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Administrative data

Description of key information

A skin sensitisation test was performed with 44PD testing different concentrations. The study was performed under GLP and scored Klimisch 2. The test substance was found to be skin sensitizing already at a induction/challenge concentration of 0.1%.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study which meets basic scientific principles.
Justification for type of information:
Key study, GLP, Klimisch score 2
Principles of method if other than guideline:
According to G. Klecak: Identification of Contact Allergens (Predictive Tests in Animals) in: Advances in Modern Toxicology Vol. 4, - Dermatotoxicology and Pharmacology -, ed. F. N. Marzulli, H.I. Maibach, pp. 321 - 324 (1977)
GLP compliance:
yes
Type of study:
open epicutaneous test
Justification for non-LLNA method:
The test was performed in 1984, before LLNA was selected as the first-choice test and before OECD 429 was adopted.
Species:
guinea pig
Strain:
other: Pirbright White, Dunkin Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagemann GmbH & Co., KG, Exertal, Germany
- Weight at study initiation: mean 258 - 428 g (1. Experiment); mean 244 - 309 g (2. Experiment)
- Housing: 4 animals in Makrolon Type IV cages
- Diet: Ssniff GK 4 mm (Standard diet for rabbits and guinea pigs), ad libitum
- Water: tap water, ad libitum (2x times a week addition of 2 g ascorbic acid to 10 L water)
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12
Route:
epicutaneous, open
Vehicle:
other: ethanol
Concentration / amount:
0.1, 0.3, 1 and 3% in ethanol
Route:
epicutaneous, open
Vehicle:
other: ethanol
Concentration / amount:
0.1, 0.3, 1 and 3% in ethanol
No. of animals per dose:
8
Details on study design:
RANGE FINDING TESTS: In a pre-test 0.025 mL/cm² of 10, 3, 1 and 0.3% test substance in ethanol and conc. ethanol were applied to the skin of guinea pigs (epicutaneous open). 1% was found to be the minimal irritating concentration and 0.3% as the maximal non-irritating concentration. Readings were conducted 24, 48 and 72 hours following application.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 20
- Exposure period: 24 h epicutaneous
- Test groups:
application of vehicle (conc. ethanol)
- Control group:
application of test substance in ethanol
- Site: right flank (for the 3% solution application site was changed from cranial to caudal due to severe irritation reactions cranial)
- Frequency of applications: daily on week days
- Duration: 26 days
- Concentrations: 0.005, 0.01 (Experiment II), 0.1, 0.3, 1, 3% (Experiment I)

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 28 and 42
- Exposure period: 72 hours
- Test groups: epicutaneous challenge with 4 (experiment I), 2 (Experiment II) test substance concentrations (1. and 2. challenge)
- Control group: epicutaneous challenge with test substance (2. challenge all groups; 1. challenge one control group was challenged with ethanol)
- Site: left flank
- Concentrations: 0.005, 0.01, 0.1, 0.3, 1, 3%
- Evaluation (hr after challenge): 0.005, 0.01 (Experiment II), 0.1, 0.3, 1, 3% (Experiment I)

Positive control substance(s):
no
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1% induction, 0.3% challenge
No. with + reactions:
8
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1% induction, 0.3% challenge. No with. + reactions: 8.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.3% induction, 0.3% challenge
No. with + reactions:
8
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.3% induction, 0.3% challenge. No with. + reactions: 8.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
1% induction, 0.3% challenge
No. with + reactions:
7
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% induction, 0.3% challenge. No with. + reactions: 7.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
3% induction, 0.3% challenge
No. with + reactions:
7
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3% induction, 0.3% challenge. No with. + reactions: 7.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1% induction, 0.1% challenge
No. with + reactions:
2
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1% induction, 0.1% challenge. No with. + reactions: 2.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.3% induction, 0.1% challenge
No. with + reactions:
0
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.3% induction, 0.1% challenge. No with. + reactions: 0.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
1% induction, 0.1% challenge
No. with + reactions:
1
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% induction, 0.1% challenge. No with. + reactions: 1.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
3% induction, 0.1% challenge
No. with + reactions:
4
Total no. in group:
8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3% induction, 0.1% challenge. No with. + reactions: 4.0. Total no. in groups: 8.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.005 or 0.01% induction, 0.005 or 0.01% challenge
No. with + reactions:
0
Total no. in group:
8
Clinical observations:
no reactions were seen for both dose groups
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.005 or 0.01% induction, 0.005 or 0.01% challenge. No with. + reactions: 0.0. Total no. in groups: 8.0. Clinical observations: no reactions were seen for both dose groups.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1% induction, 0.1% challenge
No. with + reactions:
3
Total no. in group:
8
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1% induction, 0.1% challenge. No with. + reactions: 3.0. Total no. in groups: 8.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.3% induction, 0.1% challenge
No. with + reactions:
5
Total no. in group:
8
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.3% induction, 0.1% challenge. No with. + reactions: 5.0. Total no. in groups: 8.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1% induction, 0.1% challenge
No. with + reactions:
5
Total no. in group:
8
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% induction, 0.1% challenge. No with. + reactions: 5.0. Total no. in groups: 8.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
3% induction, 0.1% challenge
No. with + reactions:
5
Total no. in group:
8
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3% induction, 0.1% challenge. No with. + reactions: 5.0. Total no. in groups: 8.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.005 or 0.01% induction, 0.005 or 0.01% challenge
No. with + reactions:
0
Total no. in group:
8
Clinical observations:
no reactions were seen for both dose groups
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.005 or 0.01% induction, 0.005 or 0.01% challenge. No with. + reactions: 0.0. Total no. in groups: 8.0. Clinical observations: no reactions were seen for both dose groups.

Table 1. Results of Experiment I

 

Concentrations of

 

Induction

Challenge

Challenge

 

 

3%

1%

0.3%

0.1%

3%

1%

0.3%

0.1%

Control 1

a

8/8

4/8

0/8

0/8

8/8

8/8

4/8

0/8

Control 2

a

0/8 a

0/8 a

0/8 a

0/8 a

8/8

7/8

0/8

0/8

Exp. 1

3%

8/8

8/8

7/8

4/8

8/8

8/8

8/8

5/8

Exp. 2

1%

8/8

8/8

7/8

1/8

8/8

8/8

8/8

5/8

Exp. 3

0.3%

8/8

8/8

8/8

0/8

8/8

8/8

8/8

5/8

Exp. 4

0.1%

8/8

8/8

8/8

2/8

8/8

8/8

8/8

3/8

a = Ethanol (vehicle control); x/y = no. of positive reactions/no. of application sites 48 hours after challenge

Control 2 was challenged using vehicle at first challenge

Table 2. Results of Experiment II

 

Concentrations of

 

Induction

Challenge

Challenge

 

 

0.01%

0.005%

0.01%

0.005%

Control 1

a

0/8

0/8

0/8

0/8

Control 2

a

0/8 a

0/8 a

0/8

0/8

Exp. 1

0.01%

0/8

0/8

0/8

0/8

Exp. 2

0.005%

0/8

0/8

0/8

0/8

a = Ethanol (vehicle control); x/y = no. of positive reactions/no. of application sites 48 hours after challenge

Control 2 was challenged using vehicle at first challenge

Interpretation of results:
sensitising
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
DSD: Xi, R43
CLP: Cat. 1, H317
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

A skin sensitisation test was performed with 44PD testing different concentrations. Clear sensitizing reactions could be identified using 0.1 and 0.3% concentrations for the first challenge. No control animal showed skin reactions but almost all test substance induced animals showed sensitizing effects. For the second challenge 4 control animals showed reactions when re-challenged with 0.3% whereas no control animal re-challenged with 0.1% showed skin reactions and therefore again sensitizing potential for all animals induced with test substance could be identified.

Induction and challenge/re-challenge with 0.005 or 0.01% did neither result in skin nor in sensitizing reactions.

Therefore, the test substance was found to be skin sensitizing already at a induction/challenge concentration of 0.1%.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the fact that the study performed with 44PD revealed that skin sensitization already occurs at a concentration of 0.1%, a classification of the substance as Skin Sensitising Cat. 1A is deemed required. The corresponding hazard statement is H317: May cause an allergic skin reaction. For the labelling the GHS07 pictogram (exclamation mark) and the signal word “Warning” are assigned to this category.

 

The classification according to Directive 67/548 (DSD) is “ May cause sensitisation by skin contact” with R43 as the corresponding risk phrase.