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Toxicological information

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:
May - August 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
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)
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Bis(neodecanoyloxy)dioctylstannane
EC Number:
269-595-4
EC Name:
Bis(neodecanoyloxy)dioctylstannane
Cas Number:
68299-15-0
Molecular formula:
C36H72O4Sn
IUPAC Name:
Neodecanoic acid, 1,1'-(dioctylstannylene) ester
Test material form:
liquid: viscous
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
16JSVA015
- Expiration date of the lot/batch:
14. September 2018
- Purity test date:
not state

RADIOLABELLING INFORMATION (if applicable)
not applicable

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Room Temperature: (20 ± 5°C), keep container tightly closed, store under inert gas
- Stability under test conditions:
assumed stable


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
none

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Specification
Species Bos primigenius Taurus (fresh bovine corneas)
Origin
Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr.
2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and
60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution
with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100
μg/mL) in a suitable cooled container. Then, the corneas were dissected and incubated in
medium at 32 ± 1 °C in an incubation chamber for 1 hour.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
750 µL
Duration of treatment / exposure:
10 min
Duration of post- treatment incubation (in vitro):
2 h post incubation, 90 min incubation with sodium fluorescein.
Number of animals or in vitro replicates:
3
Details on study design:
SELECTION AND PREPARATION OF CORNEAS
QUALITY CHECK OF THE ISOLATED CORNEAS
After the arrival of the corneas, they were examined and only corneas which were free
from damages were used. The corneas were excised with a scalpel and cut from the globe
with a 2-3 mm ring of sclera around the outside. Each cornea was transferred to a cornea
holder in which pre-warmed cMEM (32 ± 1 °C) without phenol red was filled. The holders
were then incubated for 1 hour in the incubation chamber at 32 ± 1 °C.


NUMBER OF REPLICATES
3
NEGATIVE CONTROL USED
HBSS: Hank’s Balanced Salt Solution (HBSS) 10-fold concentrated, diluted in demin. water
(1:10)
SOLVENT CONTROL USED (if applicable)
not applicable
POSITIVE CONTROL USED
Dimethylformamide, DMF, CAS-No. 68-12-2, undiluted,
APPLICATION DOSE AND EXPOSURE TIME
750 µL, 10 min
TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: yes, 2 h

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 1
- POST-EXPOSURE INCUBATION: 2h

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The change of opacity value of each treated cornea with test item, positive control and
negative control was calculated by subtracting the initial basal opacity from the post treatment
opacity reading for each cornea.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: please specify if the decision criteria as indicated in the TG was used. yes

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
Mean of three replicates
Value:
ca. 2.13
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
According to the guideline, the test is considered as valid if the positive control causes an
IVIS that falls within two standard deviations of the current historical mean.
The negative control has to show an IVIS ≤ 3.
The validity criteria and findings are given in the following table:

Parameter Criterion Found Assessment
IVIS of negative control HBSS ≤ 3 -0.64 ok
IVIS of positive control DMF undiluted 45.08 – 153.86 74.09 ok

Values for negative and positive controls were within the range of historical data of the test
facility. Therefore, the test was acceptable.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the test item Bis(neodecanoyloxy)dioctylstannane showed no effects on the cornea of the bovine eye.
Executive summary:

One valid experiment was performed.

Bovine corneas were used. They were collected from slaughtered cattle which were between

12 and 60 months old.

The test item Bis(neodecanoyloxy)dioctylstannane was brought onto the cornea of a bovine

eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour

and whose opacity had been measured.

The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of

the test item and 2 hours post-incubation, opacity and permeability values were measured.

HBSS-solution was used as negative control. The negative control showed no irritating

effect on the cornea and the calculated IVIS (in vitro irritancy score) is -0.64.

Dimethylformamide (DMF) undiluted was used as positive control. The positive control induced

serious eye damage on the cornea and falls within two standard deviations of the

current historical mean. The calculated IVIS (in vitro irritancy score) is 74.09.

Under the conditions of this study, the test item Bis(neodecanoyloxy)dioctylstannane

showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy

score) is 2.13.

According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS ≤ 3 requires

no classification for eye irritation or serious eye damage.