Registration Dossier

Administrative data

Description of key information

Acute toxicity - oral:

In an OECD 420 study, under GLP conditions, the acute toxicity LD50 (oral) of 2-Propenoic acid, 2-[[(octadecylamino)carbonyl]oxy]ethyl ester is 2000 mg/kg (Envigo, 2016d).

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:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
Batch: 08262015
Purity: >98%
Physical state/Appearance: white solid
Expiry Date: 20 June 2017
Storage Conditions: room temperature in the dark
Species:
rat
Strain:
Wistar
Remarks:
RccHan™:WIST
Sex:
female
Details on test animals and environmental conditions:
On receipt the animals were randomly allocated to cages.

The females were nulliparous and non pregnant.

After an acclimatization 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.

The body weight variation did not exceed ±20% of the mean body weight at the start of treatment.

The 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 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 12 hours continuous light and 12 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:
arachis oil
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 group to confirm the survival of the previously dosed animals.

Clinical observations were made 30 minutes, 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 at 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. 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.
Doses:
300 mg/kg

2000 mg/kg
No. of animals per sex per dose:
300 mg/kg - 1 Female

2000 mg/kg - 5 Females
Control animals:
no
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:
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.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of 2-Propenoic acid, 2-[[(octadecylamino)carbonyl]oxy]ethyl ester 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
Value:
2 000 mg/kg bw
Quality of whole database:
1 - Guideline study according to GLP

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental Phase: 01 February 2018 to 13 March 2018. Report Issued: 30 Aug 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
yes
Remarks:
Two deviations from the study plan were not ocnsidered to have affected the purpose or validity of the study.
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female WISTAR RccHan™ : WIST strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: 200 to 350g
- Fasting period before study: No
- Housing: The animals were housed in groups of up to three by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes.
- Diet (e.g. ad libitum): ad libitum (except during exposure)
- Water (e.g. ad libitum): ad libitum (except during exposure)
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness

Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
3.82 µm
Geometric standard deviation (GSD):
3.29
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Dynamic System for Nose Only Exposure of Rats ((ADG Developments Ltd, Hitchin, Herts, UK)
- Exposure chamber volume: Approximately 30 litres
- Equilibration: The theoretical chamber equilibration time (T99) was 4 minutes. The test atmosphere was generated for 9 minutes prior to animal insertion to ensure test item concentration was being achieved.
- Method of holding animals in test chamber: During the exposure period, each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber ‘O’ ring.
- Source and rate of air: Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters. The air flow rate into the chamber was 40 litres/min.
- System of generating particulates/aerosols:A dust atmosphere was produced from the test item using a SAG 410 Solid Aerosol Generator (TOPAS GmbH, Dresden, Germany) located adjacent to the exposure chamber.
- Method of particle size determination: The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period using a Marple Personal Cascade Impactor (Westech IS Ltd, Beds., UK).
- Treatment of exhaust air: The extract from the exposure chamber passed through a ‘scrubber’ trap and was connected with a high efficiency filter to a metered exhaust system.
- Temperature, humidity, pressure in air chamber: The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd, Beds., UK) located in a vacant port in the animals’ breathing zone of the chamber and recorded every 30 minutes throughout the 4-Hour exposure period.

TEST ATMOSPHERE
- Samples taken from breathing zone: yes

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Based on preliminary study using two rats.
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
4 h
Concentrations:
4.94 mg/L
No. of animals per sex per dose:
3 male and 3 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: All animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, 1 hour after termination of exposure and subsequently once daily for 14 days. Any evidence of overt toxicity was recorded at each observation.
- Weighing: Individual body weights were recorded on arrival, prior to treatment on the day of exposure (Day 0) and on Days 1, 3, 7 and 14.
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4.94 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality was observed on the study.
Clinical signs:
other: Signs of hunched posture and pilo-erection were observed following removal from the chamber. Such signs are commonly seen in animals for short periods on removal from the chamber following 4-hour inhalation studies. Wet fur was noted both during and for
Body weight:
All males showed body weight loss on Day 1 post-exposure and expected gains in body weight during the remainder of the recovery period. With the exception of two animals that showed body weight loss on Day 1 post-exposure and one animal that showed no gain in body weight from Days 1 to 3 post-exposure, all females showed expected gains in body weight during the recovery period
Gross pathology:
At necropsy macroscopic observations were seen in the lungs of all animals and consisted of: pale colour, abnormal red colour, dark patches.

Exposure Chamber Concentration

The test atmosphere was sampled nine times during the exposure period and the actual concentration of the test item calculated. The mean values obtained were as follows:

 Group Number           Atmosphere Concentration
 Mean Achieved (mg/L)  Standard Deviation  Nominal (mg/L
 1  4.94 0.15   37.99

The chamber flow rate was maintained at 40 L/min providing 80 air changes per hour.

Particle Size Distribution

The particle size analysis of the atmosphere drawn from the animals’ breathing zone was as follows:

 Group Number  Mean Achieved Atmosphere Concentration (mg/L)  Mean Mass Median Aerodynamic Diameter µm  Inhalable Fraction (%<4 µm) Geometric Standard Deviation 
 1  4.94  3.82  51.6  3.29

Mortality Data

The mortality data were summarised as follows:

 Group Number      Mean Achieved Atmosphere Concentration (mg/L)     Deaths      
 Male  Female  Total
 1 4.94  0/3  0/3  0/6

There were no deaths on the study.

Interpretation of results:
GHS criteria not met
Conclusions:
None of the animals died in a group of six rats exposed to mean achieved atmosphere concentration of 4.94 mg/L. It was therefore considered that the acute inhalation median lethal concentration (4 hour LC50) of the test item in the Wistar strain rat was greater than 4.94 mg/L.
Executive summary:

Introduction

The study was performed to assess the acute inhalation toxicity of the test item. The method used was designed to be compatible with that described in the OECD Guideline for Testing of Chemicals (2009) No. 436 “Acute Inhalation Toxicity – Acute Toxic Class Method” and Method B.52. Acute Inhalation Toxicity – Acute Toxic Class Method, 2014, of Commission Regulation (EC) No. 440/2008.

Methods

A group of six Wistar (RccHan™:WIST) strain rats (three males and three females) was exposed to a dust atmosphere of the test item. The animals were exposed for 4 hours using a nose only exposure system, followed by a 14 day observation period.

Results

The mean achieved atmosphere conctration was as follows:

 Group Number           Atmosphere Concentration
 Mean Achieved (mg/L)  Standard Deviation  Nominal (mg/L
 1  4.94 0.15   37.99

The charateristics of the achieved atmosphere were as follows:

 Group Number  Mean Achieved Atmosphere Concentration (mg/L)  Mean Mass Median Aerodynamic Diameter µm  Inhalable Fraction (%<4 µm) Geometric Standard Deviation 
 1  4.94  3.82  51.6  3.29

The mortality data were summarised as follows:

 Group Number      Mean Achieved Atmosphere Concentration (mg/L)     Deaths      
 Male  Female  Total
 1 4.94  0/3  0/3  0/6

Clinical Observations: Common abnormalities noted during the study included decreased respiratory rate, hunched posture, pilo-erection and wet fur, there were frequent instances of red/brown staining around the snout and/or eyes. All animals appeared normal on Day 1 post-exposure.

 

Body Weight: All males showed body weight loss on Day 1 post-exposure and expected gains in body weight during the remainder of the recovery period. Two females showed body weight loss on Day 1 post-exposure and one female showed no gain in body weight from Days 1 to 3 post-exposure. Females showed expected gains in body weight during the remainder of the recovery period.

Necropsy: The following macroscopic abnormalities were detected at necropsy: Lungs – Pale, abnormally red, dark patches.

Conclusion

None of the animals died in a group of six rats exposed to mean achieved atmosphere concentration of 4.94 mg/L. It was therefore considered that the acute inhalation median lethal concentration (4 hour LC50) of the test item in the Wistar strain rat was greater than 4.94 mg/L.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
4 940 mg/m³
Quality of whole database:
1 - Guideline study according to GLP

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:
Experimental Phase: 21 December 2017 to 11 January 2018. Report Issue: 28 February 2018.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
Wistar (RccHan:WIST)™ strain rats
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation: At east 200g. The weight variation did not exceed ±20% of the mean weight for each sex.
- Fasting period before study: No
- Housing: The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes. The initial two animals were housed individually throughout the study. The further group of eight animals (four male and four female) were housed individually during the 24-hour exposure period and in groups of four, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: At least five 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:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: back or flank
- % coverage: 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.

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 24 hours. Treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- For solids, paste formed: The test item, moistened with arachis oil BP.

Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
four males and four females
Control animals:
no
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.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight and skin reactions.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no unscheduled deaths on the study.
Clinical signs:
There were no signs of systemic toxicity
Body weight:
All animals showed expected gains in body weight over the observation period except for three females who showed a body weight loss or 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:
There were no signs of dermal irritation.
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 assess the acute dermal toxicity of the test item in the Wistar strain rat. The method used was designed to meet the requirements of OECD Guideline for the Testing of Chemicals No. 402 “Acute Dermal Toxicity” (adopted 24 February 1987)

 

Methods

Initially, two animals (one male and one female) were 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. Based on the results of the initial test, a further group of eight animals (four males and four females) was similarly treated. 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 Observation: There were no signs of systemic toxicity.

Dermal Irritation: There were no signs of dermal irritation.

Body Weight: All animals showed expected gains in body weight over the observation period except for three females which showed a body weight loss or no gain in body weight during the first week but expected gain in body weight during the second week.

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
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
1 - Guideline study according to GLP

Additional information

Justification for classification or non-classification

Acute toxicity - oral:

In an OECD 420 study, under GLP conditions, the acute toxicity LD50 (oral) of 2-Propenoic acid, 2-[[(octadecylamino)carbonyl]oxy]ethyl ester is 2000 mg/kg (Envigo, 2016d).

 

According to the ECHA Guidance on the Application of the CLP Criteria (version 4.1, June 2015), a substance is classified for acute toxicity (oral) if the LD50 is less than 2000 mg/kg. The LD50 of 2-Propenoic acid, 2-[[(octadecylamino)carbonyl]oxy]ethyl ester is 2000 mg/kg, therefore, is not classified according to CLP.