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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From Jun. 4, 2003 to Aug. 27, 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented, according to accepted guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report date:
2003

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
reliabilty scoring based on 2002 guideline
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
Cyclohexyl phenyl ketone
EC Number:
211-923-5
EC Name:
Cyclohexyl phenyl ketone
Cas Number:
712-50-5
Molecular formula:
C13H16O
IUPAC Name:
cyclohexyl(phenyl)methanone
Details on test material:
- Name of test material (as cited in study report): TKA 40293 Cyclohexylphenylketon
- Physical state: solid
- Analytical purity: 98.0%
- Lot/batch No.: CHPK 10/43-53
-Storage conditions: in the original container at room temperature, away from direct sunlight
- Other: H-methyl thymidine (3HTdR). Batch No. 309. Aqueous solution sterilized 74 GBq/mmol.

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
-Source: Harlan Netherlands B.V. Postbus 6174 NL-5960 AD Horst, The Netherlands
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: 18.2 to 20.8 g
- Housing: Animals were housed in groups of 4 in Makrolon type-3 cages.
- Diet (e.g. ad libitum): Pelleted standard kliba 3433, batch no. 40/03 mouse maintenance diet ad libitum
- Water (e.g. ad libitum):Community tap water ad libitum
- Acclimation period: under test conditions after health examination


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30 to 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: July 16, 2003 To: July 30, 2003

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
other: not applicable (see LLNA below)
Vehicle:
other: not applicable (see LLNA below)
Concentration / amount:
not applicable (see LLNA below)
Challengeopen allclose all
Route:
other: not applicable (see LLNA below)
Vehicle:
other: not applicable (see LLNA below)
Concentration / amount:
not applicable (see LLNA below)
No. of animals per dose:
not applicable (see LLNA below)
Details on study design:
not applicable (see LLNA below)
Challenge controls:
not applicable (see LLNA below)
Positive control substance(s):
not required
Remarks:
not applicable (see LLNA below)

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
10, 25, or 50% (w/v)
No. of animals per dose:
4 females/dose
Details on study design:
MAIN TEST
-25 microliters of test material was spread over the entire dorsal surface of each ear lobe once daily for 3 consecutive days. An additional group of mice were treated with an equivalent volume of vehicle. Five days after the first topical application, all mice were administered with 250 microliters of 3HTdR by intravenous injection via a tail vain. Five hours after treatment, all mice were euthanized by intraperitoneal injection of ventanarcol. The draining lymph nodes were excised and pooled. Single cell suspensions of pooled lymph node cells were prepared by gentle mechanical disaggregation through stainless steel gauze. After washing 2 times in buffered saline, the lymph node cells were resuspended in 5% trichloracetic acid and incubated at 4 degrees celsius for at least 18 hours for precipitation of the macromolecules. The precipitates were resuspended in 5% trichloroacetic acid and transferred to glass scintillation vials with 10 ml of ultima gold scintillation liquid and thoroughly mixed. The level of 3HTdR incorporation was measured in a beta-scintillation counter.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
Mean values and the standard deviations were calculated in the body weight tables.

Results and discussion

Positive control results:
Alpha-hexylcinnamaldehyde was found to be a skin sensitizer and an EC3 value of 9.4% was derived.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: Calculation of the EC3 was not done because no test concentrations produced a higher stimulation index (S.I.) of 3. Group 2; S.I. = 1.8 Group 3; S.I. = 0.4 Group 4: S.I. = 1.4
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Control - 366 DPM Test Concentration 10% - 667 DPM Test Concentration 25% - 136 Test Concentration 50% - 523

Any other information on results incl. tables

No test-article related clinical signs were observed in any animals in the control group or 10% group. On the 3rd application day, a slight ear erythema was observed at both dosing sights in all mice of 25 and 50% groups. Slight ear swelling was observed in the 50% group. These signs persisted for 3 days.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information