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Description of key information

The acute toxicity of 3.3.5 -trimethylcyclohexyl acrylate was evaluated by oral and dermal route in rats. No mortality was observed at the highest dose of 2000 mg/kg bw in both studies. No acute toxicity study is available by inhalation.

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:
27 November 2015 -- 13 January 2016
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
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France.
- Age/Weight: on the day of treatment, the females were 8 weeks old and had a mean body weight of 216 g (range: 198 g to 235 g).
- Fasting period before study: yes, during the night before treatment
- Housing: polycarbonate cages
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 14 December 2015 to 13 January 2016
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: in the absence of recommendation from the Sponsor, the solubility assay first started at the concentration of 200 mg/mL, and the first choice vehicle was drinking water treated by reverse osmosis.
As unsatisfactory solubility of the test item was obtained in this vehicle (i.e. heterogeneous emulsion at the concentration of 200 mg/mL), another vehicle was chosen from the following organic solvents (in order of preference): 0.5% methylcellulose aqueous solution and corn oil.

- Maximum dose-volume applied: 10 mL/kg

DOSAGE PREPARATION (if unusual): fresh dose formulations were prepared by the CiToxLAB France Pharmacy on the day of each administration and kept at room temperature prior to administration.

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose: 300 mg/kg
Doses:
300 and 2000 mg/kg
No. of animals per sex per dose:
Three nulliparous and non-pregnant female per treatment step
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment on day 1; then on days 8 and 15.
- Necropsy of survivors performed: yes (macroscopic).
Statistics:
no
Key result
Sex:
female
Dose descriptor:
LD0
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No unscheduled deaths occurred during the study.

Clinical signs:
At 300 mg/kg, no clinical signs were observed in any animals.
At 2000 mg/kg, hypoactivity, staggering gait and half-closed eyes were observed on Day 1 approximately 6 hours after treatment in 3/6 females, while another 1/6 females showed ptyalism on Day 1, 30 minutes after treatment and a slight alopecia on both forelimbs from Day 14. No clinical signs were noted in the remaining 2/6 females treated at the same dose-level.
Body weight:
At 300 mg/kg, lower mean body weight gain was noted between Day 1 and Day 8, when compared to CiToxLAB France historical control data (+38 g vs. +46 g in control data base). This was followed by higher mean body weight gain between Day 8 and Day 15 (+16 vs. +10 g in control data base). Therefore no relevant differences from CiToxLAB France historical control data were noted in the mean body weight change of test item-treated animals over the study period.

Compared to CiToxLAB France historical control data, the mean body weight of the animals was considered to be unaffected by the test item treatment.
Gross pathology:
There were no macroscopic findings at necropsy.
Other findings:
no
Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the oral LD0 of the test item was higher than 2000 mg/kg in rats.
Executive summary:

The objective of this study was to evaluate the potential acute toxicity of the test item following a single oral administration (gavage) to rats.

This study was conducted in compliance with the OECD Guideline No. 423 and the principles of Good Laboratory Practice.

 

Methods

 

The test item was administered once by the oral route (gavage) to three groups of three fasted female Sprague-Dawley rats with a dosage-volume of 10 mL/kg. The test item was prepared in corn oil.

No relevant toxicity data were available for estimation of a lethal selected dose-level. The starting dose-level selected was 300 mg/kg for ethical reasons. After the first assay, the next higher dose-level of 2000 mg/kg was tested. Then, as no toxicity was observed at this dose-level, the results were confirmed in three additional females.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. Body weight was recorded on Day 1 and then on Days 8 and 15.

On completion of the observation period, the animals were sacrificed and then submitted for a macroscopic post-mortem examination. No tissues were preserved.

 

Results

 

No unscheduled deaths occurred during the study.

At 300 mg/kg, no clinical signs were observed in any animals.

At 2000 mg/kg, hypoactivity, staggering gait and half-closed eyes were observed on Day 1 approximately 6 hours after treatment in 3/6 females, while another 1/6 females had ptyalism on Day 1, 30 minutes after treatment and slight alopecia on both forelimbs from Day 14. No clinical signs were noted in the remaining 2/6 females treated at the same dose-level.

No relevant differences from historical control data were noted in the body weight change and body weight of test item-treated animals over the study period.

There were no macroscopic findings at necropsy.

Conclusion

Under the experimental conditions of this study, the oral LD0 of the test item was higher than 2000 mg/kg in rats.

Therefore, the test item should not be classified regarding its toxic potential by oral route according to the criteria of CLP Regulation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The oral acute study is considered to be reliable (performed with OECD and GLP guidelines).

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:
05 December 2016 - 23 December 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
24th February 1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Commission Regulation (EC) No. 440/2008, Part B.3, 30 May 2008.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France.
- Age/Weight: on the day of treatment, the animals were approximately 8 weeks old. The individual body weight of each individual was within ± 20% of the mean body weight of all the study animals of the same sex (224 g for females and 358 g for males).
- Fasting period before study: yes, during the night before treatment
- Housing: polycarbonate cages
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 01 December 2016 to 23 December 2016
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 10% of body surface, dorsal site
- Type of wrap if used: hydrophilic gauze pad + adhesive hypoallergenic aerated semi-occlusive dressing + restraining bandage

REMOVAL OF TEST SUBSTANCE
- Removal of dressing: 24h post-exposure
- Washing: at 24h post-exposure, with a moistened cotton pad

TEST MATERIAL
- Amount applied: 2000 mg/kg
- Constant volume: no
Duration of exposure:
24h
Doses:
2000 mg/kg
No. of animals per sex per dose:
five males and five females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment on Day 1; then on Days 8 and 15.
- Necropsy of survivors performed: yes (macroscopic).
Statistics:
no
Key result
Sex:
male/female
Dose descriptor:
LD0
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No unscheduled deaths occurred during the study.

Clinical signs:
On Day 1, ptyalism was noted in 1/5 female and chromodacryorrhea was seen in 1/5 male. No other clinical signs were noted thereafter.

Local reactions at the injection site were recorded and consisted in very slight to well-defined erythema in all animals between Day 2 and Day 6, dryness and/or desquamation in all females and 3/5 males from Day 3 to Day 7, and scabs in 4/5 females and 3/5 males between Day 3 and Day 15.
Body weight:
Body weight of animals was unaffected by the test item treatment.
Gross pathology:
There were no macroscopic findings at necropsy.

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD0 of the test item was higher than 2000 mg/kg in rats.
Executive summary:

The objective of this study was to evaluate the potential toxicity of the test item, following a single dermal application to rats. This study was based on the international guidelines (OECD No. 402 and Council Regulation No. 440/2008 of 30 May 2008, Part B.3) and was performed in compliance with the principles of Good Laboratory Practice.

The test item was applied in its original form to the skin of five female then five male Sprague-Dawley rats at the dose-level of 2000 mg/kg. The application site was covered by an aerated hypoallergenic dressing for 24 hours. Each animal was observed at least once a day for mortality and clinical signs for 15 days. From Day 2, any local reactions at the treatment site were also noted. Body weight was recorded on Day 1 and then on Days 8 and 15. On completion of the observation period, the animals were sacrificed and then submitted for a macroscopic post-mortem examination. No tissues were preserved.

No unscheduled deaths occurred during the study. On Day 1, ptyalism was noted in 1/5 female and chromodacryorrhea was seen in 1/5 male. No other clinical signs were noted thereafter in any animals. Local reactions were recorded at the injection site and consisted in very slight to well-defined erythema, dryness and/or desquamation and scabs. 

Body weight of animals was unaffected by the test item treatment. There were no macroscopic findings at necropsy.

In conclusion, the dermal LD0 of the test item was higher than 2000 mg/kg in rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The dermal acute study is considered to be reliable (performed with OECD and GLP guidelines).

Additional information

Oral acute toxicity (Papineau 2016):

The objective of this study was to evaluate the potential acute toxicity of the test item following a single oral administration (gavage) to rats. This study was conducted in compliance with the OECD Guideline No. 423 and the principles of Good Laboratory Practice.

 No unscheduled deaths occurred during the study. 

At 300 mg/kg, no clinical signs were observed in any animals. At 2000 mg/kg, hypoactivity, staggering gait and half-closed eyes were observed on Day 1 approximately 6 hours after treatment in 3/6 females, while another 1/6 females had ptyalism on Day 1, 30 minutes after treatment and slight alopecia on both forelimbs from Day 14. No clinical signs were noted in the remaining 2/6 females treated at the same dose-level.

No relevant differences were noted in the body weight change and body weight of test item-treated animals over the study period.

There were no macroscopic findings at necropsy.

Under the experimental conditions of this study, the oral LD0 of the test item was higher than 2000 mg/kg in rats.

Dermal acute toxicity (Bouhraoua 2017):

The objective of this study was to evaluate the potential toxicity of the test item, following a single dermal application to rats. This study was based on the international guidelines (OECD No. 402 and Council Regulation No. 440/2008 of 30 May 2008, Part B.3) and was performed in compliance with the principles of Good Laboratory Practice.

The test item was applied in its original form to the skin of five female then five male Sprague-Dawley rats at the dose-level of 2000 mg/kg. The application site was covered by an aerated hypoallergenic dressing for 24 hours. Each animal was observed at least once a day for mortality and clinical signs for 15 days. From Day 2, any local reactions at the treatment site were also noted. Body weight was recorded on Day 1 and then on Days 8 and 15. On completion of the observation period, the animals were sacrificed and then submitted for a macroscopicpost-mortemexamination. No tissues were preserved.

No unscheduled deaths occurred during the study. On Day 1, ptyalism was noted in 1/5 female and chromodacryorrhea was seen in 1/5 male. No other clinical signs were noted thereafter in any animals. Local reactions were recorded at the injection site and consisted in very slight to well-defined erythema, dryness and/or desquamation and scabs. 

Body weight of animals was unaffected by the test item treatment.There were no macroscopic findings at necropsy.

In conclusion, the dermal LD0 of the test item was higher than 2000 mg/kg in rats.

Justification for classification or non-classification

According to the available data, 3.3.5-trimethylcyclohexyl acrylate should not be classified for acute toxicity according to the Regulation EC n°1272/2008 (CLP). No classification is justified because no mortality was observed in the acute oral and dermal toxicity studies on rats at the highest doses required by CLP.

No inhalation acute study was available, but inhalation absorption is not expected due to the low vapour pressure of 3.3.5-trimethylcyclohexyl acrylate.

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