N-{[4-(2,2-DICYANO-1-HYDROXYETHENYL)PHENYL]METHYL}-5-FLUORO-2-METHOXYBENZAMIDE

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30th June 2020 to 10th July 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)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-{[4-(2,2-DICYANO-1-HYDROXYETHENYL)PHENYL]METHYL}-5-FLUORO-2-METHOXYBENZAMIDE

IUPAC Name:

N-{[4-(2,2-DICYANO-1-HYDROXYETHENYL)PHENYL]METHYL}-5-FLUORO-2-METHOXYBENZAMIDE

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:

The protocol meets the requirements of the OECD guideline, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method” (TG 439)

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm Skin Model
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: approximately 25 mg

Duration of treatment / exposure:

The test articles, the positive control (5% Sodium Dodecyl Sulfate (SDS)), and the negative control (Calcium & Magnesium Free-Dulbecco’s Phosphate Buffered Saline (CMF-DPBS)), were treated in triplicate EpiDermTM tissues for a 60±1 minute exposure period.

Duration of post-treatment incubation (if applicable):

42-hour ± 2-hour post-exposure incubation

Number of replicates:

Triplicate

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of three replicates - 1 hour

Value:

99.98

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the prediction model presented in OECD TG 439, the test article was predicted to be a GHS No Category. 

Executive summary:

The EpiDerm™ Skin Model (MatTek Corporation, MA, USA) was used to assess the potential dermal irritation of the test article. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) conversion assay, which measures the NAD(P)H-dependent microsomal enzyme reduction of MTT (and to a lesser extent, the succinate dehydrogenase reduction of MTT) to a blue formazan precipitate, was used to assess cellular metabolism after exposure to a test article. The protocol was based upon theOECD guideline, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method” (TG 439). 

The test article was administered to the test system without dilution (neat). Since the test article was a powder, 25 µL of sterile Calcium & Magnesium Free Dulbecco’s Phosphate Buffered Saline (CMF-DPBS) were applied to the tissues prior to addition of approximately 25 mg of the test article, which was administered using a dosing spoon. A sterile rod was used to spread the test article over the surface of the tissues. Following a 60 ± 1 minute exposure
period, followed by a 42 ± 2-hour post-exposure incubation period, the mean percentage tissue viability was determined to be 99.98 ± 4.88. According to the prediction model presented in OECD TG 439, the test article was predicted to be a GHS No Category.