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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: Study performed under GLP regulations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Type of study:
Buehler test
Justification for non-LLNA method:
Historical study already available

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): N4-Amine (N,N-bis(3-aminopropyl)ethylenediamine)
- Analytical purity: 98.3 peak area-% (analytical report No.: 04L00300)
- Batch identification: Behaelter 1716

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Dunkin Hartley, Crl:HA
- Source: Charles River Laboratories, Kisslegg - Germany
- Age at study initiation: 5 – 8 weeks
- Weight at study initiation: 377 g – 440 g
- Housing: The animals were housed in fully air-conditioned rooms (5 animals per cage)
- Cage type: Plastic cages with perforated base; floor area: 4000 cm² (Tecniplast Deutschland GmbH)
- Diet (e.g. ad libitum): Kliba Labordiaet (Provimi Kliba SA, Kaiseraugst, Basel, Switzerland)
- Water (e.g. ad libitum): Tap water
- Acclimation period: 14 days before the first test-substance application.


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


Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Doubly distilled water
Concentration / amount:
All inductions were performed with a 10% test-substance preparation in doubly distilled water and for the challenge a 5% test-substance preparation in doubly distilled water was chosen.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Doubly distilled water
Concentration / amount:
All inductions were performed with a 10% test-substance preparation in doubly distilled water and for the challenge a 5% test-substance preparation in doubly distilled water was chosen.
No. of animals per dose:
5
Details on study design:
Pretest:
In a pretest, the irritancy profile was determined by applying 0.5 ml of various concentrations (1%, 5%, 10%, and 25%) of aqueous test substance solutions to a 2-cm² area of the shaved flanks of Guinea pigs under occlusive conditions. Test sites were visually evaluated 1 h, 24 h and 48 h after application of test solutions. In accordance with the OECD Guideline 406 a slightly irritating concentration should be used in the main test for induction whereas the maximum non-irritant concentration should be applied for challenge.

Induction:
During the induction phase of the main test, 0.5 ml of test solution were applied to an 2-cm² area of the skin of the flank of groups of 5 Guinea pigs three times at weekly intervals, on days 0, 7 and 14 on the same application site. Skin readings were performed 24 h after application. The control animals were not treated since the doubly distilled water used as formulating agent was not expected to influence the result of the study.

Challenge:
Fourteen days after the last induction treatment, each test group animal and animals from control group were challenged with 0.5 ml of 5% aqueous solution of the test substance to a 2-cm² area on the previously untreated flank. Skin reactions were read at 24 and 48 after application of the solutions.

Evaluation:
The number of animals with skin findings at 24 and/or 48 hours after the removal of the patch was taken into account for the determination of the sensitization rate. The evaluation "sensitizing" results if at least 15 per cent of the test animals exhibit skin reactions.
Positive control substance(s):
no
Remarks:
A positive control with a known sensitizer was not included in this study. However, a separate study was performed twice a year in the same laboratory.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5% test-substance preparation in doubly distilled water
No. with + reactions:
4
Total no. in group:
20
Clinical observations:
Discrete or patchy erythema (grade 1)
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5% test-substance preparation in doubly distilled water. No with. + reactions: 4.0. Total no. in groups: 20.0. Clinical observations: Discrete or patchy erythema (grade 1).
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5% test-substance preparation in doubly distilled water
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
Moderate and confluent erythema (grade 2)
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5% test-substance preparation in doubly distilled water. No with. + reactions: 8.0. Total no. in groups: 20.0. Clinical observations: Moderate and confluent erythema (grade 2).
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5% test-substance preparation in doubly distilled water
No. with + reactions:
6
Total no. in group:
20
Clinical observations:
(grade 1)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5% test-substance preparation in doubly distilled water. No with. + reactions: 6.0. Total no. in groups: 20.0. Clinical observations: (grade 1).
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5% test-substance preparation in doubly distilled water
No. with + reactions:
10
Total no. in group:
20
Clinical observations:
(grade 2)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5% test-substance preparation in doubly distilled water. No with. + reactions: 10.0. Total no. in groups: 20.0. Clinical observations: (grade 2).

Any other information on results incl. tables

Pretest:

The 5% and 1% test-substance preparations in doubly distilled water did not cause any skin reactions. After application of the 25% test-substance preparation moderate and confluent erythema was observed in all animals. The 10% preparation caused discrete or patchy erythema in 2 out of 3 animals. Therefore the 10% test-substance preparations were used for induction whereas the 5% test-substance preparations were applied for challenge.

Induction:

The inductions caused discrete or patchy to intense erythema, swelling and eczematoid skin changes in all test group animals. For the 3rd induction the test patch was moved to the middle of the flank in one test group animal because of severe skin irritation effect.

Skin findings

Inductions

Animals with findings/Animals tested

1st

2nd

3rd

Grade 0

-

-

-

Grade 1

11/20

1/20

 

Grade 2

9/20

9/20

2/20

Grade 3

-

10/20

18/20

Swelling

-

3/20

1/20

Eczematoid skin

change

-

-

17/20

Challenge:

After the challenge discrete or patchy to moderate and confluent erythema was observed in the test group animals 24 and 48 hours after removal of the patches.

Skin findings

Control group

Test group

24 h

48 h

24 h

48 h

Grade 0

10/10

10/10

8/20

4/20

Grade 1

-

-

4/20

6/20

Grade 2

-

-

8/20

10/20

Grading:

0 No visible change

1 Discrete or patchy erythema

2 Moderate and confluent erythema

3 Intense erythema and swelling

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information