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Endpoint summary

Description of key information

Oral route: The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be > 2000 mg/kg bw (OECD 420 and EU Method B.1 bis).

 

Dermal route: The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be > 2000 mg/kg bw (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:

30 August 2016 to 22 September 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

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- Female Wistar strain rats (RccHan:WIST) were supplied by Envigo RMS (UK) Limited, Oxon, UK.
- On receipt the animals were randomly allocated to cages.
- The females were nulliparous and non-pregnant.
- After an acclimatisation 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.
- Body weight variation did not exceed ± 20 % of the mean body weight at the start of treatment.

ANIMAL CARE AND HUSBANDRY
- Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with wood flakes.
- 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 RMS (UK) Limited, Oxon, UK) was allowed throughout the study.
- The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Temperature was set to achieve limits of 19 to 25 °C.
- Relative humidity was set to achieve limits of 30 to 70 %.
- 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 which were condidered 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:

TEST ITEM PREPARATION AND ANALYSIS
- The test item was used as supplied.
- Specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.

EXPOSURE TO TEST ITEM
- 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 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 (early and late) during normal working days and once daily on weekends and public holidays.
- 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 consisting of an external examination plus opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

DATA EVALUATION
- 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 were killed for humane reasons) plus determination of the nature, severity, onset and duration of toxic effects. If possible, the signs of evident toxicity were described.
- Evident toxicity refers to toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probably mortality.
- Effects on body weights and abnormalities at necropsy were also identified.

Doses:

Single dose of 2000 mg/kg bw

No. of animals per sex per dose:

Five

Control animals:

no

Details on study design:

STUDY DESIGN
- Flow charts summarising the sighting test and main test are provided in Annex 2 and Annex 3 (attached).
- Using available information on the toxicity of the test item, the starting dose was chosen as 2000 mg/kg bw.
- A single animal was treated with 2000 mg/kg bw (specific gravity 0.970; dose volume 2.07 mL/kg).
- In the absence of toxicity, an additional group of four animals was then treated with 2000 mg/kg bw (specific gravity 0.970; dose volume 2.07 mL/kg).
- A total of five animals was therefore treated at the dose level of 2000 mg/kg bw during the study.

Statistics:

- An estimate of the acute oral median lethal dose (LD50) of the test item was made using mortality data obtained during the study.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

- No animal deaths took place during the study.

Clinical signs:

other: - No signs of systemic toxicity were noted during the observation period.

Gross pathology:

- Individual necropsy findings are given in Appendix 3 (attached).
- No abnormalities were noted at necropsy.

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 found to be > 2000 mg/kg bw.

Executive summary:

GUIDELINE

The study was performed to assess acute oral toxicity of the test item in the Wistar strain rat in compliance with the OECD Guideline for Testing of Chemicals No 420 “Acute Oral Toxicity – Fixed Dose Method” (2001) and Method B.1 bis Acute Toxicity (Oral) of Commission Regulation (EC) No 440/2008.

 

METHODS

Following a sighting test at a dose level of 200 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 bw. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

RESULTS

No animal deaths took place during the study and clinical observations resulted in no reports of systemic toxicity. All animals showed expected gains in body weight and 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 found to be > 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

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:

07 September 2016 to 21 September 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:

yes

Remarks:

Day 2 dermal observations not recorded on animals 2-0 to 2-4 with no impact on study outcome (see below)

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

yes

Remarks:

Day 2 dermal observations not recorded on animals 2-0 to 2-4 with no impact on study outcome (see below)

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

RccHan:WIST

Sex:

male/female

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- Five male and five female Wistar (RccHan:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK.
- On receipt the animals were randomly allocated to cages.
- Female rats were nulliparous and non-pregnant.
- After an acclimatisation 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 animals weighed at least 200 g and were 8 to 12 weeks of age.
- The weight variation did not exceed ± 20 % of the mean weight for each sex.

ANIMAL CARE AND HUSBANDRY
- 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 (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study.
- Diet, drinking water and bedding were routinely analysed and were not considered to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 19 to °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.
- The animals were provided with environmental enrichment items which were not considered 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:

TEST SYSTEM
- 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 single group of animals was treated as shown in the table below.
- 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.
- 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.
- After the 24-hour contact period the bandage was carefully removed and the treated skin and surrounding area were wiped with cotton wool moistened with arachis oil BP to remove any residual test item.
- Animals were returned to group housing for the remainder of the study period.
- The animals were observed for deaths or overt signs of toxicity at 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days.
- After removal of the dressings and subsequently once daily for 14 days, the test sites were examined for evidence of primary irritation and scored according to the scale shown in the table below.
- If present, any other skin reactions 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 external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

TEST ITEM PREPARATION AND ANALYSIS
- The test item was used as supplied.
- Specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.
- Absorption of the test item was not determined.

MAJOR COMPUTERISED SYSTEMS
- Delta Controls ORCAview

Statistics:

DATA EVALUATION
- Data evaluations included the relationship, if any, between the 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 any other toxicological effects.
- Using the mortality data from the study, an estimate of the acute dermal median lethal dose (LD50) was made for the test item.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

- Individual mortality data are given in Appendix 1 (attached).
- No unplanned animal deaths took place during the study.

Clinical signs:

other: - Individual clinical observations are given in Appendix 1 (attached). - No signs of systemic toxicity were noted during the observation period.

Gross pathology:

- Individual necropsy findings are given in Appendix 5.
- No abnormalities were noted at necropsy.

Other findings:

- Individual dermal reactions are given in Appendices 2 and 3 (attached).
- No signs of dermal irritation were observed.

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 > 2000 mg/kg bw.

Executive summary:

GUIDELINE

The study was performed to assess the acute dermal toxicity of the test item in the Wistar strain rat in compliance with the OECD Guideline for the Testing of Chemicals No 402 “Acute Dermal Toxicity” (adopted 24 February 1987) and Method B.3 Acute Toxicity (Dermal) of Commission Regulation (EC) No 440/2008.

 

METHODS

A group of ten 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 bw. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

RESULTS

No animal deaths took place during the study. There were no signs of systemic toxicity and no signs of dermal irritation. All animals showed expected gains in body weight and 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 > 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

Oral route

The key study was performed to assess acute oral toxicity of the test item in the Wistar strain rat in compliance with the OECD Guideline for Testing of Chemicals No 420 “Acute Oral Toxicity – Fixed Dose Method” (2001) and EU Method B.1 bis Acute Toxicity (Oral) of Commission Regulation (EC) No 440/2008.

 

Following a sighting test at a dose level of 200 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 bw. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

No animal deaths took place during the study and clinical observations resulted in no reports of systemic toxicity. All animals showed expected gains in body weight and no abnormalities were noted at necropsy. The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be > 2000 mg/kg bw.

Inhalation route

The substance has been shown to have a high onset boiling point (286 °C at 101 kPa) and the vapour pressure has been determined to be 8.46 x 10E-03 Pa at 25 °C. It is therefore expected that inhalation exposure will be low under general use conditions at ambient temperature. Lack of systemic toxicity when the substance is administered via the oral and dermal routes suggests that determined vapour pressures of 1.19 x 10E-02 Pa at 40 °C (313 K) and 6.81 x 10E-02 Pa at 100 °C also give no cause for concern. The inhalation route is therefore not the most applicable method of investigating acute toxicity.

Dermal route

The key study was performed to assess the acute dermal toxicity of the test item in the Wistar strain rat in compliance with the OECD Guideline for the Testing of Chemicals No 402 “Acute Dermal Toxicity” (adopted 24 February 1987) and Method B.3 Acute Toxicity (Dermal) of Commission Regulation (EC) No 440/2008.

 

A group of ten 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 bw. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

No animal deaths took place during the study. There were no signs of systemic toxicity and no signs of dermal irritation. All animals showed expected gains in body weight and 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 > 2000 mg/kg bw.

Justification for classification or non-classification

The acute median lethal dose (LD50) values of the test item in the Wistar strain rat were reported as > 2000 mg/kg bw following oral and dermal administration. Classification for acute oral or dermal toxicity is therefore unnecessary under the terms of Regulation (EC) No 1272/2008.