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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:
key study
Study period:
06 Apr - 14 Jun 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted in 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
adopted in 1996
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
adopted in 2003
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was conducted prior to the current requirement in Regulation (EC) 1907/2006 to perform an LLNA study (OECD 429) as the preferred in vivo skin sensitisation study.

Test material

Constituent 1
Chemical structure
Reference substance name:
(5-hydroxy-1,3-dimethyl-1H-pyrazol-4-yl)[2-(methylsulfonyl)-4-(trifluoromethyl)phenyl]methanone
EC Number:
609-256-3
Cas Number:
365400-11-9
Molecular formula:
C14H13F3N2O4S
IUPAC Name:
(5-hydroxy-1,3-dimethyl-1H-pyrazol-4-yl)[2-(methylsulfonyl)-4-(trifluoromethyl)phenyl]methanone

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Crl:HA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratory Animal Breeders, Kisslegg, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 5 - 6 weeks
- Weight at study initiation: 296 - 393 g
- Housing: 5 animals (acclimation period) or 2 - 3 animals (study period) in type IV Makrolon® cages with low-dust wood shavings bedding (Rettenmeier & Söhne GmbH & Co., Rosenberg, Germany)
- Diet: Provimi Kliba 3420 - Maintenance Diet for Guinea Pigs, Provimi Kliba AG, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40 - 70%
- Air changes (per hr): ≥ 10
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
5% (40 mg/animal)
Day(s)/duration:
single injection
Adequacy of induction:
other: highest concentration used in a dose-range-finding study for intradermal induction leading to wheal after 24 and 48 h in the test animal
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
6% (30 mg/animal)
Day(s)/duration:
48 h
Adequacy of induction:
other: concentration used in a dose-range-finding study and induced slight localized redness after 48 h in 3/4 animals
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
3% (15 mg/animal)
Day(s)/duration:
24 h
Adequacy of challenge:
other: highest concentration used in a dose-range-finding study for challenge which was non-irritant
No. of animals per dose:
Range-finding study: 1 (intradermal induction), 4 (topical induction), 2 (challenge)
Main study: 10 (controls), 20 (test group)
Details on study design:
RANGE FINDING TESTS:
For the dose range-finding for intradermal induction one animal was given intradermal injections with 0.1 mL of 0, 1, 2.5 and 5% test substance in physiological saline. The evaluation of the injection sites 24 and 48 h showed no effect at the 0% site but wheal at all other sites. Thus, 5% of the test substance was selected for the intradermal induction. For topical induction two studies with four animals each and three concentration (0, 12, 25 and 50% in the first study and 0, 1, 3 and 6% in the second study respectively) were conducted. The skin reactions were evaluated 48 and 72 h after start of application and based on the findings 6% of the test substance was selected for epicutaneous induction. Two animals were administered three concentrations of the test substance (0, 0.5, 1 and 3%) under occlusive conditions for 24 h for the dose range-finding for challenge. The skin reactions were evaluated 48 and 72 h after start of application and based on the findings 3% of the test substance was selected for challenge.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Injection 1: a 1:1 mixture FCA/physiological saline solution
Injection 2: 5% test substance in physiological saline solution
Injection 3: equal amounts of 5% test substance in physiological saline solution and FCA
Epicutaneous: 0.5 mL 6% test substance in physiological saline solution
- Control group:
Injection 1: a 1:1 mixture FCA/physiological saline solution
Injection 2: undiluted physiological saline solution
Injection 3: a 1:1 mixture FCA/physiological saline solution
Epicutaneous: 0.5 mL physiological saline solution
- Site: cranial/bilateral (injection 1 and 3), medial/bilateral (injection 2)
- Frequency of applications: single injection (intradermal), single application (epicutaneous induction; one week after intradermal application)
- Duration: 9 days
- Concentrations: intradermal induction 5%, epicutaneous 6%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: three weeks after intradermal induction (day 21)
- Exposure period: 24 h
- Test groups: 3% test substance in physiological saline solution
- Control group: 3% test substance in physiological saline solution
- Site: right flank (caudal)
- Concentrations: 3%
- Evaluation (hr after challenge): 48 and 72 h after the start of the application to induce the challenge (24 and 48 h after patch removal)

OTHER:
- Clinical signs: The animals were observed for clinical signs at least once daily throughout the entire study period.
- Body weights: The body weights of the animals were recorded before the first induction and at the end of the study.
Challenge controls:
The control group is actually a challenge control.
Positive control substance(s):
yes
Remarks:
(alpha hexyl cinnamic aldehyde formulated in polyethylene glycol 400; intradermal induction: 5%, epicutaneous induction: 25%, challenge: 12%)

Results and discussion

Positive control results:
100% of the test animals exhibited dermal reactions in the challenge treatment with the positive control substance. There was no reddening of the skin to be observed on control group animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Induction: 0%; challenge: 3%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
strong effects up to encrustation at the injection sites after intradermal induction
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
Induction: 5% (intradermal) and 6% (epicutaneous); challenge: 3%
No. with + reactions:
0
Total no. in group:
19
Clinical observations:
strong effects up to encrustation at the injection sites after intradermal induction; clinical signs (poor general condition, piloerection, labored breathing, pale) and subsequent death (day 15) in 1/20 animal
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Induction: 0%, challenge: 3%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
strong effects up to encrustation at the injection sites after intradermal induction
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Induction: 5% (intradermal) and 6% (epicutaneous), challenge: 3%
No. with + reactions:
0
Total no. in group:
19
Clinical observations:
strong effects up to encrustation at the injection sites after intradermal induction; clinical signs (poor general condition, piloerection, labored breathing, pale) and subsequent death (day 15) in 1/20 animal
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

- Body weights:

At the end of the study, the mean body weight of the treatment group animals was in the same range than that of the control group animals.

Applicant's summary and conclusion

Interpretation of results:
other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/208
Conclusions:
CLP: not classified