Hexamethylene diisocyanate, trimers, reaction products with 2-hydroxyethyl acrylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 Jul 2016

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 guideline

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)

Version / remarks:

adopted in 2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Schwabach, Germany

Test material

Test material information

Constituent 1

Reference substance name:

Hexamethylene diisocyanate, trimers, reaction products with 2-hydroxyethyl acrylate

EC Number:

500-734-6

EC Name:

Hexamethylene diisocyanate, trimers, reaction products with 2-hydroxyethyl acrylate

Cas Number:

162492-01-5

Molecular formula:

C39 H60 N6 O15

IUPAC Name:

Hexamethylene diisocyanate, trimers, reaction products with 2-hydroxyethyl acrylate

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: abattoir A. Moksel AG, Buchloe, Germany
- Storage, temperature and transport conditions of ocular tissue: Fresh eyes were transported in Hanks´balanced salt solution containing penicillin/streptomycin on ice. Immediately after arrival of the eyes at the laboratory, cornea preparation was initiated.
- Indication of any existing defects or lesions in ocular tissue samples: no (visual examination)
- Indication of any antibiotics used: no

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 100%

Duration of treatment / exposure:

10 min at 32 ± 1°C

Observation period (in vivo):

Not applicable

Duration of post- treatment incubation (in vitro):

2 h at 32 ± 1°C

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
The eyes were carefully visually examined for defects and any defective eyes were discarded. The tissue surrounding the eyeball was carefully pulled way and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS and mounted in corneal holders with the endothelial side against the O-ring of the posterior chamber. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers were filled with RPMI medium (without phenol red) containing 1% fetal bovine serum and 2 mM L-glutamine (complete RPMI). The corneas were equilibrated for 1 h at at 32 ± 1°C. After the equilibration period, the medium was replaced with fresh complete RPMI.

QUALITY CHECK OF THE ISOLATED CORNEAS
An initial measurement was performed on each of the corneas using the opacitometer.

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
0.9% NaCl (physiological saline)

POSITIVE CONTROL USED
100% ethanol

APPLICATION DOSE AND EXPOSURE TIME
750 µL of the test substances and control substances for 10 min at 32 ± 1°C

TREATMENT METHOD: open chamber

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: at least 3 times

- POST-EXPOSURE INCUBATION: 2 h at 32 ± 1°C

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: illuminance measurement using a calibrated opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: decision criteria as indicated in the TG was used

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

-0.06

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:

OTHER EFFECTS:
- Visible damage on test system: no

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values if different from the ones specified in the test guideline: IVIS of the positive control (100% ethanol; 32 replicates): 46.61 (mean value, MV); 9.15 (standard deviation, SD); 28.3 (MV - 2 x SD); 64.92 (MV + 2 x SD)

In vivo

Irritant / corrosive response data:

With the negative control (0.9% (w/v) NaCl solution) neither an increase of opacity nor permeability of the corneae could be observed. The positive control (2-ethoxyethanol) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damaging. Relative to the negative control, the test item did not cause an increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score (IVIS) was -0.06.

Applicant's summary and conclusion

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

CLP: not classified