Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2nd November 2004 to 3rd January 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted in compliance with: • OECD guideline No. 406, 17th July 1992, • EC Directive No. 96/54/EEC, B.6, 30th July 1996.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
The temperature recorded in the animal room was sometimes outside of the target ranges specified in the Study plan, • the sensitivity positive control test was performed more than 6 months ago.
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
yes
Remarks:
The temperature recorded in the animal room was sometimes outside of the target ranges specified in the Study plan, • the sensitivity positive control test was performed more than 6 months ago.
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Beige powder

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
Species and sex: male and nulliparous and non-pregnant female guinea pigs.
Strain and sanitary status: Hartley Crl: (HA) BR, Caesarian obtained, Barrier sustained - Virus Antibody Free (COBS - VAF).
Reason for this choice: species generally accepted by regulatory authorities for this type of study. The strain used has been shown to produce a satisfactory sensitization response using known sensitizers.
Breeder: Charles River Laboratories France, L’Arbresle, France.
Number: . two males and two females for the preliminary test,
. 30 animals (15 males and 15 females) for the main test.
Allocation to the groups: on day -1, the animals were weighed and allocated to two groups: a control group of ten animals (five males and five
females) and a treated group of 20 animals (ten males and ten females).
Age/weight: on day 1, the animals of the main test were 1-2 months old and had a mean body weight ± standard deviation of 381 ± 22 g for the
males and 376 ± 14 g for the females.
Acclimation: at least 5 days before the beginning of the study.
Identification: by individual ear-tattoo.

The conditions in the animal room were set as follows:
• temperature: 22 ± 2°C
• relative humidity: 30 to 70%
• light/dark cycle: 12 h/12 h
• ventilation: approximately 12 cycles/hour of filtered, non-recycled air.
The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed. In addition to these
daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular
intervals.
During the acclimation period and throughout the study, the animals were housed individually in polycarbonate cages with stainless steel lid
(48 cm x 27 cm x 20 cm) equipped with a polypropylene bottle.
Each cage contained autoclaved sawdust (SICSA, Alfortville, France).
Sawdust is analyzed by the supplier for composition and contaminant levels.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
corn oil
Concentration / amount:
By intradermal route (tested concentrations: 1% and 5% (w/w))
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Concentration / amount:
By intradermal route (tested concentrations: 1% and 5% (w/w))
No. of animals per dose:
2 males and 2 females for the preliminary test,
30 animals (15 males and 15 females) for the main test.
Details on study design:
Thirty guinea pigs were allocated to two groups: a control group of five males and five females and a treated group of ten males and ten females.
On day 1, three pairs of intradermal injections were performed in the interscapular region of all animals:
• Freund's complete adjuvant (FCA) diluted to 50% (v/v) with 0.9% NaCl (both groups),
• test item at the concentration of 5% in corn oil (treated group) or vehicle alone (control group),
• test item at the concentration of 5% in a mixture FCA/0.9% NaCl (50/50, v/v) (treated group) or vehicle at the concentration of 50% (w/v) in a
mixture FCA/0.9% NaCl (50/50, v/v) (control group).
On day 8, the animals of the treated group received a topical application of the test item at the concentration of 10% (w/w) in ethanol/water (80/20, w/w) to the same test site, which was then covered by an occlusive dressing for 48 hours. The animals of the control group received an
application of vehicle under the same experimental conditions.
On day 22, all animals of both groups were challenged by a cutaneous application of the test item at the concentration of 50% (w/w) in acetone to the right flank. The test item was maintained under an occlusive dressing for 24 hours. The vehicle was applied to the left flank
under the same experimental conditions.
Skin reactions were evaluated approximately 24 and 48 hours after removal of the dressing.
At the end of the study, the animals were killed without examination of internal organs.
No skin samples were taken from the challenge application sites.
The interpretation of results was carried out according to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent
adaptations).

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: challenge
Hours after challenge:
48
Group:
other: Control group
Dose level:
50% w/w
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
Dryness of skin
Remarks on result:
other: Reading:
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
1
Total no. in group:
19
Clinical observations:
Discrete erytheme
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5%. No with. + reactions: 1.0. Total no. in groups: 19.0. Clinical observations: Discrete erytheme.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
1
Total no. in group:
19
Clinical observations:
Dryness of skin
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5%. No with. + reactions: 1.0. Total no. in groups: 19.0. Clinical observations: Dryness of skin.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations) on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labeling of dangerous substances, the
test item should not be classified as sensitizing to the skin.
Executive summary:

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations) on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labeling of dangerous substances, the test item should not be classified as sensitizing to the skin.