Rape oil, reaction products with diethylenetriamine

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 considered to be > 2000 mg/kg body weight (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 body weight (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:

07 July 2016 to 26 July 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

Remarks:

RccHan:WIST

Sex:

female

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- On receipt the animals were randomly allocated to cages.
- The female animals were nulliparous and non-pregnant.
- After an acclimatisation period of at least five days, 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 the 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 woodflakes.
- Diet and drinking water 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 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 that 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:

DMSO

Details on oral exposure:

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 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.
- At the end of the observation period animals were killed by cervical dislocation.
- All animals were subjected to gross necropsy consisting of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Five

Control animals:

no

Details on study design:

TEST ITEM PREPARATION AND ANALYSIS
- The test item was freshly prepared, as required, as a solution in dimethyl sulphoxide. This vehicle was chosen because the test item did not dissolve/suspend in distilled water or arachis oil BP.
- The test item was formulated within 2 hours of being applied to the test system and was assumed to be stable for that duration.
- No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This was an exception with regard to GLP and was reflected in the GLP compliance statement.

STUDY DESIGN
- Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose (see Annex 2 and Annex 3, attached).
- A single female animal was treated at a dose level of 2000 mg/kg (concentration 200 mg/mL; dose volume 10 mL/kg).
- In the absence of toxicity at a dose level of 2000 mg/kg, an additional four female animals were treated at a dose level of 2000 mg/kg (concentration 200 mg/mL; dose volume 10 mL/kg).

MAJOR COMPUTERISED SYSTEMS
- Delta Controls: ORCAview.

Statistics:

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 that 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 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.

Key result

Sex:

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 unscheduled animal deaths took place.

Clinical signs:

other: - Individual clinical observations are presented in Appendix 1 (attached). - Hunched posture was noted in all animals during the day of dosing. Lethargy was also noted in the initial treated animal. All animals appeared normal 1 day after dosing.

Gross pathology:

- Individual necropsy findings for the 2000 mg/kg dose level 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 dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item, as a solution in dimethyl sulphoxide, 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 unscheduled animal deaths took place during the study. Clinical Observations of lethargy and/or hunched posture were noted in all animals but the rats appeared normal one day after dosing. 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:

09 August 2016 to 23 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data were not generated for the purpose of compliance with REACH. However, acquisition of the information was necessary as part of global notification requirements, and the study result is included for completeness.

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:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- 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.
- 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 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.
- 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:

DMSO

Details on dermal exposure:

EXPOSURE TO TEST ITEM
- 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 with the test item at a dose level of 2000 mg/kg.
- The appropriate amount of test item, moistened with dimethyl sulphoxide, 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 hair were wiped with cotton wool moistened with dimethyl sulphoxide followed by distilled water to remove any residual test item.
- The animals were returned to group housing for the remainder of the study period.
- 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 below.
- Any other skin reactions, if present were also recorded.

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:

STUDY DESIGN
- 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.

MAJOR COMPUTERISED SYSTEMS
- Delta Controls: ORCA view.

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 (attached).
- No abnormalities were noted at necropsy.

Other findings:

DERMAL REACTIONS
- Individual dermal reactions are given in Appendices 2 and 3 (attached).
- There were no signs of dermal irritation noted at the test sites of all male animals.
- Very slight erythema was noted at the test sites of all females. Small superficial scattered scabs were also noted at the test sites of four females. Crust formation and/or desquamation were noted at the test sites of three females.

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 often 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

No animal deaths took place during the study and there were no signs of systemic toxicity. desquamation, small superficial scattered scabs and crust formation were noted at the test sites in female animals but no signs of dermal irritation were noted at the test sites of male animals. 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

A key study was performed to OECD 420 (fixed dose method) 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, as a solution in dimethyl sulphoxide, 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.

 

No unscheduled animal deaths took place during the study. Clinical Observations of lethargy and/or hunched posture were noted in all animals but the rats appeared normal one day after dosing. 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 considered to be > 2000 mg/kg body weight.

Dermal route

A key study was performed to OECD Guideline 402 and EU Method B.3 to assess the acute dermal toxicity of the test item in the Wistar strain rat. 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 body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

No unscheduled animal deaths took place during the study and no signs of systemic toxicity were observed. Very slight erythema, desquamation, small superficial scattered scabs and crust formation were noted at the test sites in female animals but no signs of dermal irritation were noted at the test sites of male animals. 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 body weight.

Inhalation route

Investigation of acute toxicity via the inhalation route is not required because the substance is a waxy solid with a determined melting temperature of 29 to 60 °C and determined vapour pressures of 0.0345 Pa at 25 °C, 0.0985 Pa at 55 °C and 0.891 Pa at 145 °C. Itis therefore 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 where lack of systemic toxicity has been experimentally demonstrated.

Justification for classification or non-classification

Oral route

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was considered to be > 2000 mg/kg body weight and, in accordance with Regulation (EC) No. 1272/2008, classification for acute oral toxicity is not required.

 

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 body weight and classification for acute dermal toxicity is not required under the terms of Regulation (EC) No. 1272/2008.