D-Glucitol, 1-deoxy-1-[methyl(1-oxononyl)amino]-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24-02-2017 to 12-05-2017

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)

Version / remarks:

Adopted 26 July 2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N-methyl-N-[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]nonanamide

EC Number:

617-694-1

Cas Number:

85261-19-4

Molecular formula:

C16H33NO6

IUPAC Name:

N-methyl-N-[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]nonanamide

Test material form:

solid

Details on test material:

sample no. MTDID 48740
MEGA 9: 98.1 % (GC)
Water content: 0.3%
pH (aqueous solution 1%): 7.5

Expiry Date: 2017-12
Product Nr: MTDID 48740
Batch: 549612

Test animals / tissue source

Species:

other: Bovine corneas

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The bovine eyes, in a Hanks' Balance Salt Soluation (HBSS) with penicillin-streptomycin, were received from Spear Products on 02 March 2017 and transported to the testing facility in a refrigerated container. The eyes were examined prior to use on the day of dosing. Any eye with a cornea exhibiting evidance of vascularization, pigmentation, opacity or scratches was discarded. The dissected corneas were mounted in specially designed holders that were seperated into anterior and posterior chambers and filled separately. Ech 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 filles MEM solution, ensuring contact with the epitheliu,. Each cornea was visually inspected again to ensure there were no defects. The entire holder was incubated at 32 (+/- 1)°C and allowed to equilibrate for at least one hoer, but nozt longer than two hours.

Test system

Vehicle:

physiological saline

Amount / concentration applied:

0.75 ml of the 20 % (w(v) test article formulation in saline

Duration of treatment / exposure:

After 4 hours ± 10 minutes incubation at 32 ± 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red).

Observation period (in vivo):

1 mL of a 5 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1 °C.

Duration of post- treatment incubation (in vitro):

4 hours (+/- 10 min) exposure with the test item

Number of animals or in vitro replicates:

3 corneas for the test item:
Thereof: 3 corneas for the test item for opacity measurement, 3 corneas for the test item for permeability measurement
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive controls treated with imidazolen in physiological saline 0.9% NaCl

Details on study design:

Preparation of the Corneas:
The bovine eyes (at least six months old), in a Hanks' Balance Salt Soluation (HBSS) with penicillin-streptomycin, were received from Spear Products on 02 March 2017 and transported to the testing facility in a refrigerated container. The eyes were examined prior to use on the day of dosing. Any eye with a cornea exhibiting evidance of vascularization, pigmentation, opacity or scratches was discarded. The dissected corneas were mounted in specially designed holders that were seperated into anterior and posterior chambers and filled separately. Ech 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 filles MEM solution, ensuring contact with the epithelium. Each cornea was visually inspected again to ensure there were no defects. The entire holder was incubated at 32 (+/- 1)°C and allowed to equilibrate for at least one hoer, but not longer than two hours. Following the equilibration, the MEM solution will be removed from both chambers and the chambes refilled with fresh MEM.
A pre-exposure determination of opacity will be made for each designated cornea following the equilibration period by measuring each against the blank supplied with the OP-KIT opacitometer. Any cornea with a value > 7 units will be discarded.
A 20% (200 mg/ml) solution or susspension of the solid test article in 0.9% Saline will be prepared. A volume of 0.75 ml of the dosing solution, MEM solution or imidazole solution will be applied to the epithelium in the anterior chamber in a manner that ensures that entire cornea will be covered. All corneas will be returned to the 32 (+/-1) °C incubator for 4 hours ( +/- 10 min) in a horizontal position. Following the 4-hour exposure, the test article solution, MEM or Imidazole solution will be removed from the epithelium of cornea and the anterior chamber by washing with MEM solution containing phenol red. A final rinse will be made with MEM without phenol red. Both ethe anterior and posterior chambers will be refilled with fresh MEM solution and a measurment of opacity obtained with each treated cornea compared to the blank supplied with the OP-KIT opacitometer.

Differences in light transmission through the control and treated cornea willbe determined using an OP-KIT opacitometer produced by Electro-Design Corporation of Riom, France. Each treated cornea will be scored in comparison to the blank provided with the OP-KIT opacitometer.

The In vitro IVIS Scre (IVIS) will be calculated b yadding the corrected mean opacity score to 15 times the corrected mean optical density. The calculations to obtain an IVIS for the positive control will be performed in the same manner as the test item
IVIS= Corrected mean Opacity score + (15 x Corrected Mean Optical Density score)
The IVIS for the negative control will be calculated by adding the mean opacity score to 15 times the mean optical density.
IVIS= Mean Opacity Score + ( 15 x Mean Optical Density score)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean value test item

Value:

66.31

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

mean test Item

Value:

43

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

Positive control (Imidazole)

Value:

84.15

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation parameter:

cornea opacity score

Run / experiment:

Mean positive control

Value:

68.34

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 5

Irritation parameter:

in vitro irritation score

Run / experiment:

Men vehicle control (0.9% Sodium Chloride Irrigation)

Value:

1.48

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 6

Irritation parameter:

cornea opacity score

Run / experiment:

Mean Vehicle control (0.9% Sodium Chloride Irrigation)

Value:

1.67

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 7

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean negative control (MEM)

Value:

1.72

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 8

Irritation parameter:

cornea opacity score

Run / experiment:

Mean Negative control (MEM)

Value:

1.33

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Based on an IVIS greater than 55 (IVIS= 66.31), and as defined in OECD Guidline No 437, the tets article is in UN GHS Category 1 and is considered to be a severe/ corrosive eye irritant.

Executive summary:

To determine the potential for ocular irritation of the test item the bovine corneal opacity and premeability (BCOP) was performed according to OECD 437. The bovine Corneas per group were dosed with 0.75 ml of a 20% (200mg/ml) formulation of the test item in 0.9% Sodium chloride Irrigation (saline), Minimal Essential Media (MEM, negative contl), or a 20 % formulation of imidazole in 0.9 % saline (positive control). Vehicle control were dosed with 0.75 ml of 0.9% Sodim chloride irrigation. Following a 4-hour exposure for ech group of dosed corneas, opacity measurements and sodium fluorescin permeability were determined. Based on an IVIS greater than 55 (IVIS= 66.31), and as defined in OECD Guidline No 437, the tets article is in UN GHS Category 1 and is considered to be a severe/ corrosive eye irritant.