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Administrative data

Description of key information

LD50 > 2000 mg/kg in acute toxicity studies involving administration of test material to the Wistar rat via the oral route (OECD 420 and EU Method B.1 bis) and the dermal route (OECD 402 and EU Method B.3).

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:
02 February 2016 to 08 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
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:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
ANIMALS and ANIMAL HUSBANDRY
- Female Wistar rats were supplied by Envigo RMS UK Ltd, Oxon, UK.
- The animals were randomly allocated to cages on receipt.
- Females were nulliparous and non-pregnant.
- After an acclimatisation period of at least 5 days the animals were selected at random and given a unique number within the study by indelible ink-marking on the tail and the number was written on a cage card.
- At the start of the study the animals were 8 to 12 weeks of age.

- Body weight did not exceed ± 20 % of the mean body weight at the start of treatment.
- 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 Envigo Research UK Ltd, Oxon, UK was allowed throughout the study.
- Diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70 % respectively.
- Rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
- Animals were provided with environmental enrichment items considered not to contain any contaminant at a level that might affect the purpose or integrity of the study.
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
TEST ITEM PREPARTION and EXPERIMENTAL PREPARATION
- For the purpose of the study, the test item was freshly prepared, as required, as a solution in arachis oil BP. This vehicle was chosen because the test material did not dissolve or suspend in distilled water.
- The test item was formulated within 2 hours of being applied to the test system and it was assumed that the formulation was stable for that 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 was reflected in the GLP compliance statement.
Doses:
- 300 mg/kg was chosen as the starting dose (concentration 30 mg/mL; dose volume 10 mL/kg)
- Additional dose 2000 mg/kg (concentration 200 mg/mL; dose volume 10 mL/kg)
No. of animals per sex per dose:
- Single female animal dosed at 300 mg/kg as the starting dose.
- In the absence of toxicity at 300 mg/kg, a single female animal dosed at 2000 mg/kg
- In the absence of mortality at 2000 mg/kg, an additional group of four female animals dosed at 2000 mg/kg
Control animals:
no
Details on study design:
- All animals were dosed once 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.
- Clinical observations were made 0.5, 1, 2 and 4 hours after dosing and then daily for 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.
- Animals were killed by cervical dislocation at the end of the observation period.
- All animals were subjected to gross necropsy consisting of external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded but no tissues were retained.
Statistics:
EVALUATION OF DATA
- The test item was evaluated according to Annex 3 of the OECD Guidelines for Testing of Chemicals No 420 "Acute Oral Toxicity - Fixed Dose Method (adopted 17 December 2001) as shown in the flow chart in Annex 2 (attached).
- Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality.
- Effects on body weights and abnormalities noted at necropsy were also identified.
- Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.
Preliminary study:
DOSE LEVEL 300 mg/kg
-There was no mortality.
- No signs of systemic toxicity were noted during the observation period.
- The animal showed expected gains in body weight over the observation period.
- No abnormalities were noted at necropsy.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- There were no deaths.
Clinical signs:
- Noisy respiration was noted in the initial animal, 13 and 14 days after dosing. No other signs of systemic toxicity were noted.
Body weight:
- Animals showed expected gains in body weight over the observation period; except for one animal which showed expected gain in body weight during the first week but body weight loss during the second week.
Gross pathology:
- No abnormalities were noted at necropsy.
Interpretation of results:
other: Not classifiied
Remarks:
EU
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.
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 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of the test item, as a solution in arachis oil BP, 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: Noisy respiration was noted in the initial animal, treated at a dose level of 2000 mg/kg, 13 and 14 days after dosing. No other signs of systemic toxicity were noted.

Body weight: Animals showed expected gains in body weight over the observation period, except for one animal treated at a dose level of 2000 mg/kg which showed expected gain in body weight during the first week but body weight loss during the second week.

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 estimated to be greater than 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw

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:
25 February 2016 to 10 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP guideline study
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
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
- Physical state: Yellow semi-solid
- Analytical purity:100%
- Expiration date of the lot/batch: 23 November 2017
- Storage condition of test material: room temperature in the dark
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
ANIMALS and ANIMAL HUSBANDRY
- Female Wistar rats were supplied by Envigo RMS UK Ltd, Oxon, UK.
- The animals were randomly allocated to cages on receipt.
- Females were nulliparous and non-pregnant.
- After an acclimatisation period of at least 5 days the animals were selected at random and given a unique number within the study by indelible ink-marking on the tail and the number was written on a cage card.
- At the start of the study the animals were 8 to 12 weeks of age.

- Body weight did not exceed ± 20 % of the mean body weight for each sex.
- Animals were housed individually during 24-hour exposure period and in groups of five, by sex, for the remainder of the study in suspended solid-floor polypropylene cages furnished with woodflakes.
- Free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo Research UK Ltd, Oxon, UK) was allowed throughout the study.
- Diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70 % respectively.
- Rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
- Animals were provided with environmental enrichment items considered not to contain any contaminant at a level that might affect the purpose or integrity of the study.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
PROCEDURE
- On the day before treatment, the back and flanks of each animal were clipped free of hair.
- Using available information on the toxicity of the test item, a group of five male and five female rats was treated with the test item at a dose level of 2000 mg/kg
- The appropriate amount of test item 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 they were securely in place.
- 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 test period.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
TEST ITEM PREPARATION
- The test item was weighed out according to each animal’s individual body weight.
- Absorption of the test item was not determined.


Statistics:
- Data evaluations included the relationship, if any, between exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, body weight changes, mortality and other toxicological effects.
- Using the mortality data obtained, and estimate of the acute dermal median lethal dose (LD50) of the test item was made.
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 (see Appendix 1, attached).
Body weight:
- Individual body weights and body weight changes are shown in Appendix 4 (attached).
- All animals showed expected gains in body weight over the observation period.
Gross pathology:
- Individual necropsy findings are given in Appendix 5 (attached).
- No abnormalities were noted at necropsy.
Other findings:
DERMAL REACTIONS
- Individual dermal reactions are shown in Appendix 2 and 3 (attached).
- Very slight to well-defined erythema and very slight edema were noted at the test sites of one male and all females.
- Very slight erythema, with or without very slight edema was noted at the test sites of four males.
- Crust formation was noted at the test sites of all animals.
- Small superficial scattered scabs were noted at the test sites of two females with desquamation also noted at the test sites of two females.
- Treated skin sites of males appeared normal 7 days after dosing and the treated skin sites of four females appeared normal 7 or 14 days after treatment.
- Small superficial scattered scabs were still present at the treatment site of one female at the end of the observation period on Day 14.
Interpretation of results:
other: not classified
Remarks:
EU
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 assess the acute dermal toxicity of the test item in the Wistar strain rat.

METHODS

A group of then animals (five males and five females) was 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.

RESULTS

Mortality: There were no deaths.

Clinical observations: There were no signs of systemic toxicity.

Dermal irritation: Signs of dermal irritation noted were very slight to well-defined erythema, very slight edema, crust formation, desquamation and small superficial scattered scabs. Treated skin sites of males appeared normal 7 days after dosing and the treated skin sites of four females appeared normal 7 or 14 days after treatment. Small superficial scattered scabs were still present at the treatment site of one female at the end of the observation period on Day 14.

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

Necropsy: No abnormalities were noted a 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

ORAL

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 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of the test item, as a solution in arachis oil BP, 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 necroscopy.

RESULTS

Mortality: there were no deaths.

Clinical observations: Noisy respiration was noted in the initial animal, treated at a dose level of 2000 mg/kg, 13 and 14 days after dosing. No other signs of systemic toxicity were noted.

Body weight: Animals showed expected gains in body weight over the observation period, except for one animal treated at a dose level of 2000 mg/kg which showed expected gain in body weight during the first week but body weight loss during the second week.

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 estimated to be greater than 2000 mg/kg body weight.

INHALATION

REACH Annex XIII, Section 8.5, Column 2, states that information on acute toxicity will be provided for the oral route plus at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. The test material is a semi-solid (paste) with a vapour pressure of 0.254 Pa at 25 °C, which makes generation of inhalable forms of the substance unlikely. As a result, it is considered that inhalation exposure will be low and the most likely route of exposure for workers and consumers is the dermal route. Testing for acute toxicity via the inhalation route is consequently not applicable.

 

DERMAL 

INTRODUCTION

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

METHODS

A group of then animals (five males and five females) was 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.

RESULTS

Mortality: There were no deaths.

Clinical observations: There were no signs of systemic toxicity.

Dermal irritation: Signs of dermal irritation noted were very slight to well-defined erythema, very slight edema, crust formation, desquamation and small superficial scattered scabs. Treated skin sites of males appeared normal 7 days after dosing and the treated skin sites of four females appeared normal 7 or 14 days after treatment. Small superficial scattered scabs were still present at the treatment site of one female at the end of the observation period on Day 14.

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

Necropsy: No abnormalities were noted a 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.

Justification for classification or non-classification

No adverse effect was observed during investigation of acute toxicity via the oral or dermal routes in the Wistar rat and, based on determined vapour pressure of the substance, exposure via the inhalation route is not considered to be of significance to humans. As such, classification in accordance with Regulation (EC) No 1272/2008 is not required.