Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 February 1985 to 27 March 1985
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985

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
Details on test material:
- Appearance: Light yellow liquid, Clear Liquid
- Stability: There was no apparent change in the physical state of the test material during administration
- Storage conditions of test material: Room temperature, protected from exposure to light

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 180 - 280g
- Fasting period before study: 18 hours
- Housing: Rats were housed individually in stainless steel 0.5” wire mesh cages
- Diet: ad libitum, checked daily and added or replaced as needed. Feeders were designed to reduce soiling, bridging and scattering
- Water: ad libitum fresh tap water, fit for human consumption, using an automatic watering system
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 to 70 %
- Photoperiod (hrs dark / hrs light): 12 hours of light, 12 hours of darkness

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.25 % Methylcellulose
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg (with the exception of the top dose level which was dosed as received at 4 mL/kg)
Doses:
1600, 2000, 2500, 3200, 3600 and 4000 mg/kg
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The rats were observed immediately and at one and four hours after dosing and twice daily for fourteen days for pharmacotoxicity, CNS effects and mortality. Body weights were recorded on the 14th day.
- Necropsy of survivors performed: Yes. The surviving rats were sacrificed by CO2 inhalation and a gross necropsy performed.
Statistics:
By the method of Litchfield and Wilcoxon

Results and discussion

Effect levelsopen allclose all
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 561 mg/kg bw
Based on:
test mat.
95% CL:
3 246 - 3 906
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
3 362 mg/kg bw
Based on:
test mat.
95% CL:
3 002 - 3 766
Mortality:
None of the rats died at 1600, 2000 or 2500 mg/kg; two of ten died at 3200 mg/kg, four of ten died at 3600 mg/kg and ten of ten died at 4000 mg/kg.
Clinical signs:
Signs observed included increased activity, decreased activity, nasal discharge, diarrhoea, salivation, lacrimation, ptosis, chromodacryorrhea, dyspnoea, decreased muscle tone, abnormal gait, abnormal stance and prostration.
Gross pathology:
Necropsy of animals dying on study revealed distended stomachs, lesions in the stomach mucosa and fluid-filled intestines. Congested, bright red lungs and foci on the kidneys were also observed.
No visible lesions were observed in any of the remaining animals at terminal necropsy.

Applicant's summary and conclusion

Interpretation of results:
other: Not classified in accordance with EU criteria
Conclusions:
Under the conditions of the study, the LD50 in female rats was determined to be 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.
Executive summary:

The potential of the test material to cause acute toxicity via the oral route was investigated using methodology equivalent or similar to the standardised guideline OECD 401 under GLP conditions.

Following a dose range-finding study, six groups of ten Sprague Dawley rats (five males and five females) were administered the test material at dose levels of 1600, 2000, 2500, 3200, 3600 and 4000 mg/kg in 0.25 % methylcellulose. The rats were observed immediately and at one and four hours after dosing and twice daily for fourteen days for pharmacotoxicity, CNS effects and mortality. Body weights were recorded on the 14th day. The surviving rats were sacrificed by CO2 inhalation and a gross necropsy performed. The statistical method used was the method of Litchfield and Wilcoxon.

Signs observed included episodes of increased or decreased activity, nasal discharge, diarrhoea, salivation, lacrimation, ptosis, chromodacryorrhea, dyspnoea, decreased muscle tone, abnormal gait, abnormal stance and prostration. None of the rats died at 1600, 2000 or 2500 mg/kg; two of ten died at 3200 mg/kg, four of ten died at 3600 mg/kg and ten of ten died at 4000 mg/kg. Necropsy of the animals dying on study revealed distended stomachs, lesions in the stomach mucosa and fluid-filled intestines. Congested, bright red lungs and foci on the kidneys were also observed. No visible lesions were observed in any of the remaining animals at terminal necropsy.

The calculated acute oral LD50 in male and female rats was determined to be 3561 mg/kg with 95 % confidence limits of 3246 to 3906 mg/kg and a slope of 1.11. The calculated acute oral LD50 in females was determined to be 3362 mg/kg with 95% confidence limits of 3002 to 3766 mg/kg and a slope of 1.1. An attempt was made to calculate the LD50 in males but the data generated did not lend itself to the method of Litchfield and Wilcoxon.

Under the conditions of the study, the LD50 in female rats was determined to be 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.