Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 - 31 Mar 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
adopted 24 Feb 1987
Deviations:
no
GLP compliance:
yes
Remarks:
according to Japanese GLP standard (as described on JECDB)
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in a closed bottle at room temperature
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: water solubility is 2.71%, easy soluble in acetone and DMSO

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final dilution of a dissolved solid, stock liquid or gel: dilution in olive oil

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Remarks:
Crj:CD(SD)IGS
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc., Yokohama, Japan
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 5 weeks
- Weight at study initiation: 128 - 139 g (males), 108 - 120 g (females)
- Fasting period before study: 18 h prior to administration until 3 h after administration
- Housing: 5 animals of the same sex per cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 - 26
- Humidity (%): 45 - 63
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Amount of vehicle: 10 mL/kg bw
- Lot/batch no. (if required): 8123

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
In a pre-test, doses of 250, 500 and 1000 mg/kg bw was administered to 3 males and females. No mortality occured during the 7 day observation period. Based on this result, the main study was performed at 500, 1000 and 2000 mg/kg bw.
Doses:
500, 1000 and 2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
yes
Remarks:
gavage with olive oil
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed 5 times at the first day and daily thereafter. Individual weights were determined on Day 0 prior to administration and at Day 4, 8 and 15.
- Necropsy of survivors performed: yes

Results and discussion

Preliminary study:
Doses of 250, 500 and 1000 mg/kg bw were administered to 3 males and females. A control group was included. No death occurred during the following observation period (7 days). Locomotor activity was decreased in the high-dose group at the first day. According to this result, doses of 500, 1000 and 2000 mg/kg bw were applied in the main study.
Effect levelsopen allclose all
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
> 1 000 - < 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no male died at 1000 mg/kg bw; 3/5 males died at 2000 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 2/5 females died at 2000 mg/kg bw
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: combined approach: 5/10 animals died at 2000 mg/kg bw
Mortality:
500 mg/kg bw: 0/5 males and 0/5 females died
1000 mg/kg bw: 0/5 males and 0/5 females died
2000 mg/kg bw: 3/5 males and 2/5 females
Clinical signs:
500 and 1000 mg/kg bw: decreased locomotor activity, abnormal gait, clonic convulsion and lateral or prone position in males; decreased locomotor activity, abnormal gait and clonic convulsion in femals
2000 mg/kg bw: decreased locomotor activity and abnormal gait in males; abnormal gait in femals

all dose groups, non-surviving animals: decreased locomotor activity, clonic convulsion, lateral or prone position 15 minutes after administration
Body weight:
500 and 1000 mg/kg bw: body weight gains were within the normal ranges in males and females during the whole study period
2000 mg/kg bw: slightly decreased body weight gains observed in surviving animals at Day 4
Gross pathology:
500 and 1000 mg/kg bw: no abnormal findings
2000 mg/kg bw:
- non-survivng animals: discoloration in spleen was found in 1/2 female
- surviving animals sacrificed at termination: no abnormal findings

Any other information on results incl. tables

Table 1. Summary of results

Dose
[mg/kg bw]

Toxicological results*

Duration of clinical signs

Time of death

Mortality (%)

Males

Control

0/0/5

---

---

---

500

0/4/5

15 min – 3 h

---

0

1000

0/4/5

15 min – 3h

---

0

2000

3/5/5

15 min – day 4 (surviving)

1 – 3 h

60

Females

 Control

0/0/5

---

---

---

500

0/3/5

15 min – 1 h

---

0

1000

0/5/5

15 min – 1 h

---

0

2000

2/5/5

15 min – day 3 (surviving)

1 h – day 2

40

LD50 = 2000 mg/kg bw

* first number = number of dead animals

second number = number of animals with clinical signs

third number = number of animals used

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
CLP: Acute toxicity, Cat. 4, H302