o-phenylenediamine

Skin sensitisation

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

Test material information

Constituent 1

Reference substance name:

o-phenylenediamine

EC Number:

202-430-6

EC Name:

o-phenylenediamine

Cas Number:

95-54-5

Molecular formula:

C6H8N2

IUPAC Name:

benzene-1,2-diamine

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)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: 1:1 acetone-dioxane containing 13% guinea pig fat or dimethyl phthalate

Induction 2

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.

Challenge

Challenge 1

Route:

other: epicutaneous

Vehicle:

other: 1:1 acetone-dioxane containing 13% guinea pig fat or dimethyl phthalate

Challenge 2

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)

Results

Results 1

Reading:

other: Challenge Test I

Group:

test chemical

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.

Results 2

Reading:

other: Challenge Test I

Group:

test chemical

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.

Results 3

Reading:

other: Challenge Test II

Group:

test chemical

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.