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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
Study period:
4 Jul - 9 Aug 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

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)
Version / remarks:
adopted 7. September 2009
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Landesinstitut für Arbeitsschutz und Produktsicherheit, München

Test material

Constituent 1
Chemical structure
Reference substance name:
Fatty acids, C16-18 (even numbered), aluminum salts
EC Number:
939-582-4
Cas Number:
1471315-26-0
Molecular formula:
Not applicable (UVCB substance)
IUPAC Name:
Fatty acids, C16-18 (even numbered), aluminum salts
Details on test material:
- Name of test material (as cited in study report): Fatty acids, tallow, hydrogenated, aluminium salts
- Physical state: white solid powder
- Analytical purity: 84.33%
- Impurities (identity and concentrations): ash content 9.17%; humidity 0.75%; free fatty acids 14.13%; water-soluble salts 0.61%; aluminium 4.13%
- Lot/batch No.: C200752-001
- Expiration date of the lot/batch: 14.02.2014
- Storage condition of test material: at room temperature
- Other: CAS 91770-05-7

Test animals / tissue source

Species:
other: cattle
Strain:
other: bovine corneas
Details on test animals or tissues and environmental conditions:
TEST SYSTEM PREPARATION
- Source: isolated corneas obtained as a by-product from an abattoir from freshly slaughtered animals (Attenberger Fleisch GmbH & Co. KG, München)
- cornea preparation: on test day, fresh eyes were transported in HBSS containing Pen/Strep (5%) on ice. For cornea preparation, tissue surrounding the eyeball was carefully pulled away and corneas were excised leaving a 2 to 3 mm rim of sclera. Isolated corneas were stored in petri dishes containing HBSS.
- Cornea holders: corneas were mounted with the endothelial side against the O-ring of the posterior chamber, anterior chamber is positioned on top of of the cornea and tightend with screws, chamber of the cornea holder is filled with RPMI (without Phenol Red) containing 1% FBS and 2 mM L-glutamine, the posterior chamber was filled first, corneas were preincubated for one hour.

Test system

Vehicle:
other: sesame oil
Controls:
yes, concurrent vehicle
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% in vehicle

Duration of treatment / exposure:
4 hours ± 5 min
Observation period (in vivo):
not applicable
Number of animals or in vitro replicates:
3 corneas each per treatment
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removal of the treatment medium, with MEM, once the medium was macroscopically free of test substance the corneas were rinsed with RPMI (without phenol red) and anterior chambers were filled with fresh RPMI for opacity measurements
- Time after start of exposure: 4 hours

EVALUATION OF RESULTS:
- mean opacity value: calculated by subtracting the initial opacity reading (before treatment) from the final reading (after exposure), these values were corrected by subtracting from each the average change in opacity from corresponding negative control, the mean then was calculated by averageing the corrected opacity values for each treatment group
- permeability measurements (fluorescein): mean OD490 for each treatment were calculated by substracting the blank from each value and by subtracting the average corrected negative control value from the corrected value of each treatment measurement, the mean then was calculated by averaging the corrected OD490 values by averaging the corrected values for each treatment group
- IVIS score: (In Vitro Irritation Score) - calculated by summing up the mean OD490 score multiplied by 15 with the mean opacity value for each treatment

TOOL USED TO ASSESS SCORE: fluorescein + measurement of optical density at 490 nm and visual inspection, after opacity measurement

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: IVIS Score
Basis:
other: negative control
Time point:
other: 4 hours
Score:
1.1
Irritation parameter:
other: IVIS Score
Basis:
other: positive control
Time point:
other: 4 hours
Score:
216.51
Irritation parameter:
other: IVIS Score
Basis:
other: negative control (vehicle)
Time point:
other: 4 hours
Score:
0.05
Irritation parameter:
other: IVIS Score
Basis:
other: test item
Time point:
other: 4 hours
Score:
1.32
Irritant / corrosive response data:
no effects on the corneas were observed
Other effects:
no other effects observed

Any other information on results incl. tables

Table2: In Vitro Irritation Score

Cornea No.

Test Item

Corrected Opacity Value

Corrected OD490 Value

IVIS

1

Negative Control

1.00

0.087

 

2

0.00

0.089

 

3

-2.00

0.110

 

MV

-0.33

0.095

1.10

4

Positive Control

203.33

2.007

 

5

178.33

1.710

 

6

182.33

1.986

 

MV

188.00

1.901

216.51

10

Negative Control Sesame Oil

-1.00

0.063

 

11

0.00

0.030

 

12

-1.00

0.051

 

MV

-0.67

0.048

0.05

7

Test Item

0.67

0.026

 

8

1.67

-0.045

 

9

1.67

0.017

 

MV

1.33

-0.001

1.32

Applicant's summary and conclusion

Interpretation of results:
other: non-corrosive
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
According to Regulation (EC) No. 1272/2008, a substance can be considered a severe eye irritant (Serious eye damage Category 1/R41) based on a positive result in the Bovine Corneal Opacity and Permeability (BCOP) test. Negative in vitro corrosivity responses are not conclusive with respect to non-classification or classification as irritant (Category 2/R36) and shall therefore be subject to further evaluation."