Registration Dossier

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:
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 (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Substance %
Isopropyl Jojobate 30 – 40
Jojoba Alcohols 30 – 40
Jojoba Esters 30 – 40
Specific details on test material used for the study:
Isopropyl Jojobateand Jojoba Alcohol and Jojoba Esters
Article number: 18AC47

Test animals / tissue source

Species:
human
Details on test animals or tissues and environmental conditions:
EpiOcular™ Human Cell Construct (MatTek Corporation)

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
Amount / concentration applied:
50 µL
Duration of treatment / exposure:
The EpiOcular™ tissues were treated in
duplicate with each test article, positive control, or negative control. Prior to test article or control article applications, each tissue surface was moistened with 20 μL of Ca++Mg++-free DPBS and incubated at standard culture conditions for 30 minutes. After incubation, the EpiOcular™ tissues were tested in duplicate with 50 μL of the test article, positive control, or negative control. The tissues were then placed back into the incubator after dosing and incubated at standard culture conditions for the remainder of the 30 minute exposure.
Number of animals or in vitro replicates:
EpiOcular™ tissues were treated in duplicate with the test articles, Isopropyl Jojobate and Jojoba Alcohol and Jojoba Esters, and Hydrolyzed Jojoba Esters, in one valid
definitive trial.

Results and discussion

In vitro

Results
Irritation parameter:
other: Mean cell viability (%)
Remarks:
MTT assay
Value:
ca. 99.2
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Any other information on results incl. tables

MTT Assay

The EpiOcular™ cultures were treated in duplicate with the test articles, Isopropyl Jojobate and Jojoba Alcohol and Jojoba Esters, and Hydrolyzed Jojoba Esters, for an exposure time of 30 minutes. The negative and positive controls were also exposed in duplicate for 30 minutes. Table 1 summarizes the percent viability results of the Topical Application Ocular Irritation Screening Assay Using the EpiOcular™ Human Cell Construct for the test articles and the positive control, Methyl Acetate. The raw and analyzed data for the test articles, negative control, and positive control are included in Appendix B.

The test articles were observed to reduce MTT directly in the absence of viable cells; therefore a killed control experiment was performed. Additional calculations were performed to correct for the amount of MTT reduced directly by test article residues as described in the Presentation of Data section. The test articles were not observed to be colorants in isopropanol; therefore a colorant control was not performed.

The test articles, Isopropyl Jojobate and Jojoba Alcohol and Jojoba Esters, and Hydrolyzed Jojoba Esters, resulted in relative viabilities of 99.2% and 112.8% respectively, and are predicted to not require classification or labelling for ocular irritation (GHS No Category).

Deviation

The EpiOcular™ tissues were initially incubated in assay medium for 1 hour and 23 minutes prior to transferring the tissues to fresh assay medium. This is a deviation from the protocol which states that they will be incubated at standard culture conditions for one hour. Although the transfer of the tissues was delayed, they were transferred to fresh assay medium prior to the overnight incubation of 16-24 hours. The purpose of the replacing the assay medium after 1 hour, is to replenish nutrients and growth factors used during the initial incubation which was accomplished; therefore this deviation did not have an adverse impact on the study results.

 

IIVS Test Article

Number

Sponsor’s Designation   Conc.  

Exposure Time

Mean Viability (%)

 

Ocular Irritation Prediction*

 18AC47

Isopropyl Jojobate and

Jojoba Alcohol and

Jojoba Esters

 Neat  30 min  99.2 No Category 

Positive

Control

 Methyl Acetate  Neat  30 min  30.6  Irritant

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test articles, Isopropyl Jojobate and Jojoba Alcohol and Jojoba Esters resulted in relative viabilities of 99.2% and are predicted to not require classification or labelling for ocular irritation (GHS No Category).
Executive summary:

The test articles, Isopropyl Jojobate and Jojoba Alcohol and Jojoba Esters resulted in relative viabilities of 99.2% and are predicted to not require classification or labelling for ocular irritation (GHS No Category).