Registration Dossier

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Nov 15 - Dec 07, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was conducted according to Good Laboratory Practice (GLP) and followed the OECD Guideline for Testing of Chemicals No 404.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: CC-3-V
Batch: L55101389
Purity: 99.8 % (GC)
Expiration: Dec 31, 2006
Appearance: solid (crystal)off white transparent mass with a yellowish tinge
solubility in water: insoluble
Storage: in original container, 15 - 25 øC, in the dark, protected from moisture

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Strain: Albino rabbit, Crl:KBL(NZW)BR, female (f)
Source: Charles River Wiga GmbH, Kifßlegg
Age: about 18 weeks
Initial Weight: 3,98 kg
One female animal

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: paraffin oil
Controls:
not specified
Amount / concentration applied:
0,5 g
Duration of treatment / exposure:
4 hours
Observation period:
24, 48, 72 hours after removal of patches
Number of animals:
One female animal
Details on study design:
Identification and adaptation
The animal was kept in the experimental room for more than 7 days to allow them to acclimatize.
The animal was uniquely identified by a tattoo in the ear displaying the animal number. An individual cage card was affixed to the cage displaying the study number, test material, day of treatment, and animal number.


Assignment
One female rabbit, with the animal number 23, was used for this study. The initial body weight at the start of the experimental part was 3.98 kg.


Housing and diet
The rabbit was housed in an air-conditioned room of about 28 m^2 in the Institute of Toxicology. Lighting was controlled by a timer to provide a 12-hour light and a 12-hour dark regime. They were kept separately in special rabbit cages (manufacturer: Becker; type K99/30 KU, floor area about 5400 cm^2, a shelter with an integrated sitting board of about 1820 cm^2; overall height: 60 cm) with plastic grids placed on mobile racks. The collection pans underneath the cages were cleaned at least three times a week. The cages were cleaned before the start of the study.
Temperature and relative humidity were measured using a thermohygrograph. The room temperature varied from 16 to 18°C and the relative humidity from 47 to 67 %.
The rabbit received a commercial diet for rabbits, Provimi Kliba 3418.0 ad libitum and ssniff K snack, as well as fresh community tap water from Makrolon drinking bottles ad libitum. According to the specifications given by the manufacturer, the diet had been checked by independent laboratories. Analysis included both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics. The drinking water was periodically analyzed according to the German regulations for human drinking water.




Vehicle
Name: Liquid paraffin
Manufacturer: Merck KGaA, Darmstadt
Batch No.: K32250274
Expires: July 31, 2007



Observations for clinical symptoms
The rabbit was examined for skin alterations, behavior, and general condition 1 hour after removal of the patch, after 24, 48, 72 hours, and then daily up to experimental day 22.

Skin changes at the application sites were evaluated according to the DRAIZE-, OECD-and EEC recommendations.

Grading scale for evaluation:

Erythema and eschar formation

Scores
--------------------------------------------------
No erythema 0

Very slight erythema (barely perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to eschar
formation preventing grading of erythema 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Edema formation
Scores
--------------------------------------------------
No edema 0

Very slight edema (barely perceptible) 1

Slight edema (edges of area well
defined by definite raising) 2

Moderate edema (raised approx. 1 mm) 3

Severe edema (raised more than 1 mm
and extending beyond area of exposure) 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Total possible irritation score (maximum): 8

Mean score = Mean grading for erythema or edema of all the rabbits per time point
Mean cumulative score = Mean grading of all readings for erythema and edema per time point
Mean score per animal = Mean grading for each animal of erythema or edema (24, 48, and 72 hours after removal of the patches)

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24, 48, 72 h
Score:
2
Max. score:
3
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24, 48, 72 h
Score:
1
Max. score:
2

Any other information on results incl. tables

0.5 g of the test material, CC-3-V, was administered. After a single application to the intact dorsal skin of a rabbit, for 4 hours under semiocclusive conditions, erythema (scores 1 to 4) were observed from day 1 up to day 9 and edema (scores 1 to 2) were observed from day 2 up to day 5 of the study. Furthermore brittle skin from day 6 up to day 9 and small scales from day 10 up to day 17 of the study were seen at the site of application. From day 18 up to the end of the experimental part increased growth of hair was observed.

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to EU Regulation No. 1272/2008 and CLP the test material Art. 274689 (CC-3-V) should be regarded as an irritant to the skin.
Executive summary:

Purpose

The purpose of this assay was to identify the skin irritation/corrosion potential of CC-3 -V when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions.

This study should provide a rational basis for risk assessment to the irritating potential of the test item in man.

Study Design

To test for primary skin irritation, 0.5 g of the test material, CC-3-V, was mixed with some drops of liquid paraffin to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm2 patches and applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. The study was started as an initial test with one animal. This animal showed signs of irritation (well defined erythema, edema, brittle skin, and scales) and no more animals were treated. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 21 days.

Results

Only one rabbit was used for the study. No signs of pain were observed after administration. Under the conditions of the present study erythema (scores 1 to 3) were observed from day 1 up to day 5 and edema (scores 1 to 4) were observed from day 2 up to day 9 of the study. Furthermore brittle skin from day 6 up to day 9 and small scales from day 10 up to day 17 of the study were seen at the site of application. From day 18 up to the end of the experimental part increased growth of hair was observed.

Conclusion

The test material caused irritative effects at the site of application. According to EU Regulation No. 1272/2008 and CLP the test material Art. 274689 (CC-3-V) should be regarded as an irritant to the skin.