Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 Feb 1997 - 07 Mar 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The GPMT Assay was conducted before the LLNA Assay became mandatory.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Charles River Kisslegg
- Age at study initiation: about 4 weeks
- Weight at study initiation:
364 to 421 g
- Housing:
Two guinea-pigs were housed in a Makrolon cage type IV (floor area: 55 x 33 cm = 1815 cm2 , height: 20 cm) with a shelter, placed on mobile racks
- Diet: ad libitum
- Water: ad libitum
- Acclimation period:
at least 7 days
- Indication of any skin lesions: healthy guinea-pigs were used

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
20 - 23
- Humidity (%):
51 - 76
- Photoperiod (hrs dark / hrs light):
12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
paraffin oil
Concentration / amount:
1 g/L
Day(s)/duration:
On Day 1
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
5 g/L
Day(s)/duration:
On Day 8/ 48 hours
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
0.1 g/L
Day(s)/duration:
On Day 22/ 24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals (test group)
10 animals (control group)
Details on study design:
RANGE FINDING TESTS:
To determine the concentrations suitable for the main study, a pretest with single intradermal or topical administrations of the vehicle and of the test item preparations was performed. Intradermal injections were given to one animal. Topical applications were given to another three animals, two with and one without FCA injection.
Based on the results of the preliminary study, the following concentrations were chosen:
Intradermal induction: 1.0 g/L in liquid paraffin (slightly irritant)
Topical induction: 5.0 g/L in liquid paraffin (slightly irritant)
Topical challenge: 0.1 g/L in liquid paraffin (not irritant)

MAIN STUDY
A.1 INDUCTION EXPOSURE (intradermal injection)
- No. of exposures: six (three on each side of the spinal column)
- Test groups: FCA + phys. sodium chloride solution, Test material 1.0 g/L in liquid paraffin, FCA with test + phys. sodium chloride solution (1.0 g/L completed preparation)
- Control group: FCA + phys. sodium chloride solution, liquid paraffin, FCA + phys. sodium chloride solution
- Site: shoulder region (cranial, medial, caudal)
- Frequency of applications: once
- Concentrations: 1.0 g/L test material

A.2 INDUCTION EXPOSURE (topical application, occlusive)
- No. of exposures: one
- Exposure period: 48 hours
- Test groups: 5.0 g/L test item in liquid paraffin
- Control group: liquid paraffin (undiluted)
- Site: shoulder region
- Frequency of applications: once
- Concentrations: 5.0 g/L test material

B. CHALLENGE EXPOSURE
- No. of exposures: Once
- Day(s) of challenge: On day 22
- Exposure period: 24 hours
- Test groups: 0.1 g/L test material in liquid paraffin; liquid paraffin only
- Control group: 0.1 g/L test material in liquid paraffin; liquid paraffin only
- Site: flank
- Concentrations: 0.1 g/L test material
- Evaluation (hr after challenge): 48 and 72 hours


Challenge controls:
The control group was challenged with liquid paraffin only or 0.1 g/L of the test material in liquid paraffin
Positive control substance(s):
yes
Remarks:
Hexylcinnamaldehyde

Results and discussion

Positive control results:
Positive reactions 48 hours after challenge with HCA: 6/20 animals
Positive reactions 72 hours after challenge with HCA: 6/20 animals

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1 g/L
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
none
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0.1 g/L
No. with + reactions:
11
Total no. in group:
20
Clinical observations:
none
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.1 g/L
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0.1 g/L
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
5 g/L
No. with + reactions:
6
Total no. in group:
20
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
positive control
Dose level:
5 g/L
No. with + reactions:
6
Total no. in group:
20

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item is to be considered as skin sensitizer based on the results obtained from the GPMT Assay according to OECD Guideline 406.
Executive summary:

The test item was studied for skin sensitizing properties in the guinea-pig maximization test according to OECD Guideline 406. Ten female guinea-pigs in the negative control group (group 1) and 20 females in the test material group (group 2) were investigated.

Induction included intradermal injection of test material preparation (1 g/L with and without Freund's complete adjuvant) on day 1, and topical application of test material preparation ( 5 g/L) on day 8.

Challenge by topical application with the test material preparation ( 0.1 g/L) was performed two weeks a:fter topical induction. Positive reactions (erythemas) were observed in 12 of 20 animals (60 per cent) overall. Based on the results, the test item has to be considered as skin sensitizer.