Ozonized sunflower oil

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 442E (In Vitro Skin Sensitisation assays addressing the key event on activation of dendritic cells on the Adverse Outcome Pathway for skin sensitisation)

Version / remarks:

30 June 2022

Qualifier:

according to guideline

Guideline:

other: OECD Guideline No. 497: Defined Approaches for Skin Sensitisation - Annex I. Prediction model for the individual in chemico/in vitro tests with multiple runs for use in 2o3 DA

Version / remarks:

14 June 2021

GLP compliance:

yes

Type of study:

human Cell Line Activation Test (h-CLAT)

Details of test system:

THP-1 cell line [442E]

Vehicle / solvent control:

DMSO

Negative control:

other: DMSO

Positive control:

dinitrochlorobenzene (DNCB) [442E]

Key result

Group:

test chemical

Run / experiment:

run/experiment 1

Parameter:

CV70 [442E]

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Remarks:

Since no cytotoxicity was observed either no CV75 value was calculated

Outcome of the prediction model:

negative [in vitro/in chemico]

Interpretation of results:

GHS criteria not met

Conclusions:

Based on these results, the OECD 442E and OECD 497 prediction models, the test item “Ozonia 3000 Sunflower (Ozonized Sunflower Seed Oil)” was shown to be negative and demonstrated in vitro non-sensitising potential under the experimental conditions of the human Cell Line activation Test.

Executive summary:

In the course of this study the skin sensitisation potential of “Ozonia 3000 Sunflower (Ozonized Sunflower Seed Oil)” was examined.

The extent of cytotoxicity induced on THP-1 cells by the test item was studied in two runs of the dose finding test. The maximal final concentrations used on the plates for the test item previously dissolved in DMSO were 1000.0 μg/mL (first run) and 1001.0 μg/mL (second run). Since no cytotoxicity was observed in the first run, the same concentration range was used in the second run and start from the highest allowed concentration (1000 μg/mL). Since in the second run no cytotoxicity was observed either, no CV75 value was calculated. Based on the results of two runs of the dose finding test, eight doses between 1000.0 μg/mL – 279.1 μg/mL (nominal concentrations) were used for the main test in two independent runs. Both runs were concluded valid.

The increase in CD86 marker expression (RFI) was lower than 150 % at all tested doses (with > 50 % of cell viability) compared to the respective negative controls in both valid runs. Moreover, according to the OECD Test Guideline 497 prediction model, the increase in CD86 marker expression (RFI) was below the borderline range (122-184 %) at all tested concentrations compared to the respective negative controls in all independent runs. Also, no cytotoxicity was observed at any tested concentrations in either independent run. Based on the two valid concordant negative runs, CD86 marker expression was concluded to be negative. Effective concentration for CD86 expression (EC150) was not determined, since a negative result was obtained.

The increase in CD54 marker expression (RFI) was lower than 200 % at all tested concentrations compared to the respective negative controls in both independent valid runs. Moreover, according to the OECD Test Guideline 497 prediction model, the increase in CD54 marker expression (RFI) was below the borderline range (157-255 %) at all tested concentrations compared to the respective negative controls in both independent runs. Furthermore, no cytotoxicity was observed at any tested concentrations in either independent run.

Based on the two valid concordant negative runs, CD54 marker expression was concluded to be negative. Effective concentration for CD54 expression (EC200) was not determined, since a negative result was obtained.

 

Since both CD86 and CD54 marker expressions gave negative results, the overall h-CLAT prediction was also concluded to be negative.

Reference 2

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 luciferase KeratinoSens™ test method)

Version / remarks:

30 June 2022

Qualifier:

according to guideline

Guideline:

other: OECD Guideline No. 497: Defined Approaches for Skin Sensitisation - Annex I. Prediction model for the individual in chemico/in vitro tests with multiple runs for use in 2o3 DA

Version / remarks:

14 June 2021

GLP compliance:

yes

Type of study:

ARE-Nrf2 luciferase KeratinoSens™ test method

Details of test system:

Keratinoses transgenic cell line [442D]

Vehicle / solvent control:

DMSO

Negative control:

other: DMSO

Positive control:

cinnamic aldehyde [442D]

Positive control results:

The positive control used was Trans-Cinnamaldehyde for which a series of five 100 × master concentrations ranging from 0.4 to 6.4 mM were prepared in DMSO (from a 200 mM stock solution) and diluted as described for the 4 × master solutions. The final concentration of the positive control on the treated plates ranged from 4 to 64 μM.

Key result

Group:

test chemical

Run / experiment:

run/experiment 1

Parameter:

EC 1.5 [442D]

Value:

220.25 µg/mL

Cell viability:

yes (≥70%)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of skin sensitisation

Group:

test chemical

Run / experiment:

run/experiment 2

Parameter:

IC50 [442D]

Cell viability:

no (<70%)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Remarks:

The test item induced no cytotoxicity (or viability below 70 %) compared to the solvent/vehicle control at any concentrations in KeratinoSens™ cells. Thus, IC30 and IC50 values were not calculated.

Group:

test chemical

Run / experiment:

run/experiment 3

Parameter:

EC 1.5 [442D]

Cell viability:

no (<70%)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Remarks:

The test item induced no cytotoxicity (or viability below 70 %) compared to the solvent/vehicle control at any concentrations in KeratinoSens™ cells. Thus, IC30 and IC50 values were not calculated.

Outcome of the prediction model:

negative [in vitro/in chemico]

Interpretation of results:

GHS criteria not met

Conclusions:

Based on these results and prediction models of OECD 442D and OECD 497, the test item “Ozonia 3000 Sunflower (Ozonized Sunflower Seed Oil)” is concluded borderline for skin sensitisation potential (between skin sensitization and non-skin sensitization) under the experimental conditions of KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

Executive summary:

In the course of this study the skin sensitisation potential of the test item “Ozonia 3000 Sunflower (Ozonized Sunflower Seed Oil)” was studied using the KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method). In order to derive a prediction for the test item the results of three independent tests were used. Since the results of the two tests were not concordant, a third was needed in order to derive a conclusion. The luciferase activity induction obtained with the positive control, Trans-Cinnamaldehyde, was dose-dependent and statistically significant above the threshold of 1.5-fold in all tests. Since no defined molecular weight is available for the test item, 40 mg/mL final concentration of the test item was used in the study instead of 200 mM in accordance with the test guideline. For the test item, twelve doses ranging from 400.00 μg/mL to 0.20 μg/mL were used in all valid tests. The test item induced no cytotoxicity (a substance is considered cytotoxic if the viability is below 70 %) in KeratinoSens™ cells compared to the solvent/vehicle control. Thus, in none of the tests IC30 or IC50 values could be determined. The induction values of the test item did not exceed the 1.5-fold threshold at any tested concentrations compared to the respective negative control in the second and third tests. In the first test, there was only one induction value of the test item which exceeded the 1.5-fold threshold compared to the respective negative control at the highest tested concentration (400.00 μg/mL). Moreover, according to the OECD Test Guideline 497 prediction model, there were one or two concentrations per tests where the induction values of the test item were within or above the borderline threshold range (1.35-1.67-fold) compared to the respective negative control in all three valid tests. Because of the obtained results and based on the predictions models, the test item was concluded borderline. The EC1.5 value was determined for the first test only and it was calculated as 220.25 μg/mL. Furthermore, the overall Imax value was determined as 1.65-fold. In addition, a clear dose response could be observed throughout the study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)