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Description of key information

Acute Oral Toxicity

The acute oral median lethal dose (LD50) of the test material was estimated to be in the range of 300 - 2000 mg/kg body weight therefore the test material can be classed as harmful through the oral route.

Acute Dermal Toxicity

Under the conditions of the study, 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 Toxicity

In accordance with Section 8.5.2. Column 2 of Annex VIII of the REACH Regulation, the information required under 8.5.2 (acute toxicity via the inhalation route) to 8.5.3 (via the dermal route) shall be provided for at least one route.  The choice for the second route will depend on the nature of the substance and the likely route of human exposure.  As inhalation of the material is unlikely, data via the dermal route is provided and therefore further information on toxicity via the inhalation route is not required. Furthermore the Vapor Pressure is 0.006479 Pa at 25 °C and therefore is of no concern for outdoor uses.

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:
16 February 2016 to 16 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation.
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 to 12 weeks of old.
- Weight at study initiation: Body weights ranged from 171 to 208 g. The body weight variation did not exceed ± 20 % of the mean body weight at the start of treatment.
- Fasting period: Yes. The animals had an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing.
- Housing: The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with wood flakes.
- Diet: Ad libitum.
- Water: Ad Libitum.
- Acclimation period: A period of at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C.
- Humidity (%): 30 to 70 % respectively.
- Air changes (per hr): At least fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness.
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
All animals were dosed only once by gavage using a metal cannula attached to a graduated syringe.

VEHICLE
- Concentration in vehicle: 30 mg/mL
- Amount of vehicle: The volume administered to each animal (10 mL/kg) was calculated according to the fasted body weight at the time of dosing.
- Justification for choice of vehicle: Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (2.18 mL/kg). The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.

DOSAGE PREPARATION: The test material was freshly prepared, as required, as a solution in arachis oil BP. To aid dissolution the test material formulations were warmed in a sonic bath set at 40 °C for up to 15 minutes.
The test material was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
Doses:
300 mg/kg and 2000 mg/kg.
No. of animals per sex per dose:
1 female animal at 2000 mg/kg and 5 animals at 300 mg/kg.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 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. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals.
Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
- Necropsy of survivors performed: Yes. 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 external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained
Preliminary study:
Due to mortality and signs of systemic toxicity at a dose level of 2000 mg/kg, an additional group of 4 animals was treated at 300 mg/kg. A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The animal dosed at 2000 mg/kg was killed for humane reasons, 1 day 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 associated with dosing at 300 mg/kg.
Clinical signs:
The animal dosed at 2000 mg/k experienced signs of systemic toxicity which included noisy respiration, hunched posture, pilo-erection, diarrhoea, dehydration, lethargy, hypothermia and tiptoe gait.

For the animals dosed at 300 mg/kg, no signs of systemic toxicity were noted during the observation period in the initial treated animal. Hunched posture was noted during the day of dosing in the four additional treated animals. These four animals appeared normal 2 or 4 hours after dosing.
Body weight:
The animal dosed at 2000 mg/kg experienced weight loss from 171 g fasted weight before dosing, to 147 g at death.

All animals dosed at 300 mg/kg showed expected gains in body weight over the observation period.
Gross pathology:
Abnormalities noted at necropsy were dark liver, dark kidneys, gaseous stomach and haemorrhage of the gastric mucosa and non-glandular epithelium of the stomach for the animal dosed at 2000 mg/kg.

For the animals dosed at 300 mg/kg, no abnormalities were noted at necropsy.

Individual Clinical Observations and Mortality Data - 2000 mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing

30 Minutes

1 Hour

2 Hours

4 Hours

1 Day

2000

2-0 Female

Rn

Rn H

H

0

H P D Dh Wt

(H P L Dh Ho D)*

0 = No signs of systemic toxicity

D = Diarrhea

Dh = Dehydration

H = Hunched posture

Ho = Hypothermia

L = Lethargy

P = Pilo-erection

Rn= Noisy respiration

Wt =Tiptoe gait 30 Minutes Rn ( ) = Observations noted prior to humanely killing

* = Animal killed 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.

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the conditions of this study, the acute oral median lethal dose (LD50) of the test material was estimated to be in the range of 300 - 2000 mg/kg body weight.
Executive summary:

The acute oral toxicity of the test material was assessed in the Wistar strain rat in accordance with the standardised guidelines OECD 420 and EU Method B.1bis under GLP conditions.

Following a sighting test at dose levels of 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of test material, as a solution in arachis oil BP, 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.

The animal treated at a dose level of 2000 mg/kg was killed for humane reasons, 1 day 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.

Signs of systemic toxicity noted in the animal treated at a dose level of 2000 mg/kg were noisy respiration, hunched posture, pilo-erection, diarrhoea, dehydration, lethargy, hypothermia and tiptoe gait. Hunched posture was noted in the additional group of four animals treated at a dose level of 300 mg/kg. No signs of systemic toxicity were noted in the initial animal treated at a dose level of 300 mg/kg.

Surviving animals showed expected gains in body weight.

Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg were dark liver, dark kidneys, gaseous stomach and haemorrhage of the gastric mucosa and non-glandular epithelium of the stomach. No abnormalities were noted at necropsy of animals treated at a dose level of 300 mg/kg.

Under the conditions of this study, the acute oral median lethal dose (LD50) of the test material was estimated to be in the range of 300 - 2000 mg/kg body weight therefore the test material can be classed as toxic through the oral route.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
300 mg/kg bw
Quality of whole database:
The key study was conducted under GLP conditions in accordance with the standardised guidelines OECD 420 and EU Method B.3. It was assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997).

Acute toxicity: via inhalation route

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:
16 March 2016 to 30 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation: Weight ranged between 202 - 247 g. The weight variation did not exceed ±20 % of the mean weight for each sex.
- Housing: 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.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): At least fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness.
Type of coverage:
semiocclusive
Vehicle:
arachis oil
Details on dermal exposure:
TEST MATERIAL PREPARATION
For the purpose of the study the test item was weighed out according to each animal's individual body weight and moistened with arachis oil BP prior to application.

TEST SITE
- Area of exposure: The back and flanks of each animal.
- % coverage: Applied as evenly as possible to an area of shorn skin (approximately 10 % of the total body surface area)
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage. The dressings were examined to ensure that they were securely in place shortly after dosing.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): 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.

TEST MATERIAL
- For solids, paste formed: The appropriate amount of test item was applied moistened with arachis oil BP.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
Five male and five female rats.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity 30 minutes, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days. Individual body weights were recorded prior to application of the test item on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: Yes. At the end of the study the 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.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
There were no deaths reported.
Clinical signs:
No signs of systemic toxicity were noted during the observation period.
Body weight:
Animals showed expected gains in body weight, except for one female which showed no gain in body weight during the first week but expected gain in body weight during the second week.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
DERMAL REACTIONS
Signs of dermal irritation were very slight to well-defined erythema, very slight to slight oedema, haemorrhage of dermal capillaries, light brown discoloration of the epidermis, crust formation, loss of skin leas and flexibility, hardened light brown coloured scab, scab cracking, scab lifting to reveal glossy skin, desquamation and glossy skin.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the study, 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:

The acute dermal toxicity of the test material was assessed in the Wistar strain rat in accordance with the standardized guidelines OECD 402 and EU Method B.3 under GLP conditions.

A limit test was conducted to assess the potential of the test material to cause acute dermal toxicity in the Wistar strain rat. A group of ten animals (five males and five females) were given a single, 24 hour, semi-occluded dermal application of the 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.

There were no deaths or signs of systemic toxicity associated with the test material.

Signs of dermal irritation were very slight to well-defined erythema, very slight to slight oedema, haemorrhage of dermal capillaries, light brown discoloration of the epidermis, crust formation, loss of skin elasticity and flexibility, hardened light brown coloured scab, scab cracking, scab lifting to reveal glossy skin, moderate desquamation and glossy skin. Animals showed expected gains in body weight, except for one female which showed no gain in body weight during the first week but expected gain in body weight during the second week.

No abnormalities were noted at necropsy.

Under the conditions of the study, 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
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The key study was conducted under GLP conditions in accordance with the standardised guidelines OECD 402 and EU Method B.1 bis. It was assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997).

Additional information

Acute Oral Toxicity

The acute oral toxicity of the test material was assessed in the Wistar strain rat in accordance with the standardised guidelines OECD 420 and EU Method B.1 bis under GLP conditions.

Following a sighting test at dose levels of 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of test material, as a solution in arachis oil BP, 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.

The animal treated at a dose level of 2000 mg/kg was killed for humane reasons, 1 day 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.

Signs of systemic toxicity noted in the animal treated at a dose level of 2000 mg/kg were noisy respiration, hunched posture, pilo-erection, diarrhoea, dehydration, lethargy, hypothermia and tiptoe gait. Hunched posture was noted in the additional group of four animals treated at a dose level of 300 mg/kg. No signs of systemic toxicity were noted in the initial animal treated at a dose level of 300 mg/kg.

Surviving animals showed expected gains in body weight.

Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg were dark liver, dark kidneys, gaseous stomach and haemorrhage of the gastric mucosa and non-glandular epithelium of the stomach. No abnormalities were noted at necropsy of animals treated at a dose level of 300 mg/kg.

Under the conditions of this study, the acute oral median lethal dose (LD50) of the test material was estimated to be in the range of 300 - 2000 mg/kg body weight therefore the test material can be classed as harmful through the oral route.

 

Acute Dermal Toxicity

The acute dermal toxicity of the test material was assessed in the Wistar strain rat in accordance with the standardized guidelines OECD 402 and EU Method B.3 under GLP conditions.

A limit test was conducted to assess the potential of the test material to cause acute dermal toxicity in the Wistar strain rat. A group of ten animals (five males and five females) were given a single, 24 hour, semi-occluded dermal application of the 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.

There were no deaths or signs of systemic toxicity associated with the test material.

Signs of dermal irritation were very slight to well-defined erythema, very slight to slight oedema, haemorrhage of dermal capillaries, light brown discoloration of the epidermis, crust formation, loss of skin elasticity and flexibility, hardened light brown coloured scab, scab cracking, scab lifting to reveal glossy skin, moderate desquamation and glossy skin. Animals showed expected gains in body weight, except for one female which showed no gain in body weight during the first week but expected gain in body weight during the second week.

No abnormalities were noted at necropsy.

Under the conditions of the study, 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 Toxicity

In accordance with Section 8.5.2. Column 2 of Annex VIII of the REACH Regulation, the information required under 8.5.2 (acute toxicity via the inhalation route) to 8.5.3 (via the dermal route) shall be provided for at least one route.  The choice for the second route will depend on the nature of the substance and the likely route of human exposure.  As inhalation of the material is unlikely, data via the dermal route is provided and therefore further information on toxicity via the inhalation route is not required.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance requires classification with respect to acute toxicity via the oral route (Category 4) but does not require classification via the dermal or inhalation route.

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