Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
GLP compliance:
yes (incl. certificate)
Type of study:
direct peptide binding assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Pentanol
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Tk287_20151217
- Expiration date of the lot/batch: 17.12.2016
- Purity test date: 20.04.2016
- Purity: 99.9%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: guaranteed by the sponsor
- Solubility and stability of the test substance in the solvent/vehicle: soluble, no analysis performed because the dissolved test substance was used shortly after preparation

In vitro test system

Details on study design:
The test substance is incubated with synthetic peptides for 24 hours at room temperature in the dark. The remaining non-depleted peptide concentration is determined thereafter by high performance liquid chromatography with gradient elution and UV-detection at 220 nm.
The peptide depletion of test-substance incubated samples is compared to the peptide depletion of the NC samples and expressed as relative peptide depletion.

Synthetic peptides:

Cysteine- (C-) containing peptide:
Ac-RFAACAA-COOH (MW=751.9 g/mol)

Lysine- (K-) containing peptide:
Ac-RFAAKAA-COOH (MW=776.2 g/mol)

The peptides are custom material (Supplier: GenScript, Piscataway, NJ, USA and RS Synthesis, Louisville KY, USA) containing phenylalanine to aid in detection and either cysteine or lysine as the reactive center.

Concentrations:
C-peptide: 5mM test substance, 0.5mM peptide
K-peptide: 25mM test substance, 0.5mM peptide
Test substance preparations were prepared on a weight per volume basis within 4 hours of the start of the experiment.
No. of replicates: 3

Vehicle control: acetonitrile
Reason for vehicle: solubility
Positive control: Ethylene glycol dimethacrylate (50mM in acetonitrile)

Results and discussion

In vitro / in chemico

Resultsopen allclose all
Key result
Parameter:
other: Peptide depletion
Value:
1.44
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid
Parameter:
other: C-peptide depletion
Value:
-1.46
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid
Parameter:
other: K-peptide depletion
Value:
2.88
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid
Other effects / acceptance of results:
The standard calibration curve should have an r² >0.99.
The vehicle control samples should be 0.50 mM +/- 0.05mM.
The CV of the nine vehicle controls should be < 15%.
The variability between these samples should be acceptably low (SD <14.9% for % cysteine depletion and <11.6% for % lysine depletion).
The positive control should cause depletion of both peptides comparable to historic data.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met