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Toxicological information

Eye irritation

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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: GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Principles of method if other than guideline:
The study was performed according to the methods described in the following publications:
• Lüpke N.P. (1985): Hen’s Egg Chorio allantoic Membrane Test for Irritation Potential. Fd. Chem. Toxic. 23, pp. 287 – 291.
• Spielmann, H. (1995): HET-CAM Test. In: Methods in Molecular Biology, 43 (eds.: O’Hare, S. and Atterwill, C. K.) pp. 199 – 204.
• Spielmann, H. et al. (1996): Results of a Validation Study in Germany on Two In Vitro Alternatives to the Draize Eye Irritation Test the HET-CAM Test and the 3T3 NRU Cytotoxicity Test. ATLA 24, pp. 741 – 858.
In addition the study follows the testing strategy for determination of eye irritation/corrosion as given in the following guideline: Organization for Economic Co-operation and Development (OECD), OECD Guidelines for testing of chemicals, Guideline No. 405: "Acute Eye Irritation / Corrosion", adopted April 24, 2002.
GLP compliance:
yes (incl. certificate)
Remarks:
testing lab.

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
other: in vitro test
Strain:
other: in vitro test

Test system

Vehicle:
other: Pluronic PE 6200
Controls:
no
Amount / concentration applied:
10, 20, 40%
Duration of treatment / exposure:
up to a maximum time period of 3.5 minutes
Observation period (in vivo):
210 seconds
Number of animals or in vitro replicates:
3 eggs for 10% concentration, 2 eggs for 20% concentration and 1 egg for 40% concentration
Details on study design:
Type of eggs: Fresh, fertilized hen eggs produced under controlled SPF conditions.
Strain/quality: White Leghorn, SPAFAS Inc., USA, SPF Premium.
Origin: Charles River Deutschland GmbH, Extertal.
Identification: At start of incubation period continuous numbering of the eggs with a felt pen.
Weight range at start of incubation period: 48.5 g – 61.1 g
Randomization: A randomization was not performed. For each solvent eggs of comparable weight (± 10 g) were used.
Reasons for the selection: Hen eggs are recommended as preferred type of eggs in the quoted references. Analogous to test animals, controlled SPF housing guarantees a defined microbiological status of the eggs.
Climate: Breeding in an incubator at constant temperature of 37.5°C (± 0.5°C) and a relative humidity of 62.5% (± 7.5%).
Automatic rotating device: Until including incubation day 8 and/or day 9, the eggs were rotated automatically 5 times a day. On the day before application the eggs were placed with the blunt end upward and were not rotated until preparation. The incubation conditions were checked daily. Deviations were recorded.
Candling of the eggs: The eggs were candled before the start of incubation and on the 9th and/or 10th day. Any defective or unfertilized eggs were discarded.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 210 sec
Score:
0
Max. score:
2
Remarks on result:
other: 10% in Pluronic PE 6200
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 210 sec
Score:
1 - 2
Max. score:
2
Reversibility:
other: intravascular coagulation
Remarks on result:
other: 20% in Pluronic PE 6200
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 210 sec
Score:
2
Max. score:
2
Remarks on result:
other: 40% in Pluronic PE 6200

Any other information on results incl. tables

Concentration   Egg No.  Time (seconds) until appearance of     Grading of effects     Additional findings
     Haemorrhagia  Coagulation  Haemorrhagia  Coagulation  
 10% in Pluronic PE 6200  1  210  210  0  0  none
   2  210  210  0  0  none
   3  210  210  0  0  none
 Mean:  n = 3  210  210  0  0  
             
 20% in Pluronic PE 6200  1  60  60  1  2  intravascular coagulation
   2  76  76  1  2  intravascular coagulation
 Mean:  n = 2  68  68  1  2  
             
 40% in Pluronic PE 6200  1  20  20  2  2  intravascular coagulation
 Mean:  n = 1  20  20  2  2  

Grading: 0 = no reaction; 1 = slight; 2 = moderate; 3 = severe

Applicant's summary and conclusion

Interpretation of results:
highly irritating
Remarks:
Migrated information
Conclusions:
Based on the results of this study and applying the evaluation criteria it is concluded, that the threshold concentration for effects indicating serious eye damage was > 10% < 20% for Cyclohexanone.