Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

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:
other: Well documented and reported study fully adequate for assessment. The study was conducted according to an internationally accepted technical guideline and in compliance with GLP in a recognized contract research organization.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
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.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- albino guinea pigs of the Dunkin/Hartley strain
- Source: D. Hall, Newchurch, Staffs, UK
- Age at study initiation: 5 - 8 weeks
- Weight at study initiation: 317- 397 g
- Housing: in groups of five in suspended metal cages with wire mesh floors. Environmental enrichment: autoclaved hay (3 x/week), plastic tubular pipes
- Diet (e.g. ad libitum): ad libitum. Vitamin C enriched guinea-pig diet (Harlan Teklad 9600 FD2 SQC)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Relative Humidity (%): 30 - 70 %

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction intradermal injection: 0.25% w/v in water (highest concentration causing irritation but not necrosis or give signs of toxicity)
Induction topical application: 65% w/v in water
Adequacy of induction:
other: see: Details on study design
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Topical challenge: 65 and 32.5% w/v in water
Adequacy of challenge:
other: see: Details on study design
No. of animals per dose:
5 control animals
10 test animals (all concentrations for each animal)
Details on study design:
RANGE FINDING TESTS: Preliminary study:
The animals were pre-treated with an intradermal injection of Freund's Complete Adjuvant, 50 : 50 with water for irrigation (Ph.Eur.), approximately one week prior to the start of the preliminary investigations. Procedures of investigations:
Intradermal injections: conducted with two animals, 0,1 ml/site; range 0.1 - 10% w/v in water; responses assessed appr. 24 and 72 h later.
Topical application: conducted with four animals, patches of paper (2cm x 2cm) were saturated (ca. 0.2 ml/patch); range: 20-65% w/v), applied to the clipped and shaved flanks of the animals; duration of exposure 24h, responses assessed after removal of the patches (erythema and oedema) at 0, 24 and 48 h.

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 animals
- 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 10% w/v sodium lauryl sulphate in petrolatum) 2 cm x 4 cm patch of Whatman No. 3 paper was saturated with approximately 0.4 ml of BC-300, 65% w/v in water, placed over the injection sites on the skin
- Site: 4 cm x 6 cm area of dorsal skin on the scapular region, clipped and shaved
- preparations for intradermal injection:
(1) 0.1 ml of Freund's Complete Adjuvant 50: 50 with water for irrigation (Ph.Eur.).
(2) 0.1 ml of BC-300, 0.25% w/v in water for irrigation.
(3) 0.1 ml of BC-300, 0.25% w/v in a 50 : 50 mixture of water for irrigation and Freund's
Complete Adjuvant.
- treatment of the control animals: as described above for the intradermal injections and the topical application, but without test substance


B. CHALLENGE EXPOSURE
- Day of challenge: 22
- Exposure period: 24h
- Test groups: 1 (10 animals)
- Control group: 5 animals
- Site: area on the left flank of each guineapig, clipped and shaved.
- Concentrations: 32.5% and 65% in water (posterior resp. anterior site of the flank)
- Evaluation (hr after challenge): 24 and 48
Challenge controls:
There were no dermal reactions seen in any of the control animals.
Positive control substance(s):
yes
Remarks:
hexyl cinnamic aldehyd (HCA)

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
all dose levels
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no deaths and no signs of ill health or toxicity
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
all dose levels
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no deaths and no signs of ill health or toxicity
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
all dose levels
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:
all dose levels
No. with + reactions:
0
Total no. in group:
5
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
HCA as supplied
No. with + reactions:
10
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:
HCA as supplied
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
HCA, 50% v/v in Alembicol D
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:
HCA, 50% v/v in Alembicol D
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

Necrosis was recorded at sites receiving Freund's Complete Adjuvant in test and control animals. No irritation was seen in test animals at sites receiving the test material, 0.25% w/v in water for irrigation and no irritation was observed in control animals receiving water for irrigation.

Applicant's summary and conclusion

Interpretation of results:
other: not sensitising
Conclusions:
As all of the animals gave negative responses, the test substance does not require labelling.