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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:
2000-11-13 to 2000-12-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report Date:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted July 17, 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
July 30, 1996
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
August 2000
Deviations:
no
Qualifier:
according to
Guideline:
other: Japan MAFF guideline, 59 Noh San No. 4200, January 28, 1985.
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was performed before LLNA was formally validated and available as OECD guideline in 2002.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Uvinul N 539 T
- Physical state: Liquid, slight yellowish
- Analytical purity: 99.4 area% (GC)
- Lot/batch No.: 133635; date of production: April, 2000
- Stability under test conditions: confirmed
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
female guinea pigs, Hsd Poc:DH (SPF)
- Source: Harlan Winkelmann GmbH , Borchen, FRG
- Age at study initiation: ca. 7 weeks
- Weight at study initiation: 351 - 432 g
- Housing: 5 per cage, Stainless steel wire mesh cages with plastic-coated grating, minimum floor area : 2000 cm²
- Diet (ad libitum): Labordiät (Kaninchen-Meerschweinchen-Haltungsdiät, Provimi Kliba SA, Kaiseraugst, Switzerland
- Water (ad libitum): tap water
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25°C
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: Paraffin oil or Freund's adjuvant / 0.9% aqueous NaCI-solution (1 :1 )
Concentration / amount:
5%
Day(s)/duration:
not applicable
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
48 hours
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10 (test group)
5 (control group)
Details on study design:
RANGE FINDING TESTS:
A pretest was carried out with 2 females guinea pigs for intradermal induction (5% in Freund' Adjuvant/0.9% saline and 5% in paraffin oil) and with 4 female guinea pigs for epicutaneous application (75% in Lutrol E 400 and undiluted). In the epicuataneous Pretest, animals pretreated with Freund's adjuvant / 0 .9% aqueous NaCI-solution (1 : 1) each, in the same manner as intradermal pretest without test substance about 3 weeks prior to the application of the test substance were used for detecting a possible influence on irritating effects of previous intradermal treatment with Freund's adjuvant. The test substance was applied to the skin of the flanks for 24 hours under occlusive dressing.


Based on the results of these findings, the concentrations for the main test were selected.

MAIN STUDY
A. INDUCTION EXPOSURE
1. Intradermal induction
- Test groups:
6 intradermal injections in groups of two per animal were applied to each animal.
a) front row. 2 injections each of 0. 1 ml Freund's adjuvant without test substance emulsified with 0. 9% aqueous NaCI-solution in a ratio of 1 : 1
b) middle row. 2 injections each of 0. 1 ml of a test substance formulation, 5% in paraffin oil
c) back row. 2 injections each of 0.1 m l Freund's adjuvant / 0.9% aqueous NaCI-solution (1 : 1) with test substance at 5%.
- Control group:
same treatment as test groups, but without the test substance added
- Site: shoulder
- Frequency of applications: single dose
- Concentrations: 5%
- reading: 24 h after beginning of application

2. Epicutaneous induction, one week after intradermal induction
- Test groups: application of the undiluted test substance (0.5 ml) using an occlusive patch
- Control group: untreated
- Site: shoulder, same area as in the case of the previous intradermal application
- Frequency of applications: single application
- Duration: 48 hours
- Concentrations: undiluted
- reading: 48 h after beginning of application

B. CHALLENGE EXPOSURE 14 days after epicutaneous induction
- No. of exposures: 1
- Day(s) of challenge: 1
- Exposure period: 1
- Test groups: application of the undiluted test substance (0.5 ml) using an occlusive patch
- Control group: as test group
- Site: flank, previously untreated
- Concentrations: undiluted
- Evaluation (hr after challenge): 24, 48
Positive control substance(s):
yes
Remarks:
Alpha-Hexylcinnamaldehyde; assessed in a separate study performed twice a year in the laboratory

Results and discussion

Positive control results:
Concentrations used:
Intradermal induction: 5% in Paraffin oil or Freund's adjuvant/0.9% aqueous NaCI-solution (1 :1)
Induction (epicutaneous) : 10 % in Lutrol E 400
1st+2nd challenge (epicutaneous): 5 % in Lutrol E 400

Results:
Test group: 10/10 animals with skin findings after 1st/2nd challenge with positive control substance
No skin findings after application of vehicle or in respective control groups.

Conclusion:
Based on these results, it was confirmed, that the positive control Alpha-Hexylcinnamaldehyde has a sensitizing effect on the skin of guinea pig strain used in the Maximization Test under the test conditions chosen.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
undiluted
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
undiluted
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
undiluted
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
undiluted
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none

Any other information on results incl. tables

Pretest

Intense erythema and swelling was observed in the 2 animals after intradermal application of the test substance in paraffin oil and Freund´s Adjuvans.

No skin reactions were observed in the 4 animals 1h, 24h or 48h after patch removal using 75% and 100% test substance.

Main test

Intradermal induction : Intense erythema and swelling in all test group animals.

Epicutaneous induction: Incrustation, partially open (caused by the intradermal induction) could be observed in addition to moderate and confluent erythema and swelling in all test group animals.

Challenge: No skin reactions were observed neither in the control nor in the test group animals.

BODY WEIGHTS

The expected body weight gain was generally observed in the course of the study.

Applicant's summary and conclusion