Reaction products of 4-Diazo-3,4-dihydro-7-nitro-3-oxo-1-naphthalenesulfonic acid coupled with 7-Amino-4-hydroxy-2-naphthalenesulfonic acid, subsequently chelated with Chromium(3+) and cobalt(3+), further mixed with reaction products of 2,4,6-Trichloro-1,3,5-triazine and Ethanol, 2-[(4-aminophenyl)sulfonyl]-, 1-(hydrogen sulfate), sodium salts

Endpoint summary

Description of key information

Skin irritation

Everzol SR98 was non-irritant to skin (OECD TG404).

                          

Eye irritation

Everzol SR98was non-irritant to 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 December 08, 2017 to August 24, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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: Granja RG (Suzano/SP-Brazil)
- Age at study initiation: 10 weeks old
- Weight at study initiation: 2443.8-3006.7 g
- Housing: individually in stainless steel cages
- Water: ad libitum
- Acclimation period: at least 5 days
- Temperature (°C): 22.8-23.0 °C
- Humidity (%): 50-64 %
- Photoperiod: 12hrs dark / 12hrs light

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amount(s) applied (volume or weight with unit): 0.5 g test substance was moistened with 0.5 mL water

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours after removing the bandage.

Number of animals:

Three

Details on study design:

- Area of exposure: the skin of back

Irritation parameter:

erythema score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Table 1. Results of Dermal Irritation in the Periods of Evaluation.

Animal’s data				Reactions evaluation after application						Reactions evaluation after removal of the patch
				3 min		1 h		4 h		1 h		24 h		48 h		72 h
Animal#	Initial Weight (g)	Final Weight (g)	Sex	O	E	O	E	O	E	O	E	O	E	O	E	O	E
1	2443.8	2924.4	♂	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2	3006.7	3274.4	♂	-	-	-	-	-	-	0	0	0	0	0	0	0	0
3	2852.5	2885.3	♂	-	-	-	-	-	-	0	0	0	0	0	0	0	0

O: Oedema; E: Erythema

 

Table 2. Clinical Signs.

Animal#	Clinical Signa
	After application			After removal of the patch
	3 min	1 hour	4 hours	1 hour	24 hours	48 hours	72 hours
1	0	0	0	0	0	0	0
2	-	-	-	0	0	0	0
3	-	-	-	0	0	0	0

0: None visual alterations observed.

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 404 test method test method, Everzol SR98 was non-irritant to skin. Therefore, Everzol SR98 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the BIOAGRI Study Plan for 16264.466.088.17. 0.5 g of Everzol SR98 moistened in 0.5 mL of purified water was applied to the skin of three New Zealand White rabbits and then covered with a gauze patch. After the 4 hours exposure period, the patches were removed, any residual test item removed using purified water and the animal #1 was evaluated after the application in the period of 3 minutes, 1 and 4 hours. After removing the bandage semiocclusive, three animals were examined skin reactions, behavioral and clinical alterations in approximately 1, 24, 48 and 72 hours. Adjacent untreated shaved areas of the skin were used as a negative control. Under test conditions, three rabbits do not showed any skin reactions and no treatment-related behavioral or clinical changes were observed during the observation period. On the basis of the test results given above, the response of Everzol SR98 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 December 08, 2017 to March 19, 2018

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: Granja RG Suzano/SP, Brasil
- Age at study initiation: 10 weeks old
- Weight at study initiation: 2413.6 to 2616.7 g
- Housing: individually in stainless steel cages
- Water: ad libitum
- Acclimation period: at least 5 days
- Temperature (°C): 22.8 to 23.0 °C
- Humidity (%): 50 to 64 %
- 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):

Approximately 1, 24, 48, 72 hours after the application of test substance.

Number of animals or in vitro replicates:

Three

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal: #1 and #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Irritation parameter:

chemosis score

Basis:

animal: #1, #2 and #2

Time point:

24/48/72 h

Score:

0.33

Table 1. Initial and Final Weights of the Animals and Grading of Ocular Reactions Observed during the Test.

Animal #		01				02				03
Weight (g)	Initial	2413.6				2413.8				2616.7
	Final	2775.6				2473.3				2751.7
Sex		♀				♀				♀
Observation period		Hours				Hours				Hours
		1	24	48	72	1	24	48	72	1	24	48	72
Cornea	Opacity	0	0	0	0	0	0	0	0	0	0	0	0
	Area	0	0	0	0	0	0	0	0	0	0	0	0
Iris	Irite	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctivae	Hyperemia	1	0	0	0	1	0	0	0	1	1	0	0
	Chemosis	2	1	0	0	2	1	0	0	2	1	0	0
	Secretion	-	-	-	-	-	-	-	-	-	-	-	-
Lesion	Fluorescein retention	NR	-	-	-	NR	-	-	-	NR	-	-	-

+: Presence; -: Ausence; NR: Not realized

Table 2. Clinical Signs

Animal #	Clinical Signs / Period of Evaluation
	01 hour	24 hours	48 hours	72 hours
1	0	0	0	0
2	0	0	0	0
3	0	0	0	0

0: None visual alterations observed.

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD 405 test method, Everzol SR98 showed that non-irritant to the eye. Therefore, Everzol SR98 was not met a category based on GHS criteria.

Executive summary:

This test using the procedures outlined in the BIOAGRI Study Plan for 16264.465.102.17. All the animals presented body weight gain during the observation period. The test was initially conducted using one New Zealand White rabbit. Because no severe ocular reaction was observed in the initial test, two additional New Zealand White rabbits were tested to confirm the response. Each animal received a 0.1 g dose of pure test item in the conjunctival sac of the right eye. After application the animals were examined the ocular reaction, behavioral and clinical alterations at approximately 1, 24, 48 and 72 hours. The untreated left eye was used as a negative control. The Everzol SR98 applied in tested eye of all rabbits caused hyperemia and chemosis and do not have retention of fluorescein sodium dye on the surface of the cornea. After the evaluation of 48 hours, there was regression of eyes reactions in tested eyes of all rabbits. After the evaluation of 72 hours, finalizing the study and all rabbits do not note any behavioral or clinical change related to treatment during the observation period.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation

0.5 g of Everzol SR98 moistened in 0.5 mL of purified water was applied to the skin of three New Zealand White rabbits and then covered with a gauze patch. After the 4 hours exposure period, the patches were removed, any residual test item removed using purified water and the animal #1 was evaluated after the application in the period of 3 minutes, 1 and 4 hours. After removing the bandage semiocclusive, three animals were examined skin reactions, behavioral and clinical alterations in approximately 1, 24, 48 and 72 hours. Adjacent untreated shaved areas of the skin were used as a negative control. Under test conditions, three rabbits do not showed any skin reactions and no treatment-related behavioral or clinical changes were observed during the observation period. On the basis of the test results given above, the response of Everzol SR98 was categorized as non-irritant.

 

Eye irritation

Everzol SR98 was given by a single ocular application at 0.1 g amount to NZW rabbits and followed by ocular examination within 72 hours. Three rabbits resulted in irritation of the conjunctivae, which consisted ofhyperemia and chemosis. The observed irritation had completely resolved within 48 hours in all animal. No iridial irritation or corneal opacity was observed. On the basis of the test results given above, the response of the test article was not any irritant to the eye.