Titanium tetrapropanolate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017 July to 2017 September

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 guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Zirconium tetrapropanolate

EC Number:

245-711-9

EC Name:

Zirconium tetrapropanolate

Cas Number:

23519-77-9

Molecular formula:

C12H28O4Zr

IUPAC Name:

Zirconium tetrakis(propa-1-olato)

Constituent 2

Reference substance name:

Propan-1-ol

EC Number:

200-746-9

EC Name:

Propan-1-ol

Cas Number:

71-23-8

Molecular formula:

C3H8O

IUPAC Name:

propan-2-ol

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Supplier: Borica Co., Ltd.
- Lot number: 000577Z
- Expiration date of the lot/batch: 2018-Jul-06
- CAS number: 23519-77-9

Test animals / tissue source

Species:

human

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): 50 microliter
- Concentration (if solution): neat

VEHICLE
- Not used

Duration of treatment / exposure:

30 minutes +/- 2 min

Duration of post- treatment incubation (in vitro):

12 +/- 2 min post-treatment immersion and 2 hours +/- 15 min post-treatment incubation.

Number of animals or in vitro replicates:

3 replicates per run.

Details on study design:

- Details of the test procedure used
Prior to the study, the required compatibility checks confirmed that the test item interfered with MTT and therefore additional freeze killed negative controls (NC-FK) and freeze killed test item controls (TA-FK) were included in the assay to determine the background level of interference (manifesting as altered viability) produced by the test item. The level of viability measured in the interference controls was subtracted from the standard test viability value to give an adjusted final viability. No water colouration was observed and therefore colorant control was not required.

After pre-wetting tissues with 20 microliter PBS (Sterile Dulbecco’s phosphate buffered saline) for 30 +/- 2 min, single topical application of 50 microliter of neat test item or of reference items to the surface of the EpiOcular™ models for 30 +/- 2 min, followed by a 12 +/- 2 min post-treatment immersion, and 2 hr +/- 15 min post-treatment incubation, prior to the MTT endpoint; three tissues per condition (n = 3).


- RhCE tissue construct used, including batch number
EpiOcular™ EIT (OCL-200) Reconstructed Human Ocular Epithelium
Supplier: MatTek Corporation
Keratinocyte Strain: 4F1188
Lot number: 23799
Description of the RhCE tissue: The EpiOcular™ tissue model (OCL200-MatTek Corporation) is composed of stratified human keratinocytes in a three-dimensional structure, reflecting the morphology and function of the human corneal epithelium found in vivo.


- MTT test conditions
Spectrometer: BMG LabTech FluoStar Optima
Wavelength for O.D. measurements: 570 nm


- Positive and negative control means and acceptance ranges based on historical data
1. Mean OD570 of the negative control (treated with sterile water) tissues is > 0.8 and < 2.5.
2. Mean of the positive control relative percentage viability is below 50% of negative control viability after 30 +/- 2 minutes exposure.


- Acceptable variability between tissue replicates for positive, negative controls and test items
The SD between three tissues replicates should not exceed 18% in the same run (for negative and positive control tissues and tissues of test items).

Results and discussion

In vitro

Results

Irritation parameter:

other: Tissue viability

Run / experiment:

1 run, 3 replicates per run

Value:

ca. 15.27

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Any other information on results incl. tables

Table 3. Mean and SD of viability measurements and of viability percentages after 30 +/- 2 min of application and 2 h +/- 15 min post-incubation of test and reference items.

Consition	Mean of OD	SD of OD	mean of viability %	SD of viability (%)	CV %	Classification
NC	1.657	0.016	100.00	0.938	0.938	Non-irritant
PC	0.522	0.115	31.479	6.964	22.122	Irritant
TA2	0.253	0.008	15.277	0.481	3.150	Irritant

Note:

NC: Negative control (sterile H2O)

PC: neat methyl acetate

TA2: Zirconium tetrapropanolate

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

A Test item is considered to be irritant to the eye (i.e. would require labelling as either GHS 1 or 2) if they eye model viability after exposure and post-treatment incubation is ≤ 60%.
A test item is considered as a non-irritant to the eye (i.e. would not require a warning label in the European chemical classification systems) if the eye model viability after exposure and post-treatment incubation is > 60%.
The current assay is not intended to differentiate between GHS class 1 and GHS class 2 or R36 and R41 (degree of stromal damage).
The percentage of viability obtained with test item Zirconium tetrapropanolate was 5.602%, therefore it is considered as Irritant to the eye.

Executive summary:

The eye irritation potential of Zirconium tetrapropanolate was assessed in vitro according to OECD Test Guideline 492 (Reconstructed human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labeling for eye irritation or serious eye damage).

After a 30 +/-2 minute exposure on the surface of EpiOcular™ reconstructed ocular epithelium and 2 h +/- 15 mins post-incubation time, the viability of the tissues was assessed and compared to a negative control. The percentage viability was 5.602% and therefore:

Zirconium tetrapropanolate

was classified as Irritant to the human eye.