1H-Isoindole-1,3(2H)-dione, 2-[[(5S)-2-oxo-3-[4-(3-oxo-4-morpholinyl)phenyl]-5-oxazolidinyl]methyl]-

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jan 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD 431 guideline draft

Version / remarks:

1996/ 1998

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-({(5S)-2-Oxo-3-[4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl}methyl)-1H-isoindole-1,3(2H)-dione

EC Number:

610-201-0

Cas Number:

446292-08-6

Molecular formula:

C22 H19 N3 O6

IUPAC Name:

2-({(5S)-2-Oxo-3-[4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl}methyl)-1H-isoindole-1,3(2H)-dione

Specific details on test material used for the study:

purity 99.8%

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

Corrosion of the skin is defined as an irreversible necrotic alteration of the tissue induced by a chemical. While the testing for corrosives usually involves the rabbit skin as a predictive in vivo test method, substances may be classified according to their corrosive potential by the determination of their cytotoxic effects on an in vitro reconstructed human epidermis. The test principle is based on the MTT assay reflecting the cell viability after short term exposure of the epidermal equivalent to topically applied Oxaphthalimid.

Vehicle:

physiological saline

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The experiment was carried out on a reconstructed human epidermis EST-1000 (CellSystems, St. Katharinen, Germany).

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: RT: 3 min; 37 ± 2°C: 60 min
- Temperature of post-treatment incubation (if applicable):

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 3 washing steps, volume not reported
- Observable damage in the tissue due to washing: No

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 2 h
- Spectrophotometer: (EL808, Bio-Tek; 96 well format, 200 µL)
- Wavelength: 570 nm


NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues: not reported
- Procedure used to prepare the killed tissues (if applicable): not reported
- N. of replicates : not reported
- Method of calculation used: not reported

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: None

Control samples:

yes, concurrent negative control

Amount/concentration applied:

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


NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL

Duration of treatment / exposure:

3 min and 60 min

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1/ 60 min

Value:

95.27

Vehicle controls validity:

valid

Positive controls validity:

not examined

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 / 3 min

Value:

97.26

Vehicle controls validity:

valid

Positive controls validity:

not examined

Any other information on results incl. tables

The MTT method has determined the following values of viability:

after 3 min of incubation: 97.36%

after 60 min of incubation: 95.27%

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

the test item is not corrosive to skin

Executive summary:

The study on a reconstructed human epidermis EST-1000 was carried out for detection of topically applied skin corrosives with the test item Oxaphthalimid.

A 100% concentration was tested on the skin/epidermal equivalents in triplets. For the determination of time related cytotoxic effects the incubation periods were 3 minutes (room temperature) and 60 minutes (incubator), respectively. Using the MTT (methylthiazoletetrazolium) method the cell viability after 3 minutes or after 60 minutes of incubation was determined to be 97.26% and 95.27%, respectively. Thus, Oxaphthalimid was not characterised by a significant impact on cell viability and it therefore showed no corrosive properties.