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

Endpoint summary

Description of key information

300 < Combined Oral LD50 < 2000 mg/kg bw/day (WoE OECD 423, GLP, class method in rats).

Combined Dermal LD50 > 2000 mg/kg bw (OECD 402, GLP, K, Rel. 1, limit test in rats).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

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.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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

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

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

Preliminary study:

Not applicable

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

None

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.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Between 05 October 1999 and 27 October 1999.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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: 22 September 1999 / Date of signature: 21 December 1999

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS (Sprague Dawley ICO: OFA-SD (IOPS Caw))
- Source: Iffa Crédo, 69210 L'Arbresle, Frances.
- Age at study initiation: ca. six weeks old.
- Weight at study initiation: Males: 167+/-12 g / Females: 138+/-4 g
- Fasting period before study: overnight fast (18 hours) before dosing and for approximately four hours after dosing
- Housing: in groups of three by sex in polycarbonate cages containing dust-free sawdust.
- Diet (e.g. ad libitum): free access to and food (A04C pelleted diet supplied by UAR, 91360 Villemoisson-sur-Orge, France)
- Water (e.g. ad libitum): free access to drinking water filtered by a FG Millipore membrane.
- 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): 21 +/- 2 °C
- Humidity (%): 30-70 %
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1999-10-05 To: 1999-10-27

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10 mL/kg bw
- Lot/batch no. (if required): 107H1649 (Sigma, 38297 Saint-Quentin-Fallavier, France)

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
The test substance was prepared in the vehicle. The administration was performed in a single dose by oral route using a metal gavage tube fitted to a 5 mL glass syringe (0.05 mL graduations).
The volume administered to each animal was calculated according to the bodyweight determined on the day of treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The information available suggested a starting dose at the highest dose-level and the study started by an assay at the dose-level of 2000 mg/kg bw in three males.

Doses:

2000 and 200 mg/kg bw

No. of animals per sex per dose:

3 males at 2000 mg/kg bw; 3 males at 200 mg/kg bw; 3 females at 200 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (Day 15)
- Frequency of observations: the animals were observed frequently during the hours following administration for detection of possible treatment-related clinical signs, thereafter the observation was made at least once daily.
- Frequency of weighing: prior to dosing and on Day 1, 8 and 15.
- Necropsy of survivors performed: yes, macroscopic examination (digestive tract, heart, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities)

Statistics:

None

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At the 2000 mg/kg bw dose-level, all males died between day 1 and day 2.
No mortality was observed in the animals given 200 mg/kg bw.

Clinical signs:

other: Hypactivity or sedation, dyspnea and lateral recumbery were observed prior to death at 2000 mg/kg bw/day. No clinical sign was observed in the animals given 200 mg/kg bw.

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None

None

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

200 < Oral LD50 < 2000 mg/kg bw. 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, six weeks of age Sprague-Dawley rats were given a single oral dose of test material diluted in corn oil.

A group of three fasted males was treated with the test item at a dose level of 2000 mg/kg bw bodyweight. Based on the results from this dose level, further groups of fasted males and females were treated at a dose level of 200 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.

At the 2000 mg/kg bw dose-level, all males died between day 1 and day 2. Hypactivity or sedation, dyspnea and lateral recumbery were observed prior to death at 2000 mg/kg bw/day.

No clinical signs and no mortality was observed in the animals given 200 mg/kg bw.

The body weight gain of the animals given 200 mg/kg bw was similar to that of the CIT historical control animals.

At necropsy, no apparent abnormalities were observed.

200 < Oral LD50 < 2000 mg/kg bw.

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.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
[further information is included as attachment to Iuclid section 13]

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have similar physico-chemical, toxicological and environmental fate properties because of their composition similarity

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target and the source substances are reaction mass, composed of three and two dimethylindanylpropanal isomers, respectively.

3. ANALOGUE APPROACH JUSTIFICATION
The source substance has a LD50 between 200 and 2000 mg/kg bw in the available study. The study design (OECD 423, GLP) is adequate and reliable for the purpose of the prediction based on read-across. The test material used represents the source substance as described in the hypothesis in terms of purity and impurities. The results of the studies are adequate for the purpose of classification and labelling.
The target substance has a LD50 between 300 and 2000 mg/kg bw in the available study. However, the target substance 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.
Therefore, based on the considerations above, it can be concluded that the results of the skin irritation studies conducted in the rabbit with the source substance are likely to predict the properties of the target substance and are considered as adequate to fulfil the information requirement of Annex VII, 8.5.1.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Doses:

2000 and 200 mg/kg bw

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At the 2000 mg/kg bw dose-level, all males died between day 1 and day 2.
No mortality was observed in the animals given 200 mg/kg bw.

Clinical signs:

other: Hypactivity or sedation, dyspnea and lateral recumbery were observed prior to death at 2000 mg/kg bw/day. No clinical sign was observed in the animals given 200 mg/kg bw.

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None

None

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Based on data available on the source substance: 200 < Oral LD50 < 2000 mg/kg bw. The target and source substances are therefore 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 on the source substance according to the OECD test guideline No. 423 and in compliance with GLP, groups of fasted, six weeks of age Sprague-Dawley rats were given a single oral dose of test material diluted in corn oil.

A group of three fasted males was treated with the test item at a dose level of 2000 mg/kg bw bodyweight. Based on the results from this dose level, further groups of fasted males and females were treated at a dose level of 200 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.

At the 2000 mg/kg bw dose-level, all males died between day 1 and day 2. Hypactivity or sedation, dyspnea and lateral recumbery were observed prior to death at 2000 mg/kg bw/day.

No clinical signs and no mortality was observed in the animals given 200 mg/kg bw.

The body weight gain of the animals given 200 mg/kg bw was similar to that of the CIT historical control animals.

At necropsy, no apparent abnormalities were observed.

200 < Oral LD50 < 2000 mg/kg bw.

The target and source substances are therefore 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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

A study was identified on the substance but due to potential solubilisation issues, this study cannot be considered as a key study and was used in a weight-of-evidence approach together with a study performed on another mixture of dimethylindanylpropanal isomers. Both studies were performed according to the OECD 423 and in compliance with GLP.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 26 to March 17, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, Acute dermal toxicity (2-1-2), 12 Nousan No 8147, Agricultural Production Bureau, November 24, 2000.

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Compliance Programme (inspected on July 01-03, 2014/ signed on September 15, 2014)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS: RccHan®:WIST rat
- Source: Harlan (UK) Ltd.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: ca. 8–9 weeks
- Weight at study initiation: 245-257 g for males and 174-183 g for females
- Fasting period before study: None
- Housing: Animals were housed individually from Day -1 until Day 4 when they were returned to group housing (in groups of five rats of the same sex). The cages were solid bottomed polycarbonate cages with a stainless steel mesh lid.
- Diet: Standard rodent diet (Harlan Teklad 2014C Diet), ad libitum
- Water: Drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-23 °C
- Humidity: 40-70%
- Air changes: at least 15 air changes per hour
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: February 26 to March 17, 2015

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lumbar region
- % coverage: 10% of the total body surface area
- Type of wrap if used: The test substance was applied by spreading it evenly over the prepared skin. The treatment area (approximately 50 mm x 50 mm) was covered with porous gauze held in place with a non-irritating dressing, and further covered by a waterproof dressing encircled firmly around the trunk of the animal.

REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the 24 hours exposure period the dressing was carefully removed and the treated area of skin was washed with warm water (30 - 40°C) containing a mild detergent solution, to remove any residual test substance. The treated area was blotted dry with absorbent paper.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Test item was administered, as supplied, at a dose volume of 2 mL/kg bw.
- The specific gravity, as measured by this laboratory, was 0.997 g/mL.
- Concentration (if solution): Undiluted
- Constant volume or concentration used: Yes

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
Frequency of observations:
- Mortality: Cages of rats were checked at least twice daily for any mortalities.
- Clinical observations: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1. On subsequent days, animals were observed once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only).
- Dermal reactions: Local dermal irritation at the treatment site was assessed daily for 14 days.
- Frequency of weighing: The weight of each rat was recorded on Days 1 (prior to dosing), 8 and 15. Individual weekly body weight changes and group mean body weights were calculated.
- Necropsy of survivors performed: Yes; All animals were humanely killed on Day 15 by carbon dioxide asphyxiation and were subjected to a macroscopic examination.

Statistics:

None

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality was observed.

Clinical signs:

other: No systemic response to treatment was observed.

Gross pathology:

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

Other findings:

Dermal reactions: No dermal reactions were observed in any animal during the study. However, bandage reaction/sticky residue was noted around but not on the dose site and was a result of the bandage adhesive sticking to the skin during the application period. This is a normal finding seen in this type of study and not considered to be associated with the test substance.

None

Interpretation of results:

GHS criteria not met

Conclusions:

Dermal LD50 Combined > 2000 mg/kg bw. Under the test conditions, the test material is not classified according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an acute dermal toxicity study (limit study) performed according to the OECD guideline No. 402, a group of ten RccHan^®:WIST albino rats (five males and five females) received a single topical application of the test substance at a dose level of 2000 mg/kg bw, for a duration of 24 hours. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and at the end of the study the surviving animals were sacrificed for macroscopic examination.

There were no deaths and no systemic response to treatment in any animal throughout the study. No dermal reactions were observed in any animal during the study. All animals were considered to have achieved satisfactory bodyweight gains throughout the study. No abnormalities were noted in any animal at the macroscopic examination at study termination on Day 15. 

 

Dermal LD50 Combined > 2000 mg/kg bw.

 

Under the test conditions, the test material is not classified according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The key study is GLP-compliant and of high quality (Klimisch score = 1)

Additional information

Acute toxicity: oral

A study was identified on the substance (Harlan 2002). It was performed according to OECD test guideline No. 423 and in compliance with GLP. However the substance 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 and was used in a weight-of-evidence approach together with a study performed on another mixture of dimethylindanylpropanal isomers, hereinafter referred as source substance (CIT, 2000).

In the Harlan study, 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. One other female was found dead one day after dosing. An issue with the formulation in water, e.g. floating of the substance on top of the water may explain the marked difference observed in the two groups of females treated with 2000 mg/kg bw/day.

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.

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.

In the CIT study on the source substance, also conducted according to the OECD test guideline No. 423 and in compliance with GLP, all male rats died between day 1 and day 2 at the 2000 mg/kg bw dose-level. No clinical signs and no mortality was observed in the animals given 200 mg/kg bw. Hypactivity or sedation, dyspnea and lateral recumbery were observed prior to death at 2000 mg/kg bw/day. At necropsy, no apparent abnormalities were observed.

The clinical signs observed in both studies may be sign of discomfort, particularly in the gastrointestinal tract. This is supported by the observation of sloughing of the gastric epithelium in the dead females of the Harlan study, which is probably evidence of irritation, which could give rise to the clinical signs.

No clinical signs and no mortality was observed in the animals given 200 or 300 mg/kg bw in both studies.

Based on these two studies, the LD50 of the target substance can be safely assumed to be between 300 and 2000 mg/kg bw.

Acute toxicity: dermal

A key study was identified (HLS, 2015). This acute dermal toxicity study (limit test) was performed according to the OECD guideline No. 402 and in compliance with GLP. There were no deaths and no systemic response to treatment in any rats throughout the study. No dermal reactions were observed in any animal during the study. All animals were considered to have achieved satisfactory bodyweight gains throughout the study. No abnormalities were noted in any animal at the macroscopic examination at study termination on Day 15. 

Dermal LD50 Combined > 2000 mg/kg bw.  

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self classification:

Acute toxicity via Oral route:

Based on the available data, the substance is:

- classified as Harmful if swallowed (Category 4, H302) according to the Regulation (EC) No. 1272/2008 as the LD50 is between 300 and 2000 mg/kg bw according to the acute toxic class method.

- classified as Harmful if swallowed (Category 4 according to the GHS as the LD50 is is between 300 and 2000 mg/kg bw according to the acute toxic class method.

Acute toxicity via Dermal route:

Based on the available information, the substance is:

- not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) as the dermal LD50 is higher than 2000 mg/kg bw

- not classified according to the GHS since there is no reliable evidence that indicates the LD50 to be in the range of Category 5 values (GHS criteria not met).

Acute toxicity (Inhalation): No data was available.

Specific target organ toxicity: single exposure (Oral):

The classification criteria according to the Annex VI of the Regulation (EC) No. 1272/2008 as specific target organ toxicant (STOT) – single exposure, oral are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (oral) for a Category 1 classification (C≤ 300 mg/kg bw) and at the guidance value (oral) for a Category 2 classification (2000 mg/kg bw≥C > 300 mg/kg bw). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 are not met since narcotic effects were not observed in the acute oral toxicity study.

Specific target organ toxicity: single exposure (Dermal):

The classification criteria according to the Annex VI of the Regulation (EC) No 1272/2008 as specific target organ toxicant (STOT) – single exposure, dermal are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value, dermal for a Category 1 classification (C≤ 1000 mg/kg bw) and at the guidance value, dermal for a Category 2 classification (2000 mg/kg bw ≥C > 1000 mg/kg bw). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 are not met since narcotic effects were not observed in the acute dermal toxicity study.

Specific target organ toxicity: single exposure (Inhalation): No data was available.