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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:
2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted in compliance with OECD GLP (1997) regulations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report date:
2001

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A scientifically-valid Guinea Pig Maximization Test (GPMT) was conducted prior to the implementation of the Local Lymph Node Assay (LLNA).

Test material

Constituent 1
Reference substance name:
MV31 K-Salz
IUPAC Name:
MV31 K-Salz
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): MV31 K-Salz
- Substance type: Mono-constituent
- Physical state: Solid (Grey granules)
- Analytical purity: 88%
- Purity test date: 19 September 2000
- Lot/batch No.: Lot no. 1268147/1-1268154/1
- Expiration date of the lot/batch:31 December 2001
- Stability under test conditions: Guranteed for 4 hours
- Storage condition of test material: Darkness at approximately 20 C in a fume cupboard

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright-White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winklemann, Gartenstr. 27, D-33178 Borchen SPF breeding colony
- Age at study initiation: No data
- Weight at study initiation: Mean: 312 g
- Housing: Group housed in Macrolon type IV cages on soft wood granulate
- Diet (e.g. ad libitum): Ssniff Ms-H (V2233), ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: At least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 C
- Humidity (%): 30-70%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 30 January 2001 To: 02 March 2001

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
0.2% Test article in deionized water for intradermal induction, 20% test article in deionized water for epicutaneous induction and 4% test article in deionized water for epicutaneous challenge treatment.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
0.2% Test article in deionized water for intradermal induction, 20% test article in deionized water for epicutaneous induction and 4% test article in deionized water for epicutaneous challenge treatment.
No. of animals per dose:
Control group: 5, Treatment group: 10
Details on study design:
RANGE FINDING TESTS: A pre-test rangefinder determined that 0.2% test article in deionized water was the appropriate dose for the intradermal induction and 20% test article in deionized water for epicutanous induction.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 1 (intradermal), Day 8 (epicutaneous)
- Test groups: A single treatment group opf 10 animals was exposed
- Control group: The control group was exposed to the same intradermal and epicutanous induction exposure solutions with the test article excluded
- Site: Dorsal area
- Frequency of applications: Day 1, Day 8
- Duration: Epicutaneous was covered for 48 hours
- Concentrations: Intracutaneous: 0.2% test article in deionized water, Epicutaneous: 20% test article in deionized water

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 22
- Exposure period: 24 hours
- Test groups: A single treatment group of 10 animals was exposed
- Control group: The control group was exposed to the same epicutaneous treatment with the test article exluded.
- Site: Left flank
- Concentrations: 4% test article in deionized water
- Evaluation (hr after challenge): 72 hours after beginning of challenge exposure, 48 hours after removal of the occlusive patches.

OTHER:
Challenge controls:
Negative: 0.5 mL deionized water
Positive: 50% (induction) and 25% (challenge) alpha-hexyl cinnamic aldehyde in PEG 400- tested by the lab approximately every 6 months to confirm test system.
Positive control substance(s):
yes
Remarks:
alpha-hexyl cinnamic aldehyde

Results and discussion

Positive control results:
Alpha-hexyl cinnamic aldehyde caused skin sensitization in the most recent positive control assay.

In vivo (non-LLNA)

Results
Reading:
1st reading
Hours after challenge:
72
Group:
test chemical
Dose level:
0.2% intradermal induction, 20% epicutanous induction, 4% challenge
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No erythema or edema was observed at any site in either the control or test article group.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 72.0. Group: test group. Dose level: 0.2% intradermal induction, 20% epicutanous induction, 4% challenge. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No erythema or edema was observed at any site in either the control or test article group..

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results of the study, there is no evidence of skin sensitization following treatment with the test article.
Executive summary:

The dermal sensitization potential of the test article (grey granules, purity approx. 88 Fl.%, CASRN 496805-64-2, Lot: 1268147/1-1268154/1) was evaluated in the Maximization Test using female Pribright-White guinea pigs. This study was performed in accordance with OECD GLP (1997) and the German Chemical Law (1999). The study design was based on OECD 406 (1992) and Commission Directive 96/54/EC, B.6 (1996). The test article was prepared in deionized water (vehicle) just prior to each dosing procedure. A pre-test rangefinder determined the appropriate doses for the intradermal and epidermal inductions and for the challenge phase. The test group (10 animals) received intradermal induction injections of 0.1 ml/site of a 1:1 mixture of Freund’s Complete Adjuvant (FCA): deionized water, 0.2% test article in vehicle, and 0.2% test article in 1:1 FCA/deionized water. A control group (5 animals) received the same induction treatment with the exclusion of test article. The epidermal induction of sensitization was conducted under occlusion with 20% test article (test group) or vehicle only (control group) one week after the intradermal induction. Two weeks after epidermal induction the control and test animals were challenged by epidermal application of 4% test article and vehicle under occlusive dressing. Skin reactions were evaluated at 24 and 48 hours after removal of the dressing. A positive control (alpha-hexyl cinnamic aldehyde) is tested by this contract lab approximately every 6 months and the most recent test confirmed the validity of the experimental technique.  No erythema or edema was observed at any site in either the control or test article group (0% responders).   Based on the results this study, there was no evidence of skin sensitization following treatment with the test article.