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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:
July - Aug 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
Adopted July 17, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
Adopted July 30, 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
Aug. 1998
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The substance has been tested according to the OECD Guideline 406, Guinea pig Maximization Study before the LLNA test model was selected to be the preferred test system under the REACH Regulation.

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Mol:DH (Moellegaard)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: M & B A/S, Ry, Denmark
- Females (if applicable) nulliparous and non-pregnant: yes
- Weight at study initiation:
276 g (mean)
- Housing: in transparent macrolon cages (type V) on soft wood granulate in an air-conditioned room, 5 animals per cage
- Diet (e.g. ad libitum): ssniff Ms-H (V 2233), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 50 +/- 20%
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES:
From: 1. July To: 1 Aug. 2003

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: Tylose
Concentration / amount:
5%
Day(s)/duration:
n.a.
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: Tylose
Concentration / amount:
25%
Day(s)/duration:
2
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: Tylose
Concentration / amount:
25%
Day(s)/duration:
1
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Control: 5
Treatment: 10
Details on study design:
RANGE FINDING TESTS:
Determination of the non irritating (dermal) concentration:
3 animals; 25, 5 and 1% test substance in tylose
Determination of the tolerance of the intradermal injections:
2 animals; 5, 1 and 0.2% in tylose

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2

- Exposure period:
intradermal: n.a.
epicuteous:2 days

- Test groups: 1
- Control group: 2
negative (vehicle) and positive (alpha-hexylcinnamaldehyde) control

- Site: dorsal area
- Frequency of applications:
Day 1: intradermal induction
Day 8: dermal induction

- Concentrations:
intradermal: 5% in tylose
dermal: 25% in tylose


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 1 (Study day 22)
- Exposure period: 24 h
- Test groups:
10 animals; 25% substance in tylose
- Control group:
5 animals; 25% in tylose

- Site: left flank
- Concentrations: 25%
- Evaluation (hr after challenge): 24 and 48h after removel of patches
Challenge controls:
Challenge / performed on test day 22
The test and control guinea pigs were challenged after the epidermal induction aplication and were treated in the same way.
One area of approx. 5x5 cm on the left flank was shaved mechanically. An amount of 0.5 mL of the test substance was administered to a 2x2 cm cellulose patch. The administration area was then kept for 24 h under an occlusive bandage covered with an impermeable film and an elastic badange.
On study day 23 the bandage was removed. Any remnants of the test substance were carefully washed of with warm water. The skin was examined approx. 24 and 48 h after removal of the patches.
Positive control substance(s):
yes
Remarks:
alpha-hexylcinnamaldehyde

Results and discussion

Positive control results:
After the challenge treatment nine of ten animals (90%) showed a positive reaction during the observation period.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clinical symptoms
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25%
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
no clinical symptoms
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no clinical symptoms
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25%
No. with + reactions:
0
Total no. in group:
15
Clinical observations:
no clinical symptoms
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25%
No. with + reactions:
8
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25%
No. with + reactions:
9
Total no. in group:
10

Any other information on results incl. tables

Body weight gains and clinical signs

The body weight gains of the animlas were not impaired. The treated animlas showed no clinical signs of intoxication throughout the study.

Intradermal induction treatment

Intradermal injections with Freund's Adjuvant (with and without test substance) caused severe edema. The administration sites treated with the test substance in tylose showed clear edema. Intradermal injection of the vehicle alone exhibited no signs of irritation. Due to the color of the test substance the treated skin of the animals colud not be assessed for erythema. Due to these strong irritation reactions of the skin, 10% sodium dodecylsulfate was not administered at day 7.

Dermal induction treatment

After removal of the patches at day 10, clear edema, indurated and encrusted skin were observed at the sites previously treated with Freund's Adjuvant. The administration sites treated with the test substance alone showed slight edema. THe administration sites treated with the vehicle alone showed no signs of irritation. Due to the color of the test substance the treated skin of the animals could not be assessed for erythema.

Dermal challenge treatment

No skin reactions were observed in the control and the treatment group 24 and 48 h after removal of the occlusive bandage.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of this study the test substance is considered to be non-sensitising.
Executive summary:

Skin sensitisation of the test item was performed in female guinea pigs according to the method of Magnusson&Kligman.

Intradermal induction was performed using 5% test substance in tylose, dermal induction using 25% test item in tylose. The valdity of the test system is confirmed by the periodically conducted positive control test using alpha-hexylcinnamaldehyde for maximisation test.

Based on the results of this study the test item showed no evidence for sensitising properties.