Registration Dossier

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:
other: The study was conducted according to O.E.C.D. Testing Guideline No. 437 with GLP compliance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
As per IUCLID5 Sections 1.1 - 1.4.

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
The study was conducted in vitro using isolated bovine cornea. The bovine eyes, supplied by Joseph Morris Abattoir, were excised by an abattoir employee and collected as soon after slaughter as possible. Excised eyes were maintained and transported to the laboratory, at ambient temperature, in sufficient Hanks Balanced Salt Solution (HBSS), containing 1% (v/v) Penicillin/Streptomycin solution, to cover all the eyes in the receptacle. The eyes were used within 4 hours of slaughter (incubation of mounted corneas commenced.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
750 uL undiluted
Duration of treatment / exposure:
ten minutes
Observation period (in vivo):
two hr
Number of animals or in vitro replicates:
Study was conducted in vitro, no animals were placed on study.
Details on study design:
The corneas were excised and rinsed in fresh HBSS plus 1% Penicillin/Streptomycin solution prior to mounting. The corneas were mounted in the cornea holders with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was then positioned on top of the cornea and secured with screws. Both compartments of the holder were filled with Complete Minimal Essential Medium Eagles with 5% v/v fetal calf serum (cMEM), using a syringe. The holders were then plugged and incubated, in an upright position, for 60 minutes ± 5 minutes at 32°C ± 1°C in a waterbath. The waterbath temperature remained within the limits of 32°C ± 1°C throughout the experiment. At the end of the 60 minute incubation period, the medium was removed from both the anterior and posterior compartments using a pipette tip attached to a vacuum pump. The compartments were refilled with fresh cMEM.

Corneas were treated in triplicate with either the test substance, Crude 1,3-dichloropropene, positive control ethanol (91.4%) or negative control (0.9% sodium chloride solution). Immediately prior to treatment, the medium was removed from the anterior compartment of the holder. Seven hundred and fifty μl (750 μl) of test substance, positive control or negative control was introduced into the anterior part of each holder. Following application, the anterior compartment was plugged and the holder turned to a horizontal position and slightly rotated to ensure uniform distribution of the test substance over the surface of the cornea. The test material or controls were in contact with the cornea for a total of 10 minutes (± 30 seconds). Each holder was incubated in a horizontal position at 32°C ± 1°C in a waterbath. Following incubation, the test substance, positive and negative controls were removed and the epithelial surface of the cornea washed, three times with MEM with phenol red. The anterior compartment was then filled with cMEM and compartment was re-plugged and the holders returned to the waterbath and incubated, in an upright position, for two hours ± 10 minutes at 32°C ± 1°C.

Following completion of the two hour incubation period, the medium was removed from both compartments and replaced with fresh cMEM. The posterior compartment was re-plugged and the opacity of each cornea measured and recorded. The opacitometer measured the light transmission through the centre of each mounted cornea, displaying a numerical opacity value (arbitrary unit).

Following the final opacity measurement, the medium was removed from the anterior compartment of the holder. One ml of the sodium fluorescein solution ( 4 mg/L) was added to the anterior compartment using a micropipette. Following addition of the sodium fluorescein solution to the anterior side of the holder, the compartment was plugged and the corneas incubated in a horizontal position at 32°C ± 1°C for 90 ± 5 minutes in a waterbath. Following incubation an aliquot of the mixed medium from the posterior compartment was removed and
transferred to a 1 cm path length cuvette. A spectrophotometer was adjusted to read at 490nm (OD490) and a sample of cMEM read as a blank before other sample ODs were taken.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Value:
43.2
Vehicle controls validity:
valid
Other effects / acceptance of results:
Sample Opacity ± SD Permeability ± SD In vitro irritancy Score ± SD Classification
____________________________________________________________________________________________________________________________________
Crude 1,3-Dichloropropene 11.33 ± 2.0 2.13 ± 0.69 43.2 ± 9.9 Non Corrosive/ Non Severe
Ethanol 19.33 ± 2.65 1.43 ± 0.097 40.8 ± 3.8 Non Corrosive/Non Severe t
0.9% Saline -0.333 ± 1.155 0.004 ± 0.001 Not applicable Not applicable
SD= Standard

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: O.E.C.D. Guideline for Testing No. 437.
Conclusions:
With an In Vitro Irritancy Score (IVIS) of 43.2 ± 9.9 Crude 1,3-dichloropropene is not an eye irritant under the conditions of this in vitro assay.
Executive summary:

Crude 1,3 -dichloropropene was evaluated for eye irritation potential in an O.E.C.D. Testing Guideline No. 437 " Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants" with GLP compliance. With an In Vitro Irritancy Score (IVIS) of 43.2 ± 9.9 Crude 1,3-dichloropropene is not an eye irritant under the conditions of this in vitro assay. Therefore, Classification and Labeling as an eye irritant is not required.