TEA-Esterquat

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 November 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

liquid: viscous

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: AB Schlachthof GmbH & Co. KG, 63739 Aschaffenburg, Germany
- Characteristics of donor animals (e.g. age, sex, weight): 14 month old donor cattle
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): isolated eyes were stored in HBSS containing 1% (v/v) penicillin/streptomycin (100 units/mL penicillin and 100 μg/mL streptomycin) in the cooled slaughter-house and during transportation on the same morning to the laboratory.
- Time interval prior to initiating testing: The corneae were isolated on the same day after delivery of the eyes and used in the BCOP test on the same day.
- Indication of any existing defects or lesions in ocular tissue samples: corneas presenting defects such as vascularization, pigmentation, opacity and scratches were discarded
- Selection and preparation of corneas: Only corneae with a value of the basal opacity < 7 were used

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.75 mL

Duration of treatment / exposure:

ten minutes (± 30 seconds)

Duration of post- treatment incubation (in vitro):

2 h

Number of animals or in vitro replicates:

3

Details on study design:

NEGATIVE CONTROL USED: Saline (0.9% NaCl in deionised water)

POSITIVE CONTROL USED: 2-Ethoxyethanol

TREATMENT METHOD: open chamber

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

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: opacitometer OP_KiT opacitometer (Electro Design, 63-Riom France)
- 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: as indicated in the TG

The test is acceptable if
• the positive control gives an IVIS that falls within two standard deviations of the current historical mean (updated every three months), and if
• the negative control responses result in opacity and permeability values that are less than the established upper limits for background opacity and permeability values for bovine corneae treated with the respective negative control.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

mean of 3 corneas

Value:

1.52

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

DEMONSTRATION OF TECHNICAL PROFICIENCY: the controls gave the appropriate responses

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Any other information on results incl. tables

Results after 10 Minutes Treatment Time Test Group	Opacity value = Difference (t130-t0) of Opacity	Permeability at 490 nm (OD490)	IVIS	Mean IVIS	Standard Deviation IVIS	Proposed Category
Negative Control	0	0.066	0.99	0.95	0.07	No Category
	0	0.058	0.87
	0	0.065	0.98
Positive Control	91.00*	1.319*	110.79	114.08	5.67	Category 1
	86.00*	1.656*	110.84
	94.00*	1.775*	120.63
Test item	2.00*	0.003*	2.05*	1.52	0.45	No Category
	1.00*	0.018*	1.27*
	1.00*	0.017*	1.26*

*corrected values

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, according to the current study and under the experimental conditions reported, Esterification products of triglycerides, C18 (unsaturated) with triethanolamine, dimethyl sulphate-quaternized is not categorized (GHS).

Executive summary:

This in vitro study was performed to assess the corneal damage potential of Esterification products of triglycerides, C18 (unsaturated) with triethanolamine, dimethyl sulphate-quaternized by means of the BCOP assay using fresh bovine corneae.
After a first opacity measurement of the fresh bovine corneae (t0), the neat test item, the positive, and the negative controls were applied to corneae fixed in an incubation chamber in horizontal position for 10 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae. Further, the corneae were incubated for another 120 minutes at 32 ± 1 °C in a vertical position, while the anterior chamber contained incubation medium as well. Afterwards, opacity was measured a second time (t130).
After the opacity measurements, permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.
With the negative control (0.9% (w/v) NaCl solution in deionised water), neither an increase of opacity nor permeability of the corneae was observed.
The positive control (2-Ethoxyethanol) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damaging (EU CLP/EPA/GHS (Cat 1)).
Relative to the negative control, the test item Esterification products of triglycerides, C18 (unsaturated) with triethanolamine, dimethyl sulphate-quaternized did not cause a relevant increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 1.52. According to OECD TG 437 the test item is not categorized (GHS).