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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:
2014-07-16 to 2014-10-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
2003
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
FREY-TOX GmbH, Osteroda 38, 04916 Herzberg/Elster, Germany
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The substance is surface active.

Test material

Reference
Name:
Unnamed
Test material form:
liquid: viscous

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories GmbH, NL-5800 AN Venry
- Weight at study initiation: 318-393 g
- Housing: 2-3 animals per cage, macrolon cages (2280 cm2), cleaning and change of bedding 3 times per week
- Diet: ad libitum (Altromin 3123, from Altromin D-32791 Lage, Lippe)
- Water: ad libitum, vitamin C enriched domestic quality water (pH 2.5)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
paraffin oil
Concentration / amount:
5 % (w/w)
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
1000 % (w/w)
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
50 % (w/w)
No. of animals per dose:
Control group: 5
Test group: 10
Details on study design:
RANGE FINDING TESTS:
On request of the sponsor, the skin irritating effect of the test item in the vehicle paraffin subliqu. was examined initially in four instead of two animals during a dermal preliminary investigation in order to find out the minimal irritating concentration for the dermal induction and the maximum non-irritating concentration of the test item for the challenge application.
Animals for the dermal preliminary investigation were initially pre-treated with adjuvant (FCA) by injecting the adjuvant four times intradermal in the scapular area of each animal. Three weeks after the pre-treatment with the adjuvant, a 24-hour occlusive application of four concentrations of the test item on the flanks of the four pre-treated animals took place during the dermal preliminary investigation. Appearing skin reactions were assessed after 24 hrs and 48 hrs.
On the basis of the results of the preliminary investigations the following concentration levels in the vehicle paraffin subliqu. were chosen according to the agreement with the sponsor:
- intradermal induction with 5 % (w/w) test item
- dermal induction with 100 % (w/w) test item
- challenge treatment the 50 % (w/w) test item

MAIN STUDY
A. INDUCTION EXPOSURE Intradermal
- No. of exposures: 3 simultaneous injections (dose volume 0.1 mL)
- Test groups:
Injection (1) - 1:1-mixture (v/v) of FCA and NaCI 0.9 %
Injection (2) - 5 % (w/w) test item in the vehicle paraffin subliqu.
Injection (3) - 5 % (w/w) test item in a 1:1-mixture (v/v) of FCA and NaCI 0.9 %
- Control group:
Injection (1) - 1:1-mixture (v/v) of FCA and NaCI 0.9 %
Injection (2) - vehicle paraffin subliqu.
Injection (3) - 50 % (w/v) preparation of the vehicle paraffin subliqu. in a 1:1-mixture (v/v) of FCA and NaCI 0.9 %
- Site: The first two pairs of injections were given close to each other towards the cranial part of the test area, whereas the third pair of injections was given towards the caudal part of the test area.
- Frequency of applications: 1
- Observation period: 6 days

A. INDUCTION EXPOSURE epicutaneous
- No. of exposures: 1 occlusive patch
- Exposure period: occlusive patch for 48 h
- Test groups: 0.5 mL of 100 % test substance on patch
- Control group: 0.5 mL paraffin subliqu on patch
- Site: area of dorsal skin (approx. 4 x 6 cm) in the scapular region
- Frequency of applications: 1
- Observation period: from day 7 to day 20

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day of challenge: Day 21
- Exposure period: 24 h
- Test groups: Patch 1: 0.5 mL of 50 % (w/w) test substance; Patch 2: 0.5 mL of the vehicle paraffin subliqu
- Control group: Patch 1: 0.5 mL of 50 % (w/w) test substance; Patch 2: 0.5 mL of the vehicle paraffin subliqu
- Site: Patch 1: right posterior flank of all animals; Patch 2: left posterior flank of all animals
- Evaluation: 24 and 48 h after removal of test substance
Positive control substance(s):
yes
Remarks:
alpha-Hexylcinnamaldehyde (historical control)

Study design: in vivo (LLNA)

Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
The sensitization potential of the positive reference item has been shown sufficiently by the response of 10 of 10 guinea pigs (100 %) of the test group with positive skin reactions (erythema grade 1 to 3).

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle control
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle control
No. with + reactions:
0
Total no. in group:
5
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50% (challenge concentration)
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
discrete or patchy erythema
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50% (challenge concentration)
No. with + reactions:
4
Total no. in group:
10
Clinical observations:
discrete or patchy erythema (3), moderate and confluent erythema (1)

Applicant's summary and conclusion

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria