Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From October 20, 1999 to November 24, 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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
March 1996
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity: approximately 81%
- Lot/batch No.: 876 TK
- Physical state: off white paste
- Storage condition of test material: room temperature, in the dark.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source: Stepan Company, Illinois, USA
- Batch No.of test material: 876 TK
- Date received: 10 May 1999
- Description: white paste

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Freshly prepared as a solution in distilled water
- Final dilution of a dissolved solid: The dose levels were selected using a correction factor based on the purity of the supplied test material, to ensure that the animals received equivalent to 200 or 2000 mg/kg of pure test material

FORM AS APPLIED IN THE TEST: solution

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: eight to twelve weeks old
- Weight at study initiation: males weighed 228 to 236 g, and the females 201 to 236 g
- Fasting period before study: overnight immediately before dosing
- Housing: in groups of three by sex in solid-floor polypropylene cages furnished with wood flakes
- Diet (e.g. ad libitum): ad libitum with the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): The rate of air exchange was approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): cycles of twelve hours continuous light and twelve hours darkness.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 217.4 mg/ml and 21.8 mg/ml
- Amount of vehicle (if gavage): 10 ml

DOSAGE PREPARATION: The test material solutions were prepared using a correction factor based on the purity of the test substance to ensure that the animals received 2000 and 200 mg/kg of the pure test material

MAXIMUM DOSE VOLUME APPLIED: 10 mL

CLASS METHOD
- Rationale for the selection of the starting dose: Using all available information, 2174 mg/kg bodyweight was selected as the starting dose. (This dose level was selected using a correction factor based on the purity of the supplied test material, to ensure that the animals received a dose equivalent to 2000 mg/kg bodyweight of pure test material).

OTHER:
- animals were dosed once only by gavage
Doses:
2174 mg/kg (equivalent to 2000 mg pure test material/kg) and 218 mg/kg (equivalent to 200 mg pure test material/kg)
No. of animals per sex per dose:
2174 mg/kg: a group of 3 female rats.
218 mg/kg: 6 rats, one group of 3 females and one group of 3 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Observation: Death or overt signs of toxicity were checked at 1/2, 1, 2, and four hours after dosing and subsequently once daily
- Frequency of observations and weighing: Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 500 - <= 1 000 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: Pure test substance
Mortality:
Two animals treated with 2174 mg/kg were found dead one or two days after dosing. There were no deaths noted at a dose level of 218 mg/kg.
Clinical signs:
Clinical signs in animals treated with 2174 mg/kg were hunched posture, diarrhoea and increased salivation with incidents of pallor of the extremities, emaciation, lethargy, pilo-erection, decreased respiratory rate, laboured respiration, red/brown staining around snout and tiptoe gait. The surviving female animals treated with 2174 mg/kg recovered five days after dosing. All animals treated with 218 mg/kg appeared normal throughout the study.
Body weight:
All surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
Necropsy results of animals that died during the study (treated with 2174 mg/kg) were haemorrhagic lungs, dark liver, dark kidneys, haemorragic gastric mucosa, sloughing and/or haemorrhage of the non-glandular epithelium of the stomach and haemorrhagic small and large intestines. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Any other information on results incl. tables

Mortality data

Dose level [mg/kg]

Animal number and sex

Number of animals treated

Deaths during day of dosing (hours)

Effects noted after dosing (days)

1/2

1

2

4

1

2

3

4

5

6

7

8-14

Deaths

2174

female

3

0

0

0

0

1

1

0

0

0

0

0

0

2/3

218

female

3

0

0

0

0

0

0

0

0

0

0

0

0

0/3

male

3

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Individual Clinical Observations

Dose level mg/kg

Animal Nº

and Sex

Effects Noted After Dosing (Hours)

Effects Noted During Period After Dosing (Days)

 

2174

 

 

1/2

1

2

4

1

2

3

4

5

6

7

18

9

10

11

12

13

14

1-0

Female

H

H

HD

HD

HPD

Ss

HP

H

H

 

 

 

 

 

 

 

 

 

 

1-1

Female

HS

HS

 

HSD

HSD

X

 

 

 

 

 

 

 

 

 

 

 

 

 

1-2

Female

HS

HS  

HSD

HSD

H

P

Em

Ss

Rd

RL

Wt

E

X

 

 

 

 

 

 

 

 

 

 

 

 

D: diarrhea                                                               E: pallor of extremities

Em: emaciation                                                         H: hunched posture

L: lethargy                                                               P: pilo-erection

Rd: decreased respiratory rate                                RL: labored respiration

S: increased salivation                                              Ss: red/brown stainig around snout

Wt: tiptoe gait                                                          X: animal death

 

No clinical observations were noted at the dose of 218 mg/kg.

 

Body weights

Only the animals that died during the study showed a loss of weight

 

Dose Level mg/kg

Animal Nº and sex

Body weight (g) at day

Body weight (g) at

2174

 

0

7

14

Death

1-0 Female

204

243

246

 

1-0 Female

201

 

 

175

1-0 Female

207

 

 

170

 

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral LD50 of the test substance in Sprague-Dawley rats was in the range of 500 – 1000 mg/kg of pure test substance.
Executive summary:

The Acute Oral toxicity – Acute Toxic Class method study for the test substance was performed in Sprague-Dawley CD strain rats. A stepwise procedure was used; a group of 3 females was treated with a dose of 2174 mg/kg (equivalent to 2000 mg of pure test material/kg) and 2 groups (3 females and 3 males) were treated with a dose of 218 mg/kg (equivalent to 200 mg of pure test material /kg). The test substance was administered as a solution in distilled water. An oral single dose of 10 ml/kg was given by gavage.

Clinical observations were made at 0.5, 1, 2, and 4 hours after dosing and subsequently once daily for up to 14 days. Individual body weights were recorded prior to dosing and 7 and 14 days after treatment or at death. All animals including those that died during the study were subjected to gross pathological observations. Two animals treated with 2174 mg/kg died. No mortalities were noted in animals treated with 218 mg/kg. The acute oral LD50 of the test substance in Sprague-Dawley rats was in the range of 500 – 1000 mg/kg of pure test substance.