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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation
Remarks:
other: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From April 14 to 18, 1988.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test procedures cannot be subsumed under testing guideline and according to GLP, nevertheless are well documented and scientifically acceptable.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Bovine Eye and the Chicken Chorioallantoic Membrane (BECAM ASSAY).
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Details on test animals or tissues and environmental conditions:
Calf eyes were obtained from a local slaughterhouse (Viguurs, 's-Hertogenbosch). The eyes were excised by standard procedures within 15 minutes after killing the animal. They were transported to the laboratory and examined for injuries. This was done by holding the eyes submersed in saline; eyes with corneal damage or other abnormalities were discarded. The selected eyes were preserved at ambient temperature (approximately 20 °C) under saline not longer than 4 hours.
White eggs were obtained from a chicken breeding centre.
Date of arrival: April 5, 1988.
They were transported at a temperature between 15 and 20 °C.
After approximately 6 hours of rest, the incubation was started by placing the eggs in the incubator with the air chamber upwards.
The temperature was maintained at 37-38 °C and the relative humidity between 50 and 70 %. The eggs were turned once every hour.
They were candled at the seventh day of incubation; unfertilized eggs or eggs containing a dead embryo were removed.

Test system

Amount / concentration applied:
Calf eyes: 0.1 ml
eggs: 0.2 ml
Duration of treatment / exposure:
calf eyes: 30 sec
eggs: 20 sec
Observation period (in vivo):
calf eyes: 10 min
eggs: 30 sec, 1.5 and 4.5 min
Number of animals or in vitro replicates:
5 calf eyes

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: BE Assay
Basis:
mean
Time point:
other: 0.5, 2, 5 min
Score:
0
Max. score:
9.5
Reversibility:
other: n.a.
Remarks on result:
other: not irritating
Irritation parameter:
other: CAM Assay
Basis:
mean
Time point:
other: 0.5, 2, 5 min
Score:
2.75
Max. score:
19
Reversibility:
other: n.a.
Remarks on result:
other: slightly irritating
Irritant / corrosive response data:
The test substance induced no effects in the bovine eye.
The effects on the CAM consisted of vascular injection almost instantly after application. In three eggs (no. 1, 3 and 4) haemorrhages were scored 2 minutes after application. On account of these results it is expected that eye contact with the test substance may lead to slight irritation. Acute corneal effects may be negligible.

Any other information on results incl. tables

BE ASSAY

Treatment Opacity Detachment Fluorescence Total
Neg. control 0 0 0 0
Toluene 0 0 1 1
Acetone 2 0 1.5 3.5
Eye1 0 0 0 0
Eye2 0 0 0 0
Eye3 0 0 0 0
Eye4 0 0 0 0
Eye5 0 0 0 0

Mean

0

CAM assay

30 sec 2 min 5 min Total
Time Egg I H C I H C I H C
09:37 1 + - - + - - + + (a) - 3
09:40 2 + - - + - - + - - 2
09:43 3 + - - + - - + + (a) - 3
09:46 4 + - - + - - + + (a) - 3
Mean: 2.75

a: Bleeding by 2 min

The total scores are not consistent with the above mentioned scoring system (table 2). According to this scoring in table 2 the total score for egg 1 -4 would be: 4.5, 3.5, 4.5, 4.5, respectively, which would lead to a mean score of 4.25. The score of 4.25 would lead to moderate irritation.

REFERENCE SUBSTANCES

For toluene, a score of 1.0 was obtained in the BE assay whereas the average score in the CAM assay was 1.25.

Acetone treatment resulted scores of 3.5 in the BE assay and an average score of 2.5 in the CAM assay.

As the effects, produced by acetone and toluene, fell within the historical range and the untreated eyes and eggs showed no signs of irritation, the results were considered to be valid.

To evaluate the effects observed in both assays the documented in vivo effects of the two reference substances toluene and acetone were taken into consideration. Toluene produces slight transient conjunctival irritation with negligible to slight corneal epithelial damage without opacity in the rabbit eye, while no serious effects have been reported in case of accidental exposure of human eyes.

Acetone causes temporary edema and moderate corneal opacity with epithelial damage to exposed rabbit eyes while in man reversible corneal opacity has been observed.

Applicant's summary and conclusion

Interpretation of results:
other: BE assay: not irritating (according to the CLP Regulation). CAM assay: inconclusive (according to the CLP Regulation).
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Today a validated methods and procedures protocol about the Bovine Eye assay is available. The differences between the present test and the validated protocol are not suspected of impairing the results. Therefore, it can be concluded that the substance is not irritating, based on the BE assay.
In the case of the CAM assay the protocol is not validated yet and the re-evaluation of the result obtained is not appicable. Therefore, the CAM assay can be considered as inconclusive.
Executive summary:

Method

The eye irritation potential was assessed according to the in vitro Bovine Eye and the Chicken Chorioallantoic Membrane (BECAM ASSAY) method.

Results

The test substance induced no effects in the bovine eye.

The effects on the CAM consisted of vascular injection almost instantly after application. In three eggs (no. 1, 3 and 4) haemorrhages were scored 2 minutes after application. On account of these results it is expected that eye contact with the test substance may lead to slight irritation. Acute corneal effects may be negligible.

Conclusion

Today a validated methods and procedures protocol about the Bovine Eye assay is available. The differences between the present test and the validated protocol are not suspected of impairing the results. Therefore, it can be concluded that the substance is not irritating, based on the BE assay.

In the case of the CAM assay the protocol is not validated yet and the re-evaluation of the result obtained is not appicable. Therefore, the CAM assay can be considered as inconclusive.