2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-06-29 to 2011-07-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted in accordance with the following guideline : Commission Regulation (EC) No. 440/2008/EC, Method B.46 : 'In-vitro skin irritation : Reconstructed human epidermis model test'.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No. 440/2008/EC, Method B.46; 'In-vitro skin irritation : Reconstructed human epidermis model test'

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

EC Number:

251-410-3

EC Name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

Cas Number:

33229-34-4

Molecular formula:

C12H19N3O5

IUPAC Name:

2-({4-[bis(2-hydroxyethyl)amino]-2-nitrophenyl}amino)ethan-1-ol

Details on test material:

Name : Jarocol Blue 2
Batch number : JBT03110003
Purity : 99.4%
CAS number : 33229-34-4
EC number : 251-410-3
Name : 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol

Test animals

Species:

other: In-vitro assessment using EpiDerm TM tissue

Strain:

other: In-vitro assessment using EpiDerm TM tissue

Test system

Type of coverage:

other: In vitro assessment using EpiDerm TM tissue

Preparation of test site:

other: In vitro assessment using EpiDerm TM tissue

Vehicle:

unchanged (no vehicle)

Controls:

other: In-vitro assessment using EpiDerm TM tissue

Amount / concentration applied:

10mg

Duration of treatment / exposure:

15 minutes

Observation period:

42 hours

Number of animals:

Not applicable

Details on study design:

This in-vitro study was performed in order to evaluate the potential of 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol (HC Blue 2) to evoke skin irritation in a human skin model.

The test consists of a topical exposure of the neat test item to a human reconstructed epidermis model followed by a cell viability test. Cell viability is measured by dehydrogenase conversion of MTT (3-[4,5-dimethyl thiazole 2-yl] 2,5-diphenyl-tetrazolium bromide) present in cell mitochondria into a blue formazan salt that is quantitatively measured after extraction from tissues. The percent reduction of cell viability in comparison of untreated negative controls is used to predict skin irritation potential.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: Relative individual tissue viability (%)

Value:

111.4

Remarks on result:

other:

Remarks:

Basis: other: Tissue 1. Time point: 15 minutes. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: Relative individual tissue viability (%)

Value:

107

Remarks on result:

other:

Remarks:

Basis: other: Tissue 2. Time point: 15 minutes. (migrated information)

Results 3

Irritation / corrosion parameter:

other: other: Relative individual tissue viability (%)

Value:

97.7

Remarks on result:

other:

Remarks:

Basis: other: Tissue 3. Time point: 15 minutes. (migrated information)

Any other information on results incl. tables

Mean OD540 values and percentage viabilities for the negative control item, positive control item and test item

Item	OD540 of tissues	Mean OD540 of triplicate tissues	+/- SD of OD540	Relative individual tissue viability/%	Relative mean viability/%	+/-SD of relative mean viability
Negative Control Item	0.742			106.2
Negative Control Item	0.725	0.699	0.060	103.7	100	8.6
Negative Control Item	0.631			90.3
Positive Control Item	0.036			5.2
Positive Control Item	0.043	0.035	0.009	6.2	5.0	1.3
Positive Control Item	0.026			3.7
Test Item	0.779			111.4
Test Item	0.748	0.737	0.049	107.0	105.4	7.0
Test Item	0.683			97.7

Skin irritation potential of the test item is assessed as given in the following.

Criteria for in vitro interpretation	Classification
Relative Mean Tissue Viability is <=50%	Irritant (I) R38
Relative Mean Tissue Viability is >50%	Non-irritant (NI)

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The substance 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol (HC Blue 2) was considered to be non-irritant in the Human Skin Model Test.

Executive summary:

The skin irritation potential of 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol (HC Blue 2) was assessed in the Human Skin Model Test. One valid experiment was performed. Three tissues of the human skin model Episkin TM were treated with 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol (HC Blue 2). DPBS buffer was used as negative control, 5% SDS was used as positive control. The relative mean tissue viability for the positive control treated tissues was <=40% relative to the negative control treated tissues and the standard deviation value of the percentage viability was <=18%. The positive control acceptance criterion was therefore satisfied. The mean OD540 for the negative control treated tissues was >= 0.6 and the standard deviation value of the percentage viability was <=18%. The negative control acceptance criterion was therefore satisfied. The standard deviation calculated from individual percentage tissue viabilities of the three identically treated tissues was <=18%. The test item acceptance criterion was therefore satisfied. 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol (HC Blue 2) is considered to be non-irritant in the Human Skin Model Test.