Registration Dossier

Administrative data

Description of key information

Oral: LD50, rat > 2000 mg/kg
Dermal: LD50, rat > 2000 mg/kg

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:
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.
Reason / purpose:
reference to other 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
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
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.
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.
Preliminary study:
Yes, with one female
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.

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

 


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).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
Modern GLP studies following OECD test guidelines, both Klimisch grade 1.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 June 2014 to 10 July 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: There were no deviations (unplanned changes) from the study plan.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Five male and five female Wistar 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 five 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 animais weighed at least 200 g, and were eight to twelve weeks of age. The weight variation did not exceed ±20% of the mean weight for each sex.
The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes.
The animals were housed individually during the 24-Hour exposure period and in groups of five, by sex, for the remainder of the study.
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 could 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.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
The calculated volume of test item, as received, was applied as evenly as possible to an area of shorn skin (approximately 10% of the total body surface area) using a graduated syringe. A piece of surgical gauze was placed over the treatment area and semi occluded with a piece of self adhesive bandage.
The animals were caged individually for the 24-Hour exposure period. Shortly after dosing the dressings were examined to ensure that they were securely in place.

The animals were observed for deaths or overt signs of toxicity 1/2, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.
Duration of exposure:
After the 24-Hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item. The animals were returned to group housing for the remainder of the study period.
Doses:
Test item at a dose level of 2000 mg/kg.
No. of animals per sex per dose:
a group of five male and five female rats was treated
Control animals:
no
Details on study design:
After removal of the dressings and subsequenty once daily for fourteen days, the test sites were examined for evidence of primary irritation and scored.
Any other skin reactions, if present were also recorded.
Individual body weights were recorded prior to application of the test item on Day 0 and on Days 7 and 14.
At the end of the study 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:
Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test item was made.
Preliminary study:
None
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted 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.
Other findings:
Very slight erythema was noted at the test site of one female two to seven days after dosing. Crust formation and/or glossy skin were noted at the test sites of four females five to seven days after dosing. Small superficial scattered scabs were also noted at the test site of one female three to eight days after dosing.
There were no signs of dermal irritation noted at the test sites of all males and one female.

- Refer to Tables 1 to 5 (attached)

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.
Executive summary:

Introduction

The study was performed to OECD Guideline No. 402 and EU Method B.3 to assess the acute dermal toxicity of the test item in the Wistar strain rat.

Methods

A group often animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the undiluted test item to intact skin 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: There were no signs of systemic toxicity.

Dermal Irritation: Signs of dermal irritation noted at the test sites of four females included very slight erythema, crust formation, glossy skin and small superficial scattered scabs. There were no signs of dermal irritation noted at the test sites of all males and one female.

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

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

ACUTE ORAL TOXICITY STUDY

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.

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.

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.

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.

ACUTE DERMAL TOXICITY STUDY

The study was performed to OECD Guideline No. 402 and EU Method B.3 to assess the acute dermal toxicity of the test item in the Wistar strain rat.

A group often animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the undiluted test item to intact skin 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.

Mortality: There were no deaths.

Clinical Observations: There were no signs of systemic toxicity.

Dermal Irritation: Signs of dermal irritation noted at the test sites of four females included very slight erythema, crust formation, glossy skin and small superficial scattered scabs. There were no signs of dermal irritation noted at the test sites of all males and one female.

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

Necropsy. No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.

ACUTE INHALATION

The substance is a liquid with a vapour pressure of 0.0091 Pa at 25 °C and it is expected that inhalation exposure from identified uses will be low. In addition, the most likely route of exposure for workers and consumers is the dermal route.

Justification for classification or non-classification

No adverse effects or deaths noted at 2000 mg/kg. No classification required.