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Description of key information

Key value for chemical safety assessment

Skin sensitisation

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Endpoint:
skin sensitisation: in chemico
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
Deviations:
no
GLP compliance:
yes
Type of study:
other: Direct Peptide Reactivity Assay
Details on study design:
The test item was tested by incubation with peptides containing either cysteine or lysine for 24 ± 2 h at 25 ± 2.5 °C. Subsequently samples were analysed by HPLC.
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 64.92%.
Parameter:
other: percentage depletion value of the cysteine
Run / experiment:
Mean depletion of the cysteine peptide
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of skin sensitisation
Parameter:
other: percentage deplation value of the lysine
Run / experiment:
Mean deplation of the lysine peptide
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
not determinable because of methodological limitations
Other effects / acceptance of results:
The samples containing test item and lysine peptide showed a distinct phase separation and furthermore a significant turbidity in both phases after the incubation time. Therefore, HPLC measurement of the samples was not possible and no results concerning the lysine peptide depletion of the test item are reported.
Conclusions:
In this study under the given conditions the test item showed moderate reactivity towards the cysteine peptide. The test item can be classified as “non-sensitiser” in accordance with UN GHS “Category 1”.
Executive summary:

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.

In the present study TEA Trioleate was dissolved in water.

The test item was tested by incubation with peptides containing either cysteine or lysine for 24 ± 2 h at 25 ± 2.5 °C. Subsequently samples were analysed by HPLC.

The samples containing test item and lysine peptide showed a distinct phase separation and furthermore a significant turbidity in both phases after the incubation time. Therefore, HPLC measurement of the samples was not possible and no results concerning the lysine peptide depletion of the test item are reported.

After the 24 h ± 2 h incubation period but prior to the HPLC analysis the cysteine peptide samples were inspected for precipitation, turbidity or phase separation. No precipitation, turbidity or phase separation was observed for the test item samples. A slight precipitation was observed for standard 1 and the positive control samples. Since the acceptance criteria for the linearity of the standard curve as well as for the depletion range of the positive control were fulfilled, the observed precipitations were regarded as insignificant.

No co-elution of test item with the cysteine peptide peak was observed. Since the lysine peptide samples could not be measured, sensitising potential of the test item was predicted only from the peptide depletion of the cysteine peptide by comparing the peptide concentration of the test item treated samples to the corresponding reference control C (RC C).

The test item showed no reactivity towards the synthetic peptide. The mean depletion of the cysteine peptide was < 13.89% (0.00%). Based on the prediction model 2 the test item can be considered as non-sensitiser.

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 64.92%.

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: OECD Guidelines for Testing of Chemicals, No. 442E: "In vitro Skin Sensitisation: human Cell Line Activation Test (h-CLAT)", adopted 29 July 2016.
Deviations:
no
GLP compliance:
yes
Type of study:
activation of dendritic cells
Specific details on test material used for the study:
The correlation of upregulation of immunological relevant cell surface markers with the skin sensitising potential of a chemical has been reported and represents the third key event in the skin sensitisation process as described by the AOP. This method that measures the markers of DC activation, based on DC-like cell line THP-1 is considered relevant for the assessment of the skin sensitisation potential of chemicals.
Remarks on result:
no indication of skin sensitisation
Conclusions:
In this study under the given conditions the test item did not upregulate the expression of the cell surface marker in at least two independent experiment runs. Therefore the test item considered to be no skin sensitiser.
The data generated with this method may be not sufficient to conclude on the absence of skin sensitisation potential of chemicals and should be considered in the context of integrated approach such as IATA.
Executive summary:

Thein vitro human cell line activation test (h-CLAT) enables detection of the sensitising potential of a test item by addressing the third molecular key event of the adverse outcome pathway (AOP), namely dendritic cell activation, by quantifying the expression of the cell surface markers CD54 and CD86 in the human monocytic cell line THP-1. The expression of the cell surface markers compared to the respective solvent controls is used to support discrimination between skin sensitisers and non-sensitisers.

In the present study TEA Trioleate was dissolved in THF. For the dose finding assay stock solutions with a concentration of 500 mg/mL to 3.91 mg/mL were prepared by a serial dilution of 1:2. Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained with propidium iodide and cell viability was measured by FACS analysis. No cytotoxicity was observed in the dose finding assay. Precipitation was observed for the four highest concentrations.

Based on the CV75, the main experiment was performed covering the following concentration steps:

1000; 833.33, 694.44, 578.70, 482.25, 401.88, 334.90, 279.08 µg/mL

Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained and cell surface markers CD54 and CD86 were measured by FACS analysis. Cell viability was assessed in parallel using propidium iodide staining.

No cytotoxic effects were observed for the cells treated with the test item. Relative cell viability at the highest test item concentration was 96.2% (CD86), 97.2% (CD54) and 96.9% (isotype IgG1 control) in the first experiment, and 99.5% (CD86), 96.7% (CD54) and 96.8% (isotype IgG1 control) in the second experiment.

The expression of the cell surface marker CD86 was not upregulated above the threshold of 150% in any of the experiments. The expression of cell surface marker CD54 was not upregulated above the threshold of 200% in any of the experiments. Therefore, the test item is considered to be no skin sensitiser.

The positive control (DNCB) led to an upregulation of the expression of CD54 and CD86 in both experiments. The threshold of 150% for CD86 (429% experiment 1; 640% experiment 2) and 200% for CD54 (409% experiment 1; 669% experiment 2) were clearly exceeded.

The data generated with this test should be considered in the context of integrated approached such as IATA, combining the result with other complementary information, e.g. derived from in vitro assays addressing other key events of the skin sensitisation AOP.

1.1. Conclusion

In this study under the given conditions the test item did not upregulate the expression of the cell surface marker in at least two independent experiment runs. Therefore the test item is considered to be no skin sensitiser.

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
GLP compliance:
yes
Type of study:
activation of keratinocytes
Key result
Remarks on result:
no indication of skin sensitisation

In the experiments, no significant luciferase induction of > 1.5 was found in the tested concentration range. Therefore, no EC1.5 could be calculated.

No dose response for luciferase activity induction was observed for each individual run as well as for an overall luciferase activity induction.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the condition of this study the test item is therefore considered as non sensitiser.
Executive summary:

In the present study TEA Trioleate was dissolved in tetrahydrofuran (THF). However, an oily film was noted on the surface after dissolution of the test item.

Cells were incubated with the test item for 48 h at 37°C. After exposure cells were lysed and luciferase activity was assessed by luminescence measurement.

In the experiments, no significant luciferase induction of > 1.5 was found in the tested concentration range. Therefore, no EC1.5 could be calculated.

No dose response for luciferase activity induction was observed for each individual run as well as for an overall luciferase activity induction.

In this study under the given conditions the test item did not induce the luciferase activity in the transgenic KeratinoSens™ cell line in at least two independent experiment runs. Based on these results, the test item might be considered as non sensitiser.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

According to Regualtion (EC) n. 1272/2008, TEA trioleate is not classificable as skin sensitizer.