Registration Dossier

Administrative data

Description of key information

Oral: The acute oral LD50 value of the test item in the female Sprague Dawley rat was found to be > 2000 mg/kg bw (OECD 423 and EU Method B.1.Tris).

 

Dermal: The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats (OECD 402).

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 September 2016 to 12 October 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
temperature and humidity outside planned limits with no impact on results or integrity of the study (see below)
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
temperature and humidity outside planned limits with no impact on results or integrity of the study (see below)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
temperature and humidity outside planned limits with no impact on results or integrity of the study (see below)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(SD)
Sex:
female
Details on test animals and environmental conditions:
EXPERIMENTAL ANIMALS
- Species and strain: Crl:CD(SD) Sprague Dawley rats
- Source: Charles River Laboratories, Research Models and Services GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany.
- Sex: Female, nulliparous and non-pregnant.
- Age of animals at dosing: Young healthy adult rats, 8 weeks old.
- Body weight at treatment: 189 to 195 g
- Acclimatisation period: 5 or 6 days.

HUSBANDRY
- Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
- Number of animal room: 244/1.
- Housing: Three animals per cage.
- Cage type: Type II polypropylene/polycarbonate.
- Bedding: Lignocel 3/4-S Hygienic Animal Bedding and “Arbocel crinklets natural” nesting material produced by J. Rettenmaier & Söhne GmbH & Co.KG (D-73494 Rosenberg, Germany) were available to animals during the study. A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
- Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
- Temperature: 18.4 to 24.1 °C.
- Relative humidity: 30 to 80 %.
- Temperature and relative humidity were recorded twice daily during the acclimatisation period and throughout the study.
- Ventilation: 15 to 20 air exchanges/hour.
- Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

FOOD AND WATER SUPPLY
- Animals received ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany (Batch No.: 278 5652, expiry date: 30 November 2016), ad libitum, and tap water from the municipal supply, as for human consumption from a 500 ml bottles, ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József, A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

ANIMAL IDENTIFICATION
- Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.'s Master File, for each animal allocated to the treatment groups.
- Cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Route of administration:
oral: gavage
Vehicle:
corn oil
Remarks:
Acros Organics (Batch A0348063; Expiry date 31 May 2018)
Details on oral exposure:
FORMULATION
- The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle, in the Pharmacy of CiToxLAB Hungary Ltd. on the day of administration.
- The formulation container was stirred continuously until treatment was completed.

TEST ITEM ADMINISTRATION
- The initial dose level was selected by the study director to be that which is most likely to produce mortality in some of the dosed animals. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.
- Initially, three female animals (Group 1) were treated with 2000 mg/kg bw of test item. No mortality was observed, therefore further 3 animals were treated at the dose level of 2000 mg/kg bw. As no mortality was observed in this second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris).
- A single oral gavage administration was followed by a fourteen-day observation period. On the night before treatment, the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. Food was returned three hours after treatment.

CLINICAL OBSERVATIONS
- Clinical observations were performed on all animals at 0.5, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter.
- Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

BODY WEIGHT MEASUREMENT
- The body weights of the animals were recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.

NECROPSY
- Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal 40%; Lot No.: 1409236-06, Expiry Date: September 2017, Produced by: AlfasanNederland BV, Woerden, Netherlands).
- After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Six
Control animals:
no
Statistics:
EVALUATION OF TEST RESULTS
- The method used was not intended to allow the calculation of a precise LD50 value.
- Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- The test item did not cause mortality at a dose level of 2000 mg/kg bw.
Clinical signs:
- Slightly decreased activity was observed in each animal from 1 or 2 hours after treatment for 1-3 hours. After this, there were no systemic clinical signs noted in any animal throughout the study (see Appendix 1, attached).
Body weight:
- There were no treatment related body weight changes. Body weight gains of treated animals during the study showed no indication of a test item related effect. (see Appendix 2, attached).
Gross pathology:
- There was no evidence of the macroscopic effects at a dose level of 2000 mg/kg bw (see Appendix 3 attached).
Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral LD50 value of the test item in the female Sprague Dawley rat was found to be > 2000 mg/kg bw.
Executive summary:

GUIDELINE

The single-dose oral toxicity of the test item was investigated according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Sprague Dawley rats.

 

METHODS

Two groups of three female Sprague Dawley rats were treated with the test item at a dose level of 2000 mg/kg bw (Group 1 and Group 2). A single oral treatment was carried out by gavage for each animal after withdrawal of food overnight. Food was made available again 3 hours after the treatment. The test item was administered formulated in corn oil at a concentration of 200 mg/mL at a dose volume of 10 mL/kg bw. Initially, three female animals (Group 1) were treated with 2000 mg/kg bw of test item. No mortality was observed, therefore a further three animals were treated at the dose level of 2000 mg/kg bw. No mortality was observed in the confirmatory group and no further testing was required according to the test guidelines. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

 

RESULTS

The test item did not cause mortality at a dose level of 2000 mg/kg bw. Clinical observations at the dose level of 2000 mg/kg bw were reported as Slightly decreased activity in each animal from 1 or 2 hours after treatment for 1-3 hours. After this, there were no systemic clinical signs noted in any animal throughout the study. There were no treatment related body weight changes and body weight gains of treated animals during the study did not indicate a test item-related effect. No macroscopic observations were reported at 2000 mg/kg bw.

 

CONCLUSION

The acute oral LD50 value of the test item in the female Sprague Dawley rat was found to be > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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:
08 November 2016 to 22 November 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
humidity outside expected range with no impact on results or integrity of the study (see below)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
EXPERIMENTAL ANIMALS
- Species and strain: Sprague Dawley Rat (Crl:CD(SD))
- Source: Charles River Laboratories, Research Models and Services GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany.
- Sex: Male and female, female rats were nulliparous and non-pregnant.
- Age of animals at study start: Young healthy adult rats
- Body weight range at dosing: 234 to 269 g
- Acclimatisation period: 5 days

HUSBANDRY
- Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
- Room-Box: 242/5
- Housing: Individual caging
- Cage type: Type II. polypropylene/polycarbonate
- Bedding: “Lignocel 3/4-S Hygienic Animal Bedding” and “Arbocel crinklets natural” nesting
material produced by J. Rettenmaier & Söhne GmbH & Co.KG (D-73494 Rosenberg,
Germany) were available to animals during the study. A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
- Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
- Temperature: 22.0 to 24.9 °C
- Relative humidity: 25 to 50 %
- Ventilation: 15 to 20 air exchanges/hour
- Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
- Temperature and relative humidity were recorded twice daily during the acclimatisation period and throughout the study.

FOOD AND WATER SUPPLY
- Animals received ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany (Batch number: 141 8884, expiry date: 31 January 2017)), ad libitum, and tap water from the municipal supply, as for human consumption from a 500 ml bottles, ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study and the water was considered fit for human consumption.
- Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A. u. 36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

ANIMAL IDENTIFICATION
- Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of the CiToxLAB Hungary Ltd Master File, for each animal allocated to the treatment groups.
- The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
FORMULATION
- The test item was administered as a single dose without using any vehicle.
- Sufficient water was used to dampen the test material to ensure good contact with the skin.

OTHER MATERIALS
- GSM Betasilk Surgical Silk Tape (adhesive tape); Lot number #1: 13100544; Expiry date #1: 31 October 201; Lot number #2: 14032134; Expiry date #2: 31 March 2019; Produced by: GS Medical Ltd., 83, Amiens Street 1, Dublin, Ireland.
- Pharmico Steril Mull-lap (sterile gauze pads); Lot number: 1414142; Expiry date: 31 December 2020; Produced by: Dispomedicor Zrt.,; H-4032, Debrecen, Füredi u. 98, Hungary.

TEST ITEM ADMINISTRATION
- The test item was not expected to be lethal at 2000 mg/kg bw and a limit test was therefore performed.
- A single dermal application was made and was followed by a fourteen-day observation period (male animals cages 1 to 5; female animals cages 6 to 10).

PROCEDURE
- The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment.
- Test item was applied as a single dose to the shaved skin and remained in contact with the skin for the 24-hour exposure period.
- The test material was dampened with sufficient water before application to ensure good contact with the skin.
- Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
- At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.

CLINICAL OBSERVATIONS
- Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter.
- Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

SKIN IRRITATION
- Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

MEASUREMENT OF BODY WEIGHT
- Body weights were recorded on Day 0 (before test item administration) and on Days 7 and 14 just before necropsy.

NECROPSY
- Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal 40%; Lot No.: 1409236-06, Expiry Date: September 2017, Produced by: AlfasanNederland BV, Woerden, Netherlands).
- After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw/day
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Statistics:
EVALUATION OF DATA
- The method used was not intended to allow the calculation of a precise LD50 value.
- Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- The test item did not cause mortality at the dose level of 2000 mg/kg bw.
Clinical signs:
- There were no systemic clinical signs noted in any animal throughout the study (see Appendix 1, attached).
Body weight:
- Body weight gains of treated animals during the study showed no indication of a test item-related effect (see Appendix 2, attached).
Gross pathology:
- No macroscopic observations were noted at a dose level of 2000 mg/kg bw (see Appendix 3, attached).
Other findings:
LOCAL DERMAL EFFECTS
- No local dermal signs were observed after treatment with the test item during the 14 days observation period.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats.
Executive summary:

GUIDELINE

An acute dermal toxicity study was performed with test item in Sprague Dawley rats, in compliance with OECD Guideline No 402.

 

METHODS

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 2 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

 

RESULTS

The test item did not cause mortality at the dose level of 2000 mg/kg bw and no systemic clinical signs were noted in any animal throughout the study. No local dermal signs were observed after treatment with the test item during the 14 days observation period. Body weight gains of treated animals during the study showed no indication of a test item-related effect and no macroscopic observations were noted at a dose level of 2000 mg/kg bw.

 

CONCLUSION

The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Oral route

The single-dose oral toxicity of the test item was investigated in a key study according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Sprague Dawley rats.

Two groups of three female Sprague Dawley rats were treated with the test item at a dose level of 2000 mg/kg bw (Group 1 and Group 2). A single oral treatment was carried out by gavage for each animal after withdrawal of food overnight. Food was made available again 3 hours after the treatment. The test item was administered formulated in corn oil at a concentration of 200 mg/mL at a dose volume of 10 mL/kg bw. Initially, three female animals (Group 1) were treated with 2000 mg/kgbw of test item. No mortality was observed, therefore a further three animals were treated at the dose level of 2000 mg/kg bw. No mortality was observed in the confirmatory group and no further testing was required according to the test guidelines. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

 

The test item did not cause mortality at a dose level of 2000 mg/kg bw. Clinical observations at the dose level of 2000 mg/kg bw were reported as Slightly decreased activity in each animal from 1 or 2 hours after treatment for 1-3 hours. After this, there were no systemic clinical signs noted in any animal throughout the study. There were no treatment related body weight changes and body weight gains of treated animals during the study did not indicate a test item-related effect. No macroscopic observations were reported at 2000 mg/kg bw.

The acute oral LD50 value of the test item in the female Sprague Dawley rat was found to be > 2000 mg/kg bw.

Inhalation route

The substance decomposes from approximately 232 °C at 101 kPa and the vapour pressure has been determined to be 1.75 x 10E-03 Pa at 25 °C. It is therefore expected that inhalation exposure will be low under general use conditions at ambient temperature. Lack of systemic toxicity when the substance is administered via the oral route suggests that determined vapour pressures of 1.21 Pa at 154°C also give no cause for concern. The inhalation route is therefore not the most applicable method of investigating acute toxicity.

Dermal route

An acute dermal toxicity study was performed with test item in Sprague Dawley rats, in compliance with OECD Guideline No 402.

 

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 2 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

 

The test item did not cause mortality at the dose level of 2000 mg/kg bw and no systemic clinical signs were noted in any animal throughout the study. No local dermal signs were observed after treatment with the test item during the 14 days observation period. Body weight gains of treated animals during the study showed no indication of a test item-related effect and no macroscopic observations were noted at a dose level of 2000 mg/kg bw.

 

The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats.

Justification for classification or non-classification

The acute median lethal dose (LD50) values of the test item in the Sprague Dawley rat were reported as > 2000 mg/kg bw following oral and dermal administration. Classification for acute oral or dermal toxicity is therefore unnecessary under the terms of Regulation (EC) No 1272/2008.