Registration Dossier

Administrative data

Description of key information

A key skin irritation study was conducted under GLP conditions and in accordance with EU Method B.4 (Acute Toxicity: Dermal Irritation/ Corrosion) and OECD Guideline 404 (Acute Dermal Irritation/ Corrosion). A key eye irritation study was conducted under GLP conditions in accordance with OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 March 2005 and 02 April 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
New Zealand White rabbits were supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK.; acclimatisation period of at least five days; At the start of the study the animals were in the weight range of 2.0 to 3.5 kg and
were twelve to twenty weeks old. Free access to mains drinking water and food was allowed through out the study. The temperature and relative humidity were set to achieve limits of 19 to 25 C and 30 to 70% respectively. The rate of air exchange was at least 15 changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours of darkness.
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: distilled water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.5 g of the test material, moistened with 0.5 ml of distilled water was introduced under 2.5 cm X 2.5 cm cotton gauze patch
Duration of treatment / exposure:
The patch was removed after four hours and any residual material removed from the test area by gentle swabbing with cotton wool soaked in distilled water.
Observation period:
Approximately one hour after removing the patch and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored.
Number of animals:
3
Details on study design:
The day before the test the three rabbits were clipped free of fur from the dorsal/flank area and inspected for gross abnormalities of the epidermis. Only animals with an intact healthy epidermis are selected for the study. A suitable site on the back of each rabbit was selected as test site and 0.5 g of test material moistened with 0.5 ml of distilled water was introduced under a 2.5 cm X 2.5 cm cotton gauze patch which was secured in position with surgical tape. The animals were then placed in a corset to avoid the rabbits interfering with the patch and 4 hours later the patch and the corset were removed and residual material removed. After patch removal, the test sites were evaluated after 1 hour, 24, 48, and 72 hours according to the Draize classification scheme for determination of primary irritation index.
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 24 and 72 hrs
Score:
0
Max. score:
0
Reversibility:
other: no irritation
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 1, 24,48 and 72 hr
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 1, 24, 48 and 72 hrs
Score:
0
Max. score:
0
Irritant / corrosive response data:
The individual scores for erythema/eschar and oedema were all zero for all three animals for the complete 72 hour observation period. No evidence of skin irritation was noted during the study.The Draize classification scheme for determination of primary irritation index was used
Other effects:
No evidence of skin irritation was noted during the study. There were no signs of systemic toxicity

Calculation of Primary Irritation Index and Grading of Irritancy Potential Using the Draize

Scheme: The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test

rabbits (12 values) and this total was divided by six to give the primary irritation index of the test

material.

Primary Irritation Index Classification of Irritancy

0                                   Non-irritant

>0 to2                            Mild irritant

>2 to 5                            Moderate irritant

> 5 to 8                           Severe irritant

Interpretation of results:
not irritating
Conclusions:
The test material produced a primary irritation index of 0.0 and was classified as a non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted. The test material did not meet the classification as irritant or corrosive according to the EU labelling regulations.
Endpoint:
skin corrosion: in vitro / ex vivo
Data waiving:
other justification
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 4 to July 22, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Principles of method if other than guideline:
The purpose of the study is to evaluate the potential ocular irritancy/toxicity of a test article as measured by the test article's ability to induce opacity and permeability to fluorescein in an isolated bovine cornea.
GLP compliance:
yes
Species:
other: bovine corneas
Details on test animals or tissues and environmental conditions:
Bovine eyes were obtained as by-product of freshly slaughtered animals, transported in Hank's Balanced Salt Solution supplemented with penicillin/streptomycin and sent on ice packs. The corneas are used within 24 hour of receipt. The tissue surrounding the eyeball were carefully pulled away and the corneas were excised such that a 2 to 3 mm rim of sclera was present around the cornea. The corneas are mounted in corneal folders with the endothelial side against the O-ring of the chamber and the anterior chamber secured on top. The chambers were filled with Minimum Essential Medium Eagle (MEM) without phenol red, supplemented with 1 % fetal bovine serum and incubated at 32 +- 1 C for a minimum of 1 hour.
Vehicle:
physiological saline
Controls:
yes, concurrent no treatment
Amount / concentration applied:
An aliquot of 750ug was administered as 20 % w/v ( 200 mg/ml) suspension in saline.
Duration of treatment / exposure:
Corneas were exposed to the test material suspension for approximately four hours and then the corneas are removed from the incubator. The epithelial side of each cornea was rinsed three times with complete MEM containing phenol red and a final rinse with complete MEM without phenol red. Then the solution is replaced with fresh complete MEM, supplemented with 1 % fetal bovine serum.
Observation period (in vivo):
Opacity measurements performed immediately after exposure period ends.
Number of animals or in vitro replicates:
Five corneas are tested.
Irritation parameter:
in vitro irritation score
Run / experiment:
after 4 hrs
Value:
1.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
pH6

The bovine corneal opacity and permeability assay uses an opacity measurement and a permeability measurement to determine a final in vitro score. The opacity measurement is determined by the change in opacity of each cornea by subtracting the calculated pre-treatment opacity reading from the final opacity reading. The corrected opacity value of each cornea was calculated by subtracting the average change in opacity of the negative control corneas from that of the treated corneas. The mean opacity value of each treatment group was calculated by averaging the corrected opacity value of the treated corneas from each treatment condition. The permeability measurements: the corrected OD 490 is determined by subtracting the negative controls mean OD 490 from each of the treated cornea OD 490 value. The mean OD 490 of each treatment group was calculated by averaging the corrected OD 490 values. The formula used is: in vitro score = mean opacity value + ( 15 x mean OD 490 Value). The in vitro score is then compared to a classification system established based on studies with a wide range of test materials. The classification provides a good initial guide to interpret these in vitro data: an in vitro score from 0 to 25 is mild irritant, from 25.1 to 55 is a moderate irritant and 55.1 and above is a severe irritant.

Interpretation of results:
GHS criteria not met
Conclusions:
Based on an in vitro score of 1.4 and valid positive controls, the test material was classified according to this assay's classification scheme as a mild irritant.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

The results of the key in-vivo study for skin irritation reported a score of 0.0 and was classified as a non-irritant to rabbit skin according to the Draize classification scheme.

No corrosive effects were noted. The test material did not meet the classification as irritant or corrosive according to the EU CLP regulations.

An in-vitro eye irritancy Bovine Corneal Opacity and Permeability Assay was completed and based on an in vitro score of 1.4 and valid positive controls, the test material was classified according to this assay's classification scheme as a mild irritant. However, the test item does not meet the criteria for classification according to EU CLP regulations as an eye irritant.