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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:
23 November to 17 December 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not effect the quality of the relevant results.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
Adopted 1996
Deviations:
yes
Remarks:
The low value recorded for humidity (26%) slightly below the range specified was not considered to have affected the integrity or validity of the study.
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
The low value recorded for humidity (26%) slightly below the range specified was not considered to have affected the integrity or validity of the study.
GLP compliance:
yes
Test type:
acute toxic class method

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and Female CD rats Sprague-Dawley origin (Hsd: Sprague-Dawley (CD) obtained from Harlan Laboratories UK Ltd., Bicester, Oxon, UK.
- Age at study initiation: five to seven weeks of age
- Weight at study initiation: 86 - 142 g .
- Fasting period before study: Overnight fast immediately before dosing and for approximately four hours after dosing
- Housing: The animals were housed in groups of three of the same sex in metal cages with wire mesh floors.
- Diet (e.g. ad libitum): Standard laboratory rodent diet (Special diet services RM1(E) SQC expanded pellet) was allowed throughout the study. Each batch of diet used for the study was analysed for certain nutrients, possible contaminants and micro-organisms
- Water (e.g. ad libitum): Free access to mains drinking water was allowed throughout the study. Results of routine physical and chemical examination of drinking water conducted by the supplier are made available to Huntington Life Sciences Ltd.
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to achieve limits of 20 to 23°C
- Humidity (%): Set to achieve limits of 26 - 50%
Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study.
- Air changes (per hr): At least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give twelve hours continuous light (07:00 to 19:00) and twelve hours darkness.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
The test item was formulated at a concentration of either 2% (200mg/kg) or 20% (2000mg/kg) in corn oil.

MAXIMUM DOSE VOLUME APPLIED:
10 ml/kg

DOSAGE PREPARATION (if unusual):
The test item was prepared on the day of dosing.

PROCEDURE
The appropriate dose volume of the test substance was administerd to each rat by oral gavage using a plastic syringe and catheter.
Doses:
200 and 2000 mg/kg
No. of animals per sex per dose:
2000 mg/kg 3 female
200 mg/kg 3 female
200 mg/kg 3 male
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for mortality twice daily. Animals were observed for clinical signs soon after dosing and thereafter at frequent intervals on the day of dosing. On subsequent days, surviving animals were observed at least once in the morning and again at the end of the experimental day.
- Individual bodyweights were recorded on Days 1 (prior to dosing) and eight and fifteen.
- Necropsy of survivors performed: yes; All surviving animals were killed on Day 15 by carbon dioxide asphyxiation. All animals were subjected to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The appearance of all examined organs was recorded.

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 200 - <= 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
> 200 mg/kg bw
Based on:
test mat.
Mortality:
See attached background information
Two females dosed at 2000 mg/kg died within 3 days of dosing. There were no other deaths.
Clinical signs:
See attached background information
Piloerection was observed soon after dosing in all animals treated with either 2000 or 200 mg/kg. All females treated with 2000 mg/kg also showed increased salivation, abnormal gait, ungroomed appearance, hunched posture and pink staining on muzzle and urogenital area, 2/3 also showed partially closed eyelids. Additional signs of Irritability, overactive behaviour, reduced body temperature, clear nasal discharge and deep respiration was seen in 1 of the females treated with 2000mg/kg.
In adition to piloerection 1 male and 1 female treated with 200 mg/kg showed increased salivation and all females treated with 200 mg/kg showed an abnormal gait.
Body weight:
See attached background information
All surviving animals were considered to have acheived satisfactory bodyweight gains.
Gross pathology:
See attached background information
No abnormalities were observed in animals that survived treatment and were terminated on Day 15.
Macroscopic examination of the 2 females treated with 2000 mg/kg and which died within 3 days of dosing revealed congestion of the subcutaneous tissue and brain with atrophy of the spleen and pallor of the kidneys. Congestion, gaseous distension and fluid contents were also noted in the stomach and along the alimentary tract.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the conditions of the test the acute oral median lethal dose (LD50) of the test item in the Sprague-Dawley strain rat was within the range of 200-2000 mg/kg body weight. According to the OECD 423 test guideline 1996, the LD50 cut-off value was considered to be 1000 mg/kg body weight
Executive summary:

Introduction.

The study was performed to assess the acute oral toxicity of the test item following a single oral administration in the Sprague-Dawley strain rat. The method was designed to be compatible with the following:

- OECD Guidelines for the Testing of Chemicals No. 423 “Acute Oral Toxicity – Acute Toxic Class Method” (adopted 22 March 1996)

- Method B1trisAcute Toxicity (Oral) of EEC Methods for the determination of toxicity, Annex to Directive 96/54/EEC

Method. 

A group of three fasted females was treated with a single oral gavage dose of the test item formulated in corn oil, at a dose level of 2000 mg/kg bodyweight. As a result of mortalities at this dose level indicating the acute lethal oral dose of the test item to be less than 2000 mg/kg bodyweight, a further group of three fasted females was dosed at 200 mg/kg. A group of three fasted males was then dosed at 200 mg/kg to confirm the results at 200mg/kg.

Mortality, clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. 

Two females dosed at 2000 mg/kg died within 3 days of treatment. Slight bodweight loss was observed in both animals. Macroscopic examination of both animals revealed congestion of the subcutaneous tissue and brain with atrophy of the spleen and pallor of the kidneys. Congestion, gaseous distension and fluid contents were also noted in the stomach and along the alimentary tract.

Clinical Observations. 

Clinical signs of reaction to treatment comprised piloerection, seen in all rats treated with test item. This sign was accompanied in all rats at 2000 mg/kg by increased salivation, abnormal gait, ungroomed appearance, hunched posture and pink staining on muzzle and uro/genital area with partially closed eyelids in two females at 2000 mg/kg.

Bodyweight. 

The surviving animals showed satisfactory gains in bodyweight over the study period.

Necropsy. 

No abnormalities were observed for animals that survived treatment to study termination on Day 15.

Conclusion. 

The acute oral median lethal dose (LD50) of the test item in the Sprague-Dawley strain rat was estimated to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight, (Globally Harmonised Classification System – Acute Oral Toxicity Category 4). According to the OECD 423 test guideline 1996, the LD50 cut-off value was considered to be 1000 mg/kg body weight

The test item was classified as Acute Oral Toxicity Category 4 according to the Regulation (EC) No. 1272/2008. The Signal Word ‘Warning’ and the Hazard Statement ‘H302: Harmful if swallowed’ are therefore required.