Reaction mass of 3-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal and 3-(1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal and 3-(1,1-dimethyl-2,3-dihydro-1H-inden-4-yl)propanal

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Between 08 March 2012 and 12 April 2012.

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

Study performed according to OECD test guideline No. 423 and in compliance with GLP. However the test material was formulated in water at a concentration above its water solubility limit, raising question on the accuracy of the formulations at the high dose levels. Therefore this study cannot be considered as a key study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: 19-21 July 2011 / Date of signature: 31 August 2011

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal

Cas Number:

173445-65-3

Molecular formula:

C14H18O

IUPAC Name:

3-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal

Constituent 2

Reference substance name:

3-(1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal

Cas Number:

173445-44-8

Molecular formula:

C14H18O

IUPAC Name:

3-(1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)propanal

Constituent 3

Reference substance name:

3-(1,1-dimethyl-2,3-dihydro-1H-inden-4-yl)propanal

Molecular formula:

C14H18O

IUPAC Name:

3-(1,1-dimethyl-2,3-dihydro-1H-inden-4-yl)propanal

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): ST 28 C 11
- Physical state: extremely pale yellow liquid
- Stability under test conditions:
- Storage condition of test material: room temperature in the dark under nitrogen

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS (Hsd:Sprague Dawley®™ SD®™)
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: eight to twelve weeks of age
- Weight at study initiation: Females: 167-198 g / Males: 213-229 g
- Fasting period before study: overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: in groups of three by sex in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet (e.g. ad libitum): free access to and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)
- Water (e.g. ad libitum): free access to mains drinking water
- Acclimation period: at least five days
The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 °C
- Humidity (%): 30-70 %
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2012-03-08 To: 2012-04-12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL (300 mg/kg bw) and 200 mg/mL (2000 mg/kg bw)

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
For the purpose of the study the test item was freshly prepared, as required, as a solution at the appropriate concentration in distilled water. To assure homogeneity, the test item formulations were mixed using a vortex mixer. The test item was formulated within two hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Using available information on the toxicity of the test item, 300 mg/kg bw was chosen as the starting dose.
- Treatment of animals was sequential. Sufficient time was allowed between each group and each dose level to confirm the survival of the previously dosed animals.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 females at 300 mg/kg bw; 3+3 females at 2000 mg/kg bw; 3 males at 2000 mg/kg bw.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: the animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for up to fourteen days.
- Frequency of weighing: prior to dosing and seven and fourteen days after treatment or at death.
- Necropsy of survivors performed: yes, gross pathological examination (external examination and opening of the abdominal and thoracic cavities for examination of major organs).

Statistics:

None

Results and discussion

Preliminary study:

Not applicable

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths noted at a dose level of 300 mg/kg bw and the first set of females dosed with 2000 mg/kg bw and in males dosed with 2000 mg/kg bw.
During the dosing of the second set of females with 2000 mg/kg bw, one female treated was killed for humane reasons, one 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. One other female was found dead one day after dosing.

Clinical signs:

other: Signs of systemic toxicity noted in female rats treated at a dose level of 2000 mg/kg bw were ataxia, hunched posture, lethargy and red/brown staining around the eyes and snout. In males treated at a dose level of 2000 mg/kg, ataxia was the only clinical

Gross pathology:

Abnormalities noted at necropsy of the female treated at a dose level of 2000 mg/kg bw that died during the study were dark liver, dark spleen, reddened kidneys and epithelial sloughing of the gastric mucosa. Abnormalities noted at necropsy of the female treated at a dose level of 2000 mg/kg bw that was humanely killed were pale liver, reddened kidneys and epithelial sloughing of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

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 < Oral LD50 < 2000 mg/kg bw; The LD50 cut-off is 2000 mg/kg bw according to the OECD TG 423. Under the test conditions, the test material is classified as harmful if swallowed (Category 4) according to the Regulation (EC) 1272/2008 and to the GHS.

Executive summary:

In an acute oral toxicity study performed according to the OECD test guideline No. 423 and in compliance with GLP, groups of fasted, eight to twelve weeks of age Sprague-Dawley rats were given a single oral dose of tje test material as a suspension in distilled water.

A group of three fasted females was treated with the test item at a dose level of 300 mg/kg bw bodyweight. Based on the results from this dose level, further groups of fasted females were treated at a dose level of 2000 mg/kg bw bodyweight. In order to determine if there was a difference between sexes, an additional group of three fasted males were also treated at a dose level of 2000 mg/kg bw bodyweight. Dosing was performed sequentially.

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

There were no deaths reported at the 300 mg/kg dose level, in the first set of females treated at the 2000 mg/kg bw dose level, or in males administered 2000 mg/kg bw of the test item. In the second group of females treated at a dose level of 2000 mg/kg, one female was killed for humane reasons, one 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. One other female was found dead one day after dosing.

There were no signs of systemic toxicity noted at a dose level of 300 mg/kg bw. Signs of systemic toxicity noted in females treated at a dose level of 2000 mg/kg bw were ataxia, hunched posture, lethargy and red/brown staining around the eyes and snout. In males treated at a dose level of 2000 mg/kg bw, only ataxia was observed. Surviving animals treated at a dose level of 2000 mg/kg bw appeared normal two or three days after dosing. Additional signs of systemic toxicity noted in the humanely killed female treated at a dose level of 2000 mg/kg bw were prostration, emaciation, hypothermia, pallor of the extremities and decreased and laboured respiration. Ataxia was the only clinical sign observed in the 2000 mg/kg bw female that was found dead.

Surviving animals showed expected gains in bodyweight.

Abnormalities noted at necropsy of the female treated at a dose level of 2000 mg/kg bw that died during the study were dark liver, dark spleen, reddened kidneys and epithelial sloughing of the gastric mucosa. Abnormalities noted at necropsy of the female treated at a dose level of 2000 mg/kg bw that was humanely killed were pale liver, reddened kidneys and epithelial sloughing of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

300 < Oral LD50 < 2000 mg/kg bw; the LD50 cut-off is 2000 mg/kg bw according to the OECD TG 423.

Under the test conditions, the test material is classified as harmful if swallowed:

- Category 4 (H302) according to the Regulation (EC) No. 1272/2008 (CLP) and

- Category 4 according to the GHS.

This study is considered as acceptable and satisfies the requirement for acute oral toxicity endpoint.