Reaction mass of potassium difluoro{[2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate with potassiumfluoride and potassium carbonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Study period:

2009-03-26 to 2009-06-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The experimental study on the supporting substance is guideline-conform under GLP without deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: The Bovine Corneal Opacity and Permeability Assay (BCOP) will be performed according to the INVITTOX (UK) protocol no. 98 "The Bovine Corneal Opacity and Permeability Assay", dated February 1994.

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Bovine Corneal Opacity and Permeability (BCOP) Assay, SOP of Microbiological Associates Ltd., UK, Procedure Details, April 1997.

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Draft OECD Guideline for Testing of Chemicals: The Bovine Corneal Opacity and Permeability (BCOP) Test Method for Identifying Ocular Corrosives and Severe Irritants (December, 2008).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Energie, Landwirtschaft und Verbraucherschutz

Test material

Test material information

Constituent 1

Reference substance name:

cC604

IUPAC Name:

cC604

Constituent 2

Reference substance name:

Acetic acid, 2,2-difluoro-2-[[2,2,4,5- tetrafluoro-5-(trifluoromethoxy)-1,3- dioxolan-4-yl]oxy]-, ammonium salt (1:1)

IUPAC Name:

Acetic acid, 2,2-difluoro-2-[[2,2,4,5- tetrafluoro-5-(trifluoromethoxy)-1,3- dioxolan-4-yl]oxy]-, ammonium salt (1:1)

Constituent 3

Reference substance name:

Reaction mass of ammonium difluoro {[(4S,5R)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate, ammonium difluoro {[(4R,5S)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate, ammonium difluoro {[(4S,5S)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate and ammonium difluoro {[(4R,5R)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate

EC Number:

682-238-0

Cas Number:

1190931-27-1

Molecular formula:

C6H4F9NO6

IUPAC Name:

Reaction mass of ammonium difluoro {[(4S,5R)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate, ammonium difluoro {[(4R,5S)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate, ammonium difluoro {[(4S,5S)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate and ammonium difluoro {[(4R,5R)-2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetate

Details on test material:

- Name of test material (as cited in study report): cC604 (dry salt)
- Molecular formula (if other than submission substance): C6H4F9NO6
- Molecular weight (if other than submission substance): 357
- Smiles notation (if other than submission substance): [NH4+].FC1(F)OC(F)(OC(F)(F)C([O-])=O)C(F)(OC(F)(F)F)O1
- InChl (if other than submission substance): no data
- Structural formula attached as image file (if other than submission substance): see Fig.1 (C6O4 Structural formula)
- Substance type: pure substance
- Physical state: solid
- Analytical purity: 96.2%
- Impurities (identity and concentrations): NH4F=0.021% (w/w), NH4Cl=0.31% (w/w), H2O=0.328% (w/w), the organic impurities are dioxolan based and represent the 0.8% by mol on total mol of organic phase of sample
- Composition of test material, percentage of components: not reported
- Isomers composition: not reported
- Purity test date: not reported
- Lot/batch No.: 150/31
- Expiration date of the lot/batch: 31 December 2020
- Stability under test conditions: Stable in water and saline for days
- Storage condition of test material: At room temperature

Test system

Vehicle:

physiological saline

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approx. 100 mg at a volume of 1.0 mL on the surface of the corneas
- Concentration (if solution): 100 g/l


VEHICLE
- Amount(s) applied (volume or weight with unit): 1.0 ml per applicatiom
- Concentration (if solution): 0.9% (w/v)
- Lot/batch no. (if required): no data
- Purity: no data

Duration of treatment / exposure:

240 minutes (± 5 minutes)

Observation period (in vivo):

not applicable to in vitro testing

Number of animals or in vitro replicates:

Not applicable to in vitro testing; three bovine corneas used for each test item, negative and positive controls

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After incubation, test medium was rinsed off from the application side by changing cMEM buffer
- Time after start of exposure: 240 min


SCORING SYSTEM:

The following formula was used to determine the in vitro score of the negative control:
In vitro Score = opacity value + (15 x OD490 value)
The following formula was used to determine the in vitro score of the positive control and the test item:
In vitro Score = (opacity value – opacity valuenegative control + (15 x corrected OD490 value)
The in vitro score was calculated for each individual treatment and positive control cornea.
The mean in vitro score value of each treated group was calculated from the individual in vitro score values.

In vitro score Proposed in vitro irritation scale
0-3 non eye irritant
3.1-25 mild eye irritant
25.1-55 moderate eye irritant
55.1-80 severe eye irritant
>80.1 very severe eye irritant

TOOL USED TO ASSESS SCORE: This test is designed to measure the opacity of the cornea by quantifying the ability of light of pass through it. The permeability, as a result of the irritation potential of the test item, is determined using Na-fluorescein solution.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

mean value of bovine cornea 1, 2, and 3

Run / experiment:

240 min of exposure

Value:

183.46

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

The mean "in vitro irritation score" was 183.46, thus much above the score of 80.1, which represents the minimum score for the data interpretation "very severe irritating".
maximum score observed: 243.77

Any other information on results incl. tables

Table 1: Results after 240 Minutes Incubation Time

Test Group	Opacity value = Difference (t240-t0) of Opacity *	Permeability at 490 nm (OD490)*	In vitro Score	Mean in vitro Score	Proposed in vitro Irritation Scale
Negative control (mean of 3 tests)	0	0.143		2.15	No eye irritant
Positive Control	104	-0.063	103.05
Positive Control	113	-0.034	112.49	105.73	Very severe eye irritant
Positive Control	101	0.044	101.66
cC6O4	214	1.985	243.77
cC6O4	130	2.232	163.48	183.46	Very severe eye irritant
cC6O4	109	2.276	143.14

*corrected values

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

It can be stated that in this study and under the experimental conditions reported, the test item cC6O4 ammonium salt is considered to be a very severe eye irritant.
As the observed effects caused by the test item exceeded by far the effects caused by the positive control substance, a worst case classification for C6O4 ammonium salt is suggested, i.e. EU CLP classification H318, eye damage 1.

Considering the similarity of chemical structure between F- Diox potassium salt and cC6O4 ammonium salt, the same degree of eye irritant potential of cC6O4 ammonium salt is expected for F-DIOX potassium salt. Therefore it can be concluded that F-DIOX potassium salt is a severe eye irritant.

Executive summary:

In order to evaluate the potential eye irritation of F- Diox potassium salt it was deemed appropriate to use the Read Across approach based on the experimental study performed on cC6O4 ammonium salt by virtue of similarity of chemical structure between F- Diox potassium salt and cC6O4 ammonium salt (CAS no: 1190931-27-1).

 F-Diox potassium salt and cC6O4 ammonium salt are two salts of the same caboxylic acid, they differ only for the cationic part (K+ in F-Diox potassium salt and NH4+ in cC6O4 ammonium salt). Considering the similarity of chemical structure a similar biological behaviour is expected.

This in vitro study was performed to assess the corneal irritation and damage potential of cC6O4 ammonium salt by means of the BCOP assay using fresh bovine corneas.

After a first opacity measurement of the fresh bovine corneae (t0), about 100 mg of the test item cC6O4 ammonium salt, the positive, and the negative controls were applied to corneas and incubated for 240 minutes at 32 ± 2 °C in complete medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneas and opacity was measured again (t240). After the opacity measurements permeability of the corneas was determined while application of 1 mL of a fluorescein solution for 90 minutes at 32 ± 2 °C in a horizontal position. The coming out liquid was measured spectrophotometrically.

With the negative control (0.9% NaCl solution) neither an increase of opacity nor permeability of the corneas could be observed. The in vitro score was calculated as 2.15. The positive control (10% (v/v) Benzalconium chloride) showed clear opacity and distinctive permeability of the corneas and therefore, is classified as very severe eye irritant. The in vitro score was calculated as 105.73. The test item cC6O4 ammonium salt caused opacity and permeability of the corneas compared with the results of the negative control. The calculated in vitro score was 183.46 and therefore, the test item was classified as very servere eye irritant.

As the observed effects caused by C6O4 ammonium salt exceeded by far the effects caused by the positive control substance, a worst case classification is suggested, i.e. EU CLP classification H318, eye damage 1.

Considering the similarity of chemical structure between F- Diox potassium salt and cC6O4 ammonium salt, the same degree of eye irritant potential of cC6O4 ammonium salt is expected for F-DIOX ammonium salt. Therefore it can be concluded that F-DIOX potassium salt is a severe eye irritant.