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Description of key information

Acute toxicity: Via oral route: Key study: test method according to OECD 420 and EU Method B.1. In the preliminary study, the animal treated at 300 mg/kg bw presented signs of toxicity and died. In the main study, no toxicity signs were stated at 50 mg/kg. The test substance is classified as Category III according to OECD GHS criteria.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From May 11 to July 1, 2015.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to OECD Guideline 420, EU Method B.1 bis and GLP study.
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Experimental Medicine Centre, Medical University in Bialystok.
- Age at study initiation: 9 weeks old (300 mg/kg bw dose) and 10 weeks old (50 mg/kg bw dose).
- Weight at study initiation: 193.0 g (300 mg/kg bw dose) and 198.0 g (50 mg/kg bw dose), plus 4 animals whose average body weight was 187.5 g were assigned to the 50 mg/kg bw dose group.
- Fasting period before study: 19 hours.
- Housing: Plastic cages covered with wire bar lids. (58 x 37 x 21 cm). Preliminary experiment: 1 animal/cage. Main experiment: 4 animals/cage. UV-sterilized wood shavings were used as bedding.
- Diet (e.g. ad libitum): Ad libitum access to the “Murigran” standard granulated laboratory fodder (lot numbers: 3/15 and 4/15) produced by Wytwórnia Koncentaratów i Mieszanek Paszowych AGROPOL, Motycz.
- Water (e.g. ad libitum): Tap water ad libitum.
- Acclimation period: The animals were quarantined for 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-26 ºC.
- Humidity (%): 40-80%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark.
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
1%
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 1 ml of suspension contained 10 mg of the test item.
- Amount of vehicle (if gavage): 0.5 mL/100 g bw

MAXIMUM DOSE VOLUME APPLIED: 1 mililiter.

DOSAGE PREPARATION (if unusual):The test item was carefully mixed with 1% carboxymethylcellulose. The test item (a suspensin oin 1% carboxymethylcellulose) in a volume of 0.5 mL/100 g. The suspension of the test item in 1% carboxymethylcellulose was prepared on the administration day (shortly before the administration).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No previous data were available.
Doses:
50 and 300 mg/kg bw
No. of animals per sex per dose:
Preliminary study: 1 female per dose.
Main study: 4 additional females.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: Daily.
- Necropsy of survivors performed: yes.
- Other examinations performed: clinical signs, body weight, gross necropsy examinations.
Preliminary study:
Following single administration of the test item at a dose of 300 mg/kg bw, the following signs of toxicity were observed: rounded back, wavering gait, slight and distinc decreases of locomotor activity, and tremors. The animal died during the 2nd hour after the administration of the test item.
Following single administration of the test item at a dose of 50 mg/kg bw, no signs of toxicity were stated.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 50 mg/kg bw
Based on:
test mat.
Mortality:
All the animals survived the main experiment.
Clinical signs:
No signs of toxicity were stated.
Body weight:
Body weights of all animals increased (dose of 50 mg/kg bw).
Gross pathology:
The animals did not exhibit any pathological chagnes.

Table 1. Clinical signs - overall list.

Dose (mg/kg b.w.)

Day after administration

Number of living animals

Animal number

1*

2

3

4

5

50

0

1

1

0

Signs, D

-

-

-

-

-

30

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

NC

* the female from the preliminary experiment

NC=No Changes

Signs= Clinical signs

D=Death

Table 2. Body weights of the animals (g)

Dose
(mg/kg b.w)

Animal number

Day of the experiment / Body weight (g)

Body weight gain (g)
(0-14d)

0

7

14

300

1*

193

-

-

-

50

1*
2
3
4
5

198
191
200
187
172

211
218
220
210
197

228
233
234
214
200

30
42
34
27
28

*the female from the preliminary experiment


Interpretation of results:
Category 3 based on GHS criteria
Remarks:
Migrated information
Conclusions:
In the preliminary study, the animal treated at 300 mg/kg bw presented signs of toxicity and died. In the main study, no toxicity signs were stated at 50 mg/kg. The test substance is classified as Category III according to OECD GHS criteria.
Executive summary:

This study was performed according to OECD Guideline 420 and EU method B.1 (Fixed Dose Method) and following GLP. It intended to assess the toxicity properties of the test item when administered through oral route. It was performed on female Wistar rats. Since no previous data were available, a Preliminary experiment was performed with a dose of 300 mg/kg bw to 1 animal. The test item (a suspension in 1% carboxymethylcellulose) in a volume of 0.5 mL/100 g b.w. was administered with a metal stomach tube. The animal suffered signs of toxicity: rounded back, wavering gait, slight and distinct decreases of locomotor activity and tremors, and died 2h after the treatment. Consequently, the second female rat was treated with 50 mg/kg bw of the test item without manifesting any clinical sign of toxicity. In the main experiment 4 additional females were treated with 50 mg/kg bw. The animals were observed for 14 days and then euthanized for detailed gross examination. At 50 mg/kg no toxicity signs were stated. The animals survived the experiment. Gross examinations did not reveal any pathological changes. On the grounds of the study results, the test item can be classified as Category 3 according to GHS and to Regulation (EC) No. 1272/2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
50 mg/kg bw
Quality of whole database:
Klimisch score = 1. GLP study.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Acute toxicity: via oral route: Key study: The acute oral toxicity study on rats was performed according to OECD Guideline 420 and EU Method B.1 bus (Fixed Dose Method) (GLP study). The Fixed Dose method was applied to female Wistar rats. The preliminary experiment tested a 300 mg/kg bw dose to 1 animal, which suffered evident signs of toxicity and died. Then another animal received 50 mg/kg bw without showing any sign of toxicity. In the main experiment 4 additional females received the same dose without clinical signs in the 14 days observation period. Gross examinations did not reveal any pathological changes. On the grounds of the study results, the test item can be classified as Category 3 according to GHS and to Regulation (EC) No. 1272/2008.


Justification for selection of acute toxicity – oral endpoint
One study available.

Justification for classification or non-classification

On the grounds of the study results, the test item can be classified as Category 3 according to GHS and to Regulation (EC) No. 1272/2008.