Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
other information
Study period:
December 10, 1985 to January 10, 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see "Remark"
Remarks:
Well documented and reported study, conducted equivalent to internationally accepted technical guidelines in recognized industrial research organization. A quality assurance inspection report, but not a GLP compliance statement, was included in the study report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
of 1981
Deviations:
yes
Remarks:
For the 􀂠rst induction the test substance was applied epidermally at a dose of approximately 0.4 g paste of test substance in vaseline for 24 h under occlusive dressing, instead of being administered by intradermal injection.
Qualifier:
equivalent or similar to guideline
Guideline:
other: EU Directive 79/831 (Skin Sensitisation)
Deviations:
yes
Remarks:
For the 􀂠rst induction the test substance was applied epidermally at a dose of approximately 0.4 g paste of test substance in vaseline for 24 h under occlusive dressing, instead of being administered by intradermal injection.
Principles of method if other than guideline:
The principles of the Guinea pig maximization test outlined in the OECD TGD 1981 and EU Directive 79/831 were followed, but for the 􀂠rst induction the test material was applied to the epidermis on filter patches (2 x 4cm) placed over the 4 injection sites at the neck into which 0.1
mL of freshly prepared adjuvant saline mixture/site had been injected intracutaneously. On this induction occasion the test material was applied epidrmally at a dose of approximately 0.4 g paste of test material in vaseline for 24 h under occlusive dressing, instead of being administered
by intradermal injection. This deviation from the standard Guinea pig maximization test outlined in the OECD and EU TGDs accounted for the non-injectability of final formulations or insolubility of compounds in standard vehicles.
GLP compliance:
no
Remarks:
but Quality Assurance statement was included in the report
Type of study:
other: Modified Guinea pig maximization test

Test material

Constituent 1
Details on test material:
- Name of test material (as cited in study report): PBS 2354.0
- Appearance: solid
- Expiration date of the lot/batch: December 1986
- Stability under the condition of administration: not determined
- Storage conditions: dry, room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pibright White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Toxicology GU 2.5, CIBA-GEIGY AG Basel
- Age at study initiation: Approximately 10 weeks
- Weight at study initiation: 304 – 412 g
- Housing: Individual housing in Macrolon cages (type III)
- Diet (ad libitum): Commercially available pelleted standard guinea pig diet (NAFAG No. 846,
Gossau SG, Switzerland) supplemented with
fresh carrots.
- Water (ad libitum): Fresh water
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2°C
- Humidity (%): 50 ± 10%
- Photoperiod: 12 hrs light/day

SENSITIVITY CHECK OF THE ANIMAL STRAIN
The sensitivity of the strain was controlled every six months with p - phenylenediamine.

Study design: in vivo (non-LLNA)

Induction
Route:
other: FCA/physiological saline mixture: intradermal; Test material: epidermal, occlusive
Vehicle:
other: vaseline (Demopharm SA., Bienne)
Concentration / amount:
1st Induction:
ca. 3% in vaseline, i.e. approximately 0.4 g paste of test material in vaseline epidermally (i.e.
epicutaneously) under occlusive dressing for 24 h.
2nd Induction:
ca. 3% in vaseline, i.e. approximately 0.4 g paste of test material in vaseline epidermally (i.e.
epicutaneously) under occlusive dressing for 48 h.
Challenge:
ca. 0.1% in vaseline (i.e. approximately 0.2 g paste of test material in vaseline), and vehicle
(i.e. vaseline), occlusive epicutaneous administration for 24 h on contra-lateral 􀂡anks.
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline (Demopharm SA., Bienne)
Concentration / amount:
1st Induction:
ca. 3% in vaseline, i.e. approximately 0.4 g paste of test material in vaseline epidermally (i.e.
epicutaneously) under occlusive dressing for 24 h.
2nd Induction:
ca. 3% in vaseline, i.e. approximately 0.4 g paste of test material in vaseline epidermally (i.e.
epicutaneously) under occlusive dressing for 48 h.
Challenge:
ca. 0.1% in vaseline (i.e. approximately 0.2 g paste of test material in vaseline), and vehicle
(i.e. vaseline), occlusive epicutaneous administration for 24 h on contra-lateral 􀂡anks.
No. of animals per dose:
Control group: 10 male and 10 female guinea pigs
Test group: 10 male and 10 female guinea pigs
Details on study design:
MAIN STUDY
1st Induction: Week 1
2nd Induction: Week 2
Challenge: After a rest period of 14 days.

A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: First induction 24 hours, second induction 48 hours
- Test group:
0.1 ml of 50% FCA/physiol. saline mixture injected intracutaneously at 4 sites of the animal's
neck + occlusive dressing of test material/vaseline
paste over the 4 FCA/physiol. saline mixture injection sites.
- Control group: adjuvant + vehicle (vaseline),
- Site: animal's neck
Explanatory note: FCA =Freund's complete adjuvant

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: After a rest period of 14 days.
- Exposure period: 24 hours
- Test group: Occlusive dressing of vehicle (vaseline) alone and test material/vaseline paste on
contralateral 􀂡anks.
- Control group:
during challenge: males: vehicle on one 􀂡ank, test material/vehicle paste on contralateral 􀂡ank;
females: vehicle alone
- Evaluation (h after challenge): 24 and 48 hours
The challenge application sites were chemically depilated before examination using Veet (R) for
approximately 5 minutes.
24 and 48 hours after patch removal skin reactions were evaluated for erythema/eschar
formation and edema according to Draize. In addition, scaling was recorded.
Challenge controls:
A control group receiving adjuvant and vehicle during induction and vehicle and test material during the 1st challenge was included in the study, in order to check the maximum subirritant concentration of the test material in adjuvant treated animals.
Positive control substance(s):
yes
Remarks:
p - phenylenediamine: Sensitivity of the animal strain was controlled every six months.

Results and discussion

Positive control results:
Sensitivity of the animal strain was controlled every six months usng p - phenylenediamine as positive control substance. Corresponding results were not included in this end point study report.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle vaseline alone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no findings
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle vaseline alone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no 􀂠ndings.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0.2 g of ca. 0.1% test material in vaseline
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no findings
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0.2 g of ca. 0.1% test material in vaseline. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no 􀂠ndings.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle vaseline alone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no findings
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle vaseline alone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no 􀂠ndings.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.2 g of ca. 0.1% test material in vaseline
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no findings
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.2 g of ca. 0.1% test material in vaseline. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no 􀂠ndings.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
vehicle vaseline alone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no findings
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: vehicle vaseline alone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no 􀂠ndings.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.2 g of ca. 0.1% test material in vaseline
No. with + reactions:
10
Total no. in group:
20
Clinical observations:
Erythema and/or edema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.2 g of ca. 0.1% test material in vaseline. No with. + reactions: 10.0. Total no. in groups: 20.0. Clinical observations: Erythema and/or edema.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
vehicle vaseline alone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no findings
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: vehicle vaseline alone. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no 􀂠ndings.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.2 g of ca. 0.1% test material in vaseline
No. with + reactions:
10
Total no. in group:
20
Clinical observations:
Erythema and/or edema, in 2 animals associated with scaling
Remarks on result:
other: see remarks
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.2 g of ca. 0.1% test material in vaseline. No with. + reactions: 10.0. Total no. in groups: 20.0. Clinical observations: Erythema and/or edema, in 2 animals associated with scaling.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Remarks:
Migrated information
Executive summary:

The test material was tested for skin sensitization in guinea pigs equivalent or similar to the maximization test outlined in the OECD TGD 1981 and EU Directive 79/831. The principal deviation from these test guidelines was that for the first induction the test substance was

applied epidermally (i.e. epicutaneously) for 24 h under occlusive dressing, instead of being administered by intradermal injection. For this purpose, ca. 3% test material in Vaseline, i.e. approximately 0.4 g paste, was administered on filter patches (2 x 4 cm) placed over the 4 injection sites at the neck into which 0.1 mL of freshly prepared adjuvant saline mixture/site had been injected intracutaneously. A vehicle control and a test group, each comprised of 10 male and 10 female guinea pigs, were included in the study.

Reliability grade 2 was assigned to the study. It was not conducted in compliance with GLP, but a quality assurance statement was included in the report. Sensitivity of the animal strain was controlled every six months using p – phenylenediamine as positive control agent, but corresponding results were not included in the study report.

During Week 2, the second induction was performed by occlusive, epicutaneous administration of ca. 3 % test material in Vaseline for 48 hours. After a resting period of 14 days, a challenge application was performed by occlusive, epicutaneous administration of ca. 0.1 % test material in Vaseline (approximately 0.2 g paste per filter patch of 2 x 2 cm) on one flank and, for comparison, of Vaseline alone on the contra-lateral flank for 24 hours. The vehicle control group received adjuvant saline mixture and Vaseline during the induction period. During the challenge, its females received Vaseline alone and its males received Vaseline on one flank and test material/Vaseline paste on the contra-lateral flank, in order to check the maximum sub-irritant concentration of the test material in adjuvant treated animals.

Ten of twenty animals of the test group responded to the challenge administration of test material with erythema and/or edema associated with scale formation in two of these responding animals. Skin reactions were not evident in animals of the test group on skin patches exposed to vehicle alone and also not in animals of the control group. These results led to the conclusion that the test material has a skin-sensitizing (contact allergenic) potential, moderate in degree, in albino guinea pigs.