Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Appearance/physical state: Tan waxy solid
- Storage conditions: Room temperature in the dark

Test animals

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.

Administration / exposure

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.

Results and discussion

Effect levels
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.

Applicant's summary and conclusion

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.