Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
4 Jul - 4 Aug 2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
relative humidity was higher than 70% on several days of the study, this did not effect the study outcome.
GLP compliance:
yes (incl. certificate)
Remarks:
Niedersächsisches Landesamt für Ökologie, Hildesheim, Germany
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): Only trade name given
- Substance type: organic
- Physical state: amber liquid
- Analytical purity: no data
- Lot/batch No.: ES665M0001
- Expiration date of the lot/batch: June 2007
- Storage condition of test material: room temperature (20 ± 2 °C)

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: HsdPoc.DH
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen, Germany
- Weight at study initiation: males: 396 - 444 g; females: 376 - 408 g
- Housing: group-housed for up to a maximum of 5 animals per cage in a battery of cages, equipped with a paper disposal system
- Diet: "Teklad Global Guinea Pig Diet" (pelleted diet) produced by Harlan Teklad, Blackthorn Bicester, Oxon, England (Batch No. N113), ad libitum.
- Water: Tap water as for human consumption was continuously available, ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 °C - 23 °C
- Humidity (%): 50% - 83%
- Air changes (per hr): 16 per h
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
peanut oil
Concentration / amount:
Induction: intradermal 5%, epidermal 100%
Challenge: 100%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
peanut oil
Concentration / amount:
Induction: intradermal 5%, epidermal 100%
Challenge: 100%
No. of animals per dose:
5 (controls), 10 (in test group)
Details on study design:
RANGE FINDING TESTS:
For the intradermal injection, the test article was diluted with aqua ad injectabilia and Freund's complete adjuvant (FCA; batch No. 054K8931, SIGMA, Steinheim) to a final concentration of 5.0%. Two animals were treated, skin reactions were recorded 48 h and 72 h after treatment. For the dermal application, the liquid article was used undiluted. A closed patch exposure was effected by means of an occlusive bandage using cellulose swabs (Pur-Zellin®, Hartmann AG, Heidenheim/Brenz), Leukosilk® (Beiersdorf AG, Hamburg) and non-irritating tape Elastoplast® (Beiersdorf AG, Hamburg), which enveloped the whole animal's trunk. Two animals were employed and skin reactions were recorded 48 and 72 h post application.

According to the results obtained in the preliminary test, the concentrations of the test article used in the main test were 100% (undiluted) for dermal application and 5% for intradermal injection.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/water
Injection 2: test substance in vehicle (peanut oil)
Injection 3: test substance in a 1:1 mixture (v/v) FCA/peanut oil

Epicutaneous: seven days after intradermal induction the previous injection sites were covered occlusively for 48 h with a patch carrying the test article (100%)

- Control group:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/water
Injection 2: vehicle (peanut oil)
Injection 3: vehicle (peanut oil) in FCA

Epicutaneous: seven days after intradermal induction the previous injection sites were covered occlusively for 48 h with a patch carrying the vehicle

- Site: either side of spine (intradermal + epicutaneous)
- Frequency of applications and duration:
intradermal induction: single application at test start
epidermal induction: seven days after intradermal induction, for 48 hours
- Concentrations: three pairs of intradermal injections 5%, epidermal 100%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 14 days after end of epidermal induction
- Exposure period: 24 h
- Test groups: test substance and vehicle only
- Control group: test substance and vehicle only
- Site: right flank (vehicle) and left flank (test substance)
- Concentrations: 100%
- Evaluation (hr after challenge): 24 and 48 h
Challenge controls:
The control group is actually a challenge control
Positive control substance(s):
yes
Remarks:
4-aminobenzoic acid ethyl ester (benzocaine, CAS: 94-09-7, Batch 044K0697) in vaseline

Results and discussion

Positive control results:
The positive control substance (50% 4-aminobenzoic acid in vaseline) induced positive reactions in 5/10 animals (50%), thus meeting
the reliability criteria for the GPMT test (≥ 30% positive response).

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100 %
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: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100 %
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: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100 %
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: 100 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
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: 100 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
50 %
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 50 %. No with. + reactions: 5.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
50 %
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 50 %. No with. + reactions: 5.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified