Chloromethyltrimethylsilane

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study following EU method B.47

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

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. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Chloromethyltrimethylsilane

EC Number:

219-058-5

EC Name:

Chloromethyltrimethylsilane

Cas Number:

2344-80-1

Molecular formula:

C4H11ClSi

IUPAC Name:

(chloromethyl)trimethylsilane

Test material form:

other: colourless liquid

Details on test material:

Purity 99.8%

Test animals / tissue source

Species:

other: bovine cornea

Details on test animals or tissues and environmental conditions:

The assay uses isolated corneas obtained as a by-product from an abattoir
from freshly slaughtered animals.
The eyes were carefully examined for defects and any defect eyes were
discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea
was excised leaving a 2 to 3mm rim of sclera. The isolated corneas were
stored in a petri dish containing HBSS with Ca++ and Mg++ (containing
Penicillin/Streptomycin). Corneas then were mounted in corneal holders
(MC2, Clermont, France) with the endothelial side against the a-ring of the
posterior chamber. The anterior chamber then was positioned on top of the
cornea and tightened with screws. The chambers of the corneal holder were
filled with RPMI (without phenol red) containing 1% FBS and 2mM Lglutamine
(Complete RPMI). The posterior chamber was always filled first
The corneas were incubated for one hour at 32±1 °C in a water-bath.
After the incubation period, the medium was removed from both chambers
and replaced with fresh Complete RPMI. An initial opacity measurement
was performed on each of the corneas using an opacitometer (MC2,
Clermont, France). Three corneas with opacity readings approximately
equivalent to the median opacity of all corneas were selected as negative
control corneas. The opacity of each cornea was read against an air-filled
chamber and recorded. The medium was removed from the anterior chamber
and replaced with the test item or control respectively.
750uL of the test- or control substance were introduced into the anterior
chamber. After 10 minutes incubation at 32±1 °C the test- or control
substance was removed and the epithelium washed for at least three times
with MEM (containing phenol red). Once the media were free of test
substance, the corneas were finally rinsed with Complete RPMI (without
phenol red). The anterior chamber was refilled with Complete RPMI, and an
opacity measurement was performed after 2 hours incubation at 32 ± 1 °C
After the opacity measurement had been performed, the medium was
removed .from both chambers of the holder. The posterior chamber was
refilled with fresh Complete RPMI. ImL of a 4mg/mL sodium fluorescein
solution was added to the anterior chamber and the corneas were incubated
for 90 minutes at 32±1 °C. Then the medium from the posterior chamber was
removed and its optical density at 490nm (OD490) determined, using a
spectrophotometer (Jenway 6405, MC2, Clermont, France).

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: control cornea

Amount / concentration applied:

750 uL of the test- or control substance

Duration of treatment / exposure:

After 10 minutes incubation at 32±1 °C the test- or control
substance was removed and the epithelium washed for at least three times
with MEM (containing phenol red).

Observation period (in vivo):

An opacity measurement was performed after 2 hours incubation at 32 ± 1 °C.

Number of animals or in vitro replicates:

3 corneas for the test item
3 corneas as negative controls treated with isotonic saline solution (NaCl
0.9%)
3 corneas as positive control treated with ethanol

Details on study design:

The BCOP Assay is considered to be valid if the in vitro score obtained with
the positive control falls within the range of the reference control data:
Ethanol: 36.0 to 56.0

Results and discussion

In vivo

Results

Irritation parameter:

other: mean in vitro score

Basis:

mean

Time point:

other: 220 min

Score:

ca. 18.35

Reversibility:

other: not applicable

Irritant / corrosive response data:

The following mean in vitro score was calculated: 18.35

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the evaluation criteria the test item
Chloromethyltrimethylsilane is classified as mild eye irritating.

Executive summary:

The eye irritancy potential of Chloromethyltrimethylsilane was investigated

in the bovine corneal opacity and permeability assay.

The test item was tested as provided by the sponsor.

The following mean in vitro score was calculated:

18.35

Therefore the test item was classified as mild irritant.

The controls were at the upper limit of the historical control data range, as

indicated in the INVITTOX Protocol No. 98.

According to the historical range of own control data of the testing facility,

the controls were within the historical range.

Therefore the test is considered to be sensitive.

Conclusions

According to the evaluation criteria the test item

Chloromethyltrimethylsilane is classified as mild eye irritating