Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29th November 2017 - 5th February 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of allyl (2-methylbutoxy)acetate and allyl (3-methylbutoxy)acetate
EC Number:
916-328-0
Molecular formula:
C10H18O3
IUPAC Name:
Reaction mass of allyl (2-methylbutoxy)acetate and allyl (3-methylbutoxy)acetate
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: O’Laughlin (Nantong) Fine Chemicals Co., Ltd.; NTA375
- Expiration date of the lot/batch: Sep 25, 2020
- Purity test date: CAS No. 67634-00-8: 79.45 %; CAS No.: 67634-01-9 20.28 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Immediately before application the test item was weighed, mixed with vehicle olive oil and resulting emulsion was administered to the stomach by tube. All prepared emulsions of the test item in olive oil were mixed by magnetic stirrer during administration.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Lysolajské údolí 15/53, 165 00 Prague 6, Czech Republic, RČH CZ 11760500
- Age at study initiation: 8 weeks at the time of application
- Weight at study initiation: 163-188g
- Housing: Animal room with shavings of soft wood – 3 animals of one sex in one plastic breeding cage
- Diet: Pelleted standard diet for experimental animals ad libitum
- Water: Drinking tap water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity:30 – 70 %,
- Photoperiod : Light period 12-hour light/12 hour dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage): 1 mL/100 g of animal body weight
- Justification for choice of vehicle: The test item does not make a homogenous emulsion with water, so olive oil had to be used
- Lot/batch no. (if required): 8002954001

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The dose level of 300 mg/kg bw was used as the starting dose, according to the test guideline, because there was no information about toxicity available.
Doses:
300 mg/kg bw (Group 1 and 3)
2000 mg/kg (Group 2)
No. of animals per sex per dose:
3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: After application the animals were observed individually:
- the first day: twice (30 minutes and 3 hours after application)
- the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.

Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.

- Frequency of observations and weighing: Animals were weighed before application, at the 8th day of study and at the 15th day, before euthanasia of animals. Average body weight in a group was calculated from individual body weights. Body weight increments were calculated from body weight at the start of the study,
the first week and at the end of the study.

- Necropsy of survivors performed: All test animals survived to the end of study were sacrificed on the 15th day and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Mortality:
No deaths were observed in the groups administered 300 mg/kg bw. In the group administered 2000 mg/kg bw, 2 females died and 1 female was killed for moribund status
Clinical signs:
No clinical signs of intoxication were observed in all 6 females at the dose 300 mg/kg (Group 1 and 3) during the clinical observation after the application (see Tables No. 4 and 6).
At the dose level 2000 mg/kg (Group 2) the following symptoms were observed 30 minutes after application: abdominal position in all treated females. 3 hours after application the following symptoms were observed: piloerection, apathy, decrease response to stimuli, immobility, bradypnoea in female No. 4, piloerection, abdominal position, rocking motion, decrease response to stimuli, urine containing blood in female No. 5, piloerection, abdominal position, rocking motion, decrease response to stimuli in female No. 6. This female survived till the second day morning. Piloerection, apathy, immobility, decrease response to stimuli, porphyrin secretion around nostrils (medium), bradypnoea and urine containing blood were observed.
Body weight:
Weight increments in group No. 1 and group No. 3 were adequate to species, sex and age of animals in experiment. No reduction of body weight was observed (see Tables No. 1 and 3). The animals in group No. 2 were weighed only before application (see Table No. 2), so no evaluation of weight changes is possible.
Gross pathology:
No pathological macroscopic changes were diagnosed during pathological examination of animals at the dose 300 mg/kg in group No. 1 and No. 3. (see Tables No. 7, 9). Animals in Group 1 and 3 were sacrificed on the 15th day of study. Pathological changes were observed during pathological examination at the dose level 2000 mg/kg. Stomach full of the test item and massive hemorrhages of the mucous membrane, small intestine - hyperemia of GALT (gut-associated lymphoid tissue) and liver – dark red color, were observed in group No. 2 (see Table No. 8). The necropsy was done immediately after death (4 hours after application) in female No. 4. Female No. 5 was found dead on the morning of the second day and the necropsy was done approximately 24 hours after application, but significant post-mortem rigidity indicated that the animal died on the day of application. Female No. 6 was sacrificed approximately 24 hours after application for moribund condition.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
In this acute oral toxcity test (acute toxic class method), the LD50 value of the test item, allyl amyl glycolate, for female rats is > than 300 mg/kg and < 2000 mg/ kg of body weight
Executive summary:

In an acute oral toxicity study (18-1), Wistar female rats (5/dose) were given allyl amyl glycolate in olive oil by gavage at doses of 300 and 2000 mg/kg bw and observed for 14 days.

LD50 (female): >300 < 2000 mg/kg bw.

The test item administered at the dose of 300 mg/kg bw caused no death of any animal. No serious clinical signs of intoxication were detected at this dose during the whole study. Weight increments were adequate to species, sex and age of animals in experiment. No pathological macroscopic changes were diagnosed during the pathological examination. The test item administered at the dose of 2000 mg/kg bw caused the death of two females and one female was humanely killed for moribund condition. Abdominal position was observed in all animals 30 minutes after application. Piloerection and decreased response to stimuli were observed in all animals 3 hours after application, abdominal position and rocking motion were observed in 2 animals 3 hours after application.  Apathy, bradypnoea, and immobility were observed in one female 3 hours after application. This female died 4 hours after application. Urine containing blood was observed in one female 3 hours after application. This female was found dead second day after application, but significant post-mortem rigidity indicated that the animal died on the day of application.  Piloerection, porphyrin secretion around nostrils, urine containing blood, immobility and apathy were observed in one female, on the morning of the second day after application. The animal was humanely killed for moribund condition.

This acute oral study is classified as acceptable. It does satisfy the guideline requirement for an acute oral study (OECD 423) in the rat.