Butanedioic acid, sulfo-, 4-[2-[(2-hydroxyethyl)amino]ethyl] ester, N-C18-unsatd. acyl derivs., disodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to GLP and valid methods and is considered relevant and reliable for classification. It is an in vitro study by means of BCOP, predicting severe eye irritation, which is considered adequate in combination with the in vitro irritation testing.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: ICCVAM BCOP test method, 2007

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Note for Guidance on Non-Clinical Tolerance Testing of Medicinal Products, CPMP/SWP/2145/00, adopted March 1, 2001.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of C18 unsatd. fatty acid diethanolamide and maleic anhydride, sulphonated, sodium salt

IUPAC Name:

Reaction product of C18 unsatd. fatty acid diethanolamide and maleic anhydride, sulphonated, sodium salt

Test material form:

other: Lyofilisate

Details on test material:

- Name of test material (as cited in study report): Butanedioic acid, sulfo-, 4-(2-((2-hydroxyethyl)amino)-ethyl) ester, N- C18-unsatd. acyl derivs., disodium salts ; Sulfosuccinate of Fatty acid diethanolamide, sodium salt
- Physical state: Lyophilisate
- Analytical purity: 29.9% (active ingredient)
- Impurities (identity and concentrations): See confidential details
- Composition of test material, percentage of components: See confidential details
- Purity test date: 2012-03-19
- Lot/batch No.: FL21-22-RS
- Expiration date of the lot/batch: March 2014
- Stability under test conditions: Stable
- Storage condition of test material: At +10° to +25°C (0-40°C acceptable)
- Other: Manufacturer: TFL France S.A.S., 4 rue de l’Industrie, 68333 Huningue Cedex, France

Test animals / tissue source

Species:

other: Bovine eyes from cattle in the age range of 6 to 12 months were obtained from Hubert Bahlmann GmbH & Co., Lindern.

Test system

Vehicle:

physiological saline

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL of test or control item were added to completely cover the cornea's epithelium in the anterior chamber.
- Concentration (if solution): The test item was dissolved in 0.9% NaCl-solution to a final concentration of 10% active ingredient, which complies with the requirements for surfactants.

Duration of treatment / exposure:

10 minutes

Results and discussion

In vivo

Results

Irritation parameter:

other: IVIS (in vitro irritancy score)

Score:

0.8

Irritant / corrosive response data:

The optical density (OD492) or absorbance values were measured at a wavelength of 492 nm. An opacity value of -0.336 and a permeability value of 0.075 compared to the negative control were determined. An IVIS 0.80 was calculated. Hence, the test item was not classified as a severe irritant and not corrosive, based on the results of this test.
The corneas treated with the positive control item 1% NaOH solution revealed an opacity value of 69.199 and a permeability value of 2.150 compared to the negative control. The IVIS value of 101.43 was well above the cut-off value of 55.1 and, hence, the acceptance criteria for the test were fulfilled. 1% NaOH solution was a severe irritant and corrosive to eyes.

Any other information on results incl. tables

Table 3. In vitro irritancy score (IVIS)

	Cornea No.	Opacity	Permeability	IVIS
				Per Cornea	Per Group
					Mean	SD
NaCl 0.9%	1	0.331	0.036	0.9	0.50	0.36
	2	0.125	0.008	0.3
	3	0.229	0.012	0.4
NaOH 1%	4	65.705	2.147	97.9	101.43	3.66
	5	69.863	2.091	101.2
	6	72.029	2.213	105.2
Active ingredient 1:10#	7	-0.585	0.094	0.8	0.80	0.10
	8	-0.104	0.053	0.7
	9	-0.320	0.078	0.9

NC: negative control
SD: standard deviation
#: tested as 10% concentration in 0.9% NaCl solution

Applicant's summary and conclusion

Interpretation of results:

other: predicted non-corrosive

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the present test conditions Butanedioic acid, sulfo-, 4-(2-((2-hydroxyethyl)amino)-ethyl) ester, N-C18-unsatd. acyl derivs., disodium salts, tested in the in vitro BCOP test method, had an IVIS value of < 55.1 and consequently it is not classified as a severe irritant and is not corrosive.

Executive summary:

The purpose of this study was to determine if the test item can be classified as “ocular corrosive and severe irritant” employing an in vitro system. The BCOP test method is an organotypic model that provides short-term maintenance of normal physiological and biochemical function of the bovine cornea in vitro. In this test method, damage by the test item was assessed by quantitative measurements of changes in corneal opacity and permeability in isolated corneas from bovine eyes. The corneas of the eyes were dissected and the remaining sclera was mounted in corneal holders with anterior (epithelium) and posterior (endothelium) chambers.

Corneal opacity was measured quantitatively as the amount of light transmission through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea, as detected in the medium in the posterior chamber. The measurements were used to calculate an in vitro irritancy score (IVIS), which was used to assign an in vitro irritancy hazard classification category for prediction of the in vivo ocular irritation potential of the test item.

Three corneas were used for each treatment group (test item, negative and positive controls). The test item was dissolved in 0.9% NaCl-solution to a final concentration of 10% active ingredient, which complies with the requirements for surfactants. 0.9% NaCl solution was used as the negative control and 1% NaOH solution as the positive control item. The test item and the controls were applied to the epithelial surface of the cornea by addition to the anterior chamber of the corneal holder for an exposure time of 10 minutes.

The optical density (OD₄₉₂) or absorbance values were measured at a wavelength of 492 nm. An opacity value of -0.336 and a permeability value of 0.075 compared to the negative control were determined. An IVIS 0.80 was calculated. Hence, the test item was not classified as a severe irritant and not corrosive, based on the results of this test.

The corneas treated with the positive control item 1% NaOH solution revealed an opacity value of 69.199 and a permeability value of 2.150 compared to the negative control. The IVIS value of 101.43 was well above the cut-off value of 55.1 and, hence, the acceptance criteria for the test were fulfilled. 1% NaOH solution was a severe irritant and corrosive to eyes.

Under the present test conditions Butanedioic acid, sulfo-, 4-(2-((2-hydroxyethyl)amino)-ethyl) ester, N-C18-unsatd. acyl derivs., disodium salts, tested in the in vitro BCOP test method, had an IVIS value of < 55.1 and consequently it is not classified as a severe irritant and is not corrosive.