Reactionmass of octan-2-yl prop-2-enoate and octan-3-yl prop-2-enoate and octan-4-yl prop-2-enoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

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

Remarks:

No deviations ocurred that negatively impacted the integrity of the study results.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Reactionmass of octan-2-yl prop-2-enoate and octan-3-yl prop-2-enoate and octan-4-yl prop-2-enoate

EC Number:

947-155-9

Molecular formula:

C11H20O2

IUPAC Name:

Reactionmass of octan-2-yl prop-2-enoate and octan-3-yl prop-2-enoate and octan-4-yl prop-2-enoate

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: 3M Company, Lot 11
- Purity, including information on contaminants, isomers, etc.: 99.7%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Kept in a room with controls set to maintain 18°C to 24°C

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding):
None, dosed neat.

FORM AS APPLIED IN THE TEST (if different from that of starting material)
: The test article was dosed neat.

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Bovine eyes (greater than 35 weeks of age) were obtained from an abattoir and transported to MB Research in a refrigerated container containing Hanks’ Balanced Salt Solution (HBSS) with penicillin-streptomycin on 20 Jun 2019, within 24 hours of harvest.
- Number of animals: No data
- Characteristics of donor animals (e.g. age, sex, weight): At least 35 weeks old.
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): See 'Source' field.
- Time interval prior to initiating testing: The study began the same day that the eyes were delivered to the lab.
- Indication of any existing defects or lesions in ocular tissue samples: The eyes were examined after receipt from the abattoir. Any cornea with visible evidence of neovascularization, pigmentation, opacity or scratches was discarded
- Indication of any antibiotics used: See 'source' field.
- Selection and preparation of corneas: Corneas free of visible defects were dissected from the surrounding tissues. A 2-3 mm rim of sclera was left attached to each cornea. The excised corneas were then placed in a container of fresh HBSS.
- Quality check of the isolated corneas: The dissected corneas were mounted in specially designed holders (MC2, formerly Electro-Design – the manufacturer of the Op-KIT opacitometer) that were separated into anterior and posterior chambers and filled separately. Each cornea was mounted allowing the epithelium of the cornea to project into the anterior chamber. The posterior chamber was filled with MEM solution ensuring contact with the endothelium. The anterior chamber was filled with MEM solution, ensuring contact with the epithelium. Each cornea was visually inspected again to ensure there were no defects

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

VEHICLE: None

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

3

Details on study design:

NUMBER OF REPLICATES : 3

NEGATIVE CONTROL USED : Minimal Essential Media

SOLVENT CONTROL USED: NA

POSITIVE CONTROL USED : 100% ethanol

APPLICATION DOSE AND EXPOSURE TIME : 0.75 mL for 10 minutes

TREATMENT METHOD: Closed chamber

POST-INCUBATION PERIOD: yes, 2 hours.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 2 washing steps 10 minutes after the start of exposure.

- POST-EXPOSURE INCUBATION: 2 hours.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: OP-KIT opacitometer (Electro-Design Corporation, RIOM, France)
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: The decision criteria described in OECD 437 was utilized.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

-1.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

Mean

Value:

-1.34

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

fluorescein leakage

Remarks:

Corrected Mean Optical Density (Permeability)

Run / experiment:

Mean

Value:

0.003

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:

OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: The lab is proficient in running OECD 437.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Negative control IVIS was 2.90.
- Acceptance criteria met for positive control: The In Vitro Irritancy Score (IVIS) of ethanol (positive control), 15.83, exceeded two standard deviations of the historical mean IVIS (range: 15.97 to 40.84).
Although the IVIS of the positive control exceeded two standard deviations of the historical mean range, the test is considered valid as it still demonstrated the anticipated outcome of bovine corneas exposed to 100% ethanol for a 10-minute exposure: a mild to moderately irritating, non-corrosive response.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study (IVIS = -1.30), MTDID 44428 is not irritating in the Bovine Corneal Opacity and Permeability Test (BCOP).

Executive summary:

The eye irritation potential of MTDID 44428 was evaluated in the Bovine Corneal Opacity and Permeability Test (BCOP) according to OECD 437 and in compliance with OECD GLP. MTDID 44428 (0.75 mL) was tested neat in triplicate concurrently with Minimal Essential Medium (negative control) and 100% ethanol (positive control). Corneas were exposed to the test article and controls using the closed chamber method for 10 minutes. Following a washing phase, the corneas were incubated for a 2 hour post-exposure period at 32 C. A measurement of opacity was taken with each treated cornea compared to the blank supplied with the OP-KIT. This reading was used in the final IVIS calculations. Immediately following the two-hour opacity measurement, the MEM solution was removed from the anterior chamber and replaced with 1.0 ml of 0.4% sodium fluorescein solution in Dulbecco's phosphate buffered saline (PBS). Each holder was then returned to the 32 (±1)°C incubator in a horizontal position (anterior chamber facing upward) ensuring contact of the fluorescein with the cornea. After 90 (±5) minutes, the fluid from the posterior chamber of each corneal holder was removed and the amount of dye that passed through the cornea (permeability) was measured as the optical density at 490 nm (i.e., the OD490 nm) by spectrophotometer. The In Vitro Irritancy Score (IVIS) for MTDID 44428 was -1.30, negative control was 2.90, and the positive control was 15.83. Based on the results of the study (IVIS = -1.30), MTDID 44428 is not irritating in the Bovine Corneal Opacity and Permeability Test (BCOP).