4-(2-acryloyloxy-ethoxy) benzophenone

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

2019

Deviations:

no

Remarks:

No deviations ocurred that negatively impacted the integrity of the study.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-(2-acryloyloxy-ethoxy) benzophenone

EC Number:

836-681-3

Cas Number:

22421-66-5

Molecular formula:

C18H16O4

IUPAC Name:

4-(2-acryloyloxy-ethoxy) benzophenone

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 3M Company, Lot 30029
- Expiration date of the lot/batch:
- Purity test date: 03 May, 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature.
- Stability under storage conditions: Stable
- Stability under test conditions: Stable
- Reactivity of the test substance with the solvent/vehicle /test medium (if applicable): No reactivity observed.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, tested as supplied.

FORM AS APPLIED IN THE TEST: The test article was tested as a waxy solid without further manipulation on the surface of the tissues.

In vitro test system

Test system:

human skin model

Remarks:

EpiDerm Reconstructed Human Epidermis (RhE)

Source species:

human

Cell type:

other: Reconstructed Human Epidermis - normal human keratinocytes

Cell source:

other: Keratinocyte strain 00267

Details on animal used as source of test system:

SOURCE ANIMAL
- Source: Huuman
- Sex: No data
- Age at study initiation (in days): No data

Justification for test system used:

EpiDerm RhE is recommended per OECD 439.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm RhE
- Tissue batch number(s): Lot 33722, Keratinocyte Strain 00267
- Production date: 9/16/2020
- Shipping date: 14 September, 2020
- Delivery date: 15 September, 2020
- Date of initiation of testing: 16 September, 2020

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 C
- Temperature of post-treatment incubation (if applicable): 37 C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: At the end of exposure, tissues were washed with a stream of DPBS which completely removed the control and test articles/substances from the tissue inserts. Tissues were blotted with a sterile cotton swab (inside) and on sterile paper toweling (outside) of the insert, transferred to a fresh plate containing pre-warmed medium returned to the incubator.
- Observable damage in the tissue due to washing: None observed.
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL in DMEM
- Incubation time: 3 hours
- Spectrophotometer: Molecular Devices model i3 plate reader
- Wavelength: 570 nm
- Filter: No data
- Filter bandwidth: No data
- Linear OD range of spectrophotometer: No data

NUMBER OF REPLICATE TISSUES: Each group was dosed in triplicate (n=3)

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
The test article/substance was found to be an MTT reductant capable of interference with the MTT assay. Freeze/killed (F/K) tissues were run in parallel with viable tissues as a control as described in the protocol. The F/K tissue control demonstrated that the test article/substance did not interfere with the MTT assay.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: One

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
According to OECD TG 439, an irritant is predicted if the mean relative tissue viability of three individual tissues exposed to the test substance is reduced to or below 50% of the mean viability of the negative controls.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg

VEHICLE : None
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

NEGATIVE CONTROL : DPBS
- Amount(s) applied (volume or weight): 30 uL

POSITIVE CONTROL : SDS
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): 5%

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean

Value:

105

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: None observed
- Direct-MTT reduction: The test article/substance was found to be an MTT reductant capable of interference with the MTT assay. Freeze/killed (F/K) tissues were run in parallel with viable tissues as a control as described in the protocol. The F/K tissue control demonstrated that the test article/substance did not interfere with the MTT assay.
- Colour interference with MTT: None observed.

DEMONSTRATION OF TECHNICAL PROFICIENCY: The lab that run the study is technically proficient in running the OECD 439.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes, the mean Optical Density (OD) of the negative control (NC) group was ≥ 1.0 and ≤ 2.8.
- Acceptance criteria met for positive control: Yes, the mean viability of the 60 min 5% SDS-treated positive control (PC) group tissues was ≤ 20 % of the NC.
- Acceptance criteria met for variability between replicate measurements: Yes, the standard deviation (SD) for the test substance groups (n=3) was < 18%.
- Range of historical values if different from the ones specified in the test guideline: Per the guideline.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study indicating a tissue viability of 105% compared to the negative control, AEBP is not a skin irritant when tested according to OECD 439.

Executive summary:

The skin irritation potential of AEBP was evaluated using the reconstructed human epidermal tissue model EpiDerm (EPI-200SIT). The study was conducted according to OECD 439 (2019) in compliance with OECD GLP. The test article (waxy solid) was dosed as received by covering the apical surface of each tissue with 25 mg of test article. Positive (5% SDS) and negative (DPBS) controls were run in parallel. All test article and control exposures were run in triplicate (n=3). Tissues were exposed for 60 minutes at 37 C, washed and incubated for 42 hours at 37 C. Following post-exposure incubation, viability of the tissues was determined using the MTT assay. Tissue viability was expressed as a percent of the negative control. Positive and negative control tissues performed as expected, indicating that the test system was valid. Exposure to AEBP resulted in a mean tissue viability of 105% (4% SD) compared to the negative control. The mean optical density was 2.351 for the treated tissues. Based on the results of the study indicating a tissue viability of 105% compared to the negative control, AEBP is not a skin irritant when tested according to OECD 439.