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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
not specified
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
- purity: 99.7%

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Co., Ltd. Japan
- Age at study initiation: 5-6 weeks
- Weight at study initiation: 154-171 (male) - 112-132 (female)
- Fasting period before study: from the day before administration to 3 hours after administration.
- Housing: five animals per cage (same sex)
- Diet: Lab Animals Chow E1 (MF Oriental Yeast Co., Ltd.) ad libitum.
- Water: Tap water filtered through a 5 µm filter, ad libitum. (water quality inspections performed regularly)
- Acclimation period: 6-7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25ºC
- Humidity (%): 40-70%
- Air changes (per hr): 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE: The vehicle and dose selection was based on the results of a preliminary test.
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg
Doses:
500 (only females), 700, 1000, 1400 and 2000 mg/kg.
No. of animals per sex per dose:
5 male/ 5 female per group
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations of clinical signs and mortality were performed 15, 30 minutes, 1, 2, 4, 6, 7, and 8 hours after administration on the day of administration, and once a day thereafter (up to 14 days). Body weights were recorded on days 0, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
1 110 other: mg/kg
Based on:
act. ingr.
95% CL:
>= 832 - <= 1 480
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
1 110 other: mg/kg
Based on:
act. ingr.
95% CL:
>= 877 - <= 1 404
Mortality:
60% of males and females died at concentrations of 1000 and 1400 mg/kg. At 2000 mg/kg, 60% of males and 100% of females died.
Clinical signs:
The following clinical signs were observed in both sexes. These signs were increasing in the first eight hours but the survivors recovered completely: decrease of locomotor activity was seen at 1000 mg/kg and above; hypopnoea was detected at 1400 mg/kg and above, as well as loose stool.
Body weight:
The body weight gain was lower in the 2000 mg/kg male group (females died) and in 1400 mg/kg group in both sexes.
Gross pathology:
Stomach lesions such as focal thickening of the stomach mucosa, adhesion of the stomach and other abdmoninal organs or ulcer were observed from 700 mg/kg in males and from 500 mg/kg in females at necropsy of the animals after the observation period.
In the dead animal's autopsy, the following observations were made: in the heart, there was atrial stretching; at pulmonary level, congestion, edema and/or bleeding, and endotracheal mucus retention were observed; at stomach level, hemorrhage, stomach erosion, ulcer, distension; bleeding, congestion and/or dilatation of the small intestine were observed and finally, yellowing whitening of the liver.

Any other information on results incl. tables

Cumulative mortality/No. of test animals

Sex

Dose

 

Time after administration

Days after administration

Mortality rate

 

mg/kg

15m

30m

1h

2h

4h

6h

7h

8h

1

2

3

4…

14

(%)

male

700

0/5

 

 

 

 

 

 

 

 

 

 

 

0/5

0

1000

0/5

1/5

 

 

 

 

 

 

2/5

3/5

 

 

3/5

60

1400

0/5

 

 

 

 

 

 

 

2/5

3/5

 

 

3/5

60

2000

0/5

 

 

 

 

 

1/5

 

3/5

 

 

 

3/5

60

female

500

0/5

 

 

 

 

 

 

 

 

 

 

 

0/5

0

700

0/5

 

 

 

 

 

 

 

 

 

 

 

0/5

0

1000

0/5

 

 

 

 

 

 

 

2/5

3/5

 

 

3/5

60

1400

0/5

 

 

 

 

 

 

 

3/5

 

 

 

3/5

60

2000

0/5

 

 

3/5

 

 

 

 

5/5

 

 

 

5/5

100

*m: minutes, h: hour(s), d: days.

Mean body weight

Sex

Dose

mg/kg

 

Days after administration

0

7

14

male

700

Mean
SD

Number

162

3.6

5

213

4.2

5

280

10.4

5

1000

Mean
SD

Number

163

6.5

5

210

31.8

2

282

26.2

2

1400

Mean
SD

Number

163

1.3

5

204

0.7

2

265

9.2

2

2000

Mean
SD

Number

163

5.8

5

189

14.8

2

266

15.6

2

female

500

Mean
SD

Number

120

4.7

5

160

8.6

5

187

14.7

5

700

Mean
SD

Number

120

6.6

5

157

10.2

5

181

13.0

5

1000

Mean
SD

Number

119

4.3

5

150

5.7

2

170

6.4

2

1400

Mean
SD

Number

122

6.9

5

156

19.1

2

188

17.7

2

2000

Mean
SD

Number

126

4.7

5

AD

 

*units: g; AD: all animals were dead.

Clinical signs (male): No. of animals with findings / No. of surviving animals

Dose

(mg/kg)

Clinical signs

Time after administration

15 m

30 m

1h

2h

4h

6h

7h

8h

700

Loose stool

 

 

 

 

2/5

 

 

 

 

Normal

 

5/5

5/5

5/5

3/5

5/5

5/5

5/5

5/5

1000

Decrease of locomotor activity

+

1/5

1/4

2/4

 

4/4

4/4

4/4

4/4

Hypopnoea

 

1/5

1/4

2/4

 

 

 

 

 

Salivation

+

4/5

3/4

 

 

 

 

 

 

Loose stool

 

 

 

1/4

1/4

1/4

 

1/4

1/4

Normal

 

1/5

1/4

0/4

3/4

0/4

0/4

0/4

0/4

1400

Decrease of locomotor activity

+

 

4/5

3/5

1/5

4/5

4/5

3/5

3/5

++

 

1/5

 

 

1/5

1/5

 

 

+++

 

 

 

 

 

 

1/5

1/5

Hypopnoea

 

 

3/5

1/5

 

1/5

2/5

2/5

1/5

Salivation

+

2/5

 

 

 

 

 

 

 

Loose stool

 

 

 

2/5

3/5

2/5

 

1/5

2/5

Tiptoe gait

 

 

 

 

 

 

1/5

 

 

Crouching

 

 

 

 

 

 

 

1/5

 

Prone position

 

 

 

 

 

 

 

 

1/5

Normal

 

3/5

0/5

1/5

2/5

0/5

0/5

1/5

1/5

2000

Decrease of locomotor activity

+

 

3/5

5/5

2/5

3/5

3/5

2/4

¾

++

 

 

 

1/5

2/5

2/5

1/4

 

+++

 

 

 

 

 

 

1/4

1/4

Hypopnoea

 

 

2/5

2/5

2/5

2/5

2/5

2/4

1/4

Salivation

+

1/5

1/5

 

 

 

 

 

 

Loose stool

 

 

 

1/5

3/5

3/5

 

3/4

3/4

Crouching

 

 

 

 

 

 

 

1/4

1/4

Normal

 

4/5

1/5

0/5

0/5

0/5

0/5

0/4

0/4

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

 

Clinical signs (male):No. of animals with findings / No. of surviving animals

Dose

(mg/kg)

Clinical signs

Days after administration

1

2

3

4

5

6

7

8

9

10

-14d

Normal

 

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

1000

Decrease of locomotor activity

+

2/3

1/2

1/2

1/2

 

 

1/2

1/2

 

 

 

--

1/3

 

 

 

 

 

 

 

 

 

 

Hypopnoea

 

2/3

 

 

 

 

 

1/2

1/2

 

 

 

Soiled perineal region

 

1/3

 

 

 

 

 

 

 

 

 

 

Normal

 

0/3

1/2

1/2

1/2

2/2

2/2

1/2

1/2

2/2

2/2

- 2/2

1400

Decrease of locomotor activity

+

2/3

 

 

 

 

 

 

 

 

 

 

++

1/3

 

 

 

 

 

 

 

 

 

 

Hypopnoea

 

1/3

 

½

 

 

 

 

 

 

 

 

Normal

 

0/3

2/2

½

2/2

2/2

2/2

2/2

2/2

2/2

2/2

- 2/2

2000

Decrease of locomotor activity

+

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hypopnoea

 

 

1/2

2/2

2/2

 

 

 

 

 

 

 

Soiled perineal region

 

2/2

2/2

1/2

1/2

 

 

 

 

 

 

 

Abdominal distention

 

 

2/2

 

 

 

 

 

 

 

 

 

Normal

 

0/2

0/2

0/2

0/2

0/2

0/2

0/2

0/2

2/2

2/2

- 2/2

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

 

Clinical signs (female): No. of animals with findings / No. of surviving animals

Dose

(mg/kg)

Clinical signs

Time after administration

15 m

30 m

1h

2h

4h

6h

7h

8h

500

Normal

 

5/5

5/5

5/5

3/5

5/5

5/5

5/5

5/5

700

Loose stool

 

 

 

 

2/5

3/5

 

 

 

Normal

 

5/5

5/5

5/5

3/5

2/5

5/5

5/5

5/5

1000

Decrease of locomotor activity

+

2/5

4/5

 

1/5

3/5

4/5

4/5

3/5

++

1/5

1/5

 

 

 

 

 

1/5

Hypopnoea

 

 

 

 

1/5

1/5

1/5

1/5

1/5

Irrgular respiration

 

1/5

 

 

 

 

 

 

 

Salivation

+

2/5

1/5

 

1/5

 

 

 

 

++

 

1/5

1/5

 

 

 

 

 

Crouching

 

1/5

 

 

 

 

 

 

 

Normal

 

2/5

0/5

4/5

4/5

2/5

1/5

1/5

1/5

1400

Decrease of locomotor activity

+

 

 

1/5

2/5

2/5

2/5

2/5

2/5

++

2/5

3/5

 

 

1/5

2/5

2/5

2/5

Hypopnoea

 

 

2/5

 

 

2/5

2/5

2/5

2/5

Salivation

+

1/5

3/5

 

 

 

 

 

 

Crouching

 

2/5

2/5

 

 

 

1/5

1/5

1/5

Normal

 

3/5

1/5

4/5

3/5

2/5

1/5

1/5

1/5

2000

Decrease of locomotor activity

+

2/5

2/5

2/5

1/2

2/2

2/2

1/2

1/2

++

1/5

2/5

 

1/2

 

 

1/2

1/2

+++

 

1/5

2/5

 

 

 

 

 

Hypopnoea

 

4/5

4/5

3/5

1/2

2/2

2/2

2/2

1/2

Gasping

 

 

 

1/5

 

 

 

 

 

Salivation

+

4/5

4/5

1/5

 

 

 

 

 

Tiptoe gait

 

 

 

 

 

 

 

 

1/2

Crouching

 

 

 

 

 

 

 

 

1/2

Prone position

 

 

1/5

 

 

 

 

 

 

Loose stool

 

 

 

 

1/2

1/2

 

 

1/2

Normal

 

1/5

0/5

0/5

0/2

0/2

0/2

0/2

0/2

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe.

  

Clinical signs (female): No. of animals with findings / No. of surviving animals

Dose

(mg/kg)

Clinical signs

Days after administration

1

2

3

4

5

6

7

8

9

10

-14d

500

Normal

 

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

700

Normal

 

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

- 5/5

1000

Decrease of locomotor activity

+

2/3

1/2

 

 

 

 

 

 

 

 

 

++

1/3

 

 

 

 

 

 

 

 

 

 

Hypopnoea

 

2/3

 

 

 

 

 

 

 

 

 

 

Normal

 

0/3

1/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

- 2/2

1400

Decrease of locomotor activity

+

2/2

1/2

1/2

1/2

1/2

1/2

1/2

1/2

 

 

 

Hypopnoea

 

2/2

1/2

2/2

 

 

 

 

 

 

 

 

Tiptoe gait

 

1/2

 

 

 

 

 

 

 

 

 

 

Normal

 

0/2

1/2

0/2

1/2

1/2

1/2

1/2

1/2

2/2

2/2

1/2

2000

-

 

AD

 

 

 

 

 

 

 

 

 

 

*m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Autopsy findings (male):No. of animals with findings / No. of animals examined

 

Organ

findings

Dose (mg/kg)

700

1000

1400

2000

Dead animals

Heart

Dilatation of atrium

-

1/3

0/3

3/3

Lung

Congestion/edema

-

1/3

0/3

2/3

Trachea

Mucus stagnation

-

1/3

0/3

0/3

Stomach

Focal hemorrhage (glandular area)

-

1/3

1/3

2/3

Erosion/ulcer (glandular area)

-

2/3

2/3

1/3

dilatation

-

1/3

2/3

1/3

Small intestine

Focal hemorrhage

-

1/3

0/3

0/3

Congestion

-

1/3

0/3

0/3

Dilatation

-

1/3

0/3

0/3

Liver

Yellowish change / whitish dot

-

1/3

1/3

0/3

Urinary bladder

Dark-red urine

-

1/3

0/3

0/3

Normal

 

-

0/3

0/3

0/3

Surviving animals

Stomach

Focal thickening of mucosa (non-glandular area)

5/5

2/2

2/2

2/2

Adhesion (stomach and other abdominal organs)

4/5

2/2

2/2

2/2

Kidney

Dilatation of the renal pelvis

0/5

1/2

0/2

0/2

Normal

 

0/5

0/2

0/2

0/2

 *m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Autopsy findings (female):No. of animals with findings / No. of animals examined

 

Organ

findings

 

Dose (mg/kg)

500

700

1000

1400

2000

Dead animals

Heart

Dilatation of atrium

-

-

0/3

1/3

3/5

Lung

Focal hemorrhage

-

-

0/3

0/3

1/5

Trachea

Mucus stagnation

-

-

0/3

0/3

1/5

Stomach

Focal hemorrhage (glandular area)

-

-

2/3

1/3

3/5

Focal hemorrhage (non- glandular area)

-

-

1/3

0/3

0/5

Erosion/ulcer (glandular area)

-

-

0/3

3/3

1/5

dilatation

-

-

1/3

1/3

1/5

Small intestine

Congestion

-

-

1/3

2/3

0/5

Dilatation

-

-

0/3

0/3

1/5

Liver

Yellowish change / whitish dot

-

-

1/3

1/3

1/5

Normal

 

-

-

0/3

0/3

0/5

Surviving animals

Stomach

Focal thickening of mucosa

(non-glandular area)

5/5

5/5

2/2

2/2

-

Adhesion (stomach and other abdominal organs)

4/5

4/5

2/2

2/2

-

Kidney

Dilatation of the renal pelvis

0/5

1/5

0/2

0/2

-

Normal

 

0/5

0/5

0/2

0/2

-

 *m: minutes, h: hour(s), d: days; +: slight, ++: moderate; +++: severe; AD: all animals were dead.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Remarks:
EU criteria.
Conclusions:
The LD50 for the test substance in the Acute Oral Toxicity test was found to be 1110 mg/kg bw for both sexes.
Executive summary:

The Acute Oral Toxicity Test was performed according to OECD Guideline 401 (GLP Study). Four concentrations of the test substance were assayed in five males (700, 1000, 1400 and 2000 mg/kg) and five in females (500, 700, 1000, 1400 and 2000 mg/kg) in a single dose toxicity test in Crj:CD (SD) rats; 5 male and 5 female per group. All animals were subject to daily observations and weekly determinations of body weight. Necropsy was performed on the day of their death or after sacrifice, at the end of the observation period. 60% of males and females died at concentrations of 1000 and 1400 mg/kg. At 2000 mg/kg, 60% of males and 100% of females died. Decreased locomotor activity was seen at 1000 mg/kg and above; hypopnoea and loose stool were detected at 1400 mg/kg and above. These signs increased in the first eight hours but survivors recovered completely. The body weight gain was lower in the 2000 mg/kg male group (females died) and in 1400 mg/kg group in both sexes. Autopsies showed stomach lesions at all doses tested. The LD50 for the test item in rats was found to be 1110 mg/kg bw for both sexes.