Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04.Apr.2007 - 30.Aug.2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
- During the study, the temperature in the animals house was 19-28 °C and the relative humidity 35-70%. - The animals received A04C diet instead of TEKLAD 2014 as indicated in the Study Plan. These deviations had no impact on the Study
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
Commission Directive 92/69/EEC of 31 July 1992, Annex, Part B, Method 8.3, published in the Official Journal on 29 December 1992
Qualifier:
according to guideline
Guideline:
other: Directive 2003/63/EC
Version / remarks:
Commission Directive 2003/63/EC of 25 June 2003 amending Directive 2001/83/EC of the European Parliament and of the Council on the Community code relating to medicinal products for human use.
GLP compliance:
yes
Test type:
standard acute method

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
TEST ITEM PREPARATION
Dose levels were in terms of the test item as supplied by the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Approximately 24 hours before adminsitration of the test item, the back of each animal was shaved using an electric razor, from the scapula area to the lumbar region.
The test item was applied on an area representing around 10% of the total body surface area.
The test item was placed on square of hydrophilic gauze of approximately 6 x 5 cm and applied to the corresponding test area. The gauze was then covered with a hypoallergenic microporous ahesive band. This route was chosen to study the possible toxic effects produced by skin exposition to the test item.
Finally a strip, of gauze was wrapped around the trunk of the rat in order to hold the patches in place.
The band was fixed to the body of the animal using adhesive tape.
The aim of this semioclusive bandage was to allow good skin absorption of the test item and keep animals from ingesting it.
The bandage and the patches were removed 24 hours after expousure. The remains of the product were removed with distilled water.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Interfauna Ibérica, S.L. (Ctra. Sant Miquel del Fai, km 3, 08182-Sant Feliu de Codines, Barcelona, Spain
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks
- Weight at study initiation: 202-227g
- Number of animals. 14
- Identification: By means af an ear-punch technique
- Veterinaty inspection: During acclimatization, the animals were examined by a veterinary surgeon. Only animals without any visible signs of illness were used for the treatment.
- Acclimation period: From 7 to 8 days
- Housing: in groups of maximum five rats of the same sex and treatment group, in Makrolon type-5 (59.0x 38.5 x 20.0cm) cages with Lignocel 3-4 sawdust bedding (supplied by Harlan Interfauna Ibérica, S.L.). At the time of administration, the animals will be moved to individual cages with grilled floor to avoid contact with sawdust or other alíen materials that could lead to skin irritation. Once it is checked there is no skin irritation, the grilled floor will be replaced by sawdust again.
- Fasting period before study: not specified
- Diet (e.g. ad libitum): Pelleted standard SAFE A04C rat/mouse maintenance diet (Scientific Animal Food & Engineering, Route de St.Bris, 89290-Augy, France, Batches no.60928 and 70206, Expiry dates: 28 September 2007 and 6 February 2008, respectively) ad libitum. Results of analyses for contaminants are archived at RCC CIDA S.A.
- Water (e.g. ad libitum):Tap water in bottles adbilitum. Results of bacteriological chemical and contaminant analyses are archived at RCC CIDA S.A.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 (monitored)
- Humidity (%): 35-70 (occasionally reaching 80%) (monitored)
- Air changes (per hr): 10-20
- Photoperiod (hrs dark / hrs light): 12-hour fluorescent light/12-hour dark cycle

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
other: square of hydrophilic gauze
Details on dermal exposure:
TREATMENT
Approximately 24 hours before administration of the test item, the back of each animal was
shaved using an electric razor, from the scapula area to the lumbar region.
The test item was applied on an area representing around 1 0% of the total body surface area.
The test item was placed on a square of hydrophilic gauze of approximately 6 x 5 cm and applied to the corresponding test area. The gauze was then covered with a hypoallergenic microporous adhesive band. This route was chosen to study the possible toxic effects produced by skin exposition to the test item.
Finally, a strip of gauze was wrapped around the trunk of the rat in order to hold the patches in
place.
The band was fixed to the body of the animal using adhesive tape.
The aim of this semiocclusive bandage was to allow good skin absorption of the test item and
keep animals from ingesting it.
The bandage and the patches were removed 24 hours after exposure. The remains of the product were removed with distilled water.

DOSE LEVELS
In the sighting study, the dose of 2000 mg/kg was administered.
Since no mortality was observed, this dose was administered in the main study.
Duration of exposure:
24 hours: The bandage and the patches were removed 24 hours after exposure. The remains of the product were removed with distilled water.
Doses:
In the sighting study, the dose of 2000 mg/kg was administered. Since no mortality was observed, this dose was administered in the main study.
No. of animals per sex per dose:
A single dose of test ítem was administered to five males and five females
Control animals:
not specified
Details on study design:
SIGHTING STUDY
The test ítem was administered by the dermal route on a single occasion, to two males and two females.
The rats were observed after the treatment to record the clínica! responses and mortality. They were sacrificed 14 days after the treatment, when sufficient information was obtained. No autopsies were performed nor any tissues were kept.

MAIN STUDY
A single dose of test ítem was administered to five males and five females.

OBSERVATIONS
Mortality/Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (in common with the clínica! signs) and twice daily during days 2-15.
Body weights: On test days 1 (prior to administration), 8 and 15.
Clínica! signs: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test da y 1. Once daily d uring days 2-15.
Statistics:
No statistical analysis was used.

Results and discussion

Preliminary study:
No mortality was recorded.
No clinical signs were recorded.
No dermal alterations were recorded on removing the semiocclusive patches.
The body weight of the animals was within the range commonly recorded for this strain and age.
Effect levels
Key result
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No deaths ocurred at the dose level of 2000 mg/kg during the study.
Clinical signs:
No clinical signs were observed during the course of the study.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necrospy
Other findings:
All surviving animals were killed at the end of the observation period by an intraperitoneal injection of sodium pentobarbital at the dese of 200 mg/kg body weight and at volume of 10 mL/kg. A necropsy was carried out on all the animals of the main study. This included the examination of the intact animal and all superficial tissues, followed by the in situ observation of the cranial, thoracic and abdominal cavities.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Since no mortality was recorded after administration of the test item at the dose of 2000mg/kg, the LD50 of the test item has been found to be higher than the above-mentioned dose when administered by dermal route to rats, and therefore it is not necessary to assign it a risk phrase.
Executive summary:

In the main study, one group of ten Hsd: Sprague Dawley "SD" rats were treated with the test item by dermal route at a dosage 2000mg/kg body weight.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at appoximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 -15. Mortality/viability was recorded and approximately 30minutes, 1,2, 3 and 5 hours after administration on test day 1 (with the clinical signs)and twice daily during days 2 -15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period. No clinical signs were observed during the course of the study.

No dermal alterations were recorded on removal of the semiocclusive patches after 24 hours of exposure.

The body weight of the animals was within the range conmonly recorded for this strain and age.

No macroscopic findings were recorded at necropsy.