Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
other: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
12 December, 1988, to 5 January, 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
study conducted on the analogue substance; the read across justification is detailed in section 13. The Reliability of the Source Study is 1.
Justification for type of information:
The read across justification is detailed in section 13.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1981
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
An in vitro or in chemico skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study (different from LLNA test) is available.

Test material

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

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin, AG
- Age at study initiation: males: 7 weeks females: 8 weeks
- Weight at study initiation: males: 416 - 461 g; females: 409 - 459 g
- Housing: individually in Makrolon type-3 cages with standard softwood bedding
- Diet: pelleted standard Kliba 342, Batch 47188 guinea pig breeding/maintenance diet, ad libitum
- Water: tap water, ad libitum
- Acclimation period: one week under test conditions after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C
- Humidity: 40-70 %
- Air changes: 10-15 air changes per hour
- Photoperiod: 12 hours artificial fluorescent light/12 hours dark, music/light period

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
petrolatum
Concentration / amount:
0.5 %
Day(s)/duration:
single injection
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
5 %
Day(s)/duration:
48 hours
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
3 %
Day(s)/duration:
24 hours
No. of animals per dose:
5 per sex per control
10 per sex per dose
Details on study design:
RANGE FINDING TESTS:
A.Intradermal Injection
Intradermal injections (0.1 ml/site) were made into the clipped flank of two guinea pigs at concentrations of 0.1, 0.3, 0.5, 1 3 and 5 % of the test substance in petrolatum oil. The resulting dermal reactions were assesses 24 hours later.
B. Epidermal Application
Patches of filter paper (2*2 cm ) were saturated with concentrations of 3, 5, 10 and 25 % of the test substance in petrolatum oil and applied to the clipped and shaved flanks of each of four animals.

MAIN STUDY
A.Induction Exposure: injection
- No. of exposures: 6
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 ml/ site) were made at the border of a 4 x 6 cm area in the clipped region.
- Concentrations: 0.5 % test item in petrolatum

B. Induction Exposure: epidermal application (one week after the injections)
- No. of exposures: 1
- Exposure period: 48 h
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: (6 x 8 cm)
- Type of coverage: filter paper was saturated with the test substance (5 % in petrolatum) and placed over the injection sites of the test animals. The patch was covered by aluminum foil and firmly secured by an elastic plaster wound round the trunk of the animal and secured with impervious adhesive tape.
- Duration: 24 hours
- Concentrations: 5% in petrolatum

B. Challenge Exposure: test and control guinea pigs were challenged two weeks after the epidermal induction application.
- No. of exposures: 2
- Exposure period: approximately 24 hours later
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea pig
- Type of coverage: two patches (2 x 2 cm) of filter paper
- Concentrations: 3 % in petrolatum oil
- Evaluation: 0, 24, 48 hrs

OTHER OBSERVATIONS:
Mortality/Viability: once daily
Body Weights: at acclimatization start, start of application and end of test.
Symptoms (local / systemic): daily
Necropsy: no
All animals were killed at the end of the test period with an intraperitoneal injection of T61 (Hoechst, AG) and discarded
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitro-benzol (DNCB) in an independent study performed during September 1988

Results and discussion

Positive control results:
Clear positive results were observed in the DNCB traeted animals after the epidermal challenge application: 67 % positive reactions/total animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.5 % intradermal induction dose
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.5 % intradermal induction dose
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
other: not classified according to the CLP Regulation (EC) No. 1272/2008
Conclusions:
The test item did not demonstrate any sensitising potential.
Executive summary:

In order to evaluate the skin sensitising potential of the test item, an in vivo guinea pig maximisation test (GPMT) was performed according to the OECD Guideline 406 (1981). A primary irritation experiment was performed in order to determine the test concentrations of the main study. In the main study, ten control animals (5 males, 5 females) were treated with the vehicle alone (petrolatum oil) and 20 treatment animals (10 males, 10 females) were treated with the test item. Treatments were firstly two inductions: (i) an intradermal injection of 0.5 % test item; followed by (ii) an epidermal application of 5 % test item. After two weeks, the animals were subjected to two epidermal challenge applications separated by 24 hours using doses of 3 % in petrolatum.

No treatment or control animals were positive for sensitisation, therefore, the sensitisation rate of the test item was found to be 0 % to guinea pigs after intradermal and epidermal induction.