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

Endpoint summary

Description of key information

Skin irritation

CJ305was not irritant to the skin (OECD TG439).

 

Eye irritation       

CJ305 was not irritant to the eye (OECD TG405).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

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

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

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).

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From December 23, 2016 to April 13, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: the animal health research institute, council of agriculture, executive yuan
- Age: approximately 2-3 months
- Weight at study initiation: 2189-2410 g
- Housing: individually in a stainless steel wire mesh cage
- Water: ad libitum
- Acclimation period: at least 5 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 50 ± 20%
- Photoperiod: 12-hrs dark / 12-hrs light

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amount(s) applied (volume or weight with unit): 0.1 g

Observation period (in vivo):

1, 24, 48 and 72 hours, 7 and 14 days

Number of animals or in vitro replicates:

Three

Irritation parameter:

cornea opacity score

Basis:

animal: #01

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal: #02

Time point:

24/48/72 h

Score:

0.66

Reversibility:

fully reversible within: 72hours

Remarks on result:

positive indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal: #03

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 hours

Remarks on result:

positive indication of irritation

Irritation parameter:

iris score

Basis:

animal: #01, 02 and 03

Time point:

24/48 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal: #01

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal: #02

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48hours

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal: #03

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: Day 7

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

animal: #01

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal: #02 and 03

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48hours

Remarks on result:

positive indication of irritation

Table 1. Individual Body Weights and Clinical Observations

Animal ID	Body weight (g)		Body weight change (g) (D4-D1)	Clinical observation
	D1	D4
01	2189	2262	73	Normal^1
02	2410	2483	73	Normal^1
03	2327	2406	79	Normal^2

1: D1-D7

2: D1-D14

 

Table 2. Eye Irritation Scores at Observation Point on Treated Eye

Animal I.D.	Observation point	Cornea (D/A^5)^1	Iris	Conjunctivae (R/D)^2
01	1 h^4	0/-	0	2/1
	24 h	0/-	0^3	0/0
	48 h	0/-	0^3	0/0
	72 h	0/-	0^3	0/0
	D7	-	Normal	-
02	1 h	0/-	0	2/1
	24 h	1/1	0^3	1/1
	48 h	1/1	0^3	0/0
	72 h	0/-	0^3	0/0
	D7	-	Normal	-
03	1 h	0/-	0	2/1
	24 h	1/1	0^3	1/1
	48 h	0/-	0^3	1/0
	72 h	0/-	0^3	1/0
	D7	0/-	0^3	0/0
	D14	-	Normal	-

¹(D/A) = (Degree of density/Area of opacity)                                                                

²(R/D) = (Redness/Chemosis)

³Red colored iris

⁴Eye rinsed with saline at this time point

⁵Area of corneal opacity involved: 1=less than one-quarter; 2=greater than one-quarter but less than half; 3=greater than half but less than three-quarter; 4=greater than three-quarter, up to whole area

-: Not available

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 405 test method, CJ305 showed that positive irritant to the eye. Therefore, CJ305 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the QPS Taiwan Study Plan for T65316004-IR which is based on the SOP for the OECD 405 (CTPS-TE00441) and OECD 405 (OECD, 2012). CJ305 was given by a single ocular application at 0.1 g amount to NZW female rabbits and followed by ocular examination and clinical observation no more than 14 days. There were no test article effects on body weight. No irritation reaction in iris. Reversible corneal opacity, conjunctiva redness and chemosis were observed within 7 days. On the basis of the test results given above, CJ305 showed the light irritation to eyes. However, the result has not met a category based on GHS criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation

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.

 

Eye irritation

CJ305 was given by a single ocular application at 0.1 g amount to NZW female rabbits and followed by ocular examination and clinical observation no more than 14 days. There were no test article effects on body weight. No irritation reaction in iris. Reversible corneal opacity, conjunctiva redness and chemosis were observed within 7 days. On the basis of the test results given above, CJ305 showed the light irritation to eyes. However, the result has not met a category based on GHS criteria.