Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2002
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study, OECD guideline conform and well documented, read across substance

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Ibm: GOHI; SPF-quality guinea pigs (synonym: Himalayan spotted)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Biotechnology & Animal Breeding Division, Fuellinsdorf / Switzerland
- Age at study initiation: 3 - 6 weeks
- Weight at study initiation: 379 - 426 g
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, Muttenz)
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418, batch no. 93/01, guinea pig breeding / maintenance diet, containing Vitamin C (Provimi Kliba AG, Kaiseraugst), ad libitum.
- Water (e.g. ad libitum): Community tap water from Fiillinsdorf, ad libitum
- Acclimation period: One week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Polyethylene glycol 300 (PEG 300)
Concentration / amount:
intradermal induction: 10% test item
epidermal induction: 25% test item
challenge: 1% test item
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Polyethylene glycol 300 (PEG 300)
Concentration / amount:
intradermal induction: 10% test item
epidermal induction: 25% test item
challenge: 1% test item
No. of animals per dose:
15 males for main study
5 males for pretests
Details on study design:
RANGE FINDING TESTS: To determine the different concentrations one intradermal and two epidermal pretests were performed.
Intradermal injections (0.1 ml/site) were made into the clipped flank of the same guinea pig at concentrations of 10 %, 5 % and 3% of the test item in PEG 300. The three concentrations were determined during non-GLP formulation trials performed before the study initiation date. The concentration of 10 % was considered to be the highest technically applicable concentration which could be injected into the intra-cellular space in spite of the high viscosity of the application dilution and the obstacle caused by the tissues.
For intradermal injections4 patches of filter paper (3x3 cm) were saturated with the test item at 25 % (technically the highest possible concentration to be applied sufficiently), 15 %, 10 % and 5 % in PEG 300 and applied to the clipped and shaved flanks. The amount of test item preparation applied was approximately 0.2 g for the test item at 25 % and a volume of approximately 0.2 ml was applied for the remaining test item concentrations. 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 item. The dressings were removed after an exposure period of 24 hours.
Twenty-one hours after removal of the dressing the application site was depilated with an approved depilatory cream (VEET Cream, Reckitt & Colman AG, Allschwil) in order to visualize any resulting erythema. The depilatory cream was placed on the patch sites and surrounding areas, and left on for 3-5 minutes. It was then thoroughly washed off with a stream of warm, running water. Thereafter, the animals were dried with a disposable towel, and returned to their cages. The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman.

MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal induction
Test Group:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) The test item, at 10 % in PEG 300.
3) The test item at 10 % in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
Control Group:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) PEG 300
3) 1:1 (w/w) mixture of PEG 300 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.

Epidermal induction on test day 8:
- time point of exposure: One week after the intradermal induction
- Exposure period: 48 hours
- Site: scapular area
- Exposure area: 2 x 4 cm patch of filter paper was saturated with the test item and placed over the injection sites of the test animals
- Amount applied: approximately 0.3 g
- Concentrations: 25 % test item in PEG 300

B. CHALLENGE EXPOSURE
- time point of exposure: test and control guinea pigs were challenged two weeks after the epidermal induction
- Exposure period: 24 hours
- Site: on the left and right flank of each guinea pig just prior to the application; test item was applied to the left flank, vehicle only (PEG 300) was applied to the right flank
- Exposure area: 3x3 cm; two patches of filter paper were saturated with the test item at the highest tested non-irritating concentration
- Concentrations: 1 % test item in vehicle and the vehicle only (PEG 300) using the same method as for the epidermal application.
- Amount applied: approximately 0.2 ml
- Evaluation (hr after challenge): 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman.
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No findings noted.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle alone. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No findings noted..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No findings noted.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle alone. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No findings noted..
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1% test item in PEG
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No findings noted. But animal no. 52 died.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1% test item in PEG. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No findings noted. But animal no. 52 died..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1% test item in PEG
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No findings observed. But animal no. 62 died.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% test item in PEG. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No findings observed. But animal no. 62 died..

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information