Butyl ricinoleate

Eye irritation

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Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Vertellus, Lot# 0000143574
- Expiration date of the lot/batch: 2018 March 21

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, closed container, in a dry, well ventilated location
- Stability under test conditions: assumed to be stable

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Local slaughterhouse, e.g. A. Moksel AG, Buchloe, Germany.

ENVIRONMENTAL CONDITIONS : room temperature, closed container, in a dry, well ventilated location

The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before mounting the corneas in corneal holders (BASF, Duratec) with the endothelial side against the O-ring of the posterior chamber, they were visually examined for defects and any defective cornea were discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always be filled first. The corneas were incubated for one hour at 32 ± 1 °C.

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): 750 µL

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

2 hr

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS : The assay used isolated corneas obtained as a by-product from animals freshly slaughtered at an abattoir, e.g. A. Moksel AG, Buchloe, Germany.

QUALITY CHECK OF THE ISOLATED CORNEAS : The eyes were carefully examined for defects and any defective eyes will be discarded.

NUMBER OF REPLICATES : 3

NEGATIVE CONTROL USED : Physiological saline 0.9% NaCl

SOLVENT CONTROL USED (if applicable) : N/A

POSITIVE CONTROL USED : 100% Ethanol

APPLICATION DOSE AND EXPOSURE TIME : 750 µL, 10 min

TREATMENT METHOD: Closed chamber

POST-INCUBATION PERIOD: Yes, 2 hr

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3
- POST-EXPOSURE INCUBATION: Yes, 2 hr

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: BASF-OP3.0, Duratec
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (OD490) , Jenway 6405
- Others (e.g, pertinent visual observations, histopathology): Each cornea was observed visually and pertinent observations were recorded.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: Opacity is determined using a manufacturer-specific formula, using illuminance through a holder without cornea and illuminance through a holder with cornea as independent variables. Change in opacity between treated and untreated samples is determined, and the irritation potential of the test substance is evaluated based upon the following conditions:
≤ 3: No Category
> 3; ≤ 55: No prediction can be made
> 55: Category 1

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

1.1

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

mean

Value:

0.91

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

other: permeability score

Run / experiment:

mean

Value:

0.013

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline: N/A

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance was evaluated for potential to cause eye irritation using an ex vivo BCOP assay according to OECD guideline 437. An in vitro irritation score of 1.10 was calculated based on corneal opacity and permeability following treatment of the ex vivo eye model with the test substance. The test substance did not meet GHS criteria for classification of eye irritation.