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Description of key information

Skin sensitisation: Buehler test, Not sensitising (OECD 406, GLP, K, Rel.2)

Respiratory sensitisation: no data available

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 1988-08-02 to 1988-09-22
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study performed according to the OECD test guideline No. 406 and in compliance with GLP. The dose used for the third week induction was reduced from 3% to 1% due to moderate to severe irritation (score 2 to 3) observed at induction week 2. This reduction does not affect the reliability of the result since slight erythema was still observed at induction week 3.
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
At the time of study performance (1988), the LLNA OECD TG was not adopted
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, Fuellinsdorf, Switzerland
- Age at study initiation: approximately 8 weeks
- Weight at study initiation: males: 342-408 g; females: 333-409 g
- Housing: individually in Makrolon type-3 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 342, batch 44/88 and 45/88 guinea pig breeding/maintenance diet ad libitum (Kaiseraugsr, Switzerland).
- Water (e.g. ad libitum): community tap water from Itingen, ad libitum.
- Acclimation period: 7days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15 per hr
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 1988-08-02 To: 1988-09-22
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
3 % (week 1 and 2), 1 % (week 3)
Day(s)/duration:
once a week, during 6 hours
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
0.5 and 1%
Day(s)/duration:
2 weeks after the last induction exposure
Adequacy of challenge:
highest non-irritant concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
0.5 and 1%
Day(s)/duration:
2 weeks after challenge #1
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 males and 20 females per dose. 10 control animals for the primary challenge + 10 additional untreated control animals for the rechallenge.
Details on study design:
RANGE FINDING TESTS:
- Irritation screen for induction: 8 animals, up to 4 concentration per animal: 50, 30, 10, 5, 3, 2, 1, 0.5, 0.3, 0.1 % in ethanol. Concentrations determined for the main test were as follow: induction week 1 and 2: 3 %, induction week 3: 1 %.
- Irritation screen for challenge: 4 animals (2M +2 F), test material at 3, 2, 1 or 0.5 % in ethanol. Concentrations determined for the challenge and rechallenge were 1 and 0.5 %.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Test groups: 0.3 mL of a freshly prepared test article solution in a 25 mm Hill Top Chamber. The animals were put in the restrainer and the designated patch was applied to the clipped surface as quickly as possible after the test article has been applied. The patch appliance was occluded with a rubber dam (medium gauge 12-15 cm wide) pulled snug on each side of the animal and secured with at least one large size clip on each side of the restrainer. The rubber dam was placed under the front and back metal restraining bands, and had snug contact with the animal over the entire dorsal surface.
- Control group: treated accordingly with the vehicle only
- Site: left shoulder
- Frequency of applications: once a week
- Duration: 6 hours. If necessary the treated skin was flushed with lukewarm tap water and padded dry with disposable paper towels.
- Concentrations: 3 % (week 1 and 2), 1 % (week 3)

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: approximately two weeks after the last induction exposure (the time between the last induction exposure and the primary challenge for the test group may vary from 12 to 16 days). The test group was rechallenged 15 days following primary challenge.
- Test groups: similar method than for induction
- Control group: similar method than for induction
- Site: right side
- Concentrations: 0.5 or 1 %
- Evaluation (hr after challenge): 24 and 48 h for challenge, 24 and 44 h without depilation + 48 h after depilation for rechallenge.
Challenge controls:
Not applicable
Positive control substance(s):
yes
Remarks:
DNCB
Positive control results:
In a separate test (February, 1988) the incidence to allergenic reactions of the guinea-pig strain used, was tested with a known allergen (DNCB; Dinitro-chloro-benzol).
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.5 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.5 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
0.5 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
44
Group:
test chemical
Dose level:
0.5 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
0.5 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
44
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1 %
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
2 with score 1 (well defined erythema); 6 with very slight erythema
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
4 with very slight erythema
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
44
Group:
test chemical
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
1 %
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
2 with score 1 (well defined erythema); 5 with very slight erythema
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
44
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
other: Separate study
Group:
positive control
Dose level:
DNCB
Clinical observations:
NA
Remarks on result:
positive indication of skin sensitisation
Remarks:
Historical data; no details on results available

One rechallenge control animal showed very slight sign of erythema at 48 h. No score was observed in the other vehicle control animals.

No death occurred during the entire test period. No systemic symptoms were observed in any animal during the test period. The body weight gain of all animals (control / test material) were not affected by the treatment.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test material is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In a dermal sensitization study performed according to the OECD Guideline No. 406 and in compliance with GLP, the test material was tested in Dunkin/Hartley guinea-pigs using the Buehler test (20 treated animals/sex/dose + 10 controls animals/ sex/dose).

 

The preliminary study determined the lowest irritant test substance concentration used at induction phase (3 % in ethanol) and the highest non-irritant test substance concentration used at challenge (0.5 and 1 % in ethanol). The induction concentration was lowered in week 3 to 1 % to avoid reactions which were encountered with the concentration 3 % in week 1 and 2.

 

For induction, guinea-pigs were exposed to the test material using a Top Hill Chamber once a week, during 3 weeks. Animals were topically challenged approximately two weeks after the last induction exposure. Rechallenge was performed 15 days following primary challenge.

 

The sensitivity of the guinea-pig was checked in a separate test with Dinitro-chloro-benzol.

 

No signs of ill health or toxicity were recorded in any animals during the observation period.

 

The test material did not produce evidence of skin sensitisation.

 

Under the test conditions, the test material is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for sensitisation endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

A key study was identified (RCC, 1988). In this dermal sensitization study performed according to the OECD Guideline No. 406 and in compliance with GLP, the test material was tested in Dunkin/Hartley guinea-pigs using the Buehler test (20 treated animals/sex/dose + 10 controls animals/ sex/dose).

The preliminary study determined the lowest irritant test substance concentration used at induction phase (3 % in ethanol) and the highest non-irritant test substance concentration used at challenge (0.5 and 1 % in ethanol). The induction concentration was lowered in week 3 to 1 % to avoid reactions which were encountered with the concentration 3 % in week 1 and 2. This reduction does not affect the reliability of the study since slight erythema was still observed at induction week 3.

For induction, guinea-pigs were exposed to the test material using a Top Hill Chamber once a week, during 3 weeks. Animals were topically challenged approximately two weeks after the last induction exposure. Rechallenge was performed 15 days following primary challenge.

The sensitivity of the guinea-pig was checked in a separate test with Dinitro-chloro-benzol.

No signs of ill health or toxicity were recorded in any animals during the observation period.

The test material did not produce evidence of skin sensitisation under the test conditions.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Based on the available data, no additional self-classification is proposed according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

No data was available for respiratory sensitisation. However, this substance is not a skin sensitizer, therefore according to Figure R.7.3 -2 of the Chapter R.7 (V 6 - July 2017) the chemical is not considered as a respiratory sensitizer.

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