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EC number: 821-932-1 | CAS number: 694510-10-6
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in chemico
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- June 02 - Nov 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 019
- Report date:
- 2019
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
- Qualifier:
- according to guideline
- Guideline:
- other: Direct Peptide Reactivity Assay (DPRA) for Skin Sensitization Testing, DB-ALM Protocol n°154, January 12, 2013
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Behörde für Gesundheit und Verbraucherschutz, Freie Handelsstadt Hamburg
- Type of study:
- direct peptide reactivity assay (DPRA)
Test material
- Reference substance name:
- trans-1-butyl-4-[trans-4-[(E)-prop-1-enyl]cyclohexyl]cyclohexane
- EC Number:
- 821-932-1
- Cas Number:
- 694510-10-6
- Molecular formula:
- C19 H34
- IUPAC Name:
- trans-1-butyl-4-[trans-4-[(E)-prop-1-enyl]cyclohexyl]cyclohexane
- Test material form:
- solid
Constituent 1
In chemico test system
- Details on the study design:
- The in chemico direct peptide reactivity assay (DPRA) enables detection of the sensitising potential of a test item by quantifying the reactivity of test chemicals towards synthetic peptides containing either lysine or cysteine.
Results and discussion
- Positive control results:
- The 100 mM stock solution of the positive control (cinnamic aldehyde) showed high reactivity towards the synthetic peptides. The mean depletion of both peptides was 68.60%.
In vitro / in chemico
Resultsopen allclose all
- Key result
- Run / experiment:
- other: cysteine run
- Parameter:
- other: mean peptide depletion [%]
- Value:
- 0
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: lysine run
- Parameter:
- other: mean peptide depletion [%]
- Value:
- 0
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- Acceptance Criteria
The following criteria must be met for a run to be considered valid:
a) The standard calibration curve should have an r2 > 0.99.
b) The mean percent peptide depletion value of the three replicates for the positive control cinnamic aldehyde should be between 60.8% and 100% for the cysteine peptide and between 40.2% and 69.0% for the lysine peptide and the maximum standard deviation (SD) for the positive control replicates should be < 14.9% for the percent cysteine depletion and < 11.6% for the percent lysine depletion.
c) The mean peptide concentration of reference controls A should be 0.50 ± 0.05 mM and the coefficient of variation (CV) of peptide peak areas for the nine reference controls B and C in acetonitrile should be <15.0%.
If one or more of these criteria is not met, the run would have been repeated.
The following criteria must be met for a test item’s results to be considered valid:
a) The maximum standard deviation for the test item replicates should be < 14.9% for the percent cysteine depletion and < 11.6% for the percent lysine depletion.
b) The mean peptide concentration of the three reference controls C in the appropriate solvent should be 0.50 ± 0.05 mM.
If these criteria were not met, the data would have been rejected and the run have been repeated for that specific test item.
Any other information on results incl. tables
Peptide Depletion in %
1) Cystein Depletion
| Sample |
Peak Area |
Actual Peptide [mM] |
Peak Are Ref Control C |
Peptide Depletion [%] |
Corr. peptide Depletion [%] |
| 1 |
50.9758 |
0.503 |
50.8281 |
-0.03 |
0.00 |
| 2 |
51.5350 |
0.509 |
51.0861 |
-1.13 |
0.00 |
| 3 |
51.5140 |
0.508 |
50.9661 |
-1.09 |
0.00 |
| Mean |
51.342 |
0.507 |
50.9601 |
-0.75 |
0.00 |
| Evaluation Reactivity Class. Negative |
|||||
1) Lysine Depletion
| Sample |
Peak Area |
Actual Peptide [mM] |
Peak Are Ref Control C |
Peptide Depletion [%] |
Corr. peptide Depletion [%] |
| 1 |
36.3497 |
0.464 |
35.0532 |
-2.95 |
0.00 |
| 2 |
36.4728 |
0.465 |
35.3563 |
-3.29 |
0.00 |
| 3 |
36.5649 |
0.466 |
35.5197 |
-3.55 |
0.00 |
| Mean |
36.462 | 0.465 |
35.3097 |
-3.26 |
0.00 |
| Evaluation Reactivity Class. Negative |
|||||
Applicant's summary and conclusion
- Interpretation of results:
- other: The result has to be considered in an integrated approach.
- Conclusions:
- The test material revealed a mean cysteine and lysine peptide depletion of 0.0% and, hence, the test item is considered negative and predicted to be a non-sensitiser (no or minimal reactivity) in the Direct Peptide Reactivity Assay (DPRA).
- Executive summary:
Pupose
The purpose of this study was to determine the sensitising potential of the test material in a Direct Peptide Reactivity Assay (DPRA).
Methods
The study was performed according to OECD guideline 442C. The DPRA is an in chemico method which quantifies the remaining concentration of cysteine- or lysine-containing peptide following incubation with the test item. Cysteine and lysine peptide percent depletion values are then calculated and used in a prediction model, which allows assigning the test item to one of four reactivity classes used to support the discrimination between sensitisers and nonsensitisers.
Results
The test item was dissolved at a concentration of 100 mM in acetone. No precipitate in the reaction mixture at the end of the incubation time and no co-elution were observed.
The test material-treated samples revealed a cysteine peptide depletion of 0.0% and a lysine peptide depletion of 0.0% (mean peptide depletion of 0.0%) and, hence, were well
below 6.38%. The test material is considered negative and predicted to be a nonsensitiser (no or minimal reactivity) in the Direct Peptide Reactivity Assay (DPRA).
Cinnamic aldehyde was used as positive control at a concentration of 100 mM in acetone. Treatment with the positive control item revealed a cysteine and lysine peptide depletion of
68.65% for cysteine and 59.73% for lysine peptide. These values are within the required range. Therefore, the study can be regarded as valid.
Conclusion
The test material revealed a mean cysteine and lysine peptide depletion of 0.0% and, hence, the test item is considered negative and predicted to be a non-sensitiser (no or
minimal reactivity) in the Direct Peptide Reactivity Assay (DPRA).
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