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Description of key information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
This skin sensitisation study was performed before the testing guideline of the LLNA test was adopted.
Species:
guinea pig
Strain:
other: Himalayan spotted
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC ltd, Biotechnology & Animal Breeding Division, Wölferstrasse 4, CH-4414 Füllinsdorf, Switzerland
- Age at delivery: 4 - 6 weeks
- Age at pretest start/beginning of acclimatisation period: 4 - 6 weeks
- Body weight at pretest start: 310 - 363 g (pretest group)
- Body weight at beginning of acclimatisation period: 335 - 406 g (control and test group)
- Housing: Individually in Macrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet (e.g. ad libitum): Pelleted standard Nafag Ecosan 845 25W4, batch nos. 91/98 and 103/98, guinea pig breeding / maintenance diet ("Nafag", Nähr- und Futtermittel AG, CH-9202 Gossau), ad libitum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum, once weeckly additional supply of ascorbic acid (approx. 1 g/l) via the drinking water was provided
- Acclimatization period: One week for the control and test group under test conditions after health examination. No acclimatisation for the animals of the pretest. Only animals without any visible signs of illness were used for the study.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 38 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12, music was played during the light period


DISTRIBUTION OF ANIMALS (ALL FEMALE)
Control group: 5
Test group: 10
Intradermal pretest: 1
Epidermal pretest: 2
Route:
intradermal
Vehicle:
physiological saline
Remarks:
1:1 (v/v) mixture of Freund's Complete Adjuvant
Concentration / amount:
without test substance / 0.1 ml
Day(s)/duration:
Day 1
Route:
intradermal
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
1 % test substance in PEG 400 / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
highest technically applicable concentration used
Route:
intradermal
Vehicle:
physiological saline
Remarks:
1:1 emulsion with Freund's Complete Adjuvant
Concentration / amount:
1% test substance / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
highest technically applicable concentration used
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
50%
Day(s)/duration:
Day 8 / 48 h
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
50%
Day(s)/duration:
Day 22 / 24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
5 control animals
10 test animals
Details on study design:
RANGE FINDING TESTS: PRETEST
The neck of three guinea pigs was shaved and the animals were pre-treated with four intradermal injections (0.1 ml/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline, one week prior to the start of the preliminary investigations.

Intradermal injections: 0.1 ml/site, in the clipped flank of one guinea pig at 1% and 3% of the test article in polyethylene glycol (PEG 400; the test article at 5% was technically not applicable). Dermal reactions assessed 24 h later.

Epidermal applications: conducted with two animals, 4 patches of paper (3 x 3 cm) were saturated with test article (ca. 0.2ml/patch): range 10-50% in PEG 400 and applied to both clipped and shaved flanks of each guinea pig. The patches were covered with aluminum foil and firmly secured by elastic plaster wrapped around the trunk covered by impervious adhesive tape for 24 h. The application sites were depilated with depilatory cream for 3-5 minutes, washed with warm water and dried with a disposable towel. Response assessed 24 and 48 h after bandage removal for erythema and edema.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 4 (3 preparations of interdermal injections, 1 preparation for the topical application)
- Exposure period: 48h for the topical induction application
- Test group: 10 animals
- Control group: 5 animlals
- Exposure: on day 1 three pairs of intradermal injections (0.1 ml/site), day 8 (after clipping and shaving again and preparation of the site with 0.5 ml 10% w/v sodium lauryl sulphate in paraffinum perliquidum) 2 x 4 cm patch of filter paper was saturated with the test article (50% in PEG 400), the amount of the test article was approximately 0.3 g, placed over the injection sites on the skin and covered with aluminum foil and secured with elastic plaster and impervious adhesive tape. The occlusive dressing was left in place for 48 h.
- Site: 6 x 8 cm area of dorsal skin on the scapular region, clipped and shaved
- preparations for intradermal injection:
(1) 0.1 ml 1:1 (v/v) mixture of Freund's Complete Adjuvant with physiological saline.
(2) 0.1 ml of test article, 1% in PEG 400.
(3) 0.1 ml of test article at 1% in a 1:1 (v/v) mixture of Freund's Complete Adjuvant with physiological saline.

- treatment of the control animals: as described above for the intradermal injections and the topical application, but without test article


B. CHALLENGE EXPOSURE
- Day of challenge: 22
- Exposure period: 24h
- Test groups: 1 (10 animals)
- Control group: 5 animals
- Site: 5 x 5 cm area on the left and right flank of each guinea pig, clipped and shaved.
- Concentrations: 50 and 0% of test article in PEG 400 (highest non-irritating concentration and vehicle only on left and right flank, respectively), on 3 x 3 cm filter paper using the same method as for the epidermal application. 21h after removal of the dressing, the applications sites were depilated as described in the epidermal pretest.
- Evaluation (hr after challenge): 24 and 48 for erythema and edema.
Challenge controls:
There were no dermal reactions seen in any of the control animals.
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole
Positive control results:
Positiv control results were generated in the period from 01.12.1998 to 01.01.1999.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no symptoms of systemic toxicity and no deaths were observed
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no symptoms of systemic toxicity and no deaths were observed
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
10%
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
10%
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Maximum concentration not causing irritating effects in preliminary test: 50 %

Signs of irritation during induction:
The expected and common findings, as erythema, oedema,
necrotizing dermatitis, encrustation and exfoliation of
encrustation was observed.

Evidence of sensitisation of each challenge concentration:
0% of the animals showed sensitization.

Interpretation of results:
other: not sensitising according to REGULATION (EC) 1272/2008
Conclusions:
Based on these results and according to the EC criteria for classification and labelling according to GHS/CLP (Regulation (EC) No. 1272/2008), the test item does not have to be classified and has no obligatory labelling requirement as skin sensitising.
Executive summary:

As no skin sensitisation was induced in a GLP-study acc. to OECD guideline 406, test item is considered as not skin sensitising.

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

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Since no classification criteria according to GHS/CLP (Regulation (EC) No. 1272/2008) are fulfilled, the substance is not classified for sensitisation.