5-({4-chloro-6-[ethyl(phenyl)amino]-1,3,5-triazin-2-yl}amino)-4-hydroxy-3-[(2-sulfophenyl)diazenyl]naphthalene-2,7-disulfonic acid, lithium sodium salts

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From November 29, 2016 to June 09, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

5-({4-chloro-6-[ethyl(phenyl)amino]-1,3,5-triazin-2-yl}amino)-4-hydroxy-3-[(2-sulfophenyl)diazenyl]naphthalene-2,7-disulfonic acid, lithium sodium salts

EC Number:

943-066-4

Molecular formula:

Not applicable; this UVCB substance contains: C27H19ClN7O10S3.xLi.yNa, (x + y) = 3; 0 < (x,y) < 3 with 753.9 < MW < 802.0 g/mol (UVCB substance) and traces of NaCl.

IUPAC Name:

5-({4-chloro-6-[ethyl(phenyl)amino]-1,3,5-triazin-2-yl}amino)-4-hydroxy-3-[(2-sulfophenyl)diazenyl]naphthalene-2,7-disulfonic acid, lithium sodium salts

Test material form:

solid: particulate/powder

In vitro test system

Test system:

human skin model

Remarks:

EPISKIN Small Model

Source species:

human

Cell type:

other: reconstructed epidermis of normal human keratinocytes

Cell source:

other: Adult donors

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: % mean cell viability

Value:

100.2

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Table 1. Optical Density (OD) and the calculated Non Specific Colour % (NSC_living%) of the Additional Control Tissues

Additional control	Optical Density (OD)			NSC %
		Measured	Blank corrected
Treated with CJ305	1	0.048	0.002	1.0
	2	0.059	0.013
	mean	−	0.007

Notes:

1. Mean blank value was 0.046.

2. Optical density means the mean value of the duplicated wells for each sample (rounded to three decimal places).

3. NSC %:Non Specific Colour %

Table 2. Optical Density (OD) and the calculated relative viability % of the samples

Substance	Optical Density (OD)			Viability (% RV)	SD
		Measured	Blank corrected
Negative Control: Phosphate buffered saline	1	0.826	0.780	107.1	6.1
	2	0.746	0.699	96.0
	3	0.752	0.706	96.9
	mean	−	0.728	100.0
Positive Control: 5% (w/v) SDS solution	1	0.101	0.055	7.5	1.5
	2	0.106	0.060	8.2
	3	0.085	0.039	5.3
	mean	−	0.051	7.0
Test Item: CJ305	1	0.766	0.720	98.8	1.6
	2	0.789	0.742	101.9
	3	0.773	0.726	99.7
	mean	−	0.729	100.2

Notes:

1. Mean blank value was 0.046.

2. Optical density means the mean value of the duplicated wells for each sample (rounded to three decimal places).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 439 test method, CJ305 was non-irritant to skin. CJ305 was not met classification on GHS criteria.

Executive summary:

This test using the procedures outlined in the CiToxLAB Study Plan for 16/377-043B and OECD 439 (OECD, 2015). An in vitro skin irritation test of CJ305 was performed in a reconstructed human epidermis model. EPISKIN^TM(SM) is designed to predict and classify the irritation potential of chemicals by measuring its cytotoxic effect as reflected in the MTT assay. All data met the validity criteria. The mean cell viability of CJ305 was 100.2% compared to the negative control. This is above the threshold of 50%. Therefore, CJ305 was considered as being non-irritant to skin.