Reaction mass of disodium 2-[[2-hydroxy-3-[[methyl-(2-oxido-2-oxoethyl)amino]methyl]-5-nonylphenyl]methyl-methylamino and sodium 2-[(2-hydroxy-5-nonylphenyl)methyl-methylamino]acetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental Start Date: 21 February 2019 Experimental Completion Date: 21 February 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Annex VIII Data Requirement

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

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)

Version / remarks:

09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Sodium N-[(2-hydroxy-5-nonylphenyl)methyl]-N-methylaminoacetate

EC Number:

260-483-0

EC Name:

Sodium N-[(2-hydroxy-5-nonylphenyl)methyl]-N-methylaminoacetate

Cas Number:

56968-08-2

Molecular formula:

C19H31NO3.Na

IUPAC Name:

Sodium N-[(2-hydroxy-5-nonylphenyl)methyl]-N-methylaminoacetate

Constituent 2

Reference substance name:

Disodium N,N'-[(2-hydroxy-5-nonylphen-1,3-ylene)bis(methylene)]bis[N-methylaminoacetate]

EC Number:

263-601-9

EC Name:

Disodium N,N'-[(2-hydroxy-5-nonylphen-1,3-ylene)bis(methylene)]bis[N-methylaminoacetate]

Cas Number:

62568-43-8

Molecular formula:

C23H38N2O5.2Na

IUPAC Name:

disodium N,N’-[(2-hydroxy-5-nonylphen-1,3-ylene)]bis[N-methylaminoacetate]

Test material form:

solid

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:

10 minutes

Duration of post- treatment incubation (in vitro):

120 minutes

Number of animals or in vitro replicates:

3 eyes per exposure (i.e. negative control, positive control and test item)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Value:

>= 6

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Value:

>= 12.7

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Treatment	Cornea Number	Opacity					Permeability (OD492)		In VitroIrritancy Score
		Pre-Treatment	Post Treatment	Post Incubation	Post Incubation – Pre Treatment	Corrected Value		Corrected Value
Negative Control	1	4	3	4	0		0.003
	2	2	2	2	0		0.001
	3	3	2	2	0		0.000
					0.0^*		0.001^$		0.0
Positive Control	6	3	30	31	28	28.0	0.849	0.848
	9	3	29	34	31	31.0	1.250	1.249
	10	3	26	27	24	24.0	0.998	0.997
						27.7^^		1.031^^	43.1
Test Item	11	3	7	11	8	8.0	0.656	0.655
	12	3	7	8	5	5.0	0.359	0.358
	14	2	7	7	5	5.0	0.319	0.318
						6.0^^		0.443^^	12.7

OD = Optical Density

^*= Mean of the post-incubation – pre-treatment values

^{^}= Mean corrected value

^$=Mean permeability

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

No prediction of eye irritation can be made.

Executive summary:

Introduction

 

The purpose of this test was to identify test items that can induce serious eye damage and to identify test items not requiring classification for eye irritation or serious eye damage. The Bovine Corneal Opacity and Permeability (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 is assessed by quantitative measurements of changes in corneal opacity and permeability.

 

The test method can correctly identify test items (both chemicals and mixtures) inducing serious eye damage as well as those not requiring classification for eye irritation or serious eye damage, as defined by the United Nations (UN) Globally Harmonized System of Classification and Labelling of Items (GHS) and EU Classification, Labelling and Packaging (CLP) of chemicals (Regulation (EC) No 1272/2008), and it was therefore endorsed as scientifically valid for both purposes. Test items inducing serious eye damage are classified as UN GHS and EU CLP Category 1. Items not classified for eye irritation or serious eye damage are defined as those that do not meet the requirements for classification as UN GHS/EU CLP Category 1 or 2 (2A or 2B), i.e. they are referred to as UN GHS/EU CLP No Category.

 

Methods

 

The undiluted test item was applied for 10 minutes followed by an incubation period of 120 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate anIn VitroIrritancy Score (IVIS).

 

Data Interpretation

 

The test item is classified according to the prediction model as follows:

 

IVIS	UN GHS
≤ 3	No category
> 3; ≤ 55	No prediction can be mae
> 55	Category 1

 

Results

 

TheIn Vitroirritancy scores are summarized as follows:

 

Treatment	In Vitro Irritancy Score
Test Item	12.7
Negative Control	0.0
Positive Control	43.1

 

Conclusion

 

No prediction of eye irritation can be made.