Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant and OECD/EU Method Guidelines. No restrictions, do deviations, fully valid for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): Terpenes and terpenoids, turpentine oil, alpha pinene fraction
- Substance type: UVCB
- Physical state: extremely viscous pale amber coloured liquid
- Analytical purity: 100%
- Lot/batch No.: 0910002624
- Expiration date of the lot/batch:indefinite
- Storage condition of test material: room temperature in the dark

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd. Oxon, UK
- Age at study initiation: 8-12 weeks old
- Weight at study initiation: 15-23 g
- Housing: The animals were individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): free access
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature : 19-25°C
- Humidity: 30-70%
- Air changes (per hr):
- Photoperiod: 12h dark, 12 h light

Study design: in vivo (non-LLNA)

Induction
Concentration / amount:
25%, 10%, 5% w/w butanone
Challenge
Concentration / amount:
25%, 10%, 5% w/w butanone
No. of animals per dose:
4 mice per group + control
Positive control substance(s):
yes
Remarks:
vehicle alone

Study design: in vivo (LLNA)

No. of animals per dose:
4
Details on study design:
The preliminary test showed no systemic toxicity or excessive local skin irritation at the highest suitable concetration. Daily application of 25µl of the appropriate concentration of the test item was applied to the dorsal surface of each ear for three consecutive days. Five days following the first topical application all mice were injected via the tail vein with 250 µl of phosphate buffered saline (PBS) containing 3H-methyl thymidine giving a total of 20 µCi to each mouse.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: Concentration 5% w/w in butanone: 1.31 Concentration 10% w/w in butanone: 1.04 Concentration 25% w/w in butanone: 2.94
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Vehicle: 10498.34 Concentration 5% w/w in butanone: 13729.04 Concentration 10% w/w in butanone: 10916.45 Concentration 25% w/w in butanone: 30825.74

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item is considered to be a non-sensitiser under the condition of this test.
Executive summary:

A LLNA test was performed to investigate the skin sensitisation potential of Terpenes and terpenoids, turpentine oil, alpha pinene fraction oligomers. The test item was applied onto the dorsal surface of the ear of CBA/Ca strain female mice. Following a preliminary screening test in which no systemic toxicity was recorded at concentration of 25% w/w, this concentration was selected as the highest dose investigated. The results were expressed as Simulation Index (SI) and under the condition of the study, the test item was considered to be a non-sensitiser.