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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
Study period:
June 06 2000 - June 30 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
No LLNA study was conducted as an in vivo study according to OCED 406 was conducted in 2000, before the first version of the LLNA (OECD 429) was issued in 2002.

Test material

Constituent 1
Chemical structure
Reference substance name:
(trans(trans))-4'-vinyl-4-(4-methylphenyl)bicyclohexyl
EC Number:
439-730-3
EC Name:
(trans(trans))-4'-vinyl-4-(4-methylphenyl)bicyclohexyl
Cas Number:
155041-85-3
Molecular formula:
Hill formula: C21H30 CAS formula: C21H30
IUPAC Name:
(trans(trans))-4'-Vinyl-4-(4-methylphenyl)bicyclohexyl

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: HsdPoc:DH
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Age at study initiation: about 6 weeks (beginning of treatment)
- Weight at study initiation: 372 g (range from 316 g to 415 g)
- Housing: single Makrolon cage type with a shelter placed on mobile racksbedding conventional softwood granulate bedding change twice weekly
- Diet: ad libitum, exclusive diet for guinea pigs No. 845, Eberle Nafag
- Water: ad libitum, community tap water from Makrolon drinking bottles
- Acclimation period: At least 7 days prior to the start of dosing under test conditions after health examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-27
- Humidity (%): 46-84
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
paraffin oil
Remarks:
or Sodium chloride (NaCl) solution with Freund's complete adjuvant (FCA)
Concentration / amount:
Amount: 0.1 mL
liqiud paraffin: 5 g test material/L
NaCl+FCA: 5 g test material/L

Day(s)/duration:
on day 1
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
Amount: 1 mL
50 g test material/L liquid paraffin
Day(s)/duration:
on day 8, treatement for 48 h
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
Amount: 0.5 mL
10 g test material/L liquid paraffin
Day(s)/duration:
on day 22, treatement for 24 h
No. of animals per dose:
Total: 35 females
Pre-test: 5 females
Control group: 10 females (negative control)
Test group: 20 females (test material)
Details on study design:
RANGE FINDING TESTS:
- intradermal injection: 0, 1, 5, 10, 25, 50 g/L given to one animal
- topical applications, no pretreatment: 1, 10, 50, 100, 200, 400 g/L given to four animals
- topical applications, pretreatment with FCA: 1, 5, 10, 25 g/L given to two animals

MAIN STUDY
A1. INDUCTION EXPOSURE (intradermal injection)
- No. of exposures: 6 injections of 0.1 mL each, three on each side of the spinal column)
- Test groups: 0.1 mL FCA + NaCl, 5g/L test item ad liquid paraffin, FCA + 5 g/L test item + NaCl
- Control group: 0.1 ml FCA + NaCl, 0.1 mL liquid paraffin, 0.1 mL FCA + NaCl
- Site: cranial, medial, caudal
- Frequency of applications: single
- Concentrations: 5 g/L

A2. INDUCTION EXPOSURE (topical application)
- No. of exposures: one week after injection (A1) single topical application of a filter plate soaked with either test solution, or vehicle
- Test groups: Test item (50 g/L) in liquid paraffin
- Control group: liquid paraffin
- Site: cranial, medial, caudal (8 cm^2)
- Frequency of applications: single
- Duration: 48 h, occlusive
- Concentrations: 50 g/L

B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day of challenge: day 22
- Exposure period: 24 h
- Test groups: 10 g/L test item
- Control group: liquid paraffin
- Site: cranial, medial, caudal (4 cm^2)
- Concentrations: 10 g/L
- Evaluation (hr after challenge): 48 and 72 h

Other:
During the induction phase, the guinea pigs were examined on days 2, 3, 7, 11, 12, 15 and 22 for local skin reactions.
The behavior and general condition of all animals were monitored daily.
The body weight of the guinea pigs was determined prior to the start of the experimental part, on days 8, 15, 22, and at the end of the experimental part.
Challenge controls:
Data from the GLP study with Experimental part: April 11 until May 05, 2000.
Positive control substance(s):
yes
Remarks:
Hexylcinnamaldehyde

Results and discussion

Positive control results:
Positive control data from a different GLP study showed a rate of skin sensitisation reaction of 80 %.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 g/L
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no signs of toxicity
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 g/L
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no signs of toxicity
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
10 g/L
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no signs of toxicity
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
10 g/L
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no signs of toxicity
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
10 g/L
No. with + reactions:
7
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
positive control
Dose level:
10 g/L
No. with + reactions:
4
Total no. in group:
10

Any other information on results incl. tables

Results of the pre-test:

Vehicle: Liquid paraffin

Intradermal induction: 5 g test material/L: slightly irritant

Topical induction: 50 g test material/L: slightly irritant

Topical challenge: 10 g test material/L: not irritant

Findings in the induction phase

After intradermal injection, the common signs of irritation after injection of Freund's complete adjuvant were observed. The injection sites were swollen and red, later on, scabs developed. The median with test material treated injection sites were swollen and red.

After removal of the patches, swollen application sites and scabs were observed at the application sites.

Findings after challenge

Group 1: negative control group

After challenge with liquid paraffin, no erythema or edema were observed at the readings.

Single treatment was performed with the test material (10 g/L liquid paraffin) to exclude the primary irritation potential of the test material. No positive reactions were observed in the treated areas at any reading.

Group 2: test material group

The challenge was performed on the right flank with liquid paraffin and with the test material preparation in liquid paraffin.

No positive reactions were observed in the areas treated with liquid paraffin at any reading.

After challenge with the test material (10 g/L preparation in liquid paraffin) no positive skin reactions were observed at the readings. This results in 0 % positive reactions at challenge.

Clinical findings and mortality

The clinical behavior of the guinea pigs was normal during the experimental part. All animals survived the experimental part.

Body weight

The body weight development corresponded to that of the animals of the vehicle group.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the given experimental conditions, the test material induced no reactions and showed no skin sensitisation potential.
Executive summary:

An in vivo study in guinea pigs according to OECD 406 was conducted to evaluate the skin sensitsation potential of the test substance. 10 females were treated with the vehicles liquid paraffin (group 1) and 20 females were treated with the test material (group 2). The induction included intradermal injection of test material preparation in liquid paraffin (5 g/L with and without Freund's complete adjuvant) on experimental day 1, and topical application of test material preparation in liquid paraffin (50 g/L) for 48 hours on experimental day 8. The challenge was performed by topical application of the test material preparation (10 g/L) for 24 hours, two weeks after topical induction and readings were taken at 48 hours and 72 hours after start of treatment. After challenge no positive reactions were seen 48 or 72 hours after treatment with the test material. Based on the study results the test substance showed no skin sensitization potential.