Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-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:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
none

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Centre de Production Animale (F-45160 Olivet)
- Age at study initiation: 5-6 weeks
- Weight at study initiation: 294 g - 413 g

- Temperature : between 19°C and 24°C
- Relative humidity : between 39% and 60%

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
paraffin oil
Concentration / amount:
Induction phase:
test item at 60% in liquid paraffin (Day 0)
test item at 60% in liquid paraffin, 24 hours after brushing with a solution of sodium Iauryl sulfate (Day 7)

Challenge phase: test item has been used at diluted 60% and 30% in liquid paraffin
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
Induction phase:
test item at 60% in liquid paraffin (Day 0)
test item at 60% in liquid paraffin, 24 hours after brushing with a solution of sodium Iauryl sulfate (Day 7)

Challenge phase: test item has been used at diluted 60% and 30% in liquid paraffin
No. of animals per dose:
GROUP 1 (negative control group) : 5 male guinea pigs
GROUP 2 (treated group) : 11 male guinea pigs
Details on study design:
RANGE FINDING TESTS:
1) Maximum Non Necrotizing Concentration (M.N.N.C.) determination:
The test item should be freshly prepared in view to obtain a maximum non necrotising concentration
for injection by intradermal route.
So, the different concentrations 10% to 0.3 125% have being prepared using olive oil as vehicle. At these concentrations, the test item was insoluble. Indeed, some particJes stayed in the preparation and the injections were not possible even with more suitable needles (23G x I", 0.6x25 mm). Therefore, in accordance with the experimental protocol, the first induction was made topically over 48 hours under
occJusive dressing after 4 intradermal injections of Freund's Complete Adjuvant diluted at 50%.

2) Pre-Maximum Non Irritant Concentration (pre-M.N.I.C.) determination:
2 guinea pigs identified n° C7767 & C7768. The test item was applied under an occJusive dressing during 24 hours, at the following concentrations diluted at 60%, 30%, 15 and 7.5% in liquid paraffin.

3) Maximum Non Irritant Concentration (M.N.I.C.) determination:
3 guinea pigs identified n° C7572 to C7574.
After induction by two topical applications with liquid paraffin and a 11-day rest phase, the challenge phase under occIusive dressing for 24 hours consists in a single topical application of the test item at the following concentrations: diluted at 60%, 30%, 15 and 7.5% in liquid paraffin.

MAIN STUDY
A. INDUCTION EXPOSURE (Group 1)
1st Induction:
- 4 intraderrnal injections of Freund's Complete Adjuvant diluted at 50% in a physiological saline solution.
- 3 scarifications perforrned between injection sites.
- topical application during 48 hours, on the same zone, under occlusive dressing with the test item at 60%.
- Exposure period: 48 hours
2nd induction:
topical application, on the same zone, with the test item at 60%, 24 hours after brushing with 0.5 mL of a solution of Sodium lauryl sulfate at 10%.

B. CHALLENGE EXPOSURE (Groups 1 & 2):
topical application under occlusive dressing at the following concentrations: 60% & 30%.

- Evaluation (hr after challenge): 24 hour reading time; 48-hour reading time and weighing

Challenge controls:
Overall results (readings at 24 and 48 hours)
Positive control substance(s):
yes
Remarks:
Freund's Complete Adjuvant (FCA)

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
60% Concentration
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 60% Concentration. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
30% concentration
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 30% concentration. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
60% concentration
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 60% concentration. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
30 % concentration
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 30 % concentration. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
60% concentration
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 60% concentration. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
60% concentration
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 60% concentration. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
30% concentration
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 30% concentration. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
30% concentration
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 30% concentration. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.

Any other information on results incl. tables

Weight evolution:

Not any abnormality was recorded in the body weight gain of both groups.

Mortality:

No mortality occurred during this study.

No macroscopic cutaneous reactions attributable to allergy were recorded during the examination following the removal of the occlusive dressing (challenge phase) from the animals of the treated group with the test item.

No cutaneous intolerance reaction was recorded in animals from the negative control group.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
not to be classified
Executive summary:

After induction of 11 Guinea Pigs of the treated group by 2 topical applications with the test item diluted at 60% in liquid paraffin during 48 hours, and a II-day rest phase, the challenge phase, und er occJusive dressing for 24 hours, consisted to a single topical application of the test item diluted at 60% and at 30% in liquid paraffin.

No macroscopic cutaneous reactions attributable to allergy were recorded during the examination following the removal of the occlusive dressing (challenge phase) from the animals of the treated group with the test item.

No cutaneous intolerance reaction was recorded in animals from the negative control group.

Weight evolution:

Not any abnormality was recorded in the body weight gain of both groups.

Mortality:

No mortality occurred during this study.

No macroscopic cutaneous reactionsattributable to allergy were recorded during the examination following the removal of the occlusive dressing (challenge phase) from the animals of the treated group with the test item.

No cutaneous intolerance reaction wasrecorded in animals from the negative control group.