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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:
24 Oct - 27 Dec 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

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
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No. 1907/2006. In accordance with the same Regulation, the data was included to avoid unnecessary testing.

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Crl:HA
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: approximately 4 weeks old
- Weight at study initiation: 315 - 366 g
- Housing: The animals were kept in groups in Terluran - cages on Altromin saw fibre bedding (preliminary test: lot no. 190612, main study: lot no. 290912) in a semi barrier in an air-conditioned room.
- Diet: autoclaved hay and Altromin 3122 maintenance diet for guinea pigs (preliminary test: lot no. 1114, main study: lot no. 0807), rich in crude fibre; ad libitum
- Water: tap water (drinking water, municipal residue control, microbiological controls at regular intervals); ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
other: physiological saline 0.9% NaCl (B. Braun Melsungen, lot no. 121678081, expiry date: 03/2015) for intradermal induction
Concentration / amount:
intradermal: 5%
epicutaneous: 100%
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline (Euro OTC Pharma, lot no. 1105029-01, expiry date: 05/2014) for the challenge
Concentration / amount:
50%
No. of animals per dose:
- 3 (dose range finding study)
- 10 (test group), 5 (positive control group), 5 (negative control group)
Details on study design:
RANGE FINDING TESTS:
For the justification of dose levels a preliminary test was performed.
- 1 animal was treated intradermally with concentrations of 2.5% and 5% of the test item (suspended with physiological saline 0.9% NaCl).
- 1 animal was treated topically with concentrations of 50% and 100% of the test item (suspended with vaseline) for 24 hours.
- 1 animal was treated topically with concentrations of 50% and 100% of the test item (suspended with vaseline) for 48 hours.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 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/physiological saline 0.9% NaCl
Injection 2: a 5% concentration of the test item in physiological saline 0.9% NaCl
Injection 3: a 5% concentration of the test item formulated in a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Epicutaneous: 0.5 g of 100% test item
- Control group:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Injection 2: 100% physiological saline 0.9% NaCl
Injection 3: a 50% (v/v) formulation of physiological saline 0.9% NaCl in a 1:1 (v/v) mixture FCA/physiological saline 0.9% NaCl
Epicutaneous: 0.5 g of vaseline
- Site: shoulder region
- Frequency of applications: every 7 days
- Duration: Days 0 - 8
- Concentrations: 5% intradermal, 100% epicutaneous

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20
- Exposure period: 24 h
- Test groups: 0.5 g of 50% test item in vaseline and 0.5 g vaseline only
- Control group: 0.5 g of 50% test item in vaseline and 0.5 g vaseline only
- Site: left flank (test substance) and right flank (vehicle)
- Concentrations: 50%
- Evaluation (hr after challenge): 24 and 48 h

OTHER:
- The application area was not rinsed after challenge exposure.
Challenge controls:
The control group is actually a challenge control.
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole: 2% in cottonseed oil and 50% w/v formulation of 2-mercaptobenzothiazole in a 1:1 (v/v) mixture FCA/physiological saline 0.9% NaCl

Results and discussion

Positive control results:
- 24 h after removing the patch: erythema grade 2 in 2/5, erythema grade 1 in 3/5, oedema grade 1 in 2/5, eschar in 2/5 and desquamation in 3/5 positive control animals
- 48 h after removing the patch: erythema grade 1 in 5/5, eschar in 3/5 and desquamation in 5/5 positive control animals

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
intradermal induction: 5%; epicutaneous challenge: 50%
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
intradermal induction: 5%; epicutaneous challenge: 50%
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
intradermal induction: 0%; epicutaneous challenge: 50%
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
intradermal induction: 0%; epicutaneous challenge: 50%
No. with + reactions:
0
Total no. in group:
5
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25% positive control substance
No. with + reactions:
5
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25% positive control substance
No. with + reactions:
5
Total no. in group:
5

Any other information on results incl. tables

Preliminary Test:

For each induction the highest concentration to cause mild-to-moderate skin irritation, but well-tolerated systemically, was used.

Based on the results of this preliminary test, a concentration of 5% was chosen for the intradermal application of the main test and a concentration of 100% was selected for the dermal induction.

A concentration of 50% was found to be the highest dose which did not cause any signs of irritation after a topical treatment over a period of 24 hours and therefore was chosen for the challenge application in the main test.

Main Test:

All animals of both groups survived throughout the test period.

Signs of irritation during the induction:

Intradermal Induction I (24-hour reading):

- Injection site 1: erythema grade 1 in 5/5 negative control-, in 5/5 positive control- and 10/10 test animals, oedema grade 1 in 5/5 negative control-, in 5/5 positive control- and 10/10 test animals

- Injection site 2: erythema grade 1 in 1/5 positive control animals, oedema grade 1 in 1/5 positive control animals

- Injection site 3: erythema grade 1 in 1/5 negative control-, in 5/5 positive control- and 5/10 test animals, oedema grade 1 in 1/5 negative control-, in 5/5 positive control- and 5/10 test animals

Intradermal Induction I (48-hour reading):

- Injection site 1: erythema grade 1 in 5/5 negative control-, in 5/5 positive control- and 10/10 test animals, oedema grade 1 in 5/5 negative control-, in 5/5 positive control- and 10/10 test animals

- Injection site 2: no signs of irritation

- Injection site 3: erythema grade 1 in 1/5 negative control-, in 5/5 positive control and 5/10 test animals, oedema grade 1 in 1/5 positive control- and 1/10 test animals

Dermal Induction II (48-hour exposure, occlusive):

- Immediately after removing the patch: no signs of irritation in any of the test or control animals.

- 24 hours after removing the patch: no signs of irritation in any of the test or control animals.

Applicant's summary and conclusion

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.