Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according OECD test guideline 429 under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): C-SAT 100018
- Physical state: slightly yellowish solid
- Analytical purity: 100%
- Lot/batch No.: CF00460029
- Expiration date of the lot/batch: 29.02.2010
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
mouse
Strain:
NMRI
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-97633 Sulzfeld, Germany
- Age at study initiation: 9-11 weeks
- Weight at study initiation: 26 g to 31 g
- Housing: The mice were kept in groups in transparent macrolone cages (type 150, floor area 810 cm2) with six animals in each cage. The cages were cleaned and the bedding changed at least twice a week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: >= 5 days

ENVIRONMENTAL CONDITIONS
The study took place in animal room No. 4 provided with filtered air at a temperature of 21°C ± 3°C, relative humidity being at least 30 % and preferably not exceed 70 % and air changes 10 times/ hour. The room was illuminated to give a cycle of 12 hours light and 12 hours darkness. Light was on from 6 am to 6 pm.

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
10 %, 25 % and 50 %
No. of animals per dose:
6
Details on study design:
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: IMDS
- Criteria used to consider a positive response:
The average LN cell count as a feature for the activation of the immune system (local lymph nodes) is used for the calculation of a proliferation stimulation index compared to the vehicle control:
stimulation index (SI) = LN cell count (treatment group)/LN cell count (vehicle control)
A lymph node index of 1.4 was determined as positive threshold value for the mouse strain NMRI by EHLING et al. (2005) as well as VOHR and AHR (2005).

TREATMENT PREPARATION AND ADMINISTRATION:
On day 1, all animals were weighed, the thickness of ears was measured by using a micrometer (Oditest S0247) and an amount of 25 µl of the test item or of the vehicle was applied topically on the dorsal side of the ears.
The procedure of this open application was repeated on day 2 and 3 with the different concentrations of the test item or with the vehicle.

Results and discussion

In vivo (LLNA)

Results
Parameter:
SI
Remarks on result:
other: 10%: 0.96 25%: 1.08 50%: 1.01

Any other information on results incl. tables

As compared to the negative control, no increase in ear thickness and weight and lymph node weight was observed.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
In a study according to OECD test guideline 429, the test substance was found to be not skin sensitising.
Executive summary:

The skin sensitising potential of the test substance was investigated in a study according to OECD test guideline 429 under GLP. Using the vehicle acetone/olive oil, concentrations of 10, 25 and 50% were applied topically (open) on the dorsal side of the ears of NMRI six mice each. This application was repeated twice (24 and 48h after the first application). On day 4, after humane killing of the mice, the lymph nodes were removed and the cells suspended. In addition to the primary endpoint of cell count (expressed as stimulation index), ear thickness and weight and lymph node weight were measured. As the stimulation index for all three tested concentration was clearly below 1.4, i.e. the threshold for a positive result, the test substance is not considered to be a skin sensitiser.

As also for the other endpoints no increase was observed, it can additionally be concluded that the test substance is not skin irritating.