[(3-Oxo-butyryl)-phenyl-amino]-acetic acid ethyl ester

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

n-phenyl n-acetoacetyl glycine ethyl ester

IUPAC Name:

n-phenyl n-acetoacetyl glycine ethyl ester

Test material form:

liquid

Specific details on test material used for the study:

TEST MATERIAL NAME (as stated in study report): Intermediate F37

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Valtris Specialty Chemicals / Lot No. W042272
- Expiration date of the lot/batch: November 21, 2019
- Purity: 84.5%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature In original container as supplied.

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: not specified

Justification for test system used:

Recommended by the OECD and other regulatory authorities

Vehicle:

unchanged (no vehicle)

Details on test system:

Reconstructed human epidermis tissues, SkinEthicTM RHE model, were procured from SkinEthic Laboratories, Episkin 4, Rue Alexander Fleming, 69366 Lyon Cedex 07, France.

After exposure, tissues were rinsed and then dried with cotton buds. Treated tissues were rinsed 20 times in a constant soft stream of 1 mL DPBS at a 5-8 cm distance from the insert to remove all residual test item from the epidermal surface. Mesh (applied on test item treated, negative and positive control tissues) was removed by washing for all tissues. The bottom of tissue inserts were dried on sterile absorbent paper (Kim wipes) for 1-2 seconds. The surface of the stratum corneum was gently swept up using both ends of a cotton tip (5-6 turns per end). After washing, inserts were transferred to holding plates containing 300 µL maintenance medium.

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: SkinEthicTM RHE mode
- Tissue batch number(s): Lot N° 18 RHE 029

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 min at room temperature; 60 min at 37 deg C

REMOVAL OF TEST MATERIAL AND CONTROLS
After exposure, tissues were rinsed and then dried with cotton buds. Treated tissues were rinsed 20 times in a constant soft stream of 1 mL DPBS at a 5-8 cm distance from the insert to remove all residual test item from the epidermal surface. Mesh (applied on test item treated, negative and positive control tissues) was removed by washing for all tissues. The bottom of tissue inserts were dried on sterile absorbent paper (Kim wipes) for 1-2 seconds. The surface of the stratum corneum was gently swept up using both ends of a cotton tip (5-6 turns per end). After washing, inserts were transferred to holding plates containing 300 µL maintenance medium.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
After rinsing and drying, the MTT test was performed. Tissues were placed in 300 µL of MTT (1.0 mg/mL) solution and incubated for 180 minutes at 37±1 °C in 5±1% CO2 in a 95% humidified incubator. At the end of the MTT test, tissues were observed for MTT reduction.

NUMBER OF REPLICATE TISSUES: three

PREDICTION MODEL / DECISION CRITERIA
per test guideline

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 40 µL/0.5 cm2
- Concentration (if solution): undiluted

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 40 µL/0.5 cm2
- Concentration (if solution): undiluted

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 40 µL/0.5 cm2
- Concentration (if solution): 8N

Duration of treatment / exposure:

3 minutes at room temperature
60 minutes at 37 ± 1 °C

Number of replicates:

Three

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minute exposure at room temperature

Value:

100.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not examined

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minute exposure at 37 ± 1 °C

Value:

92.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: No
- Colour interference with MTT: slight interference

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Not classified by CLP Criteria

Conclusions:

Under guideline (OECD 431) in vitro test conditions, Intermediate F37 was not corrosive to reconstructed human epidermis tissues (SkinEthicTM RHE model).