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Description of key information

Oral exposure for 28 days did not cause any relevant adverse effects in rats at a dose of 1000 mg/kg/d.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study according to GLP regulations,lbut outdated and reduced number and types of examinations
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
yes
Species:
rat
Strain:
other: WISTAR Crl rats: (WI) BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: CHARLES RIVER FRANCE; 59 rue de Ia Paix; 76140 SAINT -AUBIN-LES-ELBEUF; (FRANCE)
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: males: 198.5 ± 2.0 g; females: 161.8 ± 1.7 g

- Housing: individual, (Makrolon type 3 tall)
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± - 2,
- Humidity (%): 40 - 70%,
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hI 12 h
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was dissolved in water for injection. A series of solutions was prepared by direct addition of the test substance to water.
Solutions were prepared freshly every day.

VEHICLE
- Water
- Concentration in vehicle: 0 - 30 - 200 mg/ml
- Amount of vehicle (if gavage): 5 ml/kg
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
Administration solutions were checked analytically on day 1 of treatment. Solubility as well as stability were determined before start of treatment.
Duration of treatment / exposure:
28 days
Frequency of treatment:
1 x daily by gavage
Remarks:
Doses / Concentrations:
0 - 150 - 1000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on a dose range finding study (result: no findings) and one low dose to ensure a NOAEL
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: no satellite groups
Positive control:
not required
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: =/> 1/day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: 1/week

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight of rats was recorded upon arrival and then weekly throughout the 28 days of the study. A last weighing was done the morning before necropsy (D 29).

FOOD Consumption:
- Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY & CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 26
- Anaesthetic used for blood collection: Yes (Halothan)
- Animals fasted: Yes
- How many animals: all

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
The following parameters were statistically analyzed separately for males and females:
• mean body weights on days 1, 8, 15,22 and 28,
• mean food consumption over days 1 to 8, 8 to 15, 15 to 22 and 22 to 28,
• haematology and blood biochemistry at week 4 (Day 26),
• absolute and relative organ weights at week 4 (day 29).

The following sequence of statistical tests was used.

Homogeneity of variance between the groups was assessed with Bartlett test1 (more than 2 groups) or Fisher's test2 (2 groups). In the case of homogeneity of variances, the data were analyzed using a parametric procedure. This consisted in one way analysis of variance (ANOV A) allowing for group effect followed by Dunnett's test3 to assess the significance of any intergroup differences.
Where Bartlett's test shows a significant difference in the variances across groups, the data were analysed using non-parametric methods namely a the Kruskall-Wallis ANOVA followed by Dunn's test.
Clinical signs:
no effects observed
Description (incidence and severity):
blue staining of faeces was considered due to the color of the test substance.
Mortality:
no mortality observed
Description (incidence):
blue staining of faeces was considered due to the color of the test substance.
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
Description (incidence and severity):
A decrease of the Total White Blood Cell Count in females of group 2 and of Monocytes count in males of group 3 were observed. A decrease of the lymphocytes count was found in females of group 2. These changes were not explained and appeared as artefacts.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Decrease of cholesterol in males of group 2. But no change was observed in group 3. This decrease was not explained and appeared as an artefact. Decrease of K+ in females of group 2. This decrease was minor and had no significance.
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
Only non significant lesions were found during the necropsy of the rats. The test substance did not induce macroscopic change
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
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: no effects at all
Critical effects observed:
not specified
Conclusions:
No significant difference was observed between control and dosed rats during this 28 day study.
The no effect level was determined at 1000 mglkg/day.
The test item appears as a non toxic substance after a 28 day oral toxicity study.
Executive summary:

Objective:

The aim of the study was to assess the toxicity and to determine, if possible, a no-effect level for the test substance following daily oral (gavage) administration to rats for 4 consecutive weeks.

Method:

The study was conducted according to the following design:

 Group Nr.  Group Designation  Dose level (mg/kg/day)

 Dose Volume

(m1/kg/day)

 Dose Concentration (mg/rnl)  Males  Females
 1 Control 
 Low  150  5  30  5  5
 3  High  1000  5  200  5  5

Three groups of 5 male and 5 female Wistar rats were formed. Two doses were tested: 150 and 1000 mglkg. The high dosage was selected on the basis of a preliminary 7 day oral toxicity study. The animals received the test substance daily by oral route (gavage).

A control group received the vehicle only, under the same conditions as the treated animals.

The animals were observed for 4 weeks, during which mortality, body weight, food consumption, clinical signs, clinical pathology and pathology were recorded.

Results

No mortality was observed in the study. No dose related body weight decrease was observed in all treated groups throughout the treatment period. No significant food consumption difference with the control group was observed in treated groups. No abnormal clinical sign related to treatment, no clinical biochemistry, no blood parameter change and no significant morphological change was observed in all groups.

Conclusion

The no effect level was determined at 1000 mg/kg/day. The test substance appears as a non toxic substance after a 28 day oral toxicity study in rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
2 Reliable with restrictions due to age of study and reporting

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral exposure for 28 days did not cause any relevant adverse effects in rats at a dose of 1000 mg/kg/d.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
only available study

Justification for classification or non-classification

no classification

Oral exposure for 28 days did not cause any relevant adverse effects in rats at a dose of 1000 mg/kg/d.