Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation
Remarks:
other: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in compliance with OECD Guideline 437 without any deviation.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 437
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name of the test item (as cited in the study report): NOPYL ACETATE
IUPAC Name of the test item: (1R,5S)-2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl) ethyl acetate
Synonym: (-) Nopyl Acetate; 6,6-dimethyl -(1R,5S)-bicyclo[3.1.1]hept-2-ene-2-ethanol-2-acetate
Substance type: monoconstituent
Batch No.: 117042
Purity: 99.3%
This composition is within the specifications of the substance identity profile agreed within the SIEF.
Colour: colourless – slightly amber
Storage Conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry Date: 10 April 2013

Test animals / tissue source

Species:
other: bovine eye
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST SYSTEM
Bovine eyes (from cattle less than 12 months old) collected at the slaughterhouses of La Talaudière - France (42350) and carried in a stable medium (Hanks medium buffered with sodium bicarbonate), at room temperature, and used within 4 h after killing the animals.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: yes, 2 groups (positive and negative controls) containing 3 corneas each.
Amount / concentration applied:
TEST MATERIAL
Amount(s) applied (volume or weight with unit): 750 ± 8 µL

Concentrations:
- test material: undiluted
- negative control: 0.9 % (w/v) sodium chloride
- positive control: 10 % (w/v) sodium hydroxide
Duration of treatment / exposure:
10 ± 1 min
Observation period (in vivo):
Corneal opacity was measured after 2 h ± 10 min of incubation.
Corneal permeability was measured after 90 ± 5 min of incubation.
Number of animals or in vitro replicates:
Total: 9 corneas (3 corneas/group)
Details on study design:
Details of test procedure: bovine eyes recovered from a slaughterhouse were inspected and undamaged corneas were dissected and mounted in specially constructed BCOP corneal holders. After 1 h incubation in media, the basal opacity of each cornea was recorded using an opacitometer. 750 ± 8 µL of the test item and of each control were applied directly on the cornea using a positive-displacement micropipette and incubated in a water-bath at 32 ± 1 °C, in a vertical position (screw upward) for 10 ± 1 min. Before reading the final opacity, the corneas were rinsed and incubated for 2 h ± 10 min in fresh media to equilibrate. Then the permeability of each cornea was determined with a fluorescein solution after an incubation of 90 ± 5 min. An aliquot (360 µL) of the media from the posterior compartment was read in a plate reader at 490 nm to determine the permeability of the cornea to the fluorescein solution. The opacity and permeability values were combined to obtain an in vitro irritancy score (IVIS).

Incubation temperature: 32 ± 1 °C

Calibration information for devices used for measuring opacity and permeability (e.g., opacitometer and spectrophotometer): opacitometer (OPKIT) was calibrated with specific calibrators before each measurement of the corneal opacity. Values of opacity after three calibrations were 73-77, 147-153 and 220-240, respectively.

Controls: 0.9 % (w/v) sodium chloride and 10 % (w/v) sodium hydroxide were used as negative and positive controls, respectively.

Results and discussion

In vivo

Results
Irritation parameter:
other: In vitro irritancy score (IVIS)
Basis:
mean
Time point:
other: exposure: 10 min
Score:
0
Reversibility:
other: not applicable
Irritant / corrosive response data:
see table 7.3.2/1
Other effects:
no data

Any other information on results incl. tables

Table 7.3.2/1: Opacity and optical density

Treatment

Opacity

Optical density

IVIS

Before

treatment

(OPT0)

After 2 h

(OPT2)

Difference

(OPT2-OPT0)

Corrected

OD

Corrected

Score

Negative control

1

8

7

0.037

0

3

3

0.018

1

5

4

0.034

Mean

4.7

0.03

Standard deviation

2.1

0.01

10 % (w/v) NaOH

3

126

123

118.3

7.848

7.818

235.6

4

130

126

121.3

7.888

7.858

239.2

1

96

95

90.3

7.864

7.834

207.8

Mean

110.0

7.837

227.6

Standard deviation

17.1

0.020

17.2

Test item

0

3

3

-1.7

0.034

0.004

-1.6

1

2

1

-3.7

0.038

0.008

-3.5

3

3

0

-4.7

0.051

0.021

-4.3

Mean

-3.3

0.011

-3.2

Standard deviation

1.5

0.009

1.4

In vitro irritancy score (IVIS) = (OPT2-OPT0) + (15 x OD)

O.D. of the fluorescein solution (8 µg/mL): 1.490

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information (IVIS<55.1) Criteria used for interpretation of results: EU
Conclusions:
IVIS of nopyl acetate was 0.0. Therefore it is not classified as corrosive or severe irritant for the isolated bovine cornea, after a 10 min contact period.
Executive summary:

In an in vitro eye irritation study performed according to OECD Guideline No 437 and in compliance with GLP, 750 ± 8 µL of nopyl acetate were applied directly to isolated bovine corneas (3 corneas/group), mounted in specially constructed BCOP corneal holders, and incubated in a water-bath at 32 ± 1°C for 10 ± 1 min. Before reading the final opacity, the corneas were rinsed and incubated for 2 h in fresh media to equilibrate. Then the permeability of each cornea was determined with a fluorescein solution after an incubation of 90 min. An aliquot (360 µL) of the media from posterior compartment was read in a plate reader at 490 nm to determine the permeability of the cornea to the fluorescein solution. The opacity and permeability values were combined to obtain an in vitro irritancy score (IVIS). 0.9% (w/v) sodium chloride and 10% (w/v) sodium hydroxide were used as negative and positive controls, respectively.

The IVIS of the test item and positive control were 0.0 and 227.6 ± 17.2 respectively. Optical density values for the negative control corneas (nutritive medium) and fluorescein solution (8 µg/mL) were within the desired range and thus confirmed the validity of the test.

IVIS of nopyl acetate is 0.0. Therefore it is not classified as corrosive or severe irritant for the isolated bovine cornea, after 10 min of contact.