Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 October 2002 to 14 January 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted in accordance with EC B.6 method, OECD 406 and US EPA OPPTs 870.2600 methods in accordance with GLP
Cross-reference
Reason / purpose:
reference to same 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:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
This study has been conducted when the LLNA was not yet the standard method.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in the study report): Reaktiv Rot F00-0124

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: M & B A/s, PO Box 1079, DK-8680 Ry Denmark
- Weight at study initiation: 355 g. Minimum 324g, maximum 404g
- Housing: Macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet: ssniff Ms-H (V2233), ad libitum
- Water: Tap water in plastic bottles, ad libitum
- Acclimation period: At least 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 °C
- Humidity (%): 50±20 °C


IN-LIFE DATES: From: Oct. 29, 2002 To: Oct. 29, 2002

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
water
Remarks:
deionized
Concentration / amount:
Concentration of test material and vehicle used at induction:
a) intradermal: 5 % test substance in deionized water mixed with Freunds complete adjuvant emulsion
b) dermal: 25 % in deionizes water
Concentration of test material and vehicle used for each challenge:
25 % in deionized water
Challenge
Concentration / amount:
Concentration of test material and vehicle used at induction:
a) intradermal: 5 % test substance in deionized water mixed with Freunds complete adjuvant emulsion
b) dermal: 25 % in deionizes water
Concentration of test material and vehicle used for each challenge:
25 % in deionized water
No. of animals per dose:
Determination of tolerance of the intradermal injections: 2
Determination of the primary non iritating concentration: 3
Number of animals in test group: 10
Number of animals in negative control group: 5
Details on study design:
RANGE FINDING TESTS:
Determination of the primary non-irritant concentration:
The following test concentrations were administered to the left and right flanks of three guinea pigs respectively:
Animal 1: 25.0 % in deionized water (left flank), 5% in deionzed water (right flank)
Animal 2: 25.0 % in deionized water (left flank), 1% in deionzed water (right flank)
Animal 3: 5.0 % in deionized water (left flank), 1% in deionzed water (right flank)
0.5 mL of the test substance preparation was administered to a 2 x 2 cm cellulose patch, which was fixed to the flank and covered occlusively for 24 h with a bandage and film. 24 h after removal of the patches, the treated skin areas were examined for erythema and edema.

Determination of the tolerance of the intradermal injections:
The following preparations were administered twice by intradermal injection to 2 guinea pigs:
Site 1. 2X0.1 mL 5.0 % test substance in deionized water
Site 2. 2X0.1 mL 1.0 % test substance in deionized water
Site 3. 2X0.1 mL 0.2 % test substance in deionized water

24, 48, 72 and 96 h after administration the injection sites were examined for local tolerance.

MAIN STUDY
A. INDUCTION EXPOSURE (Intradermal)
- No. of exposures: 2
- Exposure period: 1-7 d
- Test groups: Yes
- Control group: Yes, (50 % Freund's Adjuvant)
- Site: Dorsal area
- Frequency of applications: 2 in 7 d
- Duration: 7 d
- Concentrations: 5.0% in deionized water (site 2) and 5% in 50% Freunds adjuvants

INDUCTION EXPOSURE (Dermal)
- No. of exposures: 0.5 mL of the test substance preparation as cellulose patch
- Test groups: Yes, 25.0 % test substance in deionzed water
- Control group: Yes, deionized water
- Site: Dorsal area
- Frequency of applications: Single on Day 8

B. CHALLENGE EXPOSURE (Dermal)
- No. of exposures: One
- Day(s) of challenge: Day 22
- Exposure period: 24 h under an occlusive bandage covered with an impermeable film and an elastic bandage.
- Test groups: 5.0 % test substance in deionized water
- Control group: 5% test substance in deionized water
- Site: Left flank
- Evaluation (hr after challenge): 24 and 48 h after removal of the patches

OTHER:

The substance is considered to be sensitizing if 30 % or more of the animals in the treatment group definitely showed a positive skin reaction and at the same time no irritant effects have emerged in the control group.
Positive control substance(s):
yes
Remarks:
alpha-hexylcinnamaldehyde

Results and discussion

Positive control results:
The validity of the test system is confirmed by the periodically conducted positive control test using alpha-hexylcinnamaldehyde for the maximization test.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the test conditions, none of ten animals of the treatment group showed a positives skin response after the challenge procedure.
Executive summary:

A study was conducted to assess the sensitizing potential of the test substance in female Pimol:DH Moellegaard guinea pigs, according EU Method B.6. and OECD guideline 406, using the Magnusson & Kligman method, in compliance with GLP.

Intradermal induction was performed using 5 % test substance in deionized water. Dermal induction was carried out with 25 % test substance in deionised water and the challenge test carried out with 5 % test substance in deionized water.

The validity of the test system is confirmed by the periodically conducted positive control test using alpha-hexylcinnamic aldehyde for the maximization test.

Under the test conditions, the test substance showed no evidence for sensitizing properties in guinea pigs.