Registration Dossier

Administrative data

Description of key information

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight (Globally Harmonized Classification System - Category 4). In accordance with section 2 of REACH (regulation(EC) No 1907/2006) Annex VIII, acute toxicity via dermal and inhalation routes (required in Annex VIII section 8.5) does not need to be conducted as the substance is classified as corrosive to the skin.

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:
The study was conducted between 14 July 2015 and 06 August 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
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
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Female Wistar (RccHan™:WIST) strain rats were supplied 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 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were 8 to 12 weeks of age. The body weight variation did not exceed ±20% of the mean body weight at the start of treatment.The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) 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 12 hours continuous light (06:00 to 18:00) and 12 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.JustificationRats are the preferred species of choice as historically used for safety evaluation studies and are specified in the appropriate test guidelines.
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Test Item Formulation and Experimental PreparationFor the purpose of the study the test item was freshly prepared, as required, as a solution in distilled water. The test item was formulated within 2 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.A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.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 group to confirm the survival of the previously dosed animals.
Doses:
300 and 2000 mg/kg
No. of animals per sex per dose:
A single animal was treated at 300 mg/kg.A single animal was treated at 2000 mg/kg.An additional four animals were treated at 300 mg/kg.
Control animals:
no
Details on study design:
Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 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 surviving animals were killed 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.
Statistics:
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made. 
Preliminary study:
See below
Sex:
female
Dose descriptor:
LD50
Effect level:
300 - 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose Level - 2000 mg/kgThe animal was killed for humane reasons, 6 days after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence. Dose Level - 300 mg/kgThere were no deaths.
Clinical signs:
Dose Level - 2000 mg/kgSigns of systemic toxicity noted were hunched posture, pilo-erection, labored and noisy respiration, increased respiratory rate, distended abdomen, pallor of the extremities and emaciation. Dose Level - 300 mg/kgNo signs of systemic toxicity were noted during the observation period.
Body weight:
Dose Level - 300 mg/kgAll animals showed expected gains in body weight over the observation period.
Gross pathology:
Dose Level - 2000 mg/kgAbnormalities noted at necropsy were pale liver, pale kidneys and gaseous stomach and small and large intestines. Dose Level - 300 mg/kgNo abnormalities were noted at necropsy.

Individual Clinical Observations and Mortality Data -2000mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

2000

2-0

Female

0

0

H

H

H

HP

HPRlRn

HPRlRn

HPRlRnDa

HPRlRiDaEEmX*

0 = No signs of systemic toxicity

H = Hunched posture

P = Pilo-erection

Rl = Labored respiration

Rn  = Noisy respiration

Ri = Increased respiratory rate

Da = Distended abdomen

E = Pallor of the extremities

Em = Emaciation

X* = The animal was killed for humane reasons, 6 days after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in   the UK Home Office Project Licence

Individual Body Weights and Body Weight Changes -2000mg/kg

Dose Level

mg/kg

Animal Number and Sex

Body Weight (g) at Day

Body Weight (g)
at Death

Body Weight Gain (g)
During Week

0

7

14

1

2

2000

2-0 Female

168

-

-

126

-

-

- = Animal dead

Individual Necropsy Findings -2000 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0 Female

Humanely killed Day 6

Liver: pale

Kidneys: pale

Stomach: gaseous

Small intestine: gaseous

Large intestine: gaseous

Individual Clinical Observations and Mortality Data -300mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

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

Individual Body Weights and Body Weight Changes -300mg/kg

Dose Level

mg/kg

Animal Number
and Sex

Body Weight (g) at Day

Body Weight Gain (g) During Week

0

7

14

1

2

300

1-0 Female

173

190

200

17

10

3-0 Female

151

165

178

14

13

3-1 Female

179

190

216

11

26

3-2 Female

152

163

176

11

13

3-3 Female

150

167

182

17

15

Individual Necropsy Findings-300mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

1-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Killed Day 14

No abnormalities detected

3-2 Female

Killed Day 14

No abnormalities detected

3-3 Female

Killed Day 14

No abnormalities detected

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated informationCriteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight (Globally Harmonized Classification System - Category 4).
Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

 

Methods

Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a solution in distilled water, at a dose level of 300 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

  

Results

Mortality. The animal treated at a dose level of 2000 mg/kg was killed for humane reasons, 6 days after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence. There were no deaths at a dose level of 300 mg/kg.

 

Clinical Observations. Signs of systemic toxicity noted in the animal treated at a dose level of 2000 mg/kg were hunched posture, pilo-erection, labored and noisy respiration, increased respiratory rate, distended abdomen, pallor of the extremities and emaciation. There were no signs of systemic toxicity noted at a dose level of 300 mg/kg.

 

Body Weight. Surviving animals showed expected gains in body weight.

 

Necropsy. Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg were pale liver, pale kidneys, gaseous stomach and gaseous small and large intestines. No abnormalities were noted at necropsy of animals treated at a dose level of 300 mg/kg.

 

Conclusion

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight (Globally Harmonized Classification System-Category 4).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw
Quality of whole database:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated in a GLP study conducted in accordance with GLP. The LD50 was determined to be in the range of 300 - 2000 mg/kg body weight and in accordance with the UN Globally Harmonized Classification System the test item is considered to be Acute Toxicity Category 4

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

Justification for classification or non-classification

Based on the results mentioned above, the acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was determined to be in the range of 300 - 2000 mg/kg body weight , therefore the substance is, according to CLP and GHS, classified as Acute Oral Tox. Category 4.