Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 Oct - 13 Nov 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
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)
Remarks:
The department of health of the government of the United Kingdom
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: The bodyweight variation did not exceed ± 20% of the initial/mean bodyweight of any previously dosed animal(s).
- Fasting period before study: overnight fast immediately before dosing
- Housing: The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: ad libitum (2014C Teklad Global Rodent diet supplied by Harlan Teklad, Oxon, UK)
- Water: ad libitum; drinking water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): at least 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: distilled water
Details on oral exposure:
For the purpose of the study the test item was freshly prepared, as required, as a suspension in distilled water.
The test item was formulated within two hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made ½, 1, 2, and 4 hours after dosing and subsequently once daily for 14 days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes
At the end of the observation period the animals were sacrificed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.


Results and discussion

Preliminary study:
The starting dose in the sighting test was 300 mg/kg bw administered to one female. No mortality was observed, but hunched posture was noted during the day of dosing.

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose level: 2000 mg/kg bw
Two animals were found dead during the day of dosing or two days after dosing. One animal was sacrificed for humane reasons, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence, four days after dosing (see table 1).

Dose Level: 300 mg/kg bw
There were no deaths.
Clinical signs:
Dose level: 2000 mg/kg bw:
There were no signs of systemic toxicity noted in the initial treated animal. Hunched posture was noted in 3/4 of the additional treated animals. Signs of systemic toxicity noted in one animal were lethargy, ataxia, pilo-erection, emaciation, laboured respiration, decreased respiratory rate and dehydration.

Dose Level: 300 mg/kg bw:
Hunched posture was noted in the initial treated animal during the day of dosing. No signs of systemic toxicity were noted in the four additional treated animals.
Body weight:
Dose level: 2000 mg/kg bw:
Individual bodyweights and bodyweight changes are given in Table 2.
Surviving animals showed expected gains in bodyweight over the observation period.

Dose Level: 300 mg/kg bw:
Individual bodyweights and bodyweight changes are given in Table 5.
All animals showed expected gains in bodyweight over the observation period.
Gross pathology:
Dose level: 2000 mg/kg bw:
Individual necropsy findings are given in Table 3.
Haemorrhagic, ulcerated and epithelial sloughing of the gastric mucosa were noted at necropsy of the animals that died during the study. Abnormalities noted at necropsy of the animal that was sacrificed due to humane reasons during the study were patchy pallor of the liver and gaseous stomach and small and large intestines. No abnormalities were noted in the surviving animals

Dose Level: 300 mg/kg bw:
Individual necropsy findings are given in Table 6.
No abnormalities were noted at necropsy.

Any other information on results incl. tables

Table 1. Individual clinical observations and mortality data – 2000 mg/kg bw

Dose level mg/kg bw

Animal

number

and sex

Effects noted after dosing (hours)

Effects noted during periods after doing

(days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

*

0

0

0

0

3-0

Female

H

H

H

0

0

X

 

 

 

 

 

 

 

 

 

 

 

 

3-1 Female

H

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

3-3

Female

H

HL

H

0

0

0

HPARlEm

HPALRl

RdEmDhX*

 

 

 

 

 

 

 

 

 

 

 

= No signs of systemic toxicity

H = Hunched posture

L = Lethargy

A = Ataxia

P = Pilo-erection

Em = Emaciation

Rl = Laboured respiration

Rd = Decreased respiratory rate

Dh = Dehydration

X = Animal dead

X° = Animal found dead 18 minutes after dosing

X* = Animal sacrificed for humane reasons, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home

Office Project Licence 

 

 Table 2. Individual bodyweight and bodyweight changes– 2000 mg/kg bw

Dose level mg/kg

Animal

number

and sex

Bodyweight (g) at day

 

Bodyweight (g) at

Death

Bodyweight gain (g) during week

 

0

7

14

1

2

2000

2-0

Female

169

180

190

 

11

10

3-0

Female

157

-

-

149

-

-

3-1

Female

166

194

207

 

28

13

3-2

Female

151

-

-

151

-

-

3-3

Female

154

-

-

116

-

-

- = animal dead

 

Table 3. Individual Necropsy Findings – 2000 mg/kg bw

Dose level mg/kg

Animal

number

and sex

Time of death

Macroscopic observations

2000

2-0

Female

Killed day 14

No abnormalities detected

3-0

Female

Found dead Day 2

Gastric mucosa: haemorrhagic

epithelial sloughing

3-1

Female

Killed day 14

No abnormalities detected

3-2

Female

Found dead Day 2

Gastric mucosa: haemorrhagic

ulcerated

3-3

Female

Humanely killed Day 4

Liver: patchy pallor

Stomach: gaseous

Small intestine: gaseous

Large intestine: gaseous

 

 

Table 4. Individual clinical observations and mortality data.– 300 mg/kg bw

Dose level mg/kg

Animal

number

and sex

Effects noted after dosing (hours)

Effects noted during periods after doing

(days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300 

1-0

Female

H

H

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0 = No signs of systemic toxicity

H = Hunched posture

 

 

Table 5. Individual bodyweight and bodyweight changes– 300 mg/kg bw

Dose level mg/kg

Animal

number

and sex

Bodyweight (g) at day

 

Bodyweight gain (g) during week

 

0

7

14

1

2

300

1-0

Female

158

184

200

26

16

4-0

Female

142

156

182

14

26

4-1

Female

159

164

188

5

24

4-2

Female

148

152

174

4

22

4-3

Female

155

165

182

10

17

 

 

 Table 6. Individual Necropsy Findings– 300 mg/kg bw

Dose level mg/kg

Animal

number

and sex

Time of death

Macroscopic observations

300

1-0

Female

Killed day 14

No abnormalities detected

4-0

Female

Killed day 14

No abnormalities detected

4-1

Female

Killed day 14

No abnormalities detected

4-2

Female

Killed day 14

No abnormalities detected

4-3

Female

Killed day 14

No abnormalities detected

 

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
CLP: Cat. 4, H302