Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2012-08-02 to 2013-xx-xx
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and Guideline conform study

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
no impact on the study, except unknown test item purity
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
yes
Remarks:
no impact on the study, except unknown test item purity
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
yes
Remarks:
no impact on the study, except unknown test item purity
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Constituent 1
Chemical structure
Reference substance name:
Allyl 3-cyclohexylpropionate
EC Number:
220-292-5
EC Name:
Allyl 3-cyclohexylpropionate
Cas Number:
2705-87-5
Molecular formula:
C12H20O2
IUPAC Name:
allyl 3-cyclohexylpropanoate
Test material form:
other: clear colorless liquid
Details on test material:
- Name of test material (as cited in study report): Allyl Cyclo Hexyl Propionate

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elm Hill Breeding Laboratories, Chelmsford, MA 01824
- Age at study initiation: ~7 weeks
- Weight at study initiation: 351-481 g
- Housing: individually
- Diet (e.g. ad libitum): Harlan Teklad Guinea Pig Diet (certified) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: for a minimum of 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 14-26
- Humidity (%): 20-97
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
paraffin oil
Concentration / amount:
5 % for intradermal induction
100 % for epicutaneous induction
100 % for challenge exposure
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
5 % for intradermal induction
100 % for epicutaneous induction
100 % for challenge exposure
No. of animals per dose:
20 animals for test article group
10 animals for vehicle control group
6 animals for positive control group
Details on study design:
RANGE FINDING TESTS:
Intradermal dose range: Four naive animals were exposed to four different test article concentrations by the intradermal technique. In this test, both sides of the animal were clipped and exposed to concentrations of the test article (1, 2.5, 4 and 5 %). No irritation was observed in any of the four animals.
Topical dose range: Four naive guinea pigs were exposed to four test article concentrations (25, 50, 75, and 100 %). Wrapping was removed at ~24 hours after dosing. Twenty-one hours after unwrapping the sites were depilated. Three hours later the sites were examined for skin reactions (24-hr grade). The location of each of the concentrations of test article differed in each of the four animals to compensate for any site-to-site variations. Grades of zero were observed in all the topical administrations (25, 50, 75, 100 %) of the test article.
In consultation with the Sponsor, it was decided to proceed to the main study using a test article concentration of 5 % intradermally and 100 % topically.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 pairs of injection (6 sites) for intradermal induction and 2 patches for topical induction
- Exposure period: on day 1, once for intradermal induction, on day 8, once for 48 hours for topical induction
- Test groups: one test article group
- Control group: vehicle control group and positive control group
- Site: between the shoulders (intrascapular region)
- Frequency of applications: on day 1, once for intradermal induction, on day 8, once for 48 hours for topical induction
- Concentrations: 5 % of test item for intradermal induction and 100 % of test item for topical induction

B. CHALLENGE EXPOSURE
- No. of exposures: 2 patches: on the right and left flanks
- Day(s) of challenge: Day 22
- Exposure period: 24 hours
- Test groups: one test article group
- Control group: vehicle control group and positive control group
- Site: right and left flanks
- Concentrations: 100 % of test item on the left flank, vehicle control on the right flank
- Evaluation (hr after challenge): 24 and 48 hours
Challenge controls:
The positive control animals were similarly treated with the positive control article (0.05% in petrolatum) on the left flank and the appropriate vehicle (petrolatum) on the right flank. The same occlusive technique as for topical induction was employed.
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene

Results and discussion

Positive control results:
The positive control group exhibited the anticipated positive reaction to validate the study.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
100 % test item
No. with + reactions:
13
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100 % test item. No with. + reactions: 13.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100 % test item
No. with + reactions:
13
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100 % test item. No with. + reactions: 13.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0 % test item
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 % test item. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0 % test item
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 % test item. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.05 % DNCB
No. with + reactions:
6
Total no. in group:
6
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.05 % DNCB. No with. + reactions: 6.0. Total no. in groups: 6.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.05 % DNCB
No. with + reactions:
6
Total no. in group:
6
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.05 % DNCB. No with. + reactions: 6.0. Total no. in groups: 6.0.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under conditions of this study, Allyl 3-cyclohexylpropionate revealed sensitising properties in guinea pigs in a test model according to Magnusson and Kligman. Therefore, the test item should be classified and labelled as sensitizer according to regulation (EC) No 1272/2008.
Executive summary:

The purpose of this study was to determine if the test article elicits a delayed dermal contact hypersensitivity response in guinea pigs according to Maximization Test (Magnusson-Kligman) following the test guidelines of US OPPTS 870.2600, EEC Method B.6 and OECD 406.

In the dose range finding study, no irritation was observed in any of the four animals at 1.0, 2.5, 4 or 5 % intradermally. Grades of zero were observed in all the topical administrations (25, 50, 75, 100 %) of the test article during the dose ranging finding study. In consultation with the Sponsor, it was decided to proceed to the main study using a test article concentration of 5 % intradermally and 100 % topically.

The main study was conducted with 10 animals/sex for the test article group, 5 animals/sex for the vehicle control group and 3 animals/sex for the positive control group (1-chloro-2,4-dinitrobenzene [DNCB]). On Day 1, each guinea pig received intradermal injections (0.1 mL each) at six sites between the shoulders following guideline. On Day 7, all animals in the vehicle control group were pretreated dermally with 10 % sodium lauryl sulfate (SLS) in petrolatum. On Day 8, the test article (100 %) was spread over 2 x 4 cm filter paper (0.3 mL) and applied to the injection site areas of the test group. Blenderm® tape was used to occlude the injection area. The dressings were removed following 48 hours of exposure and the areas were wiped clean with gauze and water. The vehicle and positive control groups were similarly treated with mineral oil or the DNCB positive control. Two weeks after the topical induction, the hair was removed from right and left flanks. On Day 23, all test article-treated, vehicle control and positive control animals were challenged with occluded patches for 24 hours on the left flank and right flanks. For the test groups, a 2 x 2 cm filter paper was saturated with 0.2 mL of the test article and applied to the left flank. Another 2 x 2 cm filter paper was saturated with 0.2 mL of the respective vehicle control and applied to the right flank. For the vehicle groups, a 2 x 2 cm filter paper was saturated with the respective test article and applied to the right flanks. Another 2 x 2 cm filter paper was saturated with the respective vehicle control and applied to the left flanks. The positive control animals were similarly treated with the positive control article on the left flank and the appropriate vehicle on the right flank.  The same occlusive technique was employed as for topical induction. After 24 hours, sites were unwrapped and wiped clean with gauze and water. Twenty hours after unwrapping, the sites were depilated with Nair Lotion Hair Remover®. Approximately four hours later the sites were graded for elicited skin reactions (24-hour grade). Approximately 22 hours later the sites were graded a second time (48-hour grade). No mortality was observed and all animals appeared normal throughout the course of the main study.  No responses were observed in the vehicle control animals at 24 or 48 hours following the challenge. Thirteen animals (65 %) responded to the test article at 24 and 48 hours indicating a moderate grade III sensitizer. The positive control group exhibited the anticipated positive reaction to validate the study. In conclusion, under conditions of this study, Allyl 3-cyclohexylpropionate revealed sensitising properties.