Isopropyl 2-ethylhexanoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 20, 2017 to November 09, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Isopropyl 2-ethylhexanoate

EC Number:

266-549-5

EC Name:

Isopropyl 2-ethylhexanoate

Cas Number:

67024-46-8

Molecular formula:

C11H22O2

IUPAC Name:

isopropyl 2-ethylhexanoate

Test material form:

liquid

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

The EpiOcular™ human cell construct (MatTek Corporation) is used in this assay.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Approximately 50 μL test item was applied topically onto the EpiOcular™ tissues.

Duration of treatment / exposure:

The plates with the treated tissue units were incubated for the exposure time of 30 minutes (± 2 min) at standard culture conditions (37±1°C in an incubator with 5±1% CO2, 90±10% humidified atmosphere).

Duration of post- treatment incubation (in vitro):

After the post-incubation the EpiOcular™ units were transferred into the MTT ready to use solution filled 24-well plate (300 μL of 1 mg/mL MTT per well) and then incubated for 3 hours (± 10 min) at 37±1°C in an incubator with 5±1 % CO2 protected from light, 90±10% humidified atmosphere.

Number of animals or in vitro replicates:

Two replicates were used for the test item and control(s) respectively.

Details on study design:

Disks of EpiOcular™ (two units) were treated with (50 μL/units) test item and incubated for 30 minutes (± 2 min) at standard culture conditions (37±1°C) in an incubator with 5±1 % CO2, 90±10% humidified atmosphere).
Exposure to the test item was terminated by rinsing with Ca++Mg++ Free-DPBS solution. After rinsing, the tissues were incubated for a 12 ± 2 minutes immersion incubation (Post-Soak) at room temperature. At the end of the Post-Soak immersion the test item treated tissues were incubated for 120 minutes ± 15 minutes at standard culture conditions (Post-treatment Incubation). Fresh Assay Medium was used during the Post-Soak and Post-Incubation. The viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37±1°C in an incubator with 5±1 % CO2 protected from light, 90±10% humidified atmosphere. The precipitated formazan was then extracted using isopropanol and quantified spectrophotometrically.

Results and discussion

In vitro

Results

Irritation parameter:

other: Tissue viability

Remarks:

The test item is identified as not requiring classification and labelling according to UN GHS (No Category) if the mean percent tissue viability after exposure and post-exposure incubation is more than (>) 60%.

Run / experiment:

Run 1, viability 80%; Run 2, viability 76%.

Value:

76

Vehicle controls validity:

valid

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:

The results obtained from this in vitro eye irritation test, using the EpiOcular™ model, indicated that the test item has no eye irritation potential under the testing conditions used. According to the current OECD Guideline No. 492, Isopropyl 2-ethylhexanoate is, thus, considered as non-irritant to eye (UN GHS No Category).