Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 2017 - March 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Test data is required for classification purposes.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
October 2017
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
The GLP certificate is included within the attached study report

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Specific details on test material used for the study:
Analytical purity – 97.60% (BaSeO4)
Physical state – Rhombohedral crystals
Colour – White
Molecular weight – 280.32 g/mol
Expiry date – 24th April 2019
Storage conditions – Room temperature
Stability under test conditions - Acceptable

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buhloe, Germany.

Test system

Vehicle:
other: RPMI 1640 without phenol red
Remarks:
(RPMI = Roswell Park Memorial Institute)
Amount / concentration applied:
750 μL of the test item (barium selenate) or control substance (positive control – 20% imidazole in physiological saline 0.9%; negative control – physiological saline 0.9% NaCl)
Duration of treatment / exposure:
Samples were mounted in the chambers of the corneal holder and filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for 1 hour at 32 ±1 oC.
Duration of post- treatment incubation (in vitro):
Not applicable.
Number of animals or in vitro replicates:
3 for each group (test item, negative control and positive control)

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
Test item (barium selenate)
Value:
24.13
Vehicle controls valid:
not examined
Negative controls valid:
other: The negative control responses should result in opacity and permeability values that are less than the established upper limits for the background bovine corneas treated with the respective negative control.
Positive controls valid:
yes
Other effects / acceptance of results:
The eye irritancy potential of Barium Selenate was investigated in the bovine opacity and permeability assay.
The test item was suspended with physiological saline 0.9% NaCl (see 10.3) to give a 20% concentration.
All 3 corneas treated with Barium Selenate showed opacity of the tissue
The following mean in vitro irritation score was calculated: 24.13
No prediction can be made regarding the classification of the test substance Barium Selenate according to the evaluation criteria.
The in vitro irritation score obtained with the positive control fell within two standard deviations of the current historical mean and therefore this assay is considered to be valid.
The negative control responses should result in opacity and permeability values that are less than the established upper limits for the background bovine corneas treated with the respective negative control.

Any other information on results incl. tables

Opacity of treated bovine corneas

Cornea No.

Test item

Initial opacity

Final opacity

Change of opacity value

Corrected opacity value

1

2

3

Mean value (MV)

Negative control

0.73

0.39

0.52

0.55

1.36

0.49

1.19

1.01

0.63

0.10

0.66

0.46

 

4

5

6

MV

Positive control

1.15

1.19

1.26

1.20

98.20

84.99

100.26

94.49

97.05

83.81

99.01

93.29

96.59

83.34

98.54

92.82

7

8

9

MV

Barium selenate test item

1.04

-0.02

-0.38

0.22

27.13

33.82

13.97

24.63

26.09

33.84

13.97

24.63

25.63

33.37

13.51

24.17

 

Permeability

Cornea No.

Test item

OD490

Corrected OD490 value

1

2

3

Mean value (MV)

Negative control

0.012

0.010

0.006

0.009

 

4

5

6

MV

Positive control

2.110

1.745

1.443

1.766

2.101

1.736

1.434

1.757

7

8

9

MV

Barium selenate test item

0.010

0.005

0.005

0.007

0.001

-0.004

-0.004

-0.003

 

In vitro irritation score

Cornea No.

Test item

OD490

Corrected OD490 value

IVIS

1

2

3

Mean value (MV)

Negative control

0.63

0.10

0.66

0.46

0.,012

0.010

0.006

0.009

 

 

 

0.060

4

5

6

MV

Positive control

96.59

83.34

98.54

92.82

2.101

1.736

1.434

1.757

 

 

 

119.17

7

8

9

MV

Barium selenate test item

25.63

33.37

13.51

24.17

0.001

-0.004

-0.004

-0.003

 

 

 

24.13

 

Historical mean in in vitro irritation score of the positive control

 

IVIS positive control – imidazole 20%

Mean value (MV)

Standard deviation (SD)

MV-2xSD

MV+2xSD

123.84

17.61

88.61

159.07

Number of replicates providing historical mean 30

 

 

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
Based on the results of this test, no prediction can be made regarding the classification of the test substance Barium Selenate, according to the evaluation criteria.
Executive summary:

The eye irritancy potential of Barium Selenate was investigated in the bovine corneal opacity and permeability assay. The test item (barium selenate) was suspended with physiological saline 0.9% NaCl, to give a 20% concentration.

The three corneas treated with the test item showed opacity of the tissue – the calculated mean in vitro irritation score is 24.13. No prediction could be made regarding the classification of the test substance, according to the evaluation criteria.

 

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.