α-d-Glucopyranoside, β-d-fructofuranosyl, mono, di, and tridodecanoate esters

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 11, 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:

2017

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:

yes

Remarks:

The test substance was not soluble in physiological saline, and as such was applied directly to the corneas with physiological saline used as a moistening agent.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: flakes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Mitsubishi-Kagaku Foods Corporation; 63281211
- Expiration date of the lot/batch: March 27, 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: under 15 degrees C, protected from light
- Solubility and stability of the test substance in the solvent/vehicle: The test substance was insoluble in physiological saline, and as such was applied directly to the cornea with physiological saline used as a moistening agent.

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: slaughterhouse
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Corneas were used on day of collection and transported on ice in HBSS with penicillin and streptomycin.
- Time interval prior to initiating testing: Corneas were prepared immediately upon arrival.
- indication of any existing defects or lesions in ocular tissue samples: The eyes were examined upon arrival for defects, and any with defects were discarded.
- Indication of any antibiotics used: penicillin and streptomycin

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 mg


VEHICLE
- Amount(s) applied (volume or weight with unit): Physiological saline was used to moisten the eyes.
- Concentration (if solution): 0.9% NaCl
- Lot/batch no. (if required): 1406805

Duration of treatment / exposure:

4 hrs

Duration of post- treatment incubation (in vitro):

90 minutes

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Eyes were examined for defects, and any with defects were discarded. The cornea was excised leaving 2-3 mm of sclera.

QUALITY CHECK OF THE ISOLATED CORNEAS
Isolated corneas were again checked for defects, and any with defects were discarded.

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
Yes

POSITIVE CONTROL USED
Yes

APPLICATION DOSE AND EXPOSURE TIME
750 mg test substance with 4 hr exposure

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: yes - 90 minutes

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3
- POST-EXPOSURE INCUBATION: yes

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: yes
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (OD490)
- Others (e.g, pertinent visual observations, histopathology): (please specify)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: as indicated in the test guideline

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

average of three replicates

Value:

3.22

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The in vitro irritation score was > 3 and =< 55. No predication could be made regarding the classification of the test substance for eye irritation potential of LMDT to the eye. Further testing with another method is required.

Executive summary:

The eye irritation potential of LMDT was tested in an bovine corneal opacity and permeability assay. 750 mg of test substance was applied to bovine corneas for 4 hrs and the in vitro irriation score (IVIS) was calculated. The IVIS determined was 3.22. As this value was between 3 and 55, no predication could be made regarding the classification of the test substance for eye irritation potential of LMDT to the eye.  Further testing with another method is required.