Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
8 Oct 2012 - 25 Feb 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study conducted according to GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
2003
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): L4-Ligand
- Physical state: solid, white powder
- Analytical purity: > 98% (HPLC-Fingerprint; Analytical Report 12L00037)
- Lot/batch No.: 0005473663
- Stability under test conditions: stability expiration date was given as 1st August 2013
- Storage condition of test material: at ambient temperature, under an atmosphere of Argon (instead of N2; in agreement with sponsor), protected against humidity

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, Kißlegg, Germany
- Weight at study initiation: 296 - 370 g
- Housing: 2 to 3 per cage, in Makrolon cages with a floor area of 2280 cm2; Hay bricks were added as cage enrichment
- Diet: pelleted diet "Altromin 3123", ad libitum
- Water: vitamin C enriched tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
Induction: 60%
Challenge: 50%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
Induction: 60%
Challenge: 50%
No. of animals per dose:
Control group: 10
Test group: 20
Details on study design:
RANGE FINDING TEST
In order to select suitable concentrations of test item in vehicle for the main study, a preliminary patch test was done with 2 animals. Skin reaction was assessed according to the Magnusson/Kligman grading scale.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 (day 0, day 7 and day 14)
- Exposure period: 6 hours
- Test group: treated with 0.5 mL of test item in vehicle
- Control group: vehicle only
- Site: 4 x 6 cm skin area on the left flank, clipped free of hair
- Frequency of applications: at 7 day-intervals
- Duration: day 0 to day 14
- Concentrations: 60% test item in vehicle

B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: challenge was done after 4 weeks following the first induction
- Exposure period: 6 hours
- Test groups: treated with 0.5 mL of test item in vehicle
- Control group: treated with 0.5 mL of test item in vehicle
- Site: 4 x 6 cm skin area on the right, clipped free of hair
- Concentrations: 50%
- Evaluation (hr after challenge): 24 and 48 h

READING
Skin reaction was assessed according to the Magnusson/Kligman grading scale.

FURTHER EXAMINATIONS
The animals were observed for clinical signs and reactions to treatment. They were weighed at day 0 and at the end of of the study period, i.e. on day 33.
Positive control substance(s):
yes
Remarks:
positive control test with the reference material alpha-hexylcinnamaldehyde (purity 85%) was performed from Sept to Oct 2012. 13/20 treated animals showed positive reactions, corresponding to 65%.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
60% test item in vehicle for induction, 50% test item in vehicle for challenge
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 60% test item in vehicle for induction, 50% test item in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
60% test item in vehicle for induction, 50% test item in vehicle for challenge
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
2/20 animals showed slight erythema
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 60% test item in vehicle for induction, 50% test item in vehicle for challenge. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: 2/20 animals showed slight erythema.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50% test item in vehicle for challenge
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% test item in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% test item in vehicle for challenge
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% test item in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Any other information on results incl. tables

The animals showed no clinical symptoms, and body weight gain was as expected for both, the test and the control group.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified