Pyridinium, 1-[4-[[4-[(4-amino-9,10-dihydro-9,10-dioxo-3-sulfo-1-anthracenyl)amino]cyclohexyl]amino]-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]-3-carboxy-, inner salt, sodium salt (1:3)

Endpoint summary

Description of key information

Skin irritation

The Primary Irritation Index (PII) of CR SB08 was 0 and CR SB08 was categorized as non-irritant (OECD TG404).

 

Eye irritation

CR SB08was not any 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 vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 16, 2016 to June 27, 2016

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: WEI XIN HANG
- Weight at study initiation: 2.5382 to 3.1200 kg
- Housing: individually in a stainless steel wire mesh cage
- Water: ad libitum
- Acclimation period: 7 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 55 ± 15%
- Photoperiod: 12hrs dark / 12hrs light

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

Amount / concentration applied:

- Amount(s) applied (volume or weight with unit): 0.5 g test article was moistened with 0.5 mL normal saline

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours

Number of animals:

Three

Details on study design:

- Area of exposure: upper left and lower right dorsal area
- Washing (if done): with distilled water

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Table 1. Body weight of the rabbits

Animal ID	Sex	Body weight (kg)
		Day 1	Day 4
13	Female	3.1200	3.1897
14	Female	2.5382	2.6098
15	Female	2.8504	2.9203

Table 2. Individual skin reactions

Animal I.D.	Observation time (hours)	Grading (erythema / edema)
		Test area		Control area
13	1	0/0	0/0	0/0	0/0
	24± 1	0/0	0/0	0/0	0/0
	48 ± 1	0/0	0/0	0/0	0/0
	72 ± 1	0/0	0/0	0/0	0/0
14	1	0/0	0/0	0/0	0/0
	24 ± 1	0/0	0/0	0/0	0/0
	48 ± 1	0/0	0/0	0/0	0/0
	72 ± 1	0/0	0/0	0/0	0/0
15	1	0/0	0/0	0/0	0/0
	24 ± 1	0/0	0/0	0/0	0/0
	48 ± 1	0/0	0/0	0/0	0/0
	72 ± 1	0/0	0/0	0/0	0/0

Table 3. Individual animal primary irritation scores

Animal I.D.	Scores^a		Primary Irritation Scores (PIS)^b
	Test area	Control area
13	0	0	0
14	0	0	0
15	0	0	0
Primary Irritation Index (PII)^c= 0

^aSum of scores at all time intervals on backside skin in each rabbit and divided the sum by the total number of observation.

^bPIS = (scores of test area) – (scores of control area).

^cPII = Sum of PIS÷number of animals.

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 404 test method and Evaluation table for single dermal irritation, the PII for CR SB08 was 0 and CR SB08 was categorized as non-irritant. Therefore, CR SB08 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab Study Plan for M62-151100069001EN which is based on the SOP for the OECD 404(SOPP-305) and OECD 404 (OECD, 2002). The sites were grades for erythema and edema at 1, 24, 48 and 72 hours after the patches removal. No clinical signs of toxicity were found. Body weight changes were within the normal range in all animals. All animals was no formation of erythema, eschar or edema on the test area within 72 hours after patch removal. The Primary Irritation Index for CR SB08 was calculated to be 0. On the basis of the test results given above and according to Evaluation table for single dermal irritation, the response of CR SB08 was categorized as non-irritant.

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 February 16, 2016 to June 27, 2016

Reliability:

1 (reliable without restriction)

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: WEI XIN HANG
- Weight at study initiation: 2.8282-2.9954 kg
- Housing: individually in a stainless steel wire mesh cage
- Water: ad libitum
- Acclimation period: 7 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 55 ± 15%
- 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

Duration of treatment / exposure:

1 hours

Observation period (in vivo):

1, 24, 48 and 72 hours

Number of animals or in vitro replicates:

Three

Irritation parameter:

cornea opacity score

Basis:

animal: #18, #19 and #20

Time point:

24/48/72 h

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal: #18, #19 and #20

Time point:

24/48/72 h

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: #18, #19 and #20

Time point:

24/48/72 h

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal: #18, #19 and #20

Time point:

24/48/72 h

Max. score:

0

Remarks on result:

no indication of irritation

Table 1. Body weight of the rabbits

Animal ID	Sex	Body weight (kg)
		Day 1	Day 4
18	Female	2.9954	3.0392
19	Female	2.6542	2.6986
20	Female	2.8282	2.9162

Table 2. Individual response of the test rabbits

Animal I.D.	Time^a (hour)		Grades of the irritation reaction
			Cornea		Conjunctivae			Iris
			Opacity	Area	Redness	Chemosis	Discharge
18	1	Test^b	0	0	0	1	0	0
		Control^c	0	0	0	0	0	0
	24 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	48 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	72 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
19	1	Test	0	0	0	1	0	0
		Control	0	0	0	0	0	0
	24 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	48 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	72 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
20	1	Test	0	0	0	1	0	0
		Control	0	0	0	0	0	0
	24 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	48 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0
	72 ± 1	Test	0	0	0	0	0	0
		Control	0	0	0	0	0	0

^aObservation time was started after application of the test article

^bLeft eye of the test rabbits

^cRight eye of the test rabbits

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 405 test method, CR SB08 showed that no irritant to the eye. Therefore, CR SB08 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the SuperLab Study Plan for M62-151100078001EN which is based on the SOP for the OECD 405 (SOPP-306) and OECD 405 (OECD, 2012). There were no test article effects on body weight.Reversiblechemosiswith score of 1 was observed within 1 hours after test article application.CR SB08 was given by a single ocular application at 0.1 g amount to NZW female rabbits and followed by ocular examination within 72 hoursand a 4 days clinical observation period. Conjunctivachemosiswith score 1 was determined within 24 hours and fully recovered. On the basis of the test results given above, the response of the test article was not any irritant to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation

The sites were grades for erythema and edema at 1, 24, 48 and 72 hours after the patches removal.No clinical signs of toxicity were found. Body weight changes were within the normal range in all animals. All animals was no formation of erythema, eschar or edema on the test area within 72 hours after patch removal. The Primary Irritation Index for CR SB08 was calculated to be 0. On the basis of the test results given above and according toEvaluation table for single dermal irritation, the response of CR SB08 was categorized as non-irritant.

 

Eye irritation

There were no test article effects on body weight. Reversiblechemosisof conjunctivae with score of 1 was observed within 1 hours after test article application.CR SB08 was given by a single ocular application at 0.1 g amount toNZW female rabbits and followed by ocular examination within 72 hours and a 4 days clinical observation period. Conjunctivachemosiswith score 1 was determined within 24 hours and fully recovered.On the basis of the test results given above, the response of the test article was not any irritant to the eye.