Methyl 5-deoxy-2,3-O-isopropylidene-β-D-ribofuranoside

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 December 2012 to 27 August 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Fully GLP and in accordance with current test guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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

Test material

Test material information

Constituent 1

Reference substance name:

Methylfuranoside

IUPAC Name:

Methylfuranoside

Constituent 2

Reference substance name:

Methyl 5-deoxy-2,3-O-isopropylidene-β-D-ribofuranoside

EC Number:

245-485-1

EC Name:

Methyl 5-deoxy-2,3-O-isopropylidene-β-D-ribofuranoside

Cas Number:

23202-81-5

Molecular formula:

C9H16O4

IUPAC Name:

methyl 5-deoxy-2,3-O-isopropylidene-beta-D-ribofuranoside

Test material form:

other: pale yellow liquid

Details on test material:

Name: Methylfuranoside
CAS number: 23202-81-5
Batch number: 201111082008
Quantity supplied: 1.5 kg
Purity: 99.7%
Expiry date: November 2013
Date of receipt: 19 November 2012
Appearance: pale yellow liquid
Storage details: stored in a sealed container, at room temperature (15 to 25°C) in the dark.

The negative control substance was 0.9% sodium chloride solution supplied by Baxter Healthcare Ltd, Thetford, UK.
The positive control substance was dimethyl formamide supplied by Sigma-Aldrich Co Ltd, Poole, UK.

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Bovine corneas supplied by a local abattoir. The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin at 100 IU/mL and streptomycin at 100 µg/mL) in a suitably sized container and transported on the same day to the testing facility. On arrival at the test facility the eyes were carefully examined for defects including increased opacity, scratches and neovascularisation. Only corneas free from such defects were used.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): Not applicable

VEHICLE: Not applicable

Duration of treatment / exposure:

A volume of 750 µL of the test item or negative or positive control was applied to each of three corneas followed by a ten minute incubation at 32°C.

Observation period (in vivo):

After the initial ten minute incubation, each cornea was washed with media containing phenol red (as a pH indicator) until this indicator showed no pH effect occurring (and demonstrating that the test item had been removed successfully). The corneas were then washed once in media without phenol red and the opacities measured. The corneas were incubated (horizontally) for 2 hours ± 10 minutes after which, the opacities were measured and then the anterior chamber emptied. For the permeability endpoint, 1 mL of sodium fluorescein (4 mg/mL solution) was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and held in a labelled tube. Three 350 µL aliquots of this media (per cornea) were analysed for optical density at 490 nanometers (OD490) using a spectrophotometer.

Number of animals or in vitro replicates:

Three conrneas used per group

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed once with media containing phenol red, then with media without phenol red
- Time after start of exposure: ten minutes

SCORING SYSTEM: In Vitro Irritation Score (IVIS) was determined as follows:
IVIS = mean opacity value + (15 x mean permeability value).
The test item was concluded to be a corrosive or severe irritant to the eye if it caused damage to the cornea such that the IVIS was >55.1.


TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

1

Value:

2.3

Vehicle controls validity:

valid

Remarks:

value: 0

Positive controls validity:

valid

Remarks:

value: 22.2

Results 2

Irritation parameter:

fluorescein leakage

Run / experiment:

1

Value:

-0.001

Vehicle controls validity:

valid

Remarks:

value: 0

Positive controls validity:

valid

Remarks:

value: 0.165

Results 3

Irritation parameter:

in vitro irritation score

Remarks:

IVIS

Run / experiment:

1

Value:

2.25

Vehicle controls validity:

valid

Remarks:

value: 0

Positive controls validity:

valid

Remarks:

value: 24.47

Other effects / acceptance of results:

Corneal opacity
The mean opacity reading for the test item was 2.3.
The mean opacity reading for the negative control was 0.
The mean opacity reading for the positive control was 22.0

Corneal permeability
The mean group corrected optical density for the test item was -0.005.
The mean group corrected optical density for the negative control was 0.
The mean group corrected optical density for the positive control was 0.165.

In Vitro Irritation Score
The calculated in vitro irritation score for the test item was 2.25
The calculated in vitro irritation score for the negative control was 0.
The calculated in vitro irritation score for the positive control was 24.47.

Applicant's summary and conclusion

Interpretation of results:

other: Not corrosive or severly irritating

Remarks:

Criteria used for interpretation of results: other: In Vitro Irritation Score (IVIS)

Conclusions:

The test article, Methylfuranoside, produced an IVIS score of 2.25 and was considered not to be corrosive or severely irritating to the eye.

Executive summary:

This study was conducted to determine whether the test item causes corrosion or severe irritation to excised bovine corneas using the bovine corneal opacity and permeability (BCOP) assay.

A volume of 750 µL of the test item was applied to each of three corneas followed by a ten minute incubation at 32°C. After this incubation, each cornea was washed with media containing phenol red followed by media without phenol red, the opacities measured and then the anterior chamber emptied. For the permeability endpoint, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and three 350 µL aliquots of this media (per cornea) were analysed for optical density at 490 nanometers (OD₄₉₀).

A volume of 750 µL of the negative or positive control was similarly applied to further groups of three corneas. These corneas were subject to the procedures detailed above.

The mean opacity reading for the test item was 2.3, for the negative control was 0 and for the positive control was 22.0.

The mean group corrected optical density for the test item was -0.005, for the negative control was 0 and for the positive control was 0.165.

The test item, Methylfuranoside, produced an IVIS score of 2.25 and was considered not to be corrosive or severely irritating to the eye.