(4E)-4-(3,3,4-trimethylcyclopentylidene)butanal

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2020-09-27 to 2020-12-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study performed according to OECD test guideline No. 420 and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test data for registration of agricultural chemicals, Acute oral toxicity (2-1-1), 12 Nousan No 8147, Agricultural Production Bureau

Version / remarks:

24 November 2000

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

17 December 2001

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Version / remarks:

30 May 2008

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Compliance Monitoring Programme (inspected on 2019-04-02 / issued on 2019-08-01)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

(4E)-4-(3,3,4S-trimethylcyclopentylidene)butanal

Molecular formula:

C12 H20 O

IUPAC Name:

(4E)-4-(3,3,4S-trimethylcyclopentylidene)butanal

Constituent 2

Reference substance name:

(4E)-4-(3,3,4R-trimethylcyclopentylidene)butanal

Molecular formula:

C12 H20 O

IUPAC Name:

(4E)-4-(3,3,4R-trimethylcyclopentylidene)butanal

Test material form:

liquid

Details on test material:

Appearance: Very pale yellow liquid

Test animals

Species:

rat

Strain:

Wistar

Remarks:

RccHan™:WIST albino rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Ltd.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 162-173 g
- Fasting period before study: overnight prior to and approximately four hours after dosing
- Housing: in group of one to four rats in solid bottomed polycarbonate cages with a stainless steel mesh lid.
- Diet (e.g. ad libitum): free access to a standard rodent diet (Teklad 2014C Diet)
- Water (e.g. ad libitum): ad libitum. Potable water taken from the public supply was freely available via polycarbonate bottles fitted with sipper tubes.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 40-70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Justification for choice of vehicle: The test item was not soluble in water; therefore, corn oil was used as the vehicle.

DOSE VOLUME APPLIED: 10 mL/kg

- Rationale for the selection of the starting dose: In the absence of data regarding the toxicity of the test item, 300 mg/kg body weight was chosen as the starting dose.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

1 females at 300 and 5 females at 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weight: on Days -1, 1 (prior to dosing), 8 and 15 or at death.
Mortality: at least twice daily.
Clinical Observations: Soon after dosing and at frequent intervals for the remainder of Day 1. On subsequent days, surviving animals were observed once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). Additional observations were performed as necessary when evident toxicity was observed.
- Necropsy of survivors performed: yes
All animals were subject to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The macroscopic appearance of the brain, cecum, duodenum, heart, kidneys, small and large intestine, liver, lungs and bronchi, spleen, stomach, subcutaneous tissue and urinary bladder was recorded.

Statistics:

None

Results and discussion

Preliminary study:

In the absence of data regarding the toxicity of the test item, 300 mg/kg bw was chosen as the starting dose. In the absence of mortality or evident toxicity in the animal treated with the dose level of 300 mg/kg bw, an additional animal was treated at 2000 mg/kg bw. In the absence of mortality at a dose level of 2000 mg/kg bw, an additional group of four animals was treated at 2000 mg/kg bw. A total of five animals were therefore treated at a dose level of 2000 mg/kg bw in the study. Due to mortality and signs of systemic toxicity at a dose level of 2000 mg/kg body weight, an additional group of animals of 4 animals was treated at a dose level of 300 mg/kg bw.
A total of five animals were therefore treated at a dose level of 2000 mg/kg body weight and a further five animals were treated at 300 mg/kg body weight 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 four females (Animal Numbers 7, 8, 9 and 10) dosed at 2000 mg/kg body weight in the main study were killed for welfare reasons on Day 2. Clinical signs prior to death comprised piloerection, hunched posture, elevated gait and unsteady gait in all four animals and irregular breathing, partially closed eyelids (both) and decreased activity (behavior) in two animals. These signs were seen from approximately two hours after dosing. A loss in body weight was noted for the decedents. Macroscopic examination of the animals revealed congestion (characterized by darkened tissues/organs) of the subcutaneous tissue and heart for all four females, liver and kidneys for two animals and spleen for one animal, congestion (characterized by blood vessels injected) of the brain and lungs and bronchi for all four animals, pallor of the liver and spleen for three females and the kidneys for two animals, small (atrophy) for the spleen and cecum for four females, gaseous distension of the stomach for four animals and fluid contents (yellow) for the duodenum and small and large intestines for all four animals.

Clinical signs:

other: Clinical signs observed in surviving animals were confined to loose feces, seen in the female dosed at 2000 mg/kg body weight in the sighting study. This sign was first noted approximately one hour after dosing. Recovery, as judged by external appearance

Gross pathology:

No abnormalities were noted in any animal at the macroscopic examination at study termination on Day 15.

Other findings:

None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

300 mg/kg bw < Oral LD50 Females < 2000 mg kg bw.
Under the test conditions, the test item is classified as Acute oral tox. 4 (H302: Harmful if swallowed) according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an acute oral toxicity study performed according to the OECD test guideline No. 420 and in compliance with GLP, groups of fasted, eight to twelve weeks of age Wistar (RccHanTM:WIST) female rats were given a single oral dose of test item undiluted or as a suspension in Corn oil. Sighting investigations were conducted at 300 and 2000 mg/kg body weight. Based on the results of the sighting investigations a further four fasted females were similarly dosed at 2000 mg/kg body weight. Due to the number of deaths in the main study at 2000 mg/kg body weight, a further four fasted females were similarly dosed at 300 mg/kg body weight to complete the study. During the study, clinical condition, body weight and macropathology investigations were undertaken.

 

Four females dosed at 2000 mg/kg body weight in the main study were killed for welfare reasons on Day 2. Clinical signs prior to death comprised piloerection, hunched posture, elevated gait and unsteady gait in all four animals and irregular breathing, partially closed eyelids (both) and decreased activity (behavior) in two animals. These signs were seen from approximately two hours after dosing. A loss in body weight was noted for the decedents. Macroscopic examination of the animals revealed congestion (characterized by darkened tissues/organs) of the subcutaneous tissue and heart for all four females, liver and kidneys for two animals and spleen for one animal, congestion (characterized by blood vessels injected) of the brain and lungs and bronchi for all four animals, pallor of the liver and spleen for three females and the kidneys for two animals, small (atrophy) for the spleen and cecum for four females, gaseous distension of the stomach for four animals and fluid contents (yellow) for the duodenum and small and large intestines for all four animals. Clinical signs observed in surviving animals were confined to loose feces, seen in the female dosed at 2000 mg/kg body weight in the sighting study. This sign was first noted approximately one hour after dosing. Recovery, as judged by external appearance and behavior, was complete by three hours after dosing. No clinical signs were seen in any animal dosed at 300 mg/kg body weight in the sighting or main study. A low body weight gain was seen for one female (Animal Number 14) from Day 8 to 15. All other surviving animals were considered to have achieved satisfactory body weight gains throughout the study. No abnormalities were noted in any animal at the macroscopic examination at study termination on Day 15.

 

Oral LD50 Females: in the range 300 - 2000 mg/kg bw.

Under the test conditions, the test item is classified as Acute oral tox. 4 (H302: Harmful if swallowed) according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

 

This study is considered as acceptable. It satisfies the guideline requirement for an acute oral study (OECD 420) in the rats.