Registration Dossier

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: according to guideline, GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Uvinul MC80
- Physical state: liquid, very slight yellow
- Analytical purity: purity was in the range between 97.4% and 99.4% (GC)
- Purity test date: 1993.01.25
- Lot/batch No.: LJ 25607/20
- Stability under test conditions: The stability of the test substance in olive oil DAB 9 over 4 hours was confirmed by analysis.
- Storage condition of test material: room temperature; exclusion of light and air (under nitrogen)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagemann GmbH & Co. KG, D-W4923 Extertal 1, FRG
- Age at study initiation: Young adult animals
- Weight at study initiation: 270 - 335 g
- Housing: Makrolon, type IV cages, 5 animals per cage, Bedding: Granulat Typ 3/4 (staubfrei); SSNIFF
- Diet (e.g. ad libitum): Kliba Labordiät 341 (Kaninchen-Meerschweinchen-Haltungsdiät) ad libitum
- Water (e.g. ad libitum): Water ad libitum (tap water; about 2 g of ascorbic acid per 10 l water was added to the drinking water twice a week)
- Acclimation period: 7 days before the beginning of the study in the laboratory

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 h light (6.00 - 18.00 hours) / 12 h darkness (18.00 - 6.00 hours)

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Freund's adjuvant, olive oil DAB 9
Concentration / amount:
Pretest: 5% (intradermal), 75%/ 100% (epicutaneous)

Main test:
Induction: 5% (intradermal), 75%/ 100% (epicutaneous)
Challenge: 75%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Freund's adjuvant, olive oil DAB 9
Concentration / amount:
Pretest: 5% (intradermal), 75%/ 100% (epicutaneous)

Main test:
Induction: 5% (intradermal), 75%/ 100% (epicutaneous)
Challenge: 75%
No. of animals per dose:
Pretest: 4
control group: 10
Test group: 20
Details on study design:

RANGE FINDING TESTS:
Exposure period: 2 times for 24 hours within a period of 96 hours,
Exposure: 2 x 2 cm filter paper strips (soaked in the test substance = about 0.15 g) , occlusive (rubberized linenpatches 4 x 4 cm from Russka and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG)
Site of application: flank, respective on the same area
Readings: about 24 and 48 h after the beginning of application

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal, epicutaneous)
- Exposure period: 48 hours (epicutaneous), occlusive
- Site: shoulder, same area for intradermal / epicutaneous application
- Frequency of applications: Percutaneous induction was carried out one week after intradermal induction
- Concentrations:
Intradermal:
A) 0.1 ml Freund's adjuvant, emulsified with 0.9% aqueous NaCl-solution (1 : 1)
B) 0.1 ml test substance (5% in olive oil DAB 9 )
C) 0.1 ml Freund's adjuvant in 0.9% aqueous NaC1-solution (1 : 1) containing test substance (5% in olive oil DAB 9)
Epicutaneous: approx. 0.3 g undiluted test substance
- Control group: Intradermal: test substance injections (A, B, C) but without test substance; epicutaneous: no treatment
B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21 days after intradermal induction
- Exposure period: 48 hours epicutaneous, occlusive
- Control group: treated as test group
- Site: intact flank
- Concentrations: test substance 75% in olive oil DAB 9
- Evaluation (hr after challenge): 24 and 48 h after the removal of the patch
Challenge controls:
Induction: Intradermal: test substance injections (A, B, C) but without test substance; epicutaneous: no treatment
Challenge: treated as test group
Positive control substance(s):
yes
Remarks:
1-chlor-2,4-dinitro-benzol

Results and discussion

Positive control results:
Challenge with 1% of 1-chlor-2,4-dinitro-benzol in ethanol led to visual skin findings in 20 of 20 animals
Moderate to severe erythema and very slight edema in 2 of 20 test group animals.
Well-defined erythema in 14 of 20 test group animals; in 11 additionally very slight edema.
Very slight erythema in 4 of 20 test group animals.
No skin reactions in control group animals.
The vehicle did not cause any skin reactions in all animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75 % in olive oil DAB9
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 75 % in olive oil DAB9. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75 % in olive oil DAB9
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 75 % in olive oil DAB9. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
other: reading 24 h, 48h after challenge
Group:
negative control
Dose level:
75 % in olive oil DAB9
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:
Reading:
other: reading 24 h, 48h after challenge
Group:
test group
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading:
Reading:
other: reading 24 h, 48h after challenge
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading:

Any other information on results incl. tables

Intradermal induction caused slight to well-defined signs of irritation
in the test group animals.
Percutaneous induction caused in animals of the test group 
incrustation, partially open (caused by the intradermal induction) 
in addition to slight to well-defined signs of irritation.

Challenge:
Well-defined signs of irritation (grade 2 erythema) in one test group animal.
No skin reaction was seen at skin sites of the test animals
treated with vehicle or treated with test substance solely at challenge.

According to the authors, a second challenge application was
not necessary on the basis of the inequivocal results of the
first challenge.

Applicant's summary and conclusion