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: Guideline study and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
SAT 080003
Batch No.: Huc-Sa-9370-163
Purity >99% (NMR)

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female

Study design: in vivo (LLNA)

Vehicle:
methyl ethyl ketone
Concentration:
5, 10, 25 %
No. of animals per dose:
4
Details on study design:
A solubility experiment was performed according to the recommendations given by OECD 429. The highest test item concentration, which can be
technically used, was a 25 % (w/v) solution in methyl ethyl ketone while warming at 37°C and after vortexing. Grinding in a
mortar was necessary before formulating the test item. According to the information provided by the sponsor SAT 080003 is stable in methyl
ethyl ketone for up to 4h. In acetone:olive oil (4+1), a solution of 20% could be achieved while warming and sonicating.
In all other tested vehicles (dimethylformamide, propylene glycol, dimethylsulfoxide, ethanol:deionised water (7+3) and ethanol:deionised water
(3+7)) the test item could not be formulated.To determine the highest non-irritant test concentration, a pre-test was performed in two animals.
Two mice were treated with concentrations of 10 and 25% each on three consecutive days.
In the pre-test clinical signs were recorded within 1 hour and 24 ± 4 hours after each application as well as on day 7. At the tested concentrations
the animals did not show any signs of irritation or systemic toxicity.
The test item in the main study was assayed at 5, 10, and 25%. The top dose is the highest technically achievable concentration whilst avoiding
systemic toxicity and excessive local irritation. No severe irritant effects were tolerated choosing the test concentrations.
The test item was grinded in a mortar before formulation and placed into a volumetric flask on a tared balance and methyl ethyl ketone was
quantitatively added. The different test item concentrations were prepared by serial dilution.
The preparations were made freshly before each dosing occasion. Concentrations were in terms of material as supplied.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
EC3 = 12.9% (w/v)

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: The SI values calculated for the substance concentrations 5, 10 and 25% were 1.11, 0.88 and 0.72 respectively.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Mean DPM/animal values for the experimental groups treated with test substance concentrations 5, 10 and 25% were 5292, 4192 and 3451 respectively. The mean DPM/animal value for the vehicle control group was 4769.

Any other information on results incl. tables

All treated animals survived the scheduled study period and no signs of toxicity were observed.

A test item is regarded as a sensitiser in the LLNA if the exposure to one or more test concentration resulted in 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls, as indicated by the Stimulation Index (S.I.). The estimated

concentration of test item required to produce a S.I. of 3 is referred to as the EC3 value.

In this study Stimulation Indices of 1.11, 0.88, and 0.72 were determined with the test item at concentrations of 5, 10, and 25% in methyl ethyl ketone. The EC3 value could not be calculated, since none of the tested concentrations induced an S.I. greater than 3.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The test item SAT 080003 was not a skin sensitiser under the described conditions.
Executive summary:

In the study the test item SAT 080003 dissolved in methyl ethyl ketone was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 5, 10, and 25% (highest achievable concentration).

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed.

In this study Stimulation Indices (S.I.) of 1.11, 0.88, and 0.72 were determined with the test item at concentrations of 5, 10, and 25% in methyl ethyl ketone, respectively.

The test item SAT 080003 was not a skin sensitiser in this assay.