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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:
First application (injection): 29 Mar 1978; last application (2rd rechallenge: 08 May 1978)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1978
Report Date:
1978

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
(modified grading scale; 2 rechallenges with non-irritating concentration)
GLP compliance:
no
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
LLNA study protocol was not available when study was conducted.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Citral synthetic; 2,6-Dimethyl-2,6-octadien-8-al; Substance No. 77/711
no further data

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Firma Hagemann, Lippische Versuchstierzucht, 4923 Extertal, Germany
- Weight at study initiation (injection): 507.7 g
- Diet: ad libitum; Sniff K, Standard-Diet for rabbits and guinea pigs, Intermast GmbH, Soest
- Water: ad libitum

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
other: intradermal and percutaneous
Vehicle:
paraffin oil
Concentration / amount:
1st application = intradermal induction:
- Freund´s adjuvant/aqua dest (1:1) without test substance
- 25 % test substance in paraffin oil DAB7
- 25 % test substance in paraffin oil DAB7 and Freund´s adjuvant/aqua dest. (1:1)

2nd application = Percutaneous induction:
- with 25 % test substance in paraffin oil DAB7

1st challenge: 10 % test substance in paraffin oil DAB7
1st rechallenge: 5 % test substance in paraffin oil DAB7
2nd rechallenge: 5 % test substance in paraffin oil DAB7
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
1st application = intradermal induction:
- Freund´s adjuvant/aqua dest (1:1) without test substance
- 25 % test substance in paraffin oil DAB7
- 25 % test substance in paraffin oil DAB7 and Freund´s adjuvant/aqua dest. (1:1)

2nd application = Percutaneous induction:
- with 25 % test substance in paraffin oil DAB7

1st challenge: 10 % test substance in paraffin oil DAB7
1st rechallenge: 5 % test substance in paraffin oil DAB7
2nd rechallenge: 5 % test substance in paraffin oil DAB7
No. of animals per dose:
10 per test group (10 for 1st challenge, 5 for rechallenges), 5 per control group (animals with challenge treatment only)
Details on study design:
RANGE FINDING TESTS:
A range finding test was conducted to determine the concentration of the test substance which caused no and mild-to-moderate skin irritation, respectively. No further information given.

MAIN STUDY
A. INDUCTION EXPOSURE
- 1x6 intradermal injections and one week later, treatment with SLS, followed by one percutaneous induction according to OECD 406
- Readings: 24 hours after the intradermal application and 48 hours after the percutaneous induction
- Control group: not treated during induction phase
- Site: shoulder, respective on the same area

B. CHALLENGE EXPOSURE
- No. of exposures: 3 challenges
- Day(s) of challenge: 1st challenge: 19 days after percutaneous induction, 1st rechallenge: 28 days after percutaneous induction, 2nd rechallenge: 33 days after percutaneous induction
- Site: 1st challenge: right flank, 1st rechallenge: left flank, 2nd rechallenge: right flank
- Concentrations: 1st challenge: 10 % test substance in paraffin oil DAB7, 1st and 2nd rechallenge: 5 % test substance in paraffin oil DAB7
- Evaluation (hr after challenge): 1st challenge: treated and control animals 48 and 72 hours; 1st rechallenge: treated animals only 24 h, control animals 24 and 72 h; 2nd rechallenge: treated and control animals 24, 48 and 72 h.
Challenge controls:
yes
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: 1st challenge, 1st reading
Hours after challenge:
48
Group:
test group
Dose level:
10 %
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
well-defined to moderate erythema, well-defined edema exceeding beyond the area of exposure
Remarks on result:
other: see Remark
Remarks:
Reading: other: 1st challenge, 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 % . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: well-defined to moderate erythema, well-defined edema exceeding beyond the area of exposure .
Reading:
other: 1st challenge, 2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
10 %
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
well-defined to moderate erythema and well-defined edema exceeding beyond the area of exposure
Remarks on result:
other: see Remark
Remarks:
Reading: other: 1st challenge, 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 10 % . No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: well-defined to moderate erythema and well-defined edema exceeding beyond the area of exposure.
Reading:
other: 1st challenge, both readings
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
primary irritation: slight to well-defined erythema, no edema
Remarks on result:
other: Reading:
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
5 %
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
well-defined to moderate erythema and well-defined edema
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 5 % . No with. + reactions: 5.0. Total no. in groups: 5.0. Clinical observations: well-defined to moderate erythema and well-defined edema.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 5 % . No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
other: 2nd rechallenge, 1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5 %
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
moderate erythema and well-defined edema, both beyond the area of exposure
Remarks on result:
other: see Remark
Remarks:
Reading: other: 2nd rechallenge, 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 5.0. Total no. in groups: 5.0. Clinical observations: moderate erythema and well-defined edema, both beyond the area of exposure.
Reading:
other: 2nd rechallenge, 2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5 %
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
well-defined erythema and edema
Remarks on result:
other: Reading:
Reading:
other: 2nd rechallenge, 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
5 %
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
slight to well-defined erythema and desquamation
Remarks on result:
other: Reading:
Reading:
other: 2nd rechallenge, all readings
Group:
negative control
Dose level:
5 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading:

Any other information on results incl. tables

Table: Overview of test results for different challenges and readings

Challenge
reading
timepoint

Group

Substance application

Number of animals with skin reaction

Erythema score and incidence

Edema score and incidence

Findings

1st ch
1st r
48 h

test group

10 % test substance in paraffin oil

10/10

2 in 4/10

3 in 6/10

2 in 10/10

well-defined to moderate erythema, well-defined edema exceeding beyond the area of exposure

negative control

10 % test substance in paraffin oil without induction

5/5

1 in 3/5
2 in 2/5

0 in 5/5

slight to well-defined erythema, no edema

1st ch
2nd r
72 h

test group

10 % test substance in paraffin oil

10/10

2in 9/10
3 in 1/10

2 in 10/10

well-defined to moderate erythema and well-defined edema: exceeding beyond the area of exposure

negative control

10 % test substance in paraffin oil without induction

5/5

1 in 5/5

0 in 5/5

slight erythema, no edema

2nd ch
1st r
24 h

test group

5 % test substance in paraffin oil

5/5

2 in 3/5

3 in 2/5

2 in 5/5

well-defined to moderate erythema and well-defined edema

2nd ch
both readings

negative control

5 % test substance in paraffin oil without induction

0/5

0 in 5/5

0 in 5/5

3rd ch
1st r
24 h

test group

5 % test substance in paraffin oil

5/5

3 in 5/5

2 in 5/5

moderate erythema and well-defined edema: both exceeding beyond the area of exposure

3rd ch
2nd
 r
48 h

test group

5 % test substance in paraffin oil

5/5

2 in 5/5

2 in 5/5

well-defined erythema and edema

3rd ch
3rd
 r
72 h

test group

5 % test substance in paraffin oil

5/5

1 in 4/5

2 in 1/5

0 in 5/5

Slight to well-defined erythema and desquamation

3rd ch
all readings

negative control

5 % test substance in paraffin oil without induction

0/5

0 in 5/5

0 in 5/5

Applicant's summary and conclusion