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)
Deviations:
yes
Remarks:
The relative humidity in the animal room was between aproximately 45 - 85 % and the temperature was 22 +- °C for few hours. This deviation to the study plan, however, does not affect the validity of the study.
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymphnode assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
SAT 080004
Batch No.: Huc-Sa-0370-190
Purity: > 98% (NMR)

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female

Study design: in vivo (LLNA)

Vehicle:
other: acetone : olive oil (4+1)
Concentration:
25, 50, 100 %
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 100 % of the undiluted test item. The solutions were formulated in acetone:olive oil (4+1). According to the information provided by the sponsor SAT 080004 is stable in acetone:olive oil (4+1) for up to 4h. In methyl ethyl ketone, a solution of 50% could be achieved after vortexing.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 50 and 100% 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 25, 50, and 100%. 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 melted in a drying oven at max. 60°C for homogenisation. Prior to the dose preparation it was cooled to approximately 37°C.The test item was placed into a volumetric flask on a tared balance and acetone:olive oil (4+1) 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)
Statistics:
A statistical analysis was conducted for assessment of the dose-response relationship, and the EC3 value was calculated according to the equation
EC3 = (a-c) [(3-d)/(b-d)] + c
where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a, b) and (c, d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I. value of 3 on the local lymph node assay dose response plot.

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 25, 50 and 100% were 2.11, 4.47 and 4.80 respectively.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
1137.1, 2407.8, 2588.9 per lymph node Mean DPM/animal values for the experimental groups treated with test substance concentrations 25, 50 and 100% were 9097, 19262 and 20711 respectively. The mean DPM/animal value for the vehicle control group was 4312.

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 2.11, 4.47, and 4.80 were determined with the test item at concentrations of 25, 50, and 100% in acetone:olive oil (4+1). The EC3 value calculated was 34.4%.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
The test item SAT 080004 was found to be a skin sensitiser under the described conditions.
According to the Regulation (EC) No 1272/2008 on classification, labeling and packaging (CLP) of substances and mixtures, SAT 080004 should
be classified as skin sensitiser (Category 1).
According to the Dangerous Substance Directive, 67/548/EEC, which is currently undergoing a phase-out process, SAT 080004 should be labeledas: may cause sensitisation by skin contact (R43).
According to the ECETOC classification scheme for potency (ECETOC Technical Report No. 87, Contact sensitization: Classification according to
potency, April 2003, Bruessels), SAT 080004 would be regarded as weak sensitiser.
Executive summary:

In the study the test item SAT 080004 dissolved in acetone:olive oil (4+1) was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 25, 50, and 100%.

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 2.11, 4.47, and 4.80 were determined with the test item at concentrations of 25, 50, and 100% in acetone:olive oil (4+1), respectively.

The test item SAT 080004 was found to be a skin sensitiser in this assay and an EC3 value of 34.4% was derived.

According to the Regulation (EC) No 1272/2008 on classification, labeling and packaging (CLP) of substances and mixtures, SAT 080004 should be classified as skin sensitiser (Category 1).

According to the Dangerous Substance Directive, 67/548/EEC, which is currently undergoing a phase-out process, SAT 080004 should be labeled as: may cause sensitisation by skin contact (R43).

According to the ECETOC classification scheme for potency (ECETOC Technical Report No. 87, Contact sensitization: Classification according to potency, April 2003, Bruessels), SAT 080004 would be regarded as weak sensitiser.