GAMMA-CYCLODEXTRINE, 6A,6B,6C,6D,6E,6F,6G,6H-OCTABROMO-6A,6B,6C,6D,6E,6F,6G,6H-OCTADEOXY-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD guideline 437 “Bovine corneal opacity and permeability (BCOP) test method for identifying ocular corrosives and severe irritants”

Version / remarks:

July 2013

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.47 “Bovine corneal opacity and permeability method for identifying ocular corrosives and severe irritants

Version / remarks:

December 2010

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(1R,3R,5S,6R,8S,10S,11S,13R,15S,16R,18R,20S,21R,23S,25S,26S,28R,30S,31R,33S,35S,36S,38R,40S,41R,42R,43R,44R,45R,46R,47R,48R,49R,50R,51R,52R,53R,54R,55R,56R)-5,10,15,20,25,30,35,40-octakis(bromomethyl)-2,4,7,9,12,14,17,19,22,24,27,29,32,34,37,39-hexadecaoxanonacyclo[36.2.2.2³,⁶.2⁸,¹¹.2¹³,¹⁶.2¹⁸,²¹.2²³,²⁶.2²⁸,³¹.2³³,³⁶]hexapentacontane-41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56-hexadecol

EC Number:

611-044-0

Cas Number:

53784-84-2

Molecular formula:

C48H72Br8O32

IUPAC Name:

(1R,3R,5S,6R,8S,10S,11S,13R,15S,16R,18R,20S,21R,23S,25S,26S,28R,30S,31R,33S,35S,36S,38R,40S,41R,42R,43R,44R,45R,46R,47R,48R,49R,50R,51R,52R,53R,54R,55R,56R)-5,10,15,20,25,30,35,40-octakis(bromomethyl)-2,4,7,9,12,14,17,19,22,24,27,29,32,34,37,39-hexadecaoxanonacyclo[36.2.2.2³,⁶.2⁸,¹¹.2¹³,¹⁶.2¹⁸,²¹.2²³,²⁶.2²⁸,³¹.2³³,³⁶]hexapentacontane-41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56-hexadecol

Test material form:

solid: particulate/powder

Details on test material:

- Appearance: off-white powder
- Storage condition of test material: At room temperature

Specific details on test material used for the study:

pH (1% in water, indicative range): 7.1 – 9.1 (determined by WIL Research Europe)

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Bovine eyes from young cattle were obtained from the slaughterhouse. Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: approx. 750 μl of 20% (w/v) test substance per cornea

NEGATIVE CONTROL
- Amount applied: 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount applied: 750 µl per cornea
Concentration: 20% (w/v) Imidazole

Duration of treatment / exposure:

240 ± 10 minutes

Duration of post- treatment incubation (in vitro):

none

Number of animals or in vitro replicates:

3

Details on study design:

TEST SITE
- Isolated bovine cornea
- Uniform distribution of the control or the test substance over the entire cornea was ensured by horizontal position of the corneas.
- Possible pH effects of the test substance on the corneas were recorded.
- After exposure the cornea is thoroughly rinsed to remove the test substance and the opacity of the cornea was measured. Then, corneas were incubated in sodium-fluorescein solution for 90 ± 5 minutes at 32 ± 1°C to determine the permeability.
- Any OD490 that was 1.500 or higher was diluted to bring the OD490 into the acceptable range (linearity up to OD490 of 1.500 was verified before the start of the experiment). OD490 values of less than 1.500 were used in the permeability calculation.

REMOVAL OF TEST SUBSTANCE
- Washing: at least three times with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation)

SCORING SYSTEM:
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).


TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

DATA EVALUATION:
A test substance that induces an IVIS ≤ 3 is not classified for eye irritancy (UN GHS: no category);
A test substance that induces an IVIS > 55 is defined as a corrosive or severe irritan (UN GHS: catgeory 1);
For a test substance that induces an IVIS >3 and ≥ 55, no prediction on irritant potency can be made.

The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

16.9

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The mean opacity score and mean permeability for corneas treated with Broom dex were 16.0 and 0.057, respectively. The corneas were translucent after the 240 minutes of treatment with Broomdex. No pH effect of the test item was observed on the rinsing medium.
The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 113 and within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

Broomdex induced an IVIS > 3 ≤ 55 in the Bovine Corneal Opacity and Permeability test, therefore no prediction on the classification can be made.

Executive summary:

Evaluation of the eye hazard potential of Broomdex was performed using the Bovine Corneal Opacity and Permeability Test (BCOP Test) according to OECD/EC guidelines and in compliance with GLP principles. The negative control responses of the opacity and permeability values were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 113 and within the historical positive control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. Broomdex induced ocular irritation through both endpoints, the mean opacity score was 16 and mean permeability score was 0.057, resulting in a mean in vitro irritancy score (IVIS) of 17 after 240 minutes of treatment. Since Broomdex induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.