Oligomeric reaction products of 4,4'-propane-2,2-diyldiphenol and 2-methyloxirane and 2-(chloromethyl)oxirane

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 August 2012- 15 August 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted in accordance with international guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): EP-4000S, Chemical name: Phenol, 4-4’-(1-methylethylidene)bis-, polymer with 2-(chloromethyl)oxirane and 2- methyloxirane
- Physical state: Light yellow liquid
- Analytical purity: 100%
- Lot/batch No.: 902Y1
- Expiration date of the lot/batch: 07 December 2012
- Storage condition of test material: Room temperature in the dark

Test animals / tissue source

Species:

other: bovine eyes

Details on test animals or tissues and environmental conditions:

The bovine eyes were excised and collected as soon after slaughter as possible (excised at 12.30 hours, 14 August 2012).
Excised eyes were maintained and transported to the laboratory, at ambient temperature, in sufficient HBSS, containing 1% (v/v) penicillin/streptomycin solution, to cover all the eyes in the receptacle.
The eyes were used within 4 hours of slaughter.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Concurrent positive and negative control corneas were examined

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): neat

Duration of treatment / exposure:

10 minutes (± 30 seconds)

Observation period (in vivo):

Following incubation, the positive and negative controls were removed and the epithelial surface of the cornea washed, at least three times or until the wash medium (EMEM with phenol red) was clear and there was no discolouration. The wash medium was added via the holes on the top of the holder. After each wash, the wash medium was removed using a pipette tip attached to a vacuum pump.
As the test substance, EP-4000S, was very viscous, it was not possible to remove using the vacuum pump. The anterior glass was removed and the corneas were gently washed with EMEM without phenol red until there was no discolouration of the wash medium. The anterior glass was then replaced.

Following completion of the two hour incubation period, the medium was removed from both compartments and replaced with fresh cMEM. The posterior compartment was re-plugged and the opacity of each cornea measured and recorded.
The opacity values obtained at this stage were used in calculating the final In Vitro Irritancy Score.

Throughout the assay the corneas were examined for opaque spots or other irregularities.

Number of animals or in vitro replicates:

Corneas were treated in triplicate with either the test substance, positive control (1% w/v sodium hydroxide) or negative control (0.9% sodium chloride solution).

Details on study design:

The assay uses isolated bovine corneas as a means of assessing the ocular corrosivity or severe irritancy potential of test substances in vitro.
The isolated corneas were obtained as a by-product of the meat production industry. Two endpoints, corneal opacity and permeability,
were measured and combined to give an In Vitro Irritancy Score.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: Opacity

Basis:

mean

Time point:

other: 10 min

Score:

-0.333

Remarks on result:

other: SD=1.528

Results 2

Irritation parameter:

other: Permeability OD 490

Basis:

mean

Time point:

other: 10min

Score:

-0.004

Remarks on result:

other: SD=0.013

Results 3

Irritation parameter:

other: In Vitro Irritancy

Basis:

mean

Time point:

other: 10min

Score:

-0.4

Remarks on result:

other: In Vitro Irritancy Score (IVS) was calculated as: (Opacity +(15x Corrected OD 490)

Irritant / corrosive response data:

Throughout the assay the corneas were examined for opaque spots or other irregularities.
Following treatment with test substance, EP-4000S, the corneas were noted as clear. The corneas treated with the positive control, sodium hydroxide 1% (w/v), were very opaque and the corneas treated with the negative control, 0.9% saline, were clear.

Any other information on results incl. tables

The positive control, sodium hydroxide (1%), elicited an In Vitro Irritancy Score of 131.4.

This value was within the historical range (mean ± 2 x SD = 82.3 – 144.7) for the assays performed to date.

The negative control, 0.9% saline, opacity mean change value was 1.0 which was below the maximum acceptance value of 2.0.

The permeability mean of the negative control was 0.028 which was below the maximum acceptance value of 0.1.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substance, EP-4000S, elicited an In Vitro Irritancy Score of -0.4 ± 1.7 and was predicted to be a non-corrosive/ non-severe eye irritant.