Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 5th 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification 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:

Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Molecular formula:

not applicable - UVCB substance

IUPAC Name:

Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Specific details on test material used for the study:

Test item name: Finestar 2009
CAS Number: 2225876-48-0
Water solubility: soluble in water
Molecular formula: C12 H22 O8
Molecular weight: 294.30
Batch/Lot number: 3253082031
analyzed purity: 100% active
Saponification value: 45.87
Date of manufacture: August 27, 2020
Date of expiry: August 26, 2021
Appearance: Pale yellow viscous liquid
Storage temperature: Room temperature (15 to 30°C)
Storage condition: Keep away from light, moisture, and oxidizing - reducing chemicals
Storage container: Keep in original container

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Isolated cornea from the eyes of freshly slaughtered cattle
Source : Deonar Abattoir slaughter house, Mumbai, Maharashtra
Bovine Age : Between 1 to 5 years (the age of the animals was determined based on
horn ring count in addition to the Horizontal Diameter of corneas and
central corneal thickness).
Transportation Condition : Transported (in a sealed plastic container) under cold condition in
Hanks’ Balanced Salt Solution containing antibiotics [penicillin at 100 IU/mL and streptomycin at 100 μg/mL]

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Set 1 Negative Control 750 μL Normal Saline
Set 2 Positive Control 750 μL Dimethylformamide (undiluted)
Set 3 Test Item 750 μL of Finester 2009 (undiluted)

Duration of treatment / exposure:

Corneas were exposed for approximately 10 minutes ± 30 seconds at 32 ± 1 °C

Duration of post- treatment incubation (in vitro):

After rinsing, the corneas were incubated for an additional period of approximately 2 hours ± 10
minutes at 32 ± 1 ºC.

Number of animals or in vitro replicates:

Not Specified

Details on study design:

Selection Criteria for Eyes
The eyes were used within 24 hours of slaughter. Eyes were examined prior to use and corneas
free from any visible defects were used. Corneas possessing an opacity less than seven opacity
units or equivalent for the opacitometer were used in the study.

Pre-test Procedures
A baseline reading for each cornea holder was taken with medium prior to loading of the cornea on to the cornea holder. Corneas, free from defects, were dissected so they had a 2-3 mm rim of sclera and were transferred to a container containing Hanks’ Balanced Salt Solution containing antibiotics [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL].

Selected corneas were mounted on the corneal holders with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was then placed on the top of the cornea and fixed in place. Both chambers were then filled to excess with pre-warmed phenol red free Eagle's Minimum Essential Medium (EMEM) (the posterior chamber filled first to allow the cornea to return to its natural concave position), ensuring no bubbles were present within the holders. The device was then equilibrated at 32 ± 1°C for at least one hour to allow the corneas to equilibrate with the medium and achieve normal metabolic activity, to the extent possible.

Following equilibration, the medium was removed from both chambers and fresh pre-warmed phenol red free EMEM added to both chambers. A baseline opacity reading was then taken for each cornea.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

In vitro Irritancy Score (IVIS)

Value:

1.57

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Acceptance Criteria
The following acceptance criteria should be satisfied for the assays to be considered valid:
Negative control (NC) acceptance criteria: The negative or solvent/vehicle control responses should result in opacity and permeability values that are less than the established upper limits for background opacity and permeability values for bovine corneas treated with the respective negative or solvent/vehicle control.

Positive control (PC) acceptance criteria: The mean In Vitro Irritancy Score (IVIS) of positive
control treated corneas should be within the range of two standard deviations of the mean of the
historical control data.

Any other information on results incl. tables

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in Appendix 1 (Please see attached background material).

Corneal Epithelium Condition

The condition of each cornea is given in Appendix 2 (Please see attached background material).

The corneas treated with the test item were clear post treatment. The corneas treated with the negative control item were clear post treatment. The corneas treated with the positive control item were cloudy post treatment.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The mean IVIS score for corneas treated with Finester 2009 was found to be 1.57.
Based on results of this study, the classification for Finester 2009 is as follows:
Classification (OECD 437 UN GHS) : No Category

Executive summary:

Study Guideline
This study was conducted according to:
OECD, 2020: The Organisation for Economic Co-operation and Development (OECD) Guideline for the Testing of Chemicals, 437, Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage, adopted by the Council on June 26, 2020.

RESULTS
Corneal Opacity
The mean final opacity value for Finester 2009 treated eyes (1.27) shows no observable increase in comparison to the control group (0.06). An observable marked increase in the final mean opacity was observed in corneas treated with the positive control, N,N-dimethylformamide (77.99).

Corneal Permeability
The mean final corneal permeability value for Finester 2009 treated eyes (0.020) shows no observable increase in comparison to the control group (0.005). An observable marked increase in the mean final corneal permeability was observed in corneas treated with the positive control, N,Ndimethylformamide (1.447).

In vitro Irritancy Score (IVIS)
The mean In-Vitro Irritancy Score (IVIS) of normal saline (control) and N,N-dimethylformamide (positive control) treated corneas was found to be 0.14 and 99.70 which confirmed the reliability of the test procedure.
The mean IVIS score for the corneas treated with Finester 2009 was found to be 1.57 (Refer to attached TABLE 1: In Vitro Irritation Score).

Validity of the Test
 The mean In-Vitro Irritancy Score (IVIS) of dimethylformamide (positive control) treated corneas was found to be 99.70 which is within the range of two standard deviation of the mean of the historical control data, confirming the reliability of the test procedure. The opacity and permeability values of the negative control (saline) treated corneas (0.06 and 0.005, respectively) were less than the established upper limits for background values, confirming the
reliability of the test procedure.