Registration Dossier

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 April 2010 - 30 April 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Experimental animals
Source: S&K-LAP Kft. 2173 Kartal, Császár út 135, HUNGARY
Justification of strain: The New Zealand White albino rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test.
Number of animals: 3
Age of animals at treatment: ~12 weeks old
Sex: Male
Body weight range at the beginning of the life phase: 3216-3274 g at the end of the life phase: 3315-3422 g
Date of receipt: 14 April 2010
Acclimation time: 13 days
Animal identification: The animals were identified by engraved ear tags. The cages were marked with individual identity cards with information about study code, sex, cage number, dose group and individual animal number.

Husbandry
Number of animal room: 607
Lighting periods: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20±3 °C
Relative humidity: 30-70 %
Housing/Enrichment: Rabbits were individually housed in AAALC approved metal wire rabbit cages (65x65 cm with height of 45 cm). Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
Ventilation: 15-20 air exchanges/hour
The environmental parameters were recorded twice daily during the acclimatisation and experimental phases.

Food and Feeding
Animals received PURINA Base – Lap gr. diet for rabbits produced by AGRIBRANDS Europe Hungary PLC, H-5300 Karcag, Madarasi út, Hungary, ad libitum.

Water Supply
The animals received municipal tap water, as for human consumption, ad libitum, from an automatic system.

Water Analysis
The drinking water is routinely analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. The 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). Copies of the relevant Certificates of Analysis are retained in the archives at LAB Research Ltd.

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
The test item was used undiluted, as a single dose of 0.5 ml of the test material, applied to the test area. The untreated skin of each animal served as a control.
Duration of treatment / exposure:
Duration of exposure: 4 hours. After the treatment period, the test item was removed with water at body temperature.
Observation period:
Animals were examined for signs of erythema and oedema, and the responses scored at 60 minutes and then at 24, 48 and 72 hours after patch removal. The animals were observed for 72 hours and the duration of the study was sufficient to evaluate fully the reversibility or irreversibility of the effects observed.
Number of animals:
3
Details on study design:
Approximately 24 hours prior to the test, the hair was clipped from the back and flanks of the animals. Removal of hair was performed in two steps. The majority of hair was clipped with an electronic hair clipper and the remaining hair was moistened with water and shaved with a razor. The test item was applied to an approximately 6 cm² area of intact skin as follows:
• A single layer of a fine medical gauze (open-weave with large holes) of approximately 5x5 cm was placed over the application area,
• The appropriate amount of test item was carefully spread over the application area (the gauze helped to maintain the test item in place),
• Three more layers of gauze were placed over the test item,
• These gauze patches were kept in contact with the skin by a patch of clear plastic with a surrounding adhesive hypoallergenic plaster to ensure continued good contact between the moistened test item and the shaved skin.
• The entire trunks of the animals were wrapped with plastic wrap for 4 hours.
• Medical elastic tubing was placed over the plastic to keep it in place.

An initial test was performed using one animal. One hour after application of the test item, the application site was examined. No severe irritation or corrosive effect was found in the initial test, therefore the bandage was replaced and the exposure continued for a further 3 hours (a total 4 hours exposure). Two additional animals were then included in the study. After the treatment period, the test item was removed with water at body temperature.

The dermal irritation scores were evaluated according to the scoring system by Draize (1959).

Body weights were recorded at the beginning and at the end of experiment.

At the end of the observation period, euthanasia of the animal was by intramuscular injections of CP-Ketamin 10 % and CP-Xylazine 2 % followed by i.v. Euthasol® 40% anaesthesia. Death was verified by checking pupil and cornea reflex, absence of respiration and pulse.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: Mean score from 24, 48 & 72 hours
Score:
0
Max. score:
0
Reversibility:
other: no effect seen
Irritant / corrosive response data:
At observation 1 and 24 hours after patch removal, very slight erythema (score 1) was observed in one animal (No: 00809).
At observation 48 and 72 hours after patch removal, there were no observed clinical signs noted on the skin of the treated animals
As no clinical signs were observed up to 72 hours after patch removal, the study was terminated after the 72 hour observation.
The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after patch removal) for erythema were 0.00, 0.00 and 0.33 respectively.
The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after patch removal) for oedema were 0.00, 0.00 and 0.00 respectively.
Other effects:
There was no mortality observed during the study.
There was no effect of treatment on body weight.
There were no treatment-related clinical signs noted.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to Regulation (EC) No 1272/2008, the test material does not require classification as a skin irritant.
Executive summary:

An acute skin irritation study was performed with the test material in New Zealand White rabbits. Parameters monitored during this study included mortality, body weight measurements and clinical observations. The irritancy of the test item was evaluated according to the Draize method (OECD No.: 404, 2002).

The test item was administered undiluted, at a single dose of 0.5 ml. Gauze was placed onto the hairless skin of the rabbit, test item was applied to the gauze, additional gauze was placed over the test item and an adhesive clear plastic patch applied. The trunk was wrapped in clear plastic with medical tubing used to hold the patch in place. The untreated skin of each animal served as control.

After 4 hours, the remaining test item was removed with water at body temperature.

To assess skin irritation, animals were examined at 1, 24, 48 and 72 hours after the patch removal. Additional general examinations were performed daily.

There was no mortality or systemic clinical changes related to administration of the test material.

There was no effect of treatment on body weight.

At observation 1 and 24 hours after patch removal, very slight erythema (score1) was observed in one animal.

At observation 48 and 72 hours after patch removal, there were no observed clinical signs noted on the skin of the treated animals

As no clinical signs were observed up to 72 hours after patch removal, the study was terminated after the 72 hour observation.

The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after patch removal) for erythema were 0.00, 0.00 and 0.33 respectively.

The animals’ individual mean scores (considering readings at 24, 48 and 72 hours after patch removal) for oedema were 0.00, 0.00 and 0.00 respectively.

According to Regulation (EC) No 1272/2008, the test material does not require classification as a skin irritant.