Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
09/10 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The test was carried out in compliance with Directive 96/54/EEC, Part B.6 "Acute Toxicity - Skin Sensitization", 1992, and OECD Guideline for Testing of Chemicals No. 406 "Skin Sensitization", 1992, with no deviations reported.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Swiss Federal Department of the Interior
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
test was done before new regualtion entered into force

Test material

Constituent 1
Chemical structure
Reference substance name:
(4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexan-1-one
EC Number:
700-052-0
Cas Number:
60046-50-6
Molecular formula:
C9H16O2
IUPAC Name:
(4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexan-1-one
Details on test material:
- Name of test material (as cited in study report): Actinol
- Physical state: crystalline white powder
- Analytical purity: 99.5 %
- Lot/batch No.: 410016
- Expiration date of the lot/batch: 31 July 2000
- Storage condition of test material: at room temperature, protected from sun light
- Other:

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Ibm: GOHI (SPF)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd Biotechnology & Animal Breeding Devision, Füllinsdorf, Switzerland
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 360-401 g (pretest group), 352-392 g (main test and Controls)
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding ("Lignocell", Schill-AG, Muttenz, Switzerland)
- Diet (e.g. ad libitum): standard Nafag Ecosan 845 25W4, batch nos. 64/99 and 79199, guinea pig breeding / maintenance diet (Nafag, Nahr- und Futtermittel AG, Gossau, Switzerland), ad libitum
- Water (e.g. ad libitum):Community tap water from Ftillinsdorf, ad libitum
- Acclimation period: no acclimatisation for prestes, one week for main test and Controls


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: To:

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: PEG 400 was used for the intradermal and epicutaneous pretests. It was also used for the intradermal and epicutaneous induction and the challenge in the main test. A 1:1 mixture (v/v) of Freund's Complete Adjuvant:physiological saline was used as vehicle
Concentration / amount:
Based on the results of a pretest, a test substance concentration of 5 % was selected for the intradermal induction in the main study.
Based on the results of a pretest, the test substance concentrations in the epicutaneous induction and challenge in the main study were 50 % and 10 %, respectively.
Challengeopen allclose all
Route:
epicutaneous, open
Vehicle:
other: PEG 400 was used for the intradermal and epicutaneous pretests. It was also used for the intradermal and epicutaneous induction and the challenge in the main test. A 1:1 mixture (v/v) of Freund's Complete Adjuvant:physiological saline was used as vehicle
Concentration / amount:
Based on the results of a pretest, a test substance concentration of 5 % was selected for the intradermal induction in the main study.
Based on the results of a pretest, the test substance concentrations in the epicutaneous induction and challenge in the main study were 50 % and 10 %, respectively.
No. of animals per dose:
Test group: 10 male guinea pigs
Details on study design:
RANGE FINDING TESTS:
Range finding tests were performed with one male testing animal for the intradermal injections at concentrations of 1, 3, and 5 % of Actinol in PEG 400. Epicutaneous induction was tested in two male testing animals. Four patches of filter paper (3 x 3 cm) were saturated with the test substance at 10, 15, 25 and 50 % in PEG 400 and applied to the clipped and shaved flanks of the animals. Patches were covered by a strip of aluminium foil and firmly secured with elastic plaster wrapped around the trunk and covered with impervious adhesive tape.

MAIN STUDY
A. INDUCTION EXPOSURE (intradermal injections)
- No. of exposures: three pairs of intradermal injections
- Exposure period:
- Test groups: injection of 1:1 (v/v) FCA:physiological saline, injection of test substance at 5 % in PEG 400, injection of test substance at 5 % in 1:1 (v/v) FCA:physiological saline
- Control group: injection of 1:1 (v/v) FCA:physiological saline, injection of PEG 400, injection of 1:1 (w/w) mixture of PEG 400 in 1:1 (v/v) FCA:physiological saline
- Site: dorsal skin from the scapular region
- Frequency of applications: once
- Duration:
- Concentrations: 5 %

MAIN STUDY
A. INDUCTION EXPOSURE (epicutaneous)
- No. of exposures: one
- Exposure period: starting on Test Day 8
- Test groups: a 2 x 4 cm patch of filter paper was saturated with the test substance (50 % in PEG 400) and placed over the injection sites of the test animals
- Control group: a 2 x 4 cm patch of filter paper was saturated with PEG 400 and placed over the injection sites of the test animals
- Site: dorsal skin from the scapular region
- Frequency of applications: once
- Duration: 48 hours of occlusion with aluminium foil
- Concentrations: 50 %

B. CHALLENGE EXPOSURE
- No. of exposures: two
- Day(s) of challenge: the test and Control groups were challenged two weeks after the epicutaneous induction (Test Day 22) and were treated in the same way
- Exposure period: Test Day 22
- Test groups: clipped and shaved left flank was covered with a patch (3 x 3 cm) of filter paper saturated with the test substance ath the highest non-irritating concentraiton of 10 % , clipped and shaved right flank was covered with a patch (3 x 3 cm) of filter paper saturated with PEG 400
- Control group: clipped and shaved left flank was covered with a patch (3 x 3 cm) of filter paper saturated with the test substance ath the highest non-irritating concentraiton of 10 % , clipped and shaved right flank was covered with a patch (3 x 3 cm) of filter paper saturated with PEG 400
- Site: left and right flanks
- Concentrations: 10 %
- Evaluation (hr after challenge): the reaction sites were assessed 24 and 48 hours after the removal of the bandage


OTHER:
Challenge controls:
five male guinea pigs
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 % in PEG 400 or PEG 400 only
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no signs of systemic toxicity
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10 % in PEG 400 or PEG 400 only. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no signs of systemic toxicity.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 % in PEG 400 or PEG 400 only
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no signs of systemic toxicity
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 % in PEG 400 or PEG 400 only. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no signs of systemic toxicity.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 % in PEG 400 or PEG 400 only
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no signs of systemic toxicity
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 % in PEG 400 or PEG 400 only. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no signs of systemic toxicity.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 % in PEG 400 or PEG 400 only
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no signs of systemic toxicity
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 % in PEG 400 or PEG 400 only. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no signs of systemic toxicity.

Any other information on results incl. tables

no other remarks

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Based on the above mentioned findings in an adjuvant sensitization test in guinea pigs and in accordance to Commission Directive 96/54/EEC, Actinol has not to be classified and labelled as a skin sensitizer.
Executive summary:

In order to assess the cutaneous allergenic potential of Actinol, the Maximization-Test was performed in 15 (10 test and 5 control) male albino guinea pigs, in accordance with OECD Guideline No. 406 and the Directive 96/54/EEC, B.6.

The intradermal induction of sensitization in the test group was performed in the nuchal region with a 5 % dilution of the test article in PEG 400 and in an emulsion of Freunds Complete Adjuvant (FCA) / physiological saline. The epidermal induction of sensitization

was conducted for 48 hours under occlusion with the test article at 50 % in PEG 400 one week after the intradermal induction. The animals of the control group were intradermally induced with PEG 400 and FCVphysiological saline and epidermally induced with PEG 400 underocclusion.

Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test article at 10 % in PEG 400 and PEG 400 alone under occlusive dressing. Cutaneous reactions were evaluated at 24 and 48 hours after removal of the dressing.

None of the 5 animals in the Control group and none of the 10 animals in the test group showed a positive reaction to the exposure to the test substance at 10 % in PEG 400 or to PEG 400 alone after 24 or 48 hours.

No toxic symptoms were evident in the guinea pigs of the control or test group. No deaths occurred.

Based on the above mentioned findings in an adjuvant sensitization test in guinea pigs and in accordance to Commission Directive 96/54/EEC, Actinol has not to be classified and labelled as a skin sensitizer.