Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 June 2014 to 16 July 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: There were no deviations (unplanned changes) from the study plan.
Cross-reference
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

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
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Appearance/physical state: Clear colourless liquid
- Storage conditions: Room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Female Wistar s were suppiied by Harlan Laboratories UK Ltd., Oxon, UK.
On receipt the animals were randomly allocated to cages.
The females were nulliparous and non-pregnant. After an acclimatization period of at least five days the animais were selected at random and given a number unique within the study by indelible inkmarking on the tail and a number written on a cage card.
At the start of the study the animais were eight to tweive weeks of age. The body weight variation did not exceed ±20% of the body weight of the initially dosed animal.

The animals were housed in groups of up to four in suspended solid-floor poiypropyiene cages furnished with woodflakes. With the exception of an overnight fast immediateiy before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose level to confirm the survival of the previously dosed animals.
Doses:
2000 mg/kg
No. of animals per sex per dose:
4 (main study) + 1 (preliminary) for a total of 5
Control animals:
no
Details on study design:
A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study.
Clinical observations were made 1/2, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily.
Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an extemal examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
No statistic: using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.

Results and discussion

Preliminary study:
Yes, with one female
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
There were no deaths.
Clinical signs:
Hunched posture was noted two and four hours after dosing in the initial treated animal. No signs of systemic toxicity were noted in the additional treated animals during the observation period.
Body weight:
All animals showed expected gains in body weight over the observation period.
Gross pathology:
No abnormalities were noted at necropsy.

Any other information on results incl. tables

Table1     Individual Clinical Observations and Mortality Data

Dose Level mg/kg

 

Animal Number and Sex

Effects Noted After Dosing

(Hours)

Effects Noted During Period After Dosing (Days)

Yz

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

 

 

 

 

 

 

 

2000

1-0

Female

 

0

 

0

 

H

 

H

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

2-0

Female

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

2-1

Female

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

2-2

Female

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

2-3

Female

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

0

 

---

 

0

 

0

 

0

 

0

 

0

 

0

 

0

H = hunched posture

0 = No signs of systemic toxicity

Table 2     lndividual Body Weights and Body Weight Changes

 Dose Level mg/kg

Animal Number and Sex

 

Body Weight(g) at Day

 

Body Weight Gain(g) During Week

 

0

 

7

 

14

 

1

 

2

 

 

 

 

 

 

2000

 

1-0 Female

 

167

 

181

 

191

 

14

 

10

 

2-0 Female

 

171

 

182

 

200

 

11

 

18

 

2-1 Female

 

175

 

202

 

218

 

27

 

16

 

2-2 Female

 

175

 

195

 

210

 

20

 

15

 

2-3 Female

 

 

173

 

193

 

206

--

 

20

 

13

 

1

Table3     lndividual Necropsy Findings

Dose Level mg/kg

Animal Numbe rand Sex

 Time of Death

 Macroscopic Observations

 

 

 

 

 

 

 

2000

 

1-0 Female

 

Killed Day 14

 

No abnormalities detected

 

2-0 Female

 

Killed Day 14

 

No abnormalities detected

 

2-1 Female

 

Killed Day 14

 

No abnormalities detected

 

2-2 Female

 

Killed Day 14

 

No abnormalities detected

 

2-3 Female

 

Killed Day 14

 

No abnormalities detected

 


Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was considered to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System - Unclassified).
Executive summary:

Introduction

The study was performed to OECD Guideline No. 420 and EU Method B.1 bis to assess the acute oral toxicity of the test item in the Wistar strain rat.

Methods

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

Results

Mortality: There were no deaths.

Clinical Observations: Hunched posture was noted during the day of dosing in the initial treated animal. There were no signs of systemic toxicity noted in the additional treated animals.

Body Weight: All animals showed expected gains in body weight.

Necropsy: No abnormalities were noted at necropsy.

Conclusion

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was considered to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System- Unclassified).