2,3-dihydro-2,2,6-trimethylbenzaldehyde

Endpoint summary

Description of key information

Oral: LD50 > 300 - < 2000 mg/kg bw, female rat, OECD TG 423, 2015

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:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in 2015 at recognised contract research organisation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

other: Crl:WI rats

Sex:

female

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Doses:

2000 mg/kg body weight (bw) (Group 1)
300 mg/kg bw (Group 2 and Group 3)

No. of animals per sex per dose:

Three groups of three female rats_ single oral treatment

Control animals:

yes

Details on study design:

The single-dose oral toxicity of SAFRANAL was performed according to the acute toxic class method in Crl:WI rats.
Three groups of three female rats were treated with the test item at a dose level of 2000 mg/kg body weight (bw) (Group 1) or at a dose level of 300 mg/kg bw (Group 2 and Group 3).
A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. The food was available again 3 hours after the treatment. The test item was administered formulated in Poly(ethylene glycol) 400 (PEG 400) at a concentration of 200 mg/mL or 30 mg/mL and at a dose volume of 10 mL/kg bw.
Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Since all animals died, a new group (Group 2) was treated at a dose level of 300 mg/kg bw. No mortality was observed in this group, therefore a confirmatory group (Group 3) was tested at the same dose level according to the Regulatory Guidelines.
Clinical observations were performed at 5 and 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter (or at the time points listed above, until the time point closest to the death of the animal). Body weights were measured on Day -1, and Day 0, and in surviving animals on Day 7 and Day 14 before the necropsy or at death. All animals were subjected to a necropsy and macroscopic examination.

Statistics:

no statistics required as to following top-down method

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All rats died at a dose level of 2000 mg/kg bw on Day 1 or on Day 3.
The test item did not cause mortality at a dose level of 300 mg/kg bw.

Clinical signs:

other: At a dose level of 2000 mg/kg bw on Day 0-2 the following clinical signs were recorded: decreased activity (slight) excessive digging irritability hunched back incoordination (slight to severe) continuous tremors increased salivation (slight) piloerection

Gross pathology:

At a dose level of 2000 mg/kg bw, the gastric glandular mucosa was dark red (2/3) or in the stomach yellow, liquid material was present (1/3), and was considered to be associated with administration of the test item. In the collapsed lungs of the three found dead rats, diffuse, dark red discoloration could be observed and regarded as agonal or post mortem.
In the rats at the dose level of 300 mg/kg bw, no external or internal findings were recorded at necropsy.

Interpretation of results:

Toxicity Category IV

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item SAFRANAL was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

Executive summary:

STUDY DESIGN

The single-dose oral toxicity of SAFRANAL was performed according to the acute toxic class method in Crl:WI rats.

Three groups of three female rats were treated with the test item at a dose level of 2000 mg/kg body weight (bw) (Group 1) or at a dose level of 300 mg/kg bw (Group 2 and Group 3).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. The food was available again 3 hours after the treatment. The test item was administered formulated in Poly(ethylene glycol) 400 (PEG 400) at a concentration of 200 mg/mL or 30 mg/mL and at a dose volume of 10 mL/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Since all animals died, a new group (Group 2) was treated at a dose level of 300 mg/kg bw. No mortality was observed in this group, therefore a confirmatory group (Group 3) was tested at the same dose level according to the Regulatory Guidelines.

Clinical observations were performed at 5 and 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter (or at the time points listed above, until the time point closest to the death of the animal). Body weights were measured on Day -1, and Day 0, and in surviving animals on Day 7 and Day 14 before the necropsy or at death. All animals were subjected to a necropsy and macroscopic examination.

RESULTS

Mortality

All rats died at a dose level of 2000 mg/kg bw on Day 1 or on Day 3. The test item did not cause mortality at a dose level of 300 mg/kg bw.

Clinical Observations

At a dose level of 2000 mg/kg bw on Day 0-2 the following clinical signs were recorded:

decreased activity (slight), excessive digging, irritability, hunched back, incoordination (slight to severe), continuous tremors, increased salivation (slight), piloerection and red discoloration on both fore-paws, both hind-paws and on the tail in 3 of 3 rats and red discoloration on both pinna in 2 of 3 rats.

The rats dosed to 300 mg test item/kg bw showed the following clinical signs on Day 0:

irritability, excessive digging, piloerection, hunched back, red discoloration on both fore-paws, both hind-paws and on both pinna in 6 of 6 rats, increased salivation (slight) in 4 of 6 rats, incoordination (slight) in 3 of 6 rats and red discoloration on the tail in 1 of 6 rat.

Each rat of this dose level was symptom-free from Day 1 at the latest.

Body Weight and Body Weight Gain

Body weight gains of SAFRANAL-treated surviving animals showed no indication of a test item-related effect during the study.

Macroscopic Findings

At a dose level of 2000 mg/kg bw, the gastric glandular mucosa was dark red (2/3) or in the stomach yellow, liquid material was present (1/3), and was considered to be associated with administration of the test item. In the collapsed lungs of the three found dead rats, diffuse, dark red discoloration could be observed and regarded as agonal or post mortem.

In the rats at the dose level of 300 mg/kg bw, no external or internal findings were recorded at necropsy.

CONCLUSION

Under the conditions of this study, the acute oral LD50 value of the test item SAFRANAL was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

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:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

ORAL:

Key study : OECD TG 423, 2015 : The study was performed according to OECD 423 and EU Method B1 tris Acute Toxicity under GLP to assess the acute oral toxicity of the test item following a single oral administration in the Crl: WI strain rat.

Three groups of three female rats were treated with the test item at a dose level of 2000 mg/kg body weight (bw) (Group 1) or at a dose level of 300 mg/kg bw (Group 2 and Group 3).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. The food was available again 3 hours after the treatment. The test item was administered formulated in Poly(ethylene glycol) 400 (PEG 400) at a concentration of 200 mg/mL or 30 mg/mL and at a dose volume of 10 mL/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. Since all animals died, a new group (Group 2) was treated at a dose level of 300 mg/kg bw. No mortality was observed in this group, therefore a confirmatory group (Group 3) was tested at the same dose level according to the Regulatory Guidelines.

Clinical signs and bodyweight development were monitored during the study. All animals were subjected to a necropsy and macroscopic examination.

All rats died at a dose level of 2000 mg/kg bw on Day 1 or on Day 3.

The test item did not cause mortality at a dose level of 300 mg/kg bw.

At a dose level of 2000 mg/kg bw on Day 0-2 the following clinical signs were recorded: decreased activity (slight), excessive digging, irritability, hunched back, incoordination (slight to severe), continuous tremors, increased salivation (slight), piloerection, red discoloration on both fore-paws, both hind-paws and on the tail in 3 of 3 rats and red discoloration on both pinna in 2 of 3 rats.

The rats dosed to 300 mg test item/kg bw showed the following clinical signs on Day 0: irritability, excessive digging, piloerection,  hunched back, red discoloration on both fore-paws, both hind-paws and on both pinna in 6 of 6 rats increased salivation (slight) in 4 of 6 rats, incoordination (slight) in 3 of 6 rats and red discoloration on the tail in 1 of 6 rat. Each rat of this dose level was symptom-free from Day 1 at the latest.

Body weight gains of SAFRANAL-treated surviving animals showed no indication of a test item-related effect during the study.

At a dose level of 2000 mg/kg bw, the gastric glandular mucosa was dark red (2/3) or in the stomach yellow, liquid material was present (1/3), and was considered to be associated with administration of the test item. In the collapsed lungs of the three found dead rats, diffuse, dark red discoloration could be observed and regarded as agonal or post mortem.

In the rats at the dose level of 300 mg/kg bw, no external or internal findings were recorded at necropsy.

Under the conditions of this study, the acute oral LD50 value of the test item SAFRANAL was found to be between 300 and 2000 mg/kg bw in female Crl:WI rats.

Safranal meets the criteria to be classified Category 4  according to the GHS classification.