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Diss Factsheets

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 March 2017 to 17 March 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
Principles of method if other than guideline:
None of the deviations were considered to have impacted the overall integrity of the study or the interpretation of the study results and conclusions.
See Appendix 1 in the study report for the details.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
3,7-dimethyloctanal
EC Number:
227-810-9
EC Name:
3,7-dimethyloctanal
Cas Number:
5988-91-0
Molecular formula:
C10H20O
IUPAC Name:
3,7-dimethyloctanal
Specific details on test material used for the study:
TEST MATERIAL
Name ( stated on the report) : TETRAHYDRO CITRAL
Appearance: Colourless to pale yellow liquid
Batch: SC00019145
Stable under storage conditions until: 18 March 2018 (expiry date)

Test animals / tissue source

Species:
cattle
Details on test animals or tissues and environmental conditions:
Test System: Bovine eyes were used as soon as possible after slaughter.
Rationale: In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimize the need of in vivo testing (1-6). As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed validated in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test.
Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.
Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:
unchanged (no vehicle)
Remarks:
The test item was tested neat.
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
750 microg/l (undiluted) of either the negative control, positive control (Ethanol) or test item was introduced onto the epithelium of the cornea.
Details on study design:
The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the control or the test item over the entire cornea. Corneas were incubated in a horizontal position for 10 +/-1 minutes at 32 +/- 1°CC. After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Earle’s Minimum Essential Medium, Life Technologies) and thereafter with cMEM. Possible pH effects of the test item on the corneas were recorded. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM. Subsequently the corneas were incubated for 120 +/- 10 minutes at 32 +/-1°C. After the completion of the incubation period opacity determination was performed. Each cornea was inspected visually for dissimilar opacity patterns.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Value:
>= -1.3 - <= 0.4
Vehicle controls validity:
valid
Negative controls validity:
valid
Remarks:
in vitro irritancy scores: 0.6 to 0.9
Positive controls validity:
valid
Remarks:
in vitro irritancy scores: 47 to 65
Other effects / acceptance of results:
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 (Ethanol) was 54 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.

Any other information on results incl. tables

The corneas treated with TETRAHYDRO CITRAL showed opacity values ranging from -1.2 to 0.2 and permeability values ranging from 0.000 to 0.009. The corneas were clear after the 10 minutes of treatment with TETRAHYDRO CITRAL. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -1.3 to 0.4 after 10 minutes of treatment with TETRAHYDRO CITRAL.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, since TETRAHYDRO CITRAL induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage accoridng to the CLP Regulation (EC) No. 1272/2008.
Executive summary:

The objective of this study was to evaluate the eye hazard potential of TETRAHYDROCITRAL as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea using the Bovine Corneal Opacity and Permeability test (BCOP test). This report describes the potency of chemicals to induce serious eye damage using isolated bovine corneas. The eye damage of TETRAHYDRO CITRAL was tested through topical application for 10 minutes. The study procedures described in this report were based on the most recent OECD guideline. Batch SC00019145 of TETRAHYDRO CITRAL was a colorless to pale yellow. The test item was applied as it is (750 μl) directly on top of the corneas. 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 (Ethanol) was 54 and was 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. TETRAHYDRO CITRAL did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -0.6 after 10 minutes of treatment.

In conclusion, since TETRAHYDRO CITRAL induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.