Sodium p-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]benzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 26 June 2012 and 17 July 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do no effect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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

GLP compliance:

yes (incl. QA statement)

Remarks:

The frequency of these process based inspections was changed from a monthly to a three monthly cycle. This deviation is considered not to affect the purpose or integrity of the study

Test material

Test material information

Constituent 1

Reference substance name:

Sodium p-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]benzenesulphonate

EC Number:

223-989-2

EC Name:

Sodium p-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]benzenesulphonate

Cas Number:

4156-21-2

Molecular formula:

C9H6Cl2N4O3S.Na

IUPAC Name:

sodium 4-[(4,6-dichloro-1,3,5-triazin-2-yl)amino]benzenesulfonate

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

One New Zealand White (Hsdlf:NZW) strain rabbit was supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animal weighed 2.60 kg and was twelve to twenty weeks old. After an acclimatisation period of at least five days the animal was given a number which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.The animal was housed in a suspended cage. 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. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.The animal was 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:

no

Amount / concentration applied:

A volume of 0.1 ml of the test item was placed into the conjunctival sac of the right eye

Duration of treatment / exposure:

1 hour

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

1

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbit were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. An animal free of ocular damage was used.A volume of 0.1 ml of the test item 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.After consideration of the ocular responses produced in this animal, no additional animals were treated. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation given in Appendix 2, (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).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.Additional observations were made on Days 7, 14 and 21 to assess the reversibility of the ocular effects.In order to confirm the area of corneal opacity, the treated eye was examined under ultra violet light following treatment with Sodium Fluorescein BP (Minims: Bausch & Lomb, Surrey, UK) at the 7–Day observation. The cornea, conjunctivae and iris were also examined for lesions.The animal’s bodyweight was 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:

conjunctivae score

Basis:

animal: 72171 male

Time point:

24 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Results 2

Irritation parameter:

conjunctivae score

Basis:

animal: 72171 male

Time point:

48 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 72171 male

Time point:

72 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 72171 male

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 5

Irritation parameter:

chemosis score

Basis:

animal: 72171 male

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Results 6

Irritation parameter:

chemosis score

Basis:

animal: 72171 male

Time point:

72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Results 7

Irritation parameter:

cornea opacity score

Basis:

animal: 72171 male

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 8

Irritation parameter:

cornea opacity score

Basis:

animal: 72171 male

Time point:

48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal: 72171 male

Time point:

72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Results 10

Irritation parameter:

iris score

Basis:

animal: 72171 male

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Results 11

Irritation parameter:

other: Discharge

Basis:

animal: 72171 male

Time point:

other: Highest score at 1, 24, 48 and 72 hours, 7, 14 and 21 days

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Irritant / corrosive response data:

Individual and total scores for ocular irritation are given in Table 1.Scattered or diffuse corneal opacity was noted in the treated eye one hour after treatment with translucent corneal opacity at the 24 and 48-Hour observations and scattered or diffuse corneal opacity at the 72-Hour and all subsequent observations.Iridial inflammation was noted in the treated eye one hour after treatment and at all subsequent observations.Moderate conjunctival irritation was noted in the treated eye one hour after treatment and at the 24 and 48 Hour observations with minimal conjunctival irritation noted at the 72 Hour, 7 and 14 Day observations and moderate conjunctival irritation at the 21 Day observation.The persistence of reactions in the treated eye at the 21 day observation was considered to be indicative of irreversible ocular damage.

Other effects:

BodyweightIndividual bodyweights and bodyweight change are given in Table 2.The animal showed expected gain in bodyweight during the study.

Any other information on results incl. tables

Interpretation of Results

Data was summarised in tabular form, showing the irritation scores for the designated observation time, a description of the degree and nature of irritation, the presence of serious lesions and non‑ocular effects. 

If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.

The results were interpreted according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.

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
72171Male	2	1 Hour	1	2	1	2	2	2
		24 Hour	2	3	1	2	2	2
		48 Hour	2	3	1	2	1	1
		72 Hour	1	2	1	2	1	0
		7 Days	1	3Ä	1	2	1	0
		14 Days	1	1	1	1	1	0
		21 Days	1	2	1	2	1	1

IPR=  Initial pain reaction

*=      Examinations were performed at the specified times after instillation of the test item

Ä=     Examination under ultra‑violet light following treatment with Sodium Fluorescein BP to help confirm area of corneal opacity

Table 2              Individual Bodyweights and Bodyweight Change

Rabbit Number and Sex	Individual Bodyweight (kg)		Bodyweight Change (kg)
	Day 0	Day 21
72171Male	2.60	3.02	0.42

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated informationCriteria used for interpretation of results: EU

Conclusions:

The test item was classified as Irreversible effects on the eye (Category 1) (based on one rabbit only). It is reasonable to assume that the Signal Word “Danger” and the Hazard Statement “H318: Causes serious eye damage” are therefore required.

Executive summary:

The primary eye irritation potential of Sodiump-[(4,6-dichloro- 1,3,5-triazin-2-yl)amino]benzenesulphonate) 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 one young adult New Zealand White rabbit. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, 7, 14 and 21 days after test item instillation. 

The instillation of Sodiump-[(4,6-dichloro- 1,3,5-triazin-2-yl)amino]benzenesulphonate) into the eye resulted in moderate to severe, early‑onset ocular changes, such as scattered or diffuse corneal opacity, iridial inflammation, reddening of the conjunctivae, ocular discharge and chemosis. These effects were irreversible and still evident 21 days after treatment, the end of the observation period. No clinical signs were observed.

Thus, the test item induced significant damage to the rabbit eye and induced irreversible damage to the rabbit eye.

The test item was classified asIrreversible effects on the eye (Category 1)(based on one rabbit only). It is reasonable to assume that the Signal Word “Danger” and the Hazard Statement “H318: Causes serious eye damage” are therefore required.