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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:
weight of evidence
Study period:
1989-11-20 to 1990-04-17
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
12 May, 1981
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
According to Annex XI section 1.1.2. of Regulation (EC) No 1907/2006 (REACH) testing of skin sensitisation in an in vivo LLNA is not necessary if there are data that is generated by studies not carried out according to GLP or the test methods reffered to in Article 13 (3) of the same regulation if the data is sufficcient for the purpose of classification and labelling and/or risk assessment, if the key parameters foreseen to be investigated in the recommended methods are reliably covered, the exposure duration is similar to that of the recommended methods and if the study is adequate and reliable documented. The present GPMT was conducted according to OECD guideline 406, which is reliable without restrictions and adequate for classification and labelling purposes. It is well documented, measures the recommended key parameters and exposure duration is comparable to an in vivo LLNA. Thus, due to the provided reliable and adequate data the perfomance of an in vivo LLNA is not necessary. Furthermore, the study was conducted prior to regulatory implementation of the LLNA method.

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Himalayan
Remarks:
spotted
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: males 7 weeks, females 8 weeks
- Weight at study initiation: males 357-405 g, females 345-384 g
- Housing: individually in Makrolon type-3 cages with standard soft wooding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet: ad libitum, pelleted standard Kiba 342, Batch 55/89 guinea pig breeding/maintenance diet ("Kliba", Klingentalmühle AG, CH-4303 Kaiseraugst)
- Water: ad libitum, Community tap water from Itingen
- Acclimation period: one week under test conditions after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40-70
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
other: Acetone/bi-distilled water 1:1
Concentration / amount:
intradermal injections: 3% in acetone/bi-distilled water, epicutaneous: 25% in acetone/bi-distilled water
Day(s)/duration:
intradermal: single injection followed by epicutaneous application after 7 days; epicutaneous: 48 h
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, semiocclusive
Vehicle:
other: Acetone/bi-distilled water 1:1
Concentration / amount:
left flank 15%, right flank 25%
Day(s)/duration:
24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Ten animals (5 males, 5 females) were treated as control group and twenty animals (10 males, 10 females) were treated at test group.
Details on study design:
Preliminary study:
Intradermal injections (0.1 mL/site) were made into the clipped flank of two guinea-pigs at concentrations of 5% (right flank), 3%, 1% and 0.5%
(left flank) of the test article in acetone/bi-distilled water 1:1. The resulting dermal reactions were assessed 24 hours later.
Epidermal applications:
Patches of filter paper (2 x 2 cm) were saturated with concentrations of 25%, 15%, 10% and 5% of the test article in acetone/bi-distilled water 1:1 and applied to the clipped and shaved flanks of each of four guinea-pigs. The patches were covered by a strip of aluminum foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. This procedure ensured the intensive contact of the test articleeven if it is insoluble in the vehicle used. The dressings were removed after an exposure period of 24 hours·and the reaction sites were assessed for erythema and edema on a numerical basis according to the scale described above. Further examination of the sites were performed 24 and 48 hoursafter removal of the dressings. Prior to the first reading, the epidermal application sites were gently washed off excess test article with acetone.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: One intradermal application, one epicutaneous application
- Exposure period: Intradermal: single injection followed by epicutaneous application after one week
- Test groups: Intradermal: 1) Freund´s complete adjuvant 50:50 with bi-distilled water 2) The test article, diluted to 3% with acetone/bi-distilled water 1:1 3) The test article at the concentration used in (2), emulsified in a 50:50 mixture of Freund´s adjuvant and the vehicle used in (2). Epicutaneous: A 2 x 4 cm patch of filter paper saturated with the test article (25% in acetone/bi-distilled water 1:1) placed over the injection sites
- Control group: The control groups were treated according to the test groups with the omission of the test article.
- Site: dorsal skin from the scapular region
- Frequency of applications: Once
- Duration: Intrdermal: single injection followed by epicutaneous application after one week
- Concentrations: Intradermal: 3% with acetone/bi-distilled water 1:1, Epicutaneous: 25% in acetone/bi-distilled water 1:1

B. CHALLENGE EXPOSURE
- No. of exposures: one epicutaneous application of two concentrations
- Day(s) of challenge: day 21
- Exposure period: 24 h
- Test groups: Two patches (2 x 2 cm) of filter paper were saturated with a) non-irritant concentration (15% in acetone/bi-distilled water 1:1) of the test article and b) with the acetone/bi-distilled water 1:1 only and applied to the left flank, the right flank was treated with 25% in acetone/bi-distilled water 1:1.
- Control group: Treated the same way as unter Test groups
- Site: scapular region
- Evaluation (hr after challenge): immediately, 24h and 48h after removal of the patches
Challenge controls:
The control animals were treated in the same way as described above.
Positive control substance(s):
yes
Remarks:
Formaldehyde-solution, 20% dilution in bi-distilled water for induction, 15% dilution for challenge

Results and discussion

Positive control results:
9/10 animals induced and challenged with formaldehyde-solution showed positive reactions.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
induction: 0%; challenge: 15% in acetone/bi-distilled water (1:1)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
induction: 0%; challenge: 15% in acetone/bi-distilled water (1:1)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
intradermal induction: 3% in acetone/bi-distilled water (1:1); epicutaneous induction: 25% in acetone/bi-distilled water (1:1); challenge: 15% in acetone/bi-distilled water (1:1)
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation
Remarks:
1 female animal was killed due to ethical reason on day 10 of the study
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
intradermal induction: 3% in acetone/bi-distilled water (1:1); epicutaneous induction: 25% in acetone/bi-distilled water (1:1); challenge: 15% in acetone/bi-distilled water (1:1)
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
no indication of skin sensitisation
Remarks:
1 female animal was killed due to ethical reason on day 10 of the study
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
induction 20% HCHO in bi-distilled water; challenge 15 % HCHO in bi-distilled water
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
induction 20% HCHO in bi-distilled water, challenge 15% HCHO in bi-distilled water
No. with + reactions:
4
Total no. in group:
9
Remarks on result:
positive indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, no allergenic potency of the test article was concluded. The results were interpreted according to the rating of Magnusson and Kligman (1970). According to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS), this test article is considered to be not a sensitizer and thus, does not need to be classified.