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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
Magnusson B. and Kligman A .M., 1969. The identification of contact allergens by animal assay. The guinea pig maximization test. J. Invest. Dermatol. 52: 268-276.
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Guinea pig assay was available

Test material

Constituent 1
Chemical structure
Reference substance name:
Cyclohexanol
EC Number:
203-630-6
EC Name:
Cyclohexanol
Cas Number:
108-93-0
Molecular formula:
C6H12O
IUPAC Name:
cyclohexanol
Details on test material:
- Name of test material (as cited in study report): Cyclohexanol
- Substance number: 89/843
- Batch No.: 11/tank B 207
- Analytical purity: 98.8%
- Physical state: at 25°C, sample is homogenous liquid
- Sample was stable
- Storage: RT

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright White, Dunkin Hartley HOE DHPK [SPF-LAC] BO
Sex:
female
Details on test animals and environmental conditions:
- Source: Versuchstierzucht, Hagemann GmbH & Co . KG, D-4923 Extertal 1, FRG
- Rational for choice of animals: Because of the high sensitivity of guinea pigs to a sensitization of the skin which is supposed to be most like that of man. This animal species is the test object acknowledged worldwide for sensitization studies
- Weight at study initiation: 288-340g
- Randomization: According to Salfi, R. : A Long Peri-od Random Number Generator with Application to Permutation. Compstat 1974, pp. 28 - 35.
- Housing: 5/cage
- Diet: 341 .4 mm-Kaninchen- Meerschweinchen-Haltungsdiaet; ad libitum (Supplier : Firma Klingentalmühle AG , CH-4303 Kaiseraugst, Switzerland)
- Water: ad libitum (tap water; about 2 g of ascorbic acid per 10l water was added to the drinking water twice a week
Quality of water and feed used was ascertained via analysis
- Acclimation: no data
- Clipping of the test animals: if required, about 3 to 5 hours before each reading and before each test substance application at the appropriate application sites

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
olive oil
Concentration / amount:
Induction; Intradermal: 5%
Induction; topical: 50%
Challenge: 25%
Rechallenge:25%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
Induction; Intradermal: 5%
Induction; topical: 50%
Challenge: 25%
Rechallenge:25%
No. of animals per dose:
Control I: 10
Control II: 10
Test group: 20
Details on study design:
PRETEST TESTS: (performed 4 wks before main study)
Epidermal Application
- No. of animals: 4/concentration
- Site of application: flanks, respectively on the same areas
- Route of exposure: epicuteneous, patches of filter paper (2 x 2 cm) saturated with the test substance
- Type of coverage: occlusive
- Frequency/Duration: 2X for 24h within a period of 96h
- Evaluation (hr after patch removal): 24h and 48h, scoring of findings using Draize score system for scoring skin irritation
- Result: The minimum irritant concentration was found with a 50% test substance preparation in olive oil DAB 9 and the maximum non-irritant concentration with 25% test substance preparation in olive oil DAB 9 (48 h after the beginning of application).


MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal Injection:
- Site: shoulder
- Concentration: 5% (0.1 ml/site)
- Vehicle: olive oil DAB 9
- Test group 1: Freund's complete adjuvant (FCA) 1:1 with physiological saline(0.9%).
- Test group 2: The test article in olive oil DAB 9.
- Test group 3: Freund's complete adjuvant 1:1 with physiological saline (0.9%) containing the test article
- Control group 1: Freund's complete adjuvant (FCA) 1:1 with physiological saline(0.9%).
- Control group 2: Olive oil DAB 9.
- Control group 3: Freund's complete adjuvant 1:1 with physiological saline (0.9%)
- Frequency of applications: 1x
- Evaluation (hr after injection): 24h, scoring of findings using Draize score system for scoring skin irritation

Topical Induction Exposure:
- Time Schedule: about 1 week after intradermal injections
- Site: same as intradermal injection
- Concentration: 50% cyclohexanol in Olive oil DAB 9 (test groups), olive oil DAB 9 (control groups)
- Route of exposure: epicuteneous, patches of filter paper (2 x 4 cm) saturated with the test substance
- Type of coverage: occlusive
- Duration: 48h.
- Evaluation (hr after injection): 48h, scoring of findings using Draize score system for scoring skin irritation

B. CHALLENGE EXPOSURE
1st Challenge: Test groups and control group 1 treated identically in the following manner
- Day(s) of challenge: 2 weeks after topical induction exposure
- Route of exposure: epicutaneous, applied to a 2 x 2cm piece of filter paper
- Exposure period: 24 h
- Site: intact clipped flank, right flank (test substance, 25% in vehicle), left flank (vehicle). Left flank of each control group 2 animal was treated with the vehicle. Right flank was not treated
- Vehicle: olive oil DAB 9
- Evaluation (hr after challenge): 24h, 48h and 72h, scoring of findings using Draize score system for scoring skin irritation

2nd Challenge- Test groups, control groups 1 and 2 treated identically in the following manner
- Start of challenge: 7days after 1st challenge
- Route of exposure: epicutaneous, applied to a 2 x 2cm piece of filter paper
- Exposure period: 24 h
- Site: intact clipped flank, right flank (vehicle), left flank (test substance; 25% in vehicle)
- Vehicle: olive oil DAB 9
- Evaluation (hr after challenge): 24h, 48h and 72h, scoring of findings using Draize score system for scoring skin irritation

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
slight erythema: 5/10 animals. no other findings
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: slight erythema: 5/10 animals. no other findings.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no symptom
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no symptom.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no symptom
Remarks on result:
other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no symptom.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
slight erythema: 12/20 animals. no other findings
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: slight erythema: 12/20 animals. no other findings.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
slight to moderate erythema: 14/20. no other findings
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: slight to moderate erythema: 14/20. no other findings.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: negative control. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
slight to moderate erythema: 14/20. no other findings
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: slight to moderate erythema: 14/20. no other findings.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.
Reading:
rechallenge
Hours after challenge:
72
Group:
test chemical
Dose level:
25% in olive oil DAB 9
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no symptom
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: test group. Dose level: 25% in olive oil DAB 9. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no symptom.

Applicant's summary and conclusion