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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 September 17 to October 18, 1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
(Adopted in May 1981)
Deviations:
yes
Remarks:
FCA/saline was used in the 3rd control injection instead of ethanol FCA/saline
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
(Adopted in March 1984)
Deviations:
yes
Remarks:
FCA/saline was used in the 3rd control injection instead of ethanol FCA/saline
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
LLNA was not available at time of testing

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Ibm: GoHI; SPF quality (Himalayan spotted)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4. CH-4414 Füllinsdorf.
- Age at study initiation: M: 7 weeks; F: 8 weeks.
- Weight at study initiation: M: 370-394 g; F: 374-393 g.
- Housing: individually, in Makrolon type-3 cages with standard softwood bedding.
- Diet: pelleted standard Kliba 342, Batch 59/0 guinea-pig breeding / maintenance diet, ad libitum. Not contaminated.
- Water: community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water. Not contaminated.
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12. Music during the light period.

IN-LIFE DATES: From: 1990-09-17 To: 1990-10-28

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
other: ethanol
Concentration / amount:
Intradermal induction: 5%
Epicutaneous induction: 30%
Epicutaneous challenge: 30, 10 and 3%
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: ethanol
Concentration / amount:
Intradermal induction: 5%
Epicutaneous induction: 30%
Epicutaneous challenge: 30, 10 and 3%
No. of animals per dose:
10/sex/test group, 5/sex/control group
Details on study design:
RANGE FINDING TESTS:
- Intradermal injections: 2 animals; concentrations of 5, 3 and 1% in ethanol (0.1 mL/site). Assessment of dermal reactions 24 hours later. The concentration selected for the main study was 5%.
- Epidermal application: 4 animals; concentrations of 30, 10, 3 and 1% in ethanol. Assesment of dermal reactions immediately after patch removal, 24 and 48 hours later. The concentration selected for epidermal application was 30%.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: intradermal + epicutaneous.
- Test groups:
Intradermal injections: 3 pairs of intradermal injections (0.1 mL/site) made at the border of a 4 x 6 cm area in the clipped region:
1/ FCA 50:50 with physiological saline
2/ Test article, diluted to 5% with ethanol
3/ Test article diluted to 5% by emulsion in a 50:50 mixture of FCA and physiological saline.
Epidermal applications: on day 7, the test area was pre-treated with 10% Sodiul-Lauryl-Sulfate (SLS) in petrolatum oil, because no irritation concentration could be determined in the corresponding pre-test. On day 8, patches of filter paper (2 x 2 cm) were saturated with concentration of 30% test article in ethanol and applied to the clipped and shaved flanks of each animal. The patches were covered by a strip of aluminium foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. Reactions sites were assessed immediately, 24 and 48 hours after removal of the dressing.
- Control group:
Intradermal injections: 3 pairs of intradermal injections (0.1 mL/site) made at the border of a 4 x 6 cm area in the clipped region:
1/ FCA 50:50 with physiological saline
2/ Ethanol
3/ FCA 50:50 with physiological saline.
Epidermal applications: treated as described above with the omission of test article (ethanol only).
- Site: dorsal skin from the scapular region

B. CHALLENGE EXPOSURE
- No. of exposures: 1, epicutaneous
- Day(s) of challenge: 2 weeks after the epidermal induction application
- Exposure period: 24 hours
- Test groups: patches (2 x 2 cm) of filter paper were saturated with the test article and applied to the left flank using the same method as for the epidermal application.
- Control group: patches (2 x 2 cm) of filter paper were saturated with the vehicle and applied to the right flank using the same method as for the epidermal application.
- Site: left and right flanks
- Concentrations: 0, 3, 10 and 30 %
- Evaluation (hr after challenge): 24, 48 and 72 (0, 24 and 48 after patch removal)

OTHER: Erythema and oedema were assessed using the numerical grading system of Draize.
Challenge controls:
None
Positive control substance(s):
not required
Remarks:
a control group (Formaldehyde-solution) is tested twice a year for sensitivity check of the guinea-pig.

Results and discussion

Positive control results:
The most recent test was run from April 23 to May 24, 1990. Clear positive results were observed in 7/10 Formaldehydlosung (HCHO) treated animals after the epidermal challenge application.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
30%
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
73
Group:
test group
Dose level:
30%
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
3%
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
3%
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
20

Any other information on results incl. tables

No death occurred during the study. No systemic symptoms were observed in the animals. The body weight gain of the animals was not affected during the study.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test material did not produce evidence of skin sensitization (delayed contact hypersensitivity).
Executive summary:

A dermal sensitization study wasperformed according to OECD guideline No. 406 and in compliance with GLPwith the test material diluted in ethanol. Male and female Himalayan spotted guinea-pigs were tested using the Guinea pig maximisation method (20 treated animal per concentration + 10 control animals).

The preliminary study determined the lowest irritant test substance concentration used at induction phases and the highest non-irritant test substance concentration used at challenge.

The test material diluted in ethanol at 5% was administered by injection for intradermal induction. On day 7, as the substance was not a skin irritant, the site was pre-treated with 10% sodium lauryl sulphate in petrolatum-oil. Topical induction was performed with the test material diluted in ethanol at 30 % Day 8. For the challenge, the test material was applied topically at 30, 10 and 3 % in ethanol.

 

The sensitivity of the guinea-pig was checked every 6 months at RCC with a Formaldehyde-solution, a known sensitizer.

 

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 sensitization (delayed contact hypersensitivity).

 

Under the test conditions, the test material is not classified according to the annex VI of the Regulation (EC) No. 1272/2008 (CLP) and of the Directive 67/548/EEC

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