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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 25 FEB 2004 to 10 MAR 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study (OECD TG 429)

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
in compliance with Swiss Ordinance relating to GLP
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- strain: CBA/CaOlaHsd
- Source: Harlan Netherlands
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 16-24 g
- Housing: individually, Makrolon type-2 cages
- Diet (e.g. ad libitum): pelleted standard Kliba 3433 mouse maintenance diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: yes, no further information


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hrs/12 hrs
- at least 8 hours music during the light period

Study design: in vivo (LLNA)

Vehicle:
other: ethanol:water (7:3, v/v)
Concentration:
0, 2.5, 5, 10% (w/v)
No. of animals per dose:
4 females per dose group
2 females in the pre-test
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: test item was more soluble in ethanol:water, 7:3 (v/v) than in acetone:olive oil, 4:1 (v/v) or DMSO
- 10% (w/v) was the highest technically applicable concentration
- Irritation: no irritation after single application of test item concentrations of 1, 2.5, 5, and 10 % (w/v)
- Lymph node proliferation response: no data


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response:
1: exposure to at least one concentration of the test item resulted in an incorporation of 3H-methyl thymidine at least 3-fold or greater than that recorded in control mice as indicated by the stimulation index
2: data are compatible with a conventional dose response, although allowance must be made for either local toxicity or immunological suppression

TREATMENT PREPARATION AND ADMINISTRATION:
- individual preparation of weight volume dilutions using a magnetic stirrer as homogenizer
- test item preparations were made feshly before each dosing occasion and no more than 4 hours prior to application to the ears
- 25 µl were spread over the entire dorsal surface of each ear lobe once daily for three consecutive days
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
- calculations of mean values and standard deviations for body weight

Results and discussion

Positive control results:
Stimulation indices of 1.5, 2.3 and 8.4 were determined with the positive control item at concentrations of 5%, 10% and 25% (w/v), respectively, in acetone:olive oil, 4:1 (v/v). An EC3 value of 11.7% (w/v) was calculated.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: Stimulation indices (SI) were all below 3. The following SI were calculated: 2.5% test item: 1.1 5% test item: 1.4 10% test item: 1.7
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
There was a dose dependent increase in the dpm, which were measured for the pooled lymph nodes of each treatment group (8 lymph nodes per dose group): Background: 4 or 6 dpm Control group: 1866 dpm 2.5% test item: 1960 dpm 5% test item: 2638 dpm 10% test item: 3146 dpm

Any other information on results incl. tables

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. Calculation of the EC3 value was not performed because no test concentration produced a SI of 3 or higher.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The test substance was not sensitising in the LLNA in concentrations up to 10% (the highest achieveable concentration) in ethanol:water, 7:3 (v/v).
Executive summary:

Skin sensitising potential of the test item was tested in the LLNA. The test item was not sensitising in concentrations up to 10% (the highest achieveable concentration). The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. Stimulation indices of 1.1, 1.4 and 1.7 were determined at concentrations of 2.5%, 5% and 10% (w/v), respectively, in ethanol:water, 7:3 (v/v). Therefore, the test item has not to be classified as skin sensitiser according to Regulation (EC) No 1272/2008.