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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
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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report Date:
1984

Materials and methods

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.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Kerobit BPD = N,N'-Di-sek .-butyl-p-phenylendiamin
- Physical state: red liquid
- Analytical purity: 96.92%
- Lot/batch No.: 82/175; 82/175-1
- Stability under test conditions: assured for period of study
- Storage condition of test material: under oxygen exclusion

In vivo test system

Test animals

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

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, open
Vehicle:
other: ethanol
Concentration / amount:
0.1, 0.3, 1 and 3% in ethanol
Challengeopen allclose all
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

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
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.

Any other information on results incl. tables

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

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
DSD: Xi, R43
CLP: Cat. 1, H317