Fatty acids, coco, esters with oxybis(propanediol)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, coco, esters with oxybis(propanediol)

EC Number:

288-309-9

EC Name:

Fatty acids, coco, esters with oxybis(propanediol)

Cas Number:

85711-49-5

Molecular formula:

Not possible for UVCB

IUPAC Name:

Fatty acids, coco, esters with oxybis(propanediol)

Test material form:

solid

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

- Justification of the test method and considerations regarding applicability
- Description of the cell system used: After receipt of the SkinEthicTM HCE kit, all of the corneal epithelium units used in the study were pre-incubated in maintenance medium (provided by kit supplier) at 37 ± 2°C, 5 ± 1 % CO2, ≥ 90% humidity for at least overnight before dosing. Before application, the 2 corneal epithelium units for test substance, negative control substance and positive control substance were transferred to fresh pre-warmed maintenance medium, respectively. The application was conducted by gently spreading each substance on the epithelium without touching it to ensure the substance cover all the tissue surface, 2 units per substance. A nylon mesh was used if the test substance was sticky or viscous. The incubation was made at 37 ± 2°C, 5 ± 1 % CO2, ≥ 90% humidity for 30 ± 2 minutes.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

10 ± 1 µl 0.01M PBS pH = 7.4 and 30 ± 2 µl test item

Duration of treatment / exposure:

30 ± 2 min

Duration of post- treatment incubation (in vitro):

After treatment, each treated corneal epithelium unit was rinsed with 20 ml sterile 0.01M PBS pH 7.4 (approximately 10 ml per push) to remove the residue of test chemicals from the epithelium surface. After rinsing, remaining PBS was removed by energized reversal; a cotton-swabs was used to sweep the surface carefully when necessary. The rinsed unit was transferred to 750 µl fresh pre-warmed maintenance medium. The unit was then immerged with another 750 µl fresh pre-warmed maintenance medium by applying the medium topically on the tissue. The incubation was made at 37 ± 2 °C, 5 ± 1 % CO2, ≥ 90% humidity for 30 ± 2 minutes. At the end of incubation, each unit was removed from the maintenance medium. The medium was decanted off by turning over the insert. The bottom of the insert was dried carefully by gently taping on a dry absorbent paper and cotton swab. Each corneal epithelium unit was then transferred into 300 µl MTT solution (1 mg/ml in maintenance medium protected from light) and incubated at 37 ± 2 °C, 5 ± 1 % CO2, ≥ 90% humidity for 3 hours ± 15 minutes.
After incubation, each unit was rinsed in 300 µl PBS to remove excess MTT solution or maintenance medium and then dried on absorbent paper. The unit was then transferred to a new plate with 750 µl isopropanol per well, another 750 µl isopropanol was added topically onto each tissue insert. The plate was sealed and stored in fridge overnight protected from light. After the incubation period, the plate was shaken at least for 30 minutes (~120 rpm) at room temperature to ensure the plate recover at room temperature.
The inserts were perforated using a tip on a micropipette. The extraction solution was homogenized vigorously until a homogenous solution was generated. The empty inserts were removed. Two 200 µl of solution from each well was transferred into a 96-well plate. The absorbance (OD) at 570 nm was read using microplate reader. Isopropanol was used as blank.

Number of animals or in vitro replicates:

2

Results and discussion

In vitro

Results

Irritation parameter:

other: viability (%)

Run / experiment:

mean

Value:

90.9

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the SkinEthicTM HCE EIT (for the liquid’s protocol) Validated Reference Method (VRM) and associated with UN GHS classification system, the test item - Fatty acids, coco, esters with oxybis(propanediol), Lot No: 80118 is considered No Category.