Registration Dossier

Diss Factsheets

Administrative data

Description of key information

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:
29 August 2011 - 12 October 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.
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 at study initiation: on the day of treatment, the females were approximately 8 weeks old
- Mean body weight at study initiation: 212 g (range: 195 g to 228 g)
- Fasting period before study: yes, during the night before treatment
- Housing: polycarbonate cages with stainless steel lids
- 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 6 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 (7:00 - 19:00)

IN-LIFE DATES: 14 September 2011 to 12 October 2011
Route of administration:
oral: gavage
Vehicle:
other: 0.5% methylcellulose aqueous solution
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, 0.5% methylcellulose aqueous solution, batch Nos. 107K0081 and 066K0129, supplied by Sigma (Saint-Quentin-Fallavier, France) was used. A homogenous suspension was obtained at the concentration of 200 mg/mL in 0.5% methylcellulose aqueous solution.

DOSAGE PREPARATION (if unusual):
The test item was administered as a homogenous suspension in the vehicle.
The test item was ground to a fine powder, using a mortar and pestle, and then mixed with the required quantity of vehicle.
Dosage form preparations were prepared by the CIT Pharmacy extemporaneously on the day of each administration.
The dosage forms were stored at room temperature, and delivered to the study room in brown flasks.

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose:
The starting dose-level was selected in agreement with the Sponsor. Since no relevant toxicity data were available for the estimation of a lethal dose-level and any existing data have been taken into account by the Sponsor; the starting dose level was 300 mg/kg for animal welfare reasons.
After treatment at the starting dose-level, the next dose-levels were administered in a sequential manner, under the same conditions to different
animals, according to the flow charts of OECD Guideline No. 423, 17th December 2001, which are equivalent to those of Commission Regulation (EC) No. 440/2008, B.1tris (30 May 2008).
Doses:
300 and 2000 mg/kg.
No. of animals per sex per dose:
3 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).
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No unscheduled deaths occurred in any animals.
Clinical signs:
No clincal signs at 300 mg/kg/day.
At 2000 mg/kg, hypoactivity, lateral/ventral recumbency and sedation were observed in almost all animals, accompanied in some females byhunched posture, staggering gait, half-closed eyes, ptyalism, tremors and/or dyspnea.
Body weight:
No effects on body weight were recorded, except in one animal during the first week of the study.
Gross pathology:
none.
Other findings:
none.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 of the test item was higher than 2000 mg/kg in rats.
Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.
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 principles of Good Laboratory Practice and according to OECD 423 guideline.

 The test item was administered once by oral route (gavage) to three groups of three fasted female Sprague-Dawley rats under a dosage-volume of 10 mL/kg. The test item was prepared in a 0.5% solution of methylcellulose.

Since no relevant toxicity data are available for the estimation of a lethal dose-level and any existing data have been taken into account by the Sponsor, the starting dose-level was 300 mg/kg for animal welfare reasons.After the first assay, the next higher dose-level of 2000 mg/kg was tested. Then, as no toxicity was observed at this higher dose-level, the results were confirmed in other females.

No unscheduled deaths occurred in any animals. No clinical signs were observed at 300 mg/kg.At 2000 mg/kg, hypoactivity, lateral/ventral recumbency and sedation were observed in almost all animals, accompanied in some females byhunched posture, staggering gait, half-closed eyes, ptyalism, tremors and/or dyspnea. These clinical signs were either limited to the day of administration or lasted until day 5.

No effects on body weight were recorded, except in one animal during the first week of the study and there were no macroscopic post-mortem findings.

Conclusion

The oral LD50of the test item was higher than 2000 mg/kg in rats. 

Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose

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:
03 October 2011 - 17 November 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
one female was 9-week old on the day of treatment instead of 8 week old
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
idem above
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 at study initiation: on the day of treatment, the animals were approximately 8 weeks old (except the last group 1 female which was 9 weeks old)
- Mean body weight at study initiation: a mean body weight of 335 g (range: 327 g to 344 g) and the females had a mean body weight of 231 g (range: 218 g to 256 g)
- Fasting period before study: yes, during the night before treatment
- Housing: polycarbonate cages with stainless steel lids
- 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 (7:00 - 19:00)

IN-LIFE DATES: 25 October 2011 to 17 November 2011
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(s) applied (volume or weight with unit): 2000 mg/kg
- Constant volume: no
- For solids, paste formed: the test item was placed on a gauze pad moistened with drinking water treated by reverse osmosis, which was then applied to the clipped area of skin.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
Group 1 : 5 females
Group 2 : 5 males
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).
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality.
Clinical signs:
No clinical signs indicative of systemic toxicity were observed in any animal. Very slight erythema was recorded at the application site of all females during from day 2 until days 3, 4 or 5
Body weight:
Lower body weight gain was recorded in 2/5 females and 4/5 male
Gross pathology:
None.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 of the test item was higher than 2000 mg/kg in rats.
Therefore, the test item is not classified as toxic by dermal route according to the criteria of CLP Regulation.
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 conducted in compliance with the principles of Good Laboratory Practice and accoding to OECD guideline 402.

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 a semi-occlusive dressing for 24 hours.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. From day 2, 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 a full macroscopic post-mortem examination was performed. Macroscopic lesions were preserved.

There were no unscheduled deaths during the study and no clinical signs indicative of systemic toxicity were observed in any animal.

No cutaneous reactions were observed in males.Very slight erythema was recorded at the application site of all females during from day 2 until days 3, 4 or 5. Lower body weight gain was recorded in 2/5 females and 4/5 males. There were no test item-related macroscopic findings.

Conclusion

The dermal LD50of the test item was higher than 2000 mg/kg in rats.

Therefore, the test item is not classified as toxic by dermal route according to the criteria of CLP Regulation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose

Additional information

Oral toxicity

The acute oral toxicity of the test item was evaluated in rats according to OECD(No. 423, 17th December 2001) and Commission Regulation (EC) (No. 440/2008, B.1tris, 30 May 2008) guidelines (Haag, 2011). The test item was prepared in carboxymehylcellulose and was administered by oral route (gavage), under a volume of 10 mL/kg, to groups of three fasted female Sprague-Dawley rats. Treatment was first performed at the dose-level of 300 mg/kg. As no death occurred, a second group was treated at the dose-level of 2000 mg/kg. A third group was also treated at the same dose-level to confirm the absence of mortality at this dose-level. Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy. No mortality was recorded. At 2000 mg/kg, hypoactivity, lateral decumbency, sedation, staggering gait, tremors, hunched posture, half-closed eyes and/or dyspnea, were observed after dosing in 5/6 animals. These clinical signs were either observed only the day of administration or lasted until day 3. No effects on body weight were recorded.The oral LD0of the test item was higher than 2000 mg/kg in rats.

Dermal toxicity

The acute dermal toxicity of the test item was evaluated in rats according to OECD (No. 402, 24th February 1987) and Commission Regulation (EC) (No. 440/2008, Part B.3, 30 May 2008) guidelines (Haag, 2011). The study was conducted in compliance with the principles of Good Laboratory Practice Regulations. The test item was applied to the skin of one group of ten Sprague-Dawley rats (five males and five females). The application was performed with the test item in its original form at the dose-level of 2000 mg/kg. The test site was then covered by a semi-occlusive dressing for 24 hours. Clinical signs, mortality and body weight gain were checked for a period of 14 days following the single application of the test item. All animals were subjected to necropsy. Neither mortality nor systemic clinical signs were observed during the study.

Very slight erythema was recorded at the application site of all females from day 2 until days 3, 4 or 5. Lower body weight gain was recorded in 2/5 females and 4/5 males.There were no test item-related macroscopic findings. The dermal LD50of the test item was higher than 2000 mg/kg in rats.

Therefore, the test item is not classified as toxic by dermal route according to the criteria of CLP Regulation.


Justification for selection of acute toxicity – oral endpoint
The study is a GLP study performed according to OECD 423 guideline. No mortality occured during the study. An LD50 >2000 mg/kg/day was concluded. At 2000 mg/kg, hypoactivity, lateral/ventral recumbency and sedation were observed in almost all animals, accompanied in some females byhunched posture, staggering gait, half-closed eyes, ptyalism, tremors and/or dyspnea. These clinical signs were either limited to the day of administration or lasted until day 5.

Justification for selection of acute toxicity – dermal endpoint
The study is a GLP study performed according to OECD 402 guideline. An LD50 >2000 mg/kg/day was concluded. No mortality or clinical signs occured.

Justification for classification or non-classification

No classification is warranted for oral and dermal acute toxicity under EU Dangerous Substances Directive 67/548/EEC or CLP EU Regulation 1272/2008.

Categories Display