Registration Dossier

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:
guideline study

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)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Test material form:
solid: particulate/powder
Details on test material:
It was manufactured by MOLAR CHEMICALS Ltd.

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Test System : Isolated Cornea from the eyes of freshly slaughtered cattles
Source : Deonar Abattoir slaughter house, Mumbai, Maharashtra
Bovine Age : Between 1 to 5 years (age of the animals was determined based on the teeth count and horn ring count)
Transportation Condition : Transported under cold condition in Hanks’ Balanced Salt Solution containing antibiotics [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL]

Test system

Vehicle:
other: corn oil
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
750 μL ( 20 w/v% formulation)
Duration of treatment / exposure:
4h
Observation period (in vivo):
90 min at 32C
Duration of post- treatment incubation (in vitro):
90 min at 32C
Number of animals or in vitro replicates:
3/group
Details on study design:
3 parallel treatment were performed with physiological salinates, imidazole ( 20 w/v%), corn oil, uric acid at 20 w/v% in corn oil.
Application volulme was 0.75 ml/ cornea.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
3
Value:
ca. 0.46
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes

Any other information on results incl. tables

Bovine Corneal Opacity and Permeability Test for Uric Acid

 

TABLE 1:In VitroIrritation Score

 

Group : Normal Saline, 0.75 mL

Cornea Holder N°

Io

 (LUX)

I

(Initial)

(LUX)

Initial Opacity Value

I

(Post Treatment)

(LUX)

Post Treatment Opacity

Value

Corr. Opacity

Value

OD490

Value

Corr. OD490

Value

IVIS

1

1107

1000

4.69

992

5.04

0.35

0.054

0.009

0.49

2

1112

999

4.93

1006

4.62

-0.31

0.053

0.008

-0.19

3

1098

1030

3.05

1005

4.11

1.06

0.052

0.007

1.17

Mean

0.37

-

0.008

0.49

SD

0.69

-

0.001

0.68

 

Group : Imidazole at 20% (w/v) in normal saline, 0.75 mL

Cornea Holder N°

Io

 (LUX)

I (Initial)

(LUX)

Initial Opacity Value

I

(Post

Treatment)

(LUX)

Post Treatment Opacity Value

Corr. Opacity Value

Final Opacity Value

OD490

Value

Corr. OD490Value

Final OD490Value

IVIS

4

1056

946

5.05

284

108.72

103.67

103.30

1.623

1.578

1.570

126.85

5

1070

1014

2.62

302

101.74

99.12

98.75

2.983

2.938

2.930

142.70

6

1084

1010

3.34

291

108.99

105.65

105.28

2.15

2.105

2.097

136.74

Mean

102.44

-

2.207

2.199

135.43

SD

3.35

-

0.686

0.686

8.01

 

Group : Corn Oil, 0.75 mL

Cornea Holder N°

Io

 (LUX)

I (Initial)

(LUX)

Initial Opacity Value

I

(Post

Treatment)

(LUX)

Post Treatment Opacity

Value

Corr.

Opacity

Value

OD490

Value

Corr. OD490

Value

IVIS

7

1096

1031

2.93

1023

3.27

0.34

0.064

0.019

0.63

8

1099

1011

3.89

999

4.41

0.52

0.060

0.015

0.75

9

1095

1042

2.45

1021

3.31

0.86

0.070

0.025

1.24

Mean

0.57

-

0.020

0.87

SD

0.26

-

0.005

0.32

Keys: IVIS =In VitroIrritation Score, Io =Baseline Reading (With medium but without cornea), I = LUX   Reading with Medium and Cornea, OD490= Optical Density at 490 Wave Length, - = Not Applicable, Corr.             = Corrected. Blank OD490value = 0.045.


 

TABLE 1 (Continued)

Group : Uric Acid (suspension) at 20% (w/v) in Corn Oil, 0.75 mL

Cornea Holder N°

Io (LUX)

I (Initial)

(LUX)

Initial Opacity Value

I

(Post

Treatment)

(LUX)

Post Treatment Opacity

Value

Corr. Opacity

Value

Final Opacity

Value

OD490

Value

Corr. OD490

Value

Final OD490

Value

IVIS

10

1106

1005

4.43

975

5.78

1.35

0.78

0.061

0.016

-0.004

0.72

11

1083

971

5.02

968

5.16

0.14

-0.43

0.119

0.074

0.054

0.38

12

1081

1000

3.65

981

4.48

0.83

0.26

0.066

0.021

0.001

0.28

Mean

0.20

-

0.037

0.017

0.46

SD

0.61

-

0.032

0.032

0.23

Keys: IVIS =In VitroIrritation Score, Io =Baseline Reading (With medium but without cornea), I = LUX   Reading with Medium and Cornea, OD490= Optical Density at 490 Wave Length, - = Not Applicable, Corr.             = Corrected. Blank OD490value = 0.045.

InitialOpacity Value = [((Io⁄I)-b)/a)], Where I = Initial LUX, Io = Baseline Reading

Note: a (0.0251) and b (0.9894) are constant.

Example:Initial Opacity Value = [((1107/1000)-0.9894)/0.0251] =4.69

Post TreatmentOpacity Value = [((Io⁄I)-b)/a)], Where, I = Post Treatment LUX, Io = Baseline Reading.

Note: a (0.0251) and b (0.9894) are constant.

Example:Initial Opacity Value = [((1107/992)-0.9894)/0.0251] =5.04

Corr. Opacity Value = Post Treatment Opacity Value - Initial Opacity Value

Example:Corr. Opacity Value = 5.04 – 4.69 =0.35

Final Opacity Value = Corr. Opacity Value – Mean Corr. Opacity Value of Control (Group I)

Example:Final Opacity Value= 103.67 – 0.37 =103.30

Corr. OD490Value = OD490Value – Blank OD490Value, Where Blank Value for this trial was 0.045

Example:Corr. OD490Value = 0.054 -0.045 = 0.009

Final OD490Value = Corr. OD490Value –Mean Corr.OD490Valueof Control (Group I)

Example:Final OD490Value = 1.578 -0.008 = 1.570

IVIS (Control) = Corr. Opacity Value + (15 x Corrected OD490)

Example:IVIS = 0.35 + [15 x 0.009] =0.49

IVIS (Treatment) = Final Opacity Value + (15 x Final OD490)

Example:IVIS = 103.30 + (15 x 1.570) =126.85


 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The IVIS cut-off values for identifying test item as inducing serious eye damage (UN GHS Category 1) and test item not requiring classification for eye irritation or serious eye damage (CLP no Category)
Executive summary:

test item not requiring classification for eye irritation or serious eye damage (CLP No Category)

The IVIS score for the corneas treated with 750 μLuric acid(suspension) at 20% (w/v) concentration incorn oilwere found to be 0.46.