Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, 4-[(5-chloro-2-hydroxyphenyl)azo]-4,5-dihydro-3-methyl-1-phenyl-3H-pyrazol-3-one 4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-1-phenyl-3H-pyrazol-3-one 3-[[1-[[(2-ethylhexyl)amino]carbonyl]-2-oxopropyl]azo]-2-hydroxy-5-nitrobenzoate cobaltate complexes

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Wellperformed OECD study with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, 4-[(5-chloro-2-hydroxyphenyl)azo]-4,5-dihydro-3-methyl-1-phenyl-3H-pyrazol-3-one 4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-1-phenyl-3H-pyrazol-3-one 3-[[1-[[(2-ethylhexyl)amino]carbonyl]-2-oxopropyl]azo]-2-hydroxy-5-nitrobenzoate cobaltate complexes

EC Number:

276-160-2

EC Name:

Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, 4-[(5-chloro-2-hydroxyphenyl)azo]-4,5-dihydro-3-methyl-1-phenyl-3H-pyrazol-3-one 4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-1-phenyl-3H-pyrazol-3-one 3-[[1-[[(2-ethylhexyl)amino]carbonyl]-2-oxopropyl]azo]-2-hydroxy-5-nitrobenzoate cobaltate complexes

Cas Number:

71888-93-2

Molecular formula:

C63H66ClN10O12xCo

IUPAC Name:

Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, 4-[(5-chloro-2-hydroxyphenyl)azo]-4,5-dihydro-3-methyl-1-phenyl-3H-pyrazol-3-one 4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-1-phenyl-3H-pyrazol-3-one 3-[[1-[[(2-ethylhexyl)amino]carbonyl]-2-oxopropyl]azo]-2-hydroxy-5-nitrobenzoate cobaltate complexes

Details on test material:

CAS number: 71888-93-2
Purity: 98.8% w/w

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.53 or 3.79 kg and were 12 to 20 weeks old. After an acclimatisation period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

A volume of 0.1 mL of the test item, which was found to weigh approximately 51 mg

Duration of treatment / exposure:

up to 1 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.

A volume of 0.1 mL of the test item, which was found to weigh approximately 51 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.

Eight hours after test item application, a subcutaneous injection of post dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.

After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Appendix 2.

Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Any clinical signs of toxicity, if present, were also recorded.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 74952 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: 74959 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks:

No effects observed

Results 3

Irritation parameter:

iris score

Basis:

animal: 74952 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Results 4

Irritation parameter:

iris score

Basis:

animal: 74959 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Results 5

Irritation parameter:

other: redness

Basis:

animal: 74952 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 6

Irritation parameter:

other: redness

Basis:

animal: 74959 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 7

Irritation parameter:

chemosis score

Basis:

animal: 74952 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 74959 Male

Time point:

other: Highest score at 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

Ocular Reactions
Individual and mean scores for ocular irritation are given in Table 1.

Red colored staining of the fur around the treated eye was noted in both animals during the study.

No corneal or iridial effects were noted during the study.

Moderate conjunctival irritation was noted in both treated eyes 1 and 24 hours after treatment. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 48 Hour observation. Conjunctival redness was considered not to be due to red discoloration from the test item.

Both treated eyes appeared normal at the 72-Hour observation.

No corrosive effects were noted during the study. The test item did not induce significant or irreversible damage to the rabbit eye.

Other effects:

Body weight
Individual body weights and body weight change are given in Table 2.

Both animals showed expected gain in body weight during the study.

Any other information on results incl. tables

Table 1     Eye Irritation Scores

Individual Scores for Eye Irritation

Rabbit Number and Sex	IPR	Evaluation interval	Corneal Opacity	Area of CorneaI Opacity	Iris	Conjunctivae
						Redness	Chemosis	Discharge
74952Male	0	1 Hour	0	0	0	2	1	1
74959Male	0		0	0	0	2	2	1Sf
74952Male		24 Hour	0	0	0	2	2	1Sf
74959Male			0	0	0	2	1	1Sf
74952Male		48 Hour	0	0	0	2	1	1Sf
74959Male			0	0	0	1	1	1Sf
74952Male		72 Hour	0	0	0	0	0	0Sf
74959Male			0	0	0	0	0	0Sf

 

Mean Values after 24, 48 and 72 Hours

Rabbit Number and Sex	Number of available data points	Corneal Opacity	Iris	Conjunctivae
				Redness	Chemosis
74952Male	3	0.00	0.00	1.33	1.00
74959Male	3	0.00	0.00	1.00	0.67

IPR=Initial pain reaction

Sf = Red colored staining around the treated eye

Assessment According to EC Classification Guidelines

Evaluated Intervals	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
24 Hours	Not classified	Not classified	Not classified	Not classified
48 Hours
72 Hours

Table 2     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 3
74952Male	2.53	2.57	0.04
74959Male	3.79	3.95	0.16

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the findings in this study Savinyl-Feuerrot 3GLS does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Executive summary:

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405 and Method B5. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours after test item instillation. 

 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity and iritis were 0.00 for both animals. The individual mean scores for the conjunctivae were 1.33 and 1.00 for reddening and 1.00 and 0.67 for chemosis.

 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible and were no longer evident 72 hours after treatment in both animals (end of the observation period). No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

 

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

 

According to the findings in this study the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.