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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:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1967
Report Date:
1967

Materials and methods

Principles of method if other than guideline:
The sample was used as solutions in 1:1 acetone-dioxane containing 13% guinea pig fat (f.a.d.). One drop of solution was lightly rubbed into the intact shaved skin of the animals. In the induction phase, 5 animals received 9 applications on abraded skin (1%, 5%, 10X x 3, 25% x 4, in f.a.d) and 5 animals received 4 intradermal injections (each 0.1 ml of 1% test material in dimethyl phthalate) for three weeks. After a 2 week rest period, a challenge chase was conducted with 5% and 1% solutions in f.a.d. on intact and abraded skin.
GLP compliance:
no
Type of study:
not specified
Justification for non-LLNA method:
A reliable in vivo sensitisation study in guinea pigs was available, therefore an LLNA assay was not required.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Orthophenylenediamine (OPD)
- Purity: ca 100%

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: albino
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Not reported
- Age at study initiation: Not reported
- Weight at study initiation: Not reported
- Housing: Not reported
- Diet (e.g. ad libitum): Not reported
- Water (e.g. ad libitum): Not reported
- Acclimation period: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not reported
- Humidity (%): Not reported
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): Not reported

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: 1:1 acetone-dioxane containing 13% guinea pig fat or dimethyl phthalate
Concentration / amount:
Sensitizing treatment to abraded skin: 1%, 5%, 10% x 3, 25% x 4, in 1:1 acetone-dioxane containing 13% guinea pig fat (f.a.d.)
Sensitizing treatment by intradermal injection: 1% test material in dimethyl phthalate.
Challenge Test (I): 5% and 1% solutions in f.a.d.
Challengeopen allclose all
Route:
other: epicutaneous
Vehicle:
other: 1:1 acetone-dioxane containing 13% guinea pig fat or dimethyl phthalate
Concentration / amount:
Sensitizing treatment to abraded skin: 1%, 5%, 10% x 3, 25% x 4, in 1:1 acetone-dioxane containing 13% guinea pig fat (f.a.d.)
Sensitizing treatment by intradermal injection: 1% test material in dimethyl phthalate.
Challenge Test (I): 5% and 1% solutions in f.a.d.
No. of animals per dose:
10 animals/ dose
Details on study design:
IRRITATION TEST: The sample was used as solutions in 1:1 acetone-dioxane containing 13% guinea pig fat (f.a.d.). One drop of solution was lightly rubbed into the intact shaved skin of the animals.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 5 animals received 9 applications on abraded skin (1%, 5%, 10X x 3, 25% x 4, in f.a.d).
5 animals received 4 intradermal injections (each 0.1 mL of 1% test material in dimethyl phthalate).
- Exposure period: 3 weeks
- Site: abraded skin or intradermal injection

B. CHALLENGE EXPOSURE (I)--after a 2-week rest period
- Site: intact and abraded skin
- Concentrations: 5% and 1% solutions in f.a.d.

OTHER: Two weeks after the first challenge test, a second test (II) was similarly done by repeating applications of the 5% solution as previously; at the same time these animals were tested for cross-sensitization with 5% paraphenylenediamine in 100% ethanol.
Challenge controls:
A group of previously unexposed animals was similarly tested.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: Challenge Test I
Group:
test group
Dose level:
5% OPD in f.a.d.
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
edema with erythema
Remarks on result:
other: Reading: other: Challenge Test I. Group: test group. Dose level: 5% OPD in f.a.d.. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: edema with erythema.
Reading:
other: Challenge Test I
Group:
test group
Dose level:
1% OPD in f.a.d.
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
mild-moderate erythema
Remarks on result:
other: Reading: other: Challenge Test I. Group: test group. Dose level: 1% OPD in f.a.d.. No with. + reactions: 7.0. Total no. in groups: 10.0. Clinical observations: mild-moderate erythema.
Reading:
other: Challenge Test II
Group:
test group
Dose level:
5% PPD in f.a.d.
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
edema with erythema
Remarks on result:
other: Reading: other: Challenge Test II. Group: test group. Dose level: 5% PPD in f.a.d.. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: edema with erythema.

Any other information on results incl. tables

Challenge Test and sample

Concentration (fad, solvent)

Reactions on Intact Skin

Reactions on Abraded Skin

 

 

1 Day

2 Days

1 Day

2 Days

I, oPDA

5% in f.a.d

3++++, 4+++, 3++

1++++, 5++, 4+

1++, 9+

10+

 

1% in f.a.d.

1+++, 3++, 6+

10+

10+

10+

II, oPDA

5% in f.a.d

2++++, 1+++, 6++, 1+

1++, 9+

4++, 6+

10+

II, pPDA

5% in ethanol

3+, 7 negative

3+, 7 negative

5+, 5 negative

10 negative

Reactions for individual animals are coded: +, ++, +++ = mild, moderate, strong erythema, ++++ = edema with erythema

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
Mild to moderate sensitization was demonstrated in seven of ten guinea pigs at the first challenge test. Only three of these seven animals were positive at the second challenge test. No evidence of cross-sensitization to paraphenylenediamine was observed.
Executive summary:

Orthophenylenediamine as a solution in fat-acetone-dioxane solvent produced variable strong to mild erythema at 25% and 10% concentrations on intact guinea pig skin. Five percent applications to intact skin resulted in mild erythema with occasional moderate erythema on younger animals. One percent solution produced only mild erythema on both intact and abraded skin of young and older animals. Mild to moderate sensitization was demonstrated in seven of ten guinea pigs at the first challenge test. Only three of these seven animals were positive at the second challenge test. No evidence of cross-sensitization to paraphenylenediamine was observed.