1-Propanaminium, 2-hydroxy-N,N,N-trimethyl-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)thio]-, chloride (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 April, 2015 - 09 April, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

(2013)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

(2010)

Deviations:

no

Principles of method if other than guideline:

- The Ocular Toxicity Working Group (OTWG) of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the National Interagency Centre for the Evaluation of Alternative Toxicological Methods (NICEATM), Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006.
- In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.
- Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

EC Number:

811-523-6

Cas Number:

88992-45-4

Molecular formula:

C14 H19 F13 N O S . Cl

IUPAC Name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): NS-4000
- Appearance: Off-white powder (determined by WIL Research Europe)
- Storage condition of test material: At room temperature desiccated

Test system

Vehicle:

physiological saline

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 750 µL 20% (w/v) test substance per cornea

NEGATIVE CONTROL
- Amount applied (volume or weight with unit): 750 µL of physiological saline per cornea

POSITIVE CONTROL
Amount applied (volume or weight with unit): 750 µL per cornea
Concentration (if solution): 20% (w/v) Imidazole

Duration of treatment / exposure:

240 minutes

Details on study design:

TEST SITE
- Isolated bovine cornea
- Three corneas were used for each treatment group
- The corneas were incubated at 32 ± 1°C

REMOVAL OF TEST SUBSTANCE
- Washing: yes (with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation))
- Time after start of exposure: 240 minutes

SCORING SYSTEM:
- After exposure, the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement
- Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Merck) was evaluated (90 minutes incubation at 32°C)
- After the incubation period, permeability evaluation of the cornea was performed
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate the mean in vitro score:
Mean in vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
- Opacity and permeability values were also evaluated individually

TOOL USED TO ASSESS SCORE:
- Opacitymeter and microplate reader

DATA EVALUATION:
- A test substance that induces a mean IVIS >55 is classified with Eye Irr. Cat. 1 in accordance with the CLP Regulation
- A test substance that induces a mean IVIS ≤ 3 is not classified for Eye irritation in accordance with the CLP Regulation
- For a test substance that induces a mean IVIS of >3 - ≤55, it cannot be concluded whether the substance needs to be classified or not and for these substances, more (in vivo) information is needed.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

mean; 240 minutes

Value:

11.7

Negative controls validity:

other: 0

Positive controls validity:

other: 117

Any other information on results incl. tables

Summary of opacity, permeability and in vitro scores:

Treatment	Mean Opacity	Mean Permeability	Mean In vitro Irritation Score
Negative control	0.0	0.000	0.0
Positive control	83.3	2.271	117.4
NS-4000	6.0	0.380	11.7

Individual in vitro scores:

Eye	Negative control correctedFinal Opacity	Negative control corrected Final OD490	In vitro Irritancy Score
Negative control
1	1.0	0.004	1.1
2	-1.0	-0.008	-1.1
3	0.0	0.004	0.1
Positive control
4	72.0	2.219	105.3
5	106.0	2.561	144.4
6	72.0	2.031	102.5
NS-4000
16	5.0	0.434	11.5
17	4.0	0.230	7.4
19	9.0	0.477	16.2

Applicant's summary and conclusion

Interpretation of results:

other: no prediction on the classification can be made.

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Based on a mean in vitro irritancy score of 11.7 in a Bovine Corneal Opacity and Permeability test with NS-4000, no prediction on the classification can be made.

Executive summary:

Using the Bovine Corneal Opacity and Permeability test (BCOP test) NS-4000 was screened for its eye irritancy potential in accordance with OECD 437 and according to GLP principles. The substance diluted in physiological saline was applied as such directly on top of the corneas in such a way that the cornea was completely covered (750 µL 20% (w/v) per cornea). Adequate negative and positive controls were included. NS-4000 induced ocular irritation through both endpoints (opacity/permeability), resulting in a mean in vitro irritancy score of 11.7 after 240 minutes of treatment. Since NS-4000 induced an IVIS > 3 ≤ 55, no prediction on the classification can be made for eye irritating properties.