1,4-Benzenedicarboxylic acid, 1,4-diisononyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02.09.2016 - 01.11.2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

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:

1,4-Benzenedicarboxylic acid, 1,4-diisononyl ester

EC Number:

700-453-0

Cas Number:

59802-05-0

Molecular formula:

C26H42O4

IUPAC Name:

1,4-Benzenedicarboxylic acid, 1,4-diisononyl ester

Test material form:

liquid

Test animals / tissue source

Species:

cattle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL

Duration of treatment / exposure:

The incubation time lasted ten minutes.

Duration of post- treatment incubation (in vitro):

two hours

Number of animals or in vitro replicates:

3 corneae per groups

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Freshly isolated bovine eyes of at least 9 month old donor cattle were collected from the
abattoir. Excess tissue was removed from the excised eyes. The isolated eyes were
transported to the laboratory in HBSS at ambient temperature. The corneae were isolated on the same day after delivery of the eyes and were directly used in the BCOP test.The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2 mm of tissue (sclera) was left for stability and handling of the isolated cornea.

QUALITY CHECK OF THE ISOLATED CORNEAS
All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
Saline (0.9% NaCl in deionised water)


POSITIVE CONTROL USED
2-Ethoxyethanol (purity: 99%)

APPLICATION DOSE AND EXPOSURE TIME
The anterior compartment received the test item or negative or positive control at a volume of 0.75 mL on the surface of the corneae. The corneae were incubated in a horizontal position at 32 ± 1 °C in the water-bath.
The incubation time lasted ten minutes.


POST-INCUBATION PERIOD: no


REMOVAL OF TEST SUBSTANCE
After the test item or control items, respectively, were rinsed off from the application side
with saline, fresh cMEM was added into the anterior compartment.
- Number of washing steps after exposure period: once
- POST-EXPOSURE INCUBATION:
Then the corneae were incubated at 32 ± 1 °C for further two hours in a vertical position, followed by a second opacity reading (t130).

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity:The opacitometer determines changes in the light transmission passing through the corneae, and displays a numerical opacity value.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry] (OD490)


SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
≤ 3 No Category (according to GHS)
> 3; ≤ 55 No prediction can be made
> 55 Serious eye damage according to CLP/EPA/GHS (Cat 1)

DECISION CRITERIA:
The test will be acceptable if
• the positive control gives an IVIS that falls within two standard deviations of the
current historical mean (updated every three months), and if
• the negative control responses result in opacity and permeability values that are less
than the established upper limits for background opacity and permeability values for
bovine corneae treated with the respective negative control.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 0.43

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 test item Benzol-1,4-dicarbonsäure-diisononylester was tested undiluted. Relative to the
negative control, the test item Benzol-1,4-dicarbonsäure-diisononylester did not cause an
increase of the corneal opacity or permeability. The calculated mean IVIS was 0.43
(threshold for serious eye damage: IVIS ≥ 55). According to OECD 437 the test item is not
categorized.