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EC number: 301-097-5 | CAS number: 93981-14-7
Eye irritation
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: Study was conducted by a GLP accredited laboratory using OECD Testing Guideline 437.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 013
- Report date:
- 2013
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Tosyl salt
- IUPAC Name:
- Tosyl salt
- Reference substance name:
- 6-[(p-tosyl)amino]hexanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)
- EC Number:
- 301-097-5
- EC Name:
- 6-[(p-tosyl)amino]hexanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)
- Cas Number:
- 93981-14-7
- Molecular formula:
- C13H19NO4S.C6H15NO3
- IUPAC Name:
- 2-hydroxy-N,N-bis(2-hydroxyethyl)ethanaminium 6-{[(4-methylphenyl)sulfonyl]amino}hexanoate
- Test material form:
- other: Waxy solid
- Details on test material:
- Appearance: Orange/brown waxy solid
Identity: 6-[(p-tosyl)amino]hexanoic acid, compound with 2,2',2''-nitrilotriethanol (1:1)
Storage conditions: 15 to 30°C in a sealed container, protected from the light
Purity: 98%
Constituent 1
Constituent 2
Test animals / tissue source
- Species:
- other: Bovine Cornea
- Strain:
- other: Not applicable
- Details on test animals or tissues and environmental conditions:
- Bovine corneas were supplied by a local abattoir. The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin at 100 IU/ml and streptomycin at 100 µg/ml) in a suitably sized container and transported on the same day to the testing facility. On arrival at the test facility the eyes were carefully examined for defects including increased opacity, scratches and neovascularisation. Only corneas free from such defects were used.
Test system
- Vehicle:
- other: sodium chloride
- Controls:
- yes
- Amount / concentration applied:
- The test article was tested as a 20% w/v suspension in 0.9% sodium chloride solution. A volume of 750 µl of the test article formulation was applied to each of three corneas followed by a four hour incubation at 32°C.
- Duration of treatment / exposure:
- Four hour incubation
- Observation period (in vivo):
- After the incubation period the cornea were washed once in media without phenol red and incubated for a further 1.5 hours ± 5 minutes
- Number of animals or in vitro replicates:
- Three bovine corneas were tested.
- Details on study design:
- Fresh corneas, mounted onto specifically designed holders were treated topically with the test material. Eye corrosion/severe irritation potential was based on the combined effect of the test substance on the opacity of the cornea following the treatment and the cornea’s ability to resist penetration of a fluorescent dye through the tissue.
Results and discussion
In vivo
Resultsopen allclose all
- Irritation parameter:
- other: corneal opacity
- Basis:
- mean
- Time point:
- other: 4 hour incubation after application of test substance
- Score:
- 30
- Max. score:
- 34.7
- Reversibility:
- not specified
- Remarks on result:
- other: score for test substance
- Irritation parameter:
- other: corneal opacity
- Basis:
- mean
- Time point:
- other: 4 hour incubation after application of negative control
- Score:
- 0
- Max. score:
- 1.7
- Reversibility:
- not specified
- Remarks on result:
- other: score for negative control
- Irritation parameter:
- other: corneal opacity
- Basis:
- mean
- Time point:
- other: 4 hour incubation after application of positive control
- Score:
- 60
- Max. score:
- 69.7
- Reversibility:
- not specified
- Remarks on result:
- other: score for positive control
- Irritation parameter:
- other: corneal permeability
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours after application of the test substance
- Score:
- 2.031
- Max. score:
- 2.037
- Reversibility:
- not specified
- Remarks on result:
- other: score for test substance
- Irritation parameter:
- other: corneal permeability
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours after application of negative control
- Score:
- 0
- Max. score:
- 0.129
- Reversibility:
- not specified
- Remarks on result:
- other: score for negative control
- Irritation parameter:
- other: corneal permeability
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours after application of positive control
- Score:
- 1.78
- Max. score:
- 1.986
- Reversibility:
- not specified
- Remarks on result:
- other: score for positive control
- Irritation parameter:
- overall irritation score
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours
- Score:
- 60.47
- Reversibility:
- not specified
- Remarks on result:
- other: score for test substance
- Irritation parameter:
- overall irritation score
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours
- Score:
- 0
- Reversibility:
- not specified
- Remarks on result:
- other: score for negative control
- Irritation parameter:
- overall irritation score
- Basis:
- mean
- Time point:
- other: Total incubation of 5.5 hours
- Score:
- 86.7
- Reversibility:
- not specified
- Remarks on result:
- other: score for positive control
- Irritant / corrosive response data:
- The test substance produced an In Vitro Irritation Score of 60.47.
Any other information on results incl. tables
Corneal Opacity
Substance |
Cornea number |
Initial opacity |
Post incubation opacity |
Change in opacity |
Mean change in opacity |
Corrected opacity |
Mean corrected opacity |
Test substance |
15 |
1 |
36 |
35 |
N/A |
33.7 |
30 |
16 |
1 |
37 |
36 |
34.7 |
|||
35 |
1 |
24 |
23 |
21.7 |
|||
Negative control |
3 |
0 |
3 |
3 |
1.33 |
1.7 |
0 |
9 |
1 |
2 |
1 |
-0.3 |
|||
10 |
0 |
0 |
0 |
-1.3 |
|||
Positive control |
19 |
0 |
58 |
58 |
N/A |
56.7 |
60 |
20 |
0 |
71 |
71 |
69.7 |
|||
21 |
0 |
55 |
55 |
53.7 |
Corneal Permeability
Substance |
Cornea number |
Mean blank OD490 |
OD490 |
Corrected OD490 |
Mean corrected OD490 |
Final corrected OD490 |
Mean group corrected OD490 |
Test substance |
15 |
- |
2.128 |
2.128 |
N/A |
2.030 |
2.031 |
16 |
2.136 |
2.136 |
2.037 |
||||
35 |
2.126 |
2.126 |
2.027 |
||||
Negative control |
3 |
0 |
0.035 |
0.035 |
0.099 |
-0.063 |
0 |
9 |
0.228 |
0.228 |
0.129 |
||||
10 |
0.033 |
0.033 |
-0.066 |
||||
Positive control |
19 |
- |
2.085 |
2.085 |
N/A |
1.986 |
1.780 |
20 |
1.794 |
1.794 |
1.695 |
||||
21 |
1.758 |
1.758 |
1.660 |
OD490 = Optical Density at 490 nm
Calculated In Vitro Irritation Score (IVIS)
Substance |
Mean opacity |
Mean permeability |
IVIS (mean opacity + (15 x mean permeability)) |
Negative control |
0 |
0 |
0 |
Positive control |
60 |
1.78 |
86.7 |
Test substance |
30 |
2.031 |
60.47 |
Applicant's summary and conclusion
- Interpretation of results:
- Category 1 (irreversible effects on the eye)
- Remarks:
- Migrated information Criteria used for interpretation of results: EU
- Conclusions:
- The test substance produced an In Vitro Irritation Score (IVIS) of 60.47 and was considered to be corrosive or severely irritating to the eye. The test substance is classified in Category 1 (Irreversible effects on the eye) for severe eye damage, in accordance with EU CLP criteria.
- Executive summary:
The eye irritation potential of the test substance was determined in accordance with the OECD Guideline for Testing of Chemicals 437. The test substance was applied (in vitro) to three bovine corneas, with a positive and a negative control applied to further groups of three bovine cornea. Corneal opacity was recorded after a 4 hour incubation period. The cornea were washed after the initial 4 hour incubation and incubated for a further 1.5 hours before corneal permeability was determined. The mean corneal opacity and mean corneal permeability scores were used to calculate a mean In Vitro Irritation Score (IVIS). The test substance produced an IVIS of 60.47 and was considered to be corrosive or severely irritating to the eye (an IVIS >55.1 indicates a corrosive or severe irritant to eyes). All three cornea exposed to the test material had a corneal opacity score of >20, with an average score of 30. It is not expected that the effects to the cornea would be fully reversible, the test substance is therefore classified in Category 1 (Irreversible effects on the eye) for severe eye damage, in accordance with EU CLP criteria.
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