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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:
01 June 1988 to 28 June 1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
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:
- Appearance: Brown liquid
- Storage conditions of test material: Ambient conditions

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Sprague-Dawley Crl:CD®BR
- Age at study initiation: Young adults. Males were 63 and 70 days old; females were 70 and 77 days old.
- Weight at study initiation: Males weighed between 272 and 353 g; females weighed between 186 and 217 g.
- Fasting period before study: Yes. Overnight prior to dosing.
- Housing: The animals were individually housed in stainless-steel wire-bottom cages in an air conditioned room.
- Diet: ad libitum access to certified rodent feed
- Water: ad libitum
- Acclimation period: 14 days for males, 21 days for females

ENVIRONMENTAL CONDITIONS
- Temperature: 20.2 to 20.6 °C
- Humidity: 47.9 to 76.4 % (relative)
- Photoperiod: 12-hour light/dark cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: No data on dose volume
The mean (± SD) weight of the test material administered was 0.57 to 0.87 (± 0.001 to 0.002) g/kg for the males and 0.27 to 0.59 (± 0.01 to 0.02) g/kg for the females.
Doses:
2.0, 2.5 and 3.0 g/kg for males and 1.3, 2.0 and 3.0 g/kg for females
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
yes
Remarks:
Five fasted un-dosed animals of each sex served as the controls for the 2.0 and 3.0 g/kg animals; however, in order to conserve additional animals, controls were not used for the 1.3 g/kg females and 2.5 g/kg males
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed frequently on the day of dosing for any physiological or behavioural abnormalities and at least once each day for 14 days after treatment. The animals were weighed immediately before dosing and at 2, 7, and 14 days after treatment.
- Necropsy of survivors performed: Yes. A necropsy was performed on all animals. This included an examination of the external body surface, all orifices, the thoracic and abdominal cavities, and the contents of these cavities. The animals were sacrificed by carbon dioxide.
- Other examinations performed: Histopathology. Abnormal tissues and necessary comparison tissues were collected and fixed in 10 % neutral buffered formalin by immersion. Tissues were trimmed and embedded in paraffin, sectioned and stained with haematoxylin and eosin. Glandular stomach, non-glandular stomach and thymus gland were examined from the control animals (5 males, 5 females). Tissues with abnormal macroscopic findings were examined from all animals, including the controls.
Statistics:
The LD50, slope, and 95 % confidence limits were determined using the method of Berkson.
Mean body weights of the 3.0 and 2.0 g/kg dose groups were compared to concurrent controls using one way analysis of variance (ANOVA) when there were three or more survivors per dose group.

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
2 100 mg/kg bw
Based on:
test mat.
95% CL:
>= 1 100 - <= 3 400
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
2 700 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 300 - <= 3 500
Mortality:
No animals died in the control group or the 1.3 g/kg female dose group and the 2.0 g/kg male dose group.
For the females, 2 animals died in the 2.0 g/kg dose group on days 1 to 2 and all 5 animals died in the 3.0 g/kg dose group on days 1 to 3.
For the males, 1 animal died in the 2.5 g/kg dose group on day 2 and 4 animals died in the 3.0 g/kg dose group on days 2 to 3.
Clinical signs:
Salivation, anogenital, nasal and ocular discharge, decreased motor activity, soft faeces and reduced food consumption were frequently observed signs of toxicity.
Signs of toxicity observed infrequently included: collapse, closed eyes, decreased body temperature, oral discharge, unkempt appearance, and abnormal respiration. No signs of toxicity were evident in surviving animals after Day 5. Other clinical observations were not considered to be signs of toxicity.
Body weight:
Mean body weights were significantly less than controls in males at 2.0 g/kg through Day 14 and females at 2.0 g/kg on Day 2. Depressed body weights were observed in 2.5 g/kg males through Day 7 and in 1.3 g/kg females on Day 2.
Gross pathology:
MACROSCOPIC PATHOLOGY
The following macroscopic findings were observed in the tissues of animals in this study: red discoloration or foci in the thymus, red discoloration of the glandular or non-glandular stomach, friable areas in the non-glandular stomach, tan foci in the liver, degeneration in a testicle, an enlarged mandibular lymph node and alopecia.

MICROSCOPIC PATHOLOGY
The most common findings were congestion and/or haemorrhage in the thymus gland, and congestion in the stomach (glandular and non-glandular portions). Autolysis was common in animals dying during the study period.
In one animal of the 2.5 g/kg dose group and one from the 3.0 g/kg dose group, there was epithelial necrosis in the glandular portion of the stomach. Oedema, inflammation, ulceration, and haemorrhage were present in the non-glandular portion of the stomach of one animal from the 3.0 g/kg dose group.
Ulceration and inflammation of the non-glandular stomach also occurred in the animal from the 2.5 g/kg dose group that exhibited epithelial necrosis.
The gastric necrosis (in both glandular and non-glandular portions) was considered to be related to the oral exposure to the compound. The other findings were likely secondary lesions associated with the loss of integrity in the gastric mucosa or secondary to vascular shock associated with an agonal death.

Any other information on results incl. tables

Table 1: Mean Body Weights

Sex

Dose (g/kg)

 

Weight (g)

Day 0

Day 2

Day 7

Day 14

M

0

Mean

N

287 ± 9

5

322 ± 12

5

357 ± 14

5

397 ± 15

5

2.0

Mean

N

286 ± 10

5

260 ± 23**

5

312 ± 18**

5

369 ± 19*

5

2.5

Mean

N

335 ± 11

5

288 ± 3

5

335 ± 19

4

395 ± 22

5

3.0

Mean

N

289 ± 3

5

239 ± 2

2

284†

1

356†

1

F

0

Mean

N

199 ± 8

5

214 ± 9

5

228 ± 9

5

240 ± 10

5

1.3

Mean

N

206 ± 9

5

201 ± 18

5

227 ± 13

5

244 ± 16

5

2.0

Mean

N

198 ± 7

5

171 ± 15**

3

217 ± 1

3

246 ± 10

3

3.0

Mean

N

197 ± 6

5

161†

1

-

-

†Not statistically analysed

 - = Complete mortality

*Significantly less than the controls at p=0.05

**Significantly less than the controls at p=0.01

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Not classified in accordance with EU criteria
Conclusions:
Under the conditions of the study, the LD50 and 95 % confidence limits were determined to be 2700 (2300 to 3500) mg/kg for males and 2100 (1100 to 3400) mg/kg for females.
Executive summary:

A study was conducted to determine the acute oral toxicity potential of the test material under GLP conditions using methodology equivalent to the standardised guideline OECD 401.

Single doses of the undiluted test material were administered via gavage to fasted male and female Sprague-Dawley rats. Doses administered were 2.0, 2.5 and 3.0 g/kg for males and 1.3, 2.0 and 3.0 g/kg for females. Animals were observed frequently on the day of dosing for any physiological or behavioural abnormalities and at least once each day for 14 days after treatment. The animals were weighed immediately before dosing and at 2, 7, and 14 days after treatment. A necropsy was performed on all animals and any abnormal tissues and necessary comparison tissues were collected, fixed and processed.   

No animals died in the control group or the 1.3 g/kg female dose group and the 2.0 g/kg male dose group. For the females, 2 animals died in the 2.0 g/kg dose group on days 1 to 2 and all 5 animals died in the 3.0 g/kg dose group on days 1 to 3. For the males, 1 animal died in the 2.5 g/kg dose group on day 2 and 4 animals died in the 3.0 g/kg dose group on days 2 to 3.

Salivation, anogenital, nasal and ocular discharge, decreased motor activity, soft faeces and reduced food consumption were frequently observed signs of toxicity. Signs of toxicity observed infrequently included: collapse, closed eyes, decreased body temperature, oral discharge, unkempt appearance, and abnormal respiration. No signs of toxicity were evident in surviving animals after Day 5. Other clinical observations were not considered to be signs of toxicity.

Mean body weights were significantly less than controls in males at 2.0 g/kg through Day 14 and females at 2.0 g/kg on Day 2. Depressed body weights were observed in 2.5 g/kg males through Day 7 and in 1.3 g/kg females on Day 2. Test material-related pathological changes were observed in the stomach.

Under the conditions of the study, the LD50 and 95 % confidence limits were determined to be 2700 (2300 to 3500) mg/kg for males and 2100 (1100 to 3400) mg/kg for females.