2-methylenepropane-1,3-diyl diacetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 Jul 2017 - 27 Jul 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-methylenepropane-1,3-diyl diacetate

EC Number:

223-225-8

EC Name:

2-methylenepropane-1,3-diyl diacetate

Cas Number:

3775-29-9

Molecular formula:

C8H12O4

IUPAC Name:

2-[(acetyloxy)methyl]prop-2-en-1-yl acetate

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 20160308(-88)
- Expiration date of the lot/batch:
- Physical Appearance: clear colourless liquid
- Purity: 99.7%
- Storage: room temperature in the dark
- Expiration date: 31 Mar 2019

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: local abattoir
- Number of animals: 3
- Characteristics of donor animals - age: 12 – 60 months
- Storage, temperature and transport conditions of ocular tissue: eyes were excised after slaughter, placed in HBSS supplemented with penicillin (100 IU/ mL and streptomycin at 100µg/ mL), transported to the test facility over packs of ice on the same day of slaughter. Corneas prepared immediately on arrival. Isolated corneas were mounted in BCOP holders which were filled with EMEM without phenol red and incubated at 32 ± °C for 60 min.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.75 mL

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

120 min at 32°C ±1°C

Number of animals or in vitro replicates:

3 per control group

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Eyes were excised after slaughter, placed in HBSS supplemented with penicillin (100 IU/ mL and streptomycin at 100µg/ mL), transported to the test facility over packs of ice on the same day of slaughter. Corneas prepared immediately on arrival. Isolated corneas were mounted in BCOP holders which were filled with EMEM without phenol red and incubated at 32 ± °C for 60 min.

QUALITY CHECK OF THE ISOLATED CORNEAS
Each cornea was examined, only corneas free of damage were used.

NUMBER OF REPLICATES:
3 per control group

NEGATIVE CONTROL USED
Identification: Sodium Chloride 0.9% w/v
Batch: 3011424
Purity: 0.9%
Expiry Date: 01 Jan 2017
Storage: room temperature in the dark

POSITIVE CONTROL USED
Identification: Ethanol
Batch: STBD7546V
Purity: >99.8%
Expiry Date: 01 Apr 2018
Storage: room temperature in the dark


APPLICATION DOSE AND EXPOSURE TIME
0.75ml, undiluted, applied under safety lightening, exposure for 10 min.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period:
After exposure time the test item and the control item were removed from the anterior chamber and the each cornea was rinsed 3 times with fresh EMEM containing phenol red before a final rinse with complete EMEM without phenol red.

- POST-EXPOSURE INCUBATION: The BCOP holders were incubated, anterior chamber facing forward, at 32 ± 1° for 120 min.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity:
- Corneal permeability: optical density 492nm (OD492) was measured using the Anthos 2001 microplate reader.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
In Vitro Irritancy Score = mean opacity value + (15 x mean permeability OD492 value)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

mean test item

Value:

8.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

mean negative control

Value:

2.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

mean positive control

Value:

42.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

No prediction of eye irritation can be made.