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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:
From Jul. 29, 1981 to Oct. 23, 1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report Date:
1981

Materials and methods

Principles of method if other than guideline:
Method: Test material was applied epicutaneously to intact skin of guinea pigs during the induction, challenge and rechallenge phases and skin irritation was scored by the Draize system.
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buhler study was available before REACH came into force, therefore no additional LLNA study was conducted.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 11583B15
- Physical state: Clear, colorless liquid
- Analytical purity: 91.58 %
- Density at 25 °C: 1.06749 g/cm3 (equivalent to 1067.49 mg/mL)
- Stability under test conditions: Stable at room temperature or below and slowly reacts with water
- Storage condition of test material: Refrigerator

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elm Hill Breeding Laboratories, 71 Elm Street, Chelmsford, Massachusetts 01824
- Weight at study initiation: 327-498 g
- Housing: Individually housed in stainless steel cages with wire mesh floors
- Diet (e.g. ad libitum): Purina Guinea Pig Chow, ad libitum
- Water (e.g. ad libitum): Filtered tap water, ad libitum
- Acclimation period: 7 d


ENVIRONMENTAL CONDITIONS
- Air changes (per hr): 14/ h
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light


IN-LIFE DATES: From: Jul. 07, 1981 To: Sep. 10, 1981

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, open
Vehicle:
olive oil
Concentration / amount:
- Primary skin irritation phase: 0 (control), 0.10, 0.05, 0.025, 0.0125 and 0.00625 % molar equivalents of test or positive control article (25 µL) in olive oil
- Induction phase: Single applications of 0.36 molar concentrations in olive oil (25 µL)
- Challenge and rechallenge phase: 0, 0.10, 0.05, 0.025, 0.0125 and 0.00625 % molar equivalents of test or positive control article (25µL) in olive oil
Challengeopen allclose all
Route:
epicutaneous, open
Vehicle:
olive oil
Concentration / amount:
- Primary skin irritation phase: 0 (control), 0.10, 0.05, 0.025, 0.0125 and 0.00625 % molar equivalents of test or positive control article (25 µL) in olive oil
- Induction phase: Single applications of 0.36 molar concentrations in olive oil (25 µL)
- Challenge and rechallenge phase: 0, 0.10, 0.05, 0.025, 0.0125 and 0.00625 % molar equivalents of test or positive control article (25µL) in olive oil
No. of animals per dose:
Primary Skin Irritation Phase: 5 animals/dose
Induction phase: 10 animals/dose (two sites per animal)
Challenge Phase: 10 animals/dose
Details on study design:
RANGE FINDING TESTS:
- Five animals each were exposed to 25 µL of molar dilutions (0, 0.10, 0.05, 0.025, 0.0125, 0.00625 %) of either the test or positive control article in olive oil
- Route: Epicutaneous; no patch was applied

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Single
- Test groups: Yes
- Control group: Yes, olive oil (vehicle control) and IPDI (positive control)
- Site: Flank to trunk along both sides of each animal
- Frequency of applications: Once
- Duration: 5 d
- Concentrations: 0.36 molar concentration


B. CHALLENGE EXPOSURE
- No. of exposures: Single
- Day(s) of challenge: 9 d
- Test groups: Yes
- Control group: Yes
- Site: Applied to untreated site, flank to trunk along both sides of each animal
- Concentrations: 25 µL of 0, 0.10, 0.05, 0.025, 0.0125 and 0.00625 % molar concentration
- Evaluation (hr after challenge): 28 and 48 h


OTHER:
Rechallenge Phase: 9 d after the initial challenge
Procedure: Same as challenge
Challenge controls:
Not applicable
Positive control substance(s):
yes
Remarks:
Isophorene diisocyanate (IPDI)

Results and discussion

Positive control results:
- Primary skin irritation phase: At 24 h one male exhibited a grade 1 erythema at the dose levels of 0.1 %; one female exhibited a grade 2 erythema at 0.1 and 0.05 %, and a grade 1 erythema with 0, 0.025, 0.0125 and 0.00625 % (olive oil only). By 48 h, both grade 2 erythemas had decreased to a grade 1 and the site treated with olive oil returned to normal. All other test sites appeared normal.
- Induction phase: Exhibited grades of 1, 2 and 3 for erythema and no edema at 24-hour interval. Scores had decreased slightly but were considered comparable at the 48-h interval
- Challenge phase: Mean skin irritation scores were higher at challenge than at the skin irritation phase
- Rechallenge phase: Mean skin irritation scores were less or comparable at rechallenge than at the skin irritation phase

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.1 and 0.05 %
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.1 and 0.05 %. No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.025 %
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.025 %. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.0125 %
No. with + reactions:
9
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.0125 %. No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.00625 %
No. with + reactions:
5
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.00625 %. No with. + reactions: 5.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1, 0.05, 0.025 and 0.0125 %
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1, 0.05, 0.025 and 0.0125 %. No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.00625 %
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.00625 %. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
0.1, 0.05, 0.025, 0.0125 and 0.00625 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 0.1, 0.05, 0.025, 0.0125 and 0.00625 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
0.1, 0.05, 0.025, 0.0125 and 0.00625 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1, 0.05, 0.025, 0.0125 and 0.00625 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: -.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.1, 0.05, 0.025, 0.0125%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.00625%
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.1, 0.05, 0.025, 0.0125%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.00625%
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
0.1%
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
0.05
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
0.025, 0.0125 and 0.00625%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
0.1%
No. with + reactions:
6
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
0.05
No. with + reactions:
5
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
0.025
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
0.0125 and 0.00625%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
-
Remarks on result:
other: -
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
-
Remarks on result:
other: No data
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
-
Remarks on result:
other: No data
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
-
Remarks on result:
other: no data
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
-
Remarks on result:
other: No data

Any other information on results incl. tables

Results with test material:

- Primary skin irritation phase: Two females exhibited a grade 1 erythema with 0.1 %, only at the 24 h interval. One male exhibited grade 2 erythema with 0.1 % and grade 1 erythema with 0.05 % and 0.025 % at both 24 and 48 h intervals. No skin irritation was observed at concentrations of 0.0125 % or below.

- Induction Phase: Exhibited grades 2 and 3 for erythema and no edema at the 24 h interval. By the 48 h interval, scores for erythema were either 0, 1 or 2.

- Challenge Phase: Mean skin irritation scores for all concentrations were higher at challenge than at the primary skin irritation phase.

- Rechallenge Phase: Mean skin irritation data were considered to be less than or comparable to the mean primary skin irritation phase

Table 1: Mean Skin Irritation Scores

Primary Skin Irritation Phase:

Concentration

0.1

0.05

0.025

0.0125

0.00625

0.0*

 

Er

Ed

Er

Ed

Er

Ed

Er

Ed

Er

Ed

Er

Ed

IPDI (24 h)

0.6

0.0

0.4

0.0

0.2

0.0

0.2

0.0

0.2

0.0

0.2

0.0

IPDI (48 h)

0.2

0.0

0.2

0.0

0.2

0.0

0.2

0.0

0.2

0.0

0.0

0.0

11583B15 (24 h)

0.8

0.0

0.2

0.0

0.2

0.0

0.0

0.0

0.0

0.0

0.0

0.0

11583B15 (48 h)

0.4

0.0

0.2

0.0

0.2

0.0

0.0

0.0

0.0

0.0

0.0

0.0

Challenge Phase:

IPDI (24 h)

2.7

0.5

2.1

0.0

1.5

0.0

1.1

0.0

0.9

0.0

0.0

0.0

IPDI (48 h)

1.9

0.0

1.9

0.0

1.7

0.0

1.2

0.0

0.9

0.0

0.0

0.0

11583B15 (24 h)

2.3

0.2

2.1

0.2

0.7

0.0

1.1

0.0

0.5

0.0

0.0

0.0

11583B15 (48 h)

2.1

0.0

2.0

0.0

1.0

0.0

1.2

0.0

0.8

0.0

0.2

0.0

Rechallenge Phase:

A-IPDI (24 h)

0.9

0.0

0.8

0.0

0.0

0.0

0.1

0.0

0.0

0.0

0.0

0.0

A-IPDI (48 h)

0.7

0.0

0.6

0.0

0.1

0.0

0.0

0.0

0.0

0.0

0.0

0.0

B-IPDI (24 h)

0.5

0.0

0.3

0.0

0.1

0.0

0.0

0.0

0.0

0.0

0.0

0.0

B-IPDI (48 h)

0.4

0.0

0.1

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

11583B15 (24 h)

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

11583B15 (48 h)

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

 

A - Animals treated with IPDI during induction

B- Animals treated with 11583B15 during induction

* -Vehicle (olive oil) only

Er - Erythema

Ed - Edema

Applicant's summary and conclusion

Interpretation of results:
Category 1A (indication of significant skin sensitising potential) based on GHS criteria
Conclusions:
Under the test conditions, the test material was considered to be a contact sensitizer in guinea pig.

Executive summary:

A study was conducted to assess the dermal contact sensitization potential of the test substance in guinea pig according to a Buehler protocol, in compliance with GLP. Test substance and isophorene diisocyanate (positive control substance) at concentrations of 0, 0.10, 0.05, 0.025, 0.0125 and 0.00625% were applied epicutaneously (non-occluded) to the intact skin of guinea pigs. Prior to the induction application, the primary irritation potential was determined. Challenge and rechallenge were performed 5 and 14 days after a single induction application, respectively. Clinical signs and body weight were recorded during the study and necropsies were conducted on animals at termination. Evidence of dermal contact sensitization including skin reactions were observed at sites treated with non-irritating concentrations and enhanced skin reactions were seen at sites treated with irritating concentrations. Contact sensitization was evident for both substances at initial challenge (5 day post-induction). Evidence of sensitization for both substances was negligible upon rechallenge (14 day post-induction). Under the study conditions, the test substance was considered to be a contact sensitizer in guinea pig (Calkins, 1981).