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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-12-09 to 2009-03-09
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
- supplier: Elevage Charles RIver (F-69592 L'ARBRESLE)
- acclimatation period: 5 days
- weight: 302 and 385 g at the beginning of the test

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
olive oil
Concentration / amount:
(1) Preliminary study
-determination by intrademal injection of the Maximal Non Necrotizing Concentration (MNNC): 0.78% ; 1.56% ; 3.125% ; 6.25% ; 12.5% ; 25% in olive oil
-determination by topical application of the Pre-Maximal Non Irritant Concentration (Pre-MNIC): 12.5% ; 25% ; 50% in liquid paraffin and 100% (undiluted)
-determination by topical application of the Maximal Non Irritant Concentration (MNIC): 12.5% ; 25% ; 50% in liquid paraffin and 100% (undiluted)
(2) Main study
- intradermal induction with the product at 3.125% in olive oil
- topical induction with the product at 100% (undiluted)
- challenge with the product at100% and 50% in liquid paraffin
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
(1) Preliminary study
-determination by intrademal injection of the Maximal Non Necrotizing Concentration (MNNC): 0.78% ; 1.56% ; 3.125% ; 6.25% ; 12.5% ; 25% in olive oil
-determination by topical application of the Pre-Maximal Non Irritant Concentration (Pre-MNIC): 12.5% ; 25% ; 50% in liquid paraffin and 100% (undiluted)
-determination by topical application of the Maximal Non Irritant Concentration (MNIC): 12.5% ; 25% ; 50% in liquid paraffin and 100% (undiluted)
(2) Main study
- intradermal induction with the product at 3.125% in olive oil
- topical induction with the product at 100% (undiluted)
- challenge with the product at100% and 50% in liquid paraffin
No. of animals per dose:
negative controls (vehicle): 6 animals
treated: 11 animals
Details on study design:
(1) Preliminary study
- for the determination of the MNNC, two animals received increasing concentration of the substance by intradermal injection on both sides of the spine. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours later.
- for the determination of the pre-MNIC, the product was applied on the dorso-lumbar zone of 2 animals shorn beforhand, with occlusive dressing for 24 hours. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after removal of the dressing.
- for the determination of the MNIC, 3 animals were treated according to the same treatment as negative controls for the induction phase (ie 2 intradermic injection (ID) of Freund's Complete Adjuvant (FCA) diluted at 50% in a physiological saline solution + 2 ID of physiological saline solution+ 2 ID a mixture with equal volumes v/v FCA at 50% and physiological saline solution.
A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after removal of the occlusive dressing.
(2) Main study
A- Induction
1st Intradermal Induction:
Day 0
After shearing the scapular zone, three (3) pairs of intradermal injections (ID) of 0.1 ml were performed on the scapular zone in such a way as an injection on each pair is placed to either side of the spine as follows :
GROUP 1 (Negative control):
• 2 ID: Freund’s Complete Adjuvant diluted at 50 % in isotonic sodium chloride
• 2 ID: olive oil
• 2 ID: a mixture with equal volumes v/v :
- Freund’s Complete Adjuvant at 50% and olive oil
GROUP 2 (Treated):
• 2 ID: Freund’s Complete Adjuvant diluted by 50 % in oilve oil,
• 2 ID: test item at 3.125% in olive oil,
• 2 ID a test mixture in equal volumes v/v :
- Freund’s Complete Adjuvant at 50% and the test item at 6.25%.
2nd Topical Induction:
Day 6
The scapular zone of all the animals in each group, shorn beforehand, was brushed with a solution of sodium lauryl sulfate at 10% in thick vaseline, in
order to create a local irritation.
Day 8
A topical application under occlusive dressing for 48 hours was performed on the injection sites of each animal.
GROUP 1 (Negative control): 0.5 ml of liquid parafiin
GROUP 2 (treated): 0.5 ml of the test item at 100%
B - Rest phase
The animals of both groups were left for 10 days.
C - Challenge phase
Day 21
The experimental procedure of this phase was identical for both groups GROUP 1 (Negative control) and GROUP 2 (Treated) submitted to this experimentation: on the previously shorn dorso-lumbar zone, an application on either side of the spine, under occlusive dressing, was performed during 24 hours:
- 1 sample cup containing the test item at 100% and 50% in liquid paraffin
Challenge controls:
negative controls: 11
Positive control substance(s):
yes
Remarks:
alpha-hexylcinnamaldehyde

Results and discussion

Positive control results:
alpha hexylcinnamaldehyde at 50% : between 60% and 100% of animals sensitized after 24h
alpha hexylcinnamaldehyde at 50% : between 50% and 90% of animals sensitized after 48h
alpha hexylcinnamaldehyde at 50% : between 51% and 100% of animals sensitized after 24h
alpha hexylcinnamaldehyde at 25% : between 60% and % of animals sensitized after 48h

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50%
No. with + reactions:
0
Total no. in group:
6
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 6.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50%
No. with + reactions:
0
Total no. in group:
6
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 6.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
11
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 11.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
11
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 11.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100%
No. with + reactions:
0
Total no. in group:
6
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 6.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100%
No. with + reactions:
0
Total no. in group:
6
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 6.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
11
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 11.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
11
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 11.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
In view of the results, under the experimental conditions described, the test product must no be classified in accordance with the criteria for classification, packaging and labelling of dangerous substances of the E.E.C Directives 67/548, 2001/59 and 99/45. In accordance with the Globally Harmonized System (Regulation EC N° 1272/2008), the test product must not be classified in category 1..
Executive summary:

The aim of the study was to evaluate the possible allergenic activity of the test substance after intradermic injection and topical administration in guinea pigs. After induction (intradermic injection at 3.125% and topical application at 50 and 100%) of 11 Guinea Pigs of treated group with the test product and a 10-day rest phase, the challenge phase, under occlusive dressing for 24 hours, consisted to a single topical application of the test product diluted at 50% and at 100%. The experimental protocol was established according the O.E.C.D. guideline n°406 dated July 17th, 1992 and the method B.6 of the council regulation N° 440/2008.

No macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase) from the animals of the treated group with the test item. It was recorded a slight erythema in 9% (1/11) of the animals from the treated group, 24 and 48 hours after the challenge phase, on the treated area at 100%. No cutaneous intolerance reaction was recorded in animals from the negative control group. In conclusion, in view of these results, under these experimental conditions, the test substance needs not be classified, in accordance with the criteria for classification, packaging and labelling of dangerous substances and preparations of the E.E.C. Directives 67/548, 2001/59 and 99/45. No symbol and risk phrase are required. In accordance with the Globally Harmonized System (COM(2007)355 final), the test item must not be classified in category 1. No signal word and hazard statement are required.