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

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental Start Date: 16 January 2019 Experimental Completion Date: 18 January 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

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

29 July 2016

Deviations:

no

GLP compliance:

yes

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

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other:

Cell source:

other:

Source strain:

not specified

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm™ Reconstructed Human Epidermis Model Kit

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

25mg

Duration of treatment / exposure:

3 and 60 minutes

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 Minute

Value:

>= 94

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 Minute

Value:

>= 58.6

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Exposure Period	Percentage Viability
	Negative Control	Positive Control	Test Item
3 Minute	100	3.8	94.0
60 Minute	100	3.1	58.6

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was considered to be non-corrosive to the skin.

Executive summary:

Introduction

 

The purpose of this test is to evaluate the corrosivity potential of the test item using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes.

 

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue. Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control. The results are used to make a prediction of the corrosivity potential of the test item.

 

Methods

 

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. Negative and positive control groups were treated for each exposure period. The test item was found to directly reduce MTT and therefore additional non-viable tissues were incorporated into the testing for correction purposes. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT-loading. After MTT-loading each tissue was placed in 2 mL of Isopropanol for MTT extraction.

 

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200mL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density (OD) was measured at 570 nm (OD570).

 

Data are presented in the form of percentage viability (MTT reduction in the test item treated

tissues relative to negative control tissues).

 

Results

 

The relative mean viabilities for each treatment group were as follows:

 

Exposure Period	Percentage Viability
	Negative Control	Positive Control	Test Item
3 Minute	100	3.8	94.0
60 Minute	100	3.1	58.6

 

The quality criteria required for acceptance of results in the test were satisfied.

 

Conclusion

 

The test item was considered to be non-corrosive to the skin.