bw/day group.


GROSS PATHOLOGY: Enlargement of the liver was observed in both sexes of the 1000 mg/kg bw/day group at the end of the dosing period. There was no such finding at the end of the recovery period.


HISTOPATHOLOGY: NON-NEOPLASTIC: In histopathological examination at the terminal autopsy of the dosing period, swelling of the hepatocytes in the liver was observed in both sexes of the 1000 mg/kg bw/day group. Increased eosinophilic bodies of the kidney was observed in males of the 1000 mg/kg bw/day group.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at any dose level.

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

In a 28-day oral gavage study (Hita Research Laboratories, 1998) conducted to OECD Test Guideline 407 and in compliance with GLP, the NOAEL was at least 1000 mg/kg bw/day as no adverse effects were observed.