2-Pyridinesulfonamide, 3-(2,2,2-trifluoroethoxy)-, sodium salt (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to internationally accepted test guidance and good laboratory practices guidance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

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

Principles of method if other than guideline:

The study was performed in accordance with Study Protocol No. 20699 TAL and subsequent amendments, with the following deviations from the agreed Study Protocol:
. the relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the protocol,
. during the study, the animals had free access to 110 pelleted diet instead of 112 C pelleted diet.

These minor deviations were not considered to have compromised the validity or integrity of the study.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

sodium 2-(azanidylsulfonyl)-3-(2,2,2-trifluoroethoxy)pyridine

EC Number:

700-926-1

Cas Number:

227605-94-9

Molecular formula:

C7H6F3N2NaO3S

IUPAC Name:

sodium 2-(azanidylsulfonyl)-3-(2,2,2-trifluoroethoxy)pyridine

Constituent 2

Reference substance name:

Sodium {[3-(2,2,2-trifluoroethoxy)-2-pyridinyl]sulfonyl}azanide

IUPAC Name:

Sodium {[3-(2,2,2-trifluoroethoxy)-2-pyridinyl]sulfonyl}azanide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): CA 3105 A (intermediate of CGA 362622)
- Substance type: Organic mono constituent substance
- Physical state: White powder
- Analytical purity: 93.9%
- Lot/batch No.: EZ001002
- Expiration date of the lot/batch: July 2002 (reanalysis date)
- Storage condition of test material: Room temperature
- Other: The pH of the test substance at the concentration of 10% in purified water, measured at CIT, was approximately 10.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
- Age at study initiation: No data provided.
- Weight at study initiation: On the day of treatment, the animals weighed 2.9, 3.1 and 3.3 kg, respectively.
- Housing: The animals were housed individually in polystyrene cages ( 48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
- Diet (e.g. ad libitum): During the study, the animals had free access to 110 pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France). Food is analysed regularly by the supplier for composition and contaminant levels.
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipare membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analyses of water are performed regularly by external laboratories. These analyses include the detection of possible contaminants (pesticides, heavy metals and nitrosamines). No contaminants were known to have been present in the diet or drinking water at levels which may be expected to have interfered with or prejudiced the outcome of the study. The results of the appropriate diet and water analyses are maintained in the test facility archive.
- Acclimation period: At least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 30 to 70%
- Air changes (per hr): Approximately 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: From: 26 September 2000 (experimental starting date) To: 7 November 2000 (end of the observation period)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye, which remained untreated, served as control.

Amount / concentration applied:

- Amount(s) applied (volume or weight with unit): A single dose of 100 mg of the test substance in its original form was introduced into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball. The lower and upper eyelids were held together for about one second to avoid any loss of test substance.

Duration of treatment / exposure:

Single dose: the lower and upper eyelids were held together for about one second to avoid any loss of test substance.

Observation period (in vivo):

The eyes were examined approximately 1 hour, 24, 48 and 72 hours after administration of the test substance. Following the OECD and EC guidelines, because there was persistent ocular irritation after 72 hours, the observation period was extended to 21 or 36 days in order to determine the progress of the lesions and their reversibility.

Number of animals or in vitro replicates:

As possible irritant effects were anticipated, the test substance was first administered to a single animal (No. 874). Since the test substance was not severely irritant on this first animal, it was then evaluated on two other animals (Nos. 900 and 885).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No. The eyes were not rinsed after administration of the test substance.

SCORING SYSTEM:
Ocular reactions were evaluated for each animal according to the following numerical scale:

CONJUNCTIVAL LESIONS AND DISCHARGE
Chemosis (Iids and/or nictitating membranes)
. no swelling : 0
. any swelling above normal (includes nictitating membranes): 1
. obvious swelling with partial eversion of lids: 2
. swelling with lids about half-closed: 3
. swelling with lids more than half-closed: 4

Redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
. blood vessels normal: 0
. a number of blood vessels definitely hyperemic (injected): 1
. diffuse, crimson colour, individual vessels not easily discernible: 2
. diffuse, beefy red: 3

Discharge
. absence of discharge: 0
. slight discharge (does not include small amounts normally found in inner canthus): 1
. discharge with moistening of lids and hairs adjacent to lids: 2
. discharge with moistening of lids and hairs on wide area around the eye: 3

IRIS LESIONS
. normal: 0
. markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive): 1
. no reaction to light, haemorrhage, gross destruction (any or all of these): 2

CORNEAL LESIONS
Cornea (direct examination or, if necessary, with an Ultra-Violet lamp)
Opacity (degree of intensity: area most dense taken for reading)
. no ulceration or opacity: 0
. scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible: 1
. easily discernible translucent area, details of iris slightly obscured: 2
. nacrous areas, no details of iris visible, size of pupil barely discernible: 3
. opaque cornea, iris not discernible through the opacity: 4
Area of opacity
. one quarter ( or less) but not zero: 1
. greater than one quarter but less than a half: 2
. greater than one half but less than three quarters: 3
. greater than three quarters up to whole area: 4

Any other lesions observed were noted.
Conjunctival discharge and area of corneal opacity were scored but were not used in the calculation of indices.

TOOL USED TO ASSESS SCORE: To determine the presence or absence of corneal opacification and to evaluate the affected area, the eyes were examined on day 2 and whenever necessary under an UV lamp after instillation of one or two drops of 0.5% sodium fluorescein solution.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 4 days

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.8

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

2.9

Max. score:

4

Remarks on result:

other: Chemosis still present in two animals on day 21

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

3

Max. score:

3

Remarks on result:

other: Redness still present in two animals on day 21

Any other information on results incl. tables

Very slight to moderate conjunctival reactions including very slight to moderate chemosis (grades 1 to 3), very slight to moderate redness of the conjunctiva (grades 1 to 3) and clear to whitish purulent discharge were observed in all animals from day 1: the chemosis and redness were still present in two animals on day 21; the discharge cleared no later than day 10.

A slight iritis (grade 1) was noted in all animals, from day 1 up to day 7 (two animals) or on day 2 only ( one animal).

A very slight or slight corneal opacity (grade 1 or 2) was also recorded in all animals from day 2; it persisted up to day 4 at the latest.

A black area on the conjunctiva was seen in two animals from the 24-hour examination through day 14 or 17.

Mean scores calculated for each animal over 24, 48 and 72 hours were 1, 2 and 2 for corneal opacity, 0.3, 1 and 1 for iris lesions, 2.7, 3 and 3 for chemosis and 3, 3 and 3 for redness of the conjunctiva.

Mean scores calculated for the three animals were 1.7 for corneal opacity, 0.8 for iris lesions, 2.9 for chemosis and 3 for redness of the conjunctiva.

Table 1. Individual ocular examinations and summary indices

Rabbit number	Region of eye	Description of ocular reactions	Scores										Mean score
			1h	24h	48h	72h	Day 7	Day 10	Day 14	Day 17	Day 21	Day 36	24-72 h
874	Cornea	Opacity	0	2	1	0	0	0	0	0	0	-	1
	Iris		0	1	0	0	0	0	0	0	0	-	0.3
	Conjunctivae	Chemosis	2	3	3	2	1	1	1	0	0	-	2.7
		Redness	2	3	3	3	2	1	1	1	0	-	3
900	Cornea	Opacity	0	2	2	2	0	0	0	0	0	0	2
	Iris		1	1	1	1	1	0	0	0	0	0	1
	Conjunctivae	Chemosis	2	3	3	3	2	1	1	1	1	0	3
		Redness	2	3	3	3	3	3	3	2	1	0	3
885	Cornea	Opacity	0	2	2	2	0	0	0	0	0	0	2
	Iris		1	1	1	1	1	0	0	0	0	0	1
	Conjunctivae	Chemosis	2	3	3	3	2	2	2	2	1	0	3
		Redness	2	3	3	3	3	2	2	2	1	0	3
All													Grand mean 24-72 h
	Cornea	Opacity											1.7
	Iris												0.8
	Conjunctivae	Chemosis											2.9
		Redness											3

- = Not performed

 

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under our experimental conditions, the test substance is irritant when administered by ocular route in rabbits.
According to the classification criteria laid down in Commission Directive 93/21/EEC, CA 3105 A (Intermediate of CGA 362622) should be classified as R 36: “Irritating to eyes”.

Executive summary:

The potential of the test substance CA 3105 A (Intermediate of CGA 362622) to induce ocular irritation was evaluated in rabbits according to OECD (No. 405, 24th February 1987) and EC (92/69/EEC, B.5, 31st July 1992) guidelines. The study was conducted in comp1iance with the principles of Good Laboratory Practice Regulations.

The test substance was first administered to a single male New Zealand White Rabbit. Since the test substance was not severely irritant on this first animal, it was then evaluated in two other animals. A single dose of 100 mg of the test substance in its original form was introduced into the left conjunctival sac. The right eye served as control. The eyes were not rinsed after administration of the test substance.

Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration/ and then on days 7, 10, 14, 17, 21 and 36.

Very slight to moderate chemosis, very slight to moderate redness of the conjunctiva and clear to whitish purulent discharge were observed in all animals from the 1-hour observation. The chemosis and redness were still present in two animals on day 21 ; the discharge cleared no later than day 10. A slight iritis was noted in two animals from 1 hour through 7 days after treatment; in the third animal iritis was seen only at 24 hours. A slight corneal opacity was recorded in all animals at the 24-hour examination and persisted through 48 or 72 hours. A black area on the conjunctiva was seen in two animals from the 24-hour examination through day 14 or 17. All reactions were clear in all animals by day 36.

Mean scores calculated for each animal over 24, 48 and 72 hours were 1, 2 and 2 for corneal opacity, 0.3, 1 and 1 for iris lesions, 2.7, 3 and 3 for chemosis and 3, 3 and 3 for redness of the conjunctiva. Mean scores calculated for the three animals were 1. 7 for corneal opacity, 0.8 for iris lesions, 2.9 for chemosis and 3 for redness of the conjunctiva.

Under our experimental conditions, the test substance is irritant when administered by ocular route in rabbits.According to the classification criteria laid down in Commission Directive 93/21/EEC, CA 3105 A (Intermediate of CGA 362622) should be classified as R 36: “Irritating to eyes”.