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Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guidelineopen allclose all
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

Test material

1
Chemical structure
Reference substance name:
2-(tetrapropenyl)succinic acid, monoester with propane-1,2-diol
EC Number:
257-836-6
EC Name:
2-(tetrapropenyl)succinic acid, monoester with propane-1,2-diol
Cas Number:
52305-09-6
Molecular formula:
C17H30O5 - C21H38O5
IUPAC Name:
2-[2-(2-hydroxy-1-methylethoxy)-2-oxoethyl]tetradecanoic acid
Test material form:
solid
Details on test material:
- Apperance: Yellow solid block
- Storage: Room temperature in the dark
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.

Test animals

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.

Administration / exposure

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

Results and discussion

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.
Effect levels
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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.