Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 November 2015 - 04 November 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
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)
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
Name: DAILUBE IS-30
Chemical Name: 1, 3-bis (2, 4, 4-trimethylpentan-2yl) trisulfane
Use: Lubricant Additive
Molecular Formula: CH3-C(CH3)2-CH2-C(CH3)2-SSS-C(CH3)2-CH2-C(CH3)2-CH3
Molecular Weight: 322 g/mol
Appearance Pale yellow
Physical State: Liquid
Batch Number: L002
Purity: 95.4% by weight
Storage: Room temperature, in the dark
Expiry Date: 2016/8/15
Specific details on test material used for the study:
Batch: L002
Purity: 95.4%
Physical state/Appearance: pale yellow liquid
Expiry Date: 15 August 2016
Storage Conditions: room temperature in the dark

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: local abattoir
- Age of animals: 12 to 60 months old
- Storage: refrigerated on arrival and used within 24 hours of receipt

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
0.75 mL of the test item or control items
Duration of treatment / exposure:
10 minutes
Duration of post- treatment incubation (in vitro):
120 minutes
Number of animals or in vitro replicates:
3
Details on study design:
SELECTION AND PREPARATION OF CORNEAS: Only corneas free of damage were used.

QUALITY CHECK OF THE ISOLATED CORNEAS: macroscopically examined before and after dissection

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: sodium chloride solution

POSITIVE CONTROL USED: Ethanol

APPLICATION DOSE AND EXPOSURE TIME: 0.75 mL of the test item or control items for 10 minutes.

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: yes, 120 minutes

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 4
- POST-EXPOSURE INCUBATION: 120 minutes

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Each cornea was visually observed
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microplate reader (OD492)
- Histopathology: possible conduct of histopathology

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA:
IVIS CLASSIFICATION
≤ 3 No category. Not requiring classification to UN GHS or EU CLP
> 3; ≤55 No prediction of eye irritation can be made
> 55 Category 1. UN GHS or EU CLP Causes serious eye damage

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
Main
Value:
1.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Corneal Opacity and Permeability Measurement: Table 1
Corneal Epithelium Condition: Table 2

Any other information on results incl. tables

Table 1     Individual and Mean Corneal Opacity and Permeability Measurements

 

 

Treatment

 

CorneaNumber

 

Opacity

Permeability(OD)

InVitroIrritancyScore

Pre-Treatment

Post-Treatment

PostIncubation

Post-Incubation

-Pre-Treatment

CorrectedValue

 

CorrectedValue

 

Negative Control

19

4

5

5

1

 

0.006

 

 

22

4

4

6

2

 

0.004

 

 

23

4

4

6

2

 

0.001

 

 

 

 

 

 

1.7*

 

0.004¨

 

1.7

 

Positive Control

5

3

35

33

30

28.3

0.446

0.442

 

18

3

35

34

31

29.3

1.308

1.304

 

24

4

31

29

25

23.3

0.880

0.876

 

 

 

 

 

 

27.0·

 

0.874·

40.1

 

 

Test Item

26

4

4

4

0

0.0

0.065

0.061

 

27

3

4

4

1

0.0

0.006

0.002

 

28

4

3

9

5

3.3

0.008

0.004

 

 

 

 

 

 

1.1·

 

0.023·

1.5

 

 OD=Opticaldensity       *=Meanofthepost-incubation-pre-treatmentvalues      ¨=Meanpermeability                      ·=Meancorrectedvalue

Table 2     Corneal Epithelium Condition Post Treatment and Post Incubation

 

 

Treatment

 

CorneaNumber

Observation

PostTreatment

PostIncubation

 

Negative Control

19

clear

clear

22

clear

clear

23

clear

clear

 

Positive Control

5

cloudy

cloudy

18

cloudy

cloudy

24

cloudy

cloudy

 

Test Item

26

clear

clear

27

clear

clear

28

clear

clear

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No category. Not requiring classification to UN GHS or EU CLP.