3-Pyridinesulfonyl chloride, 2-methoxy-4-(trifluoromethyl)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 January 2005 - 10 February 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-methoxy-4-(trifluoromethyl)pyridine-3-sulfonyl chloride

EC Number:

606-871-9

Cas Number:

219715-41-0

Molecular formula:

C7H5ClF3NO3S

IUPAC Name:

2-methoxy-4-(trifluoromethyl)pyridine-3-sulfonyl chloride

Test material form:

other: solid

Details on test material:

2-methoxy-4-(trifluoromethyl)pyridine-3-sulfonyl chloride
Appearance: pale tan solid
Storage: room temperature
pH: 7-8 (by wetted pH paper)
Solubility: soluble in methanol, ethanol and acetone
Stability: test material was expected to be stable for the duration of testing

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
-Strain: New Zealand albino
- Source: Robinson Services Inc., Clemmons, NC
- Age at study initiation: Young adult
- Housing: The animals were singly housed in suspended stainless steel caging with mesh floors. Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet (e.g. ad libitum): Pelleted Purina Certified Rabbit Chow #5322
- Water (e.g. ad libitum): Filtered tap water was supplied ad libitum by an automatic water dispensing system.
Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels which would have interfered with the results of this study. Analysis of the water is conducted at least once a year and the records are kept on file at Product Safety Laboratories. The most recent analysis was conducted in December 2004. Purina Certified Rabbit Diet, PMI #5322, Lot Number: JUL 06 04 3A, was analyzed in July 2004.
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22 °C
- Humidity (%): 45 - 58 % relative
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Prior to use, the test substance was ground to a powder with a mortar and pestle. One tenth of a millilitre (0.07g) was instilled into the conjunctival sac of the right eye of each rabbit.

Duration of treatment / exposure:

single exposure

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3

Details on study design:

-Preparation and Selection of Animals
Prior to instillation, both eyes of a number of animals were examined using a fluorescein dye procedure. One drop of 2% ophthalmic fluorescein sodium was instilled into both eyes of each rabbit. The eyes were rinsed with physiological saline (0.9% NaCl) approximately 30 seconds after instillation of the fluorescein. Using an ultraviolet light source, the eyes were checked for gross abnormalities according to the "Scale for Scoring Ocular Lesions" (Draize). Three healthy animals without pre-existing ocular irritation were selected for test.

-Instillation
Prior to use, the test substance was ground to a powder with a mortar and pestle. One-tenth of a millilitre (0.07 grams) of the test substance was instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for about one second before releasing to minimize loss of the test substance. The other eye of each rabbit remained untreated with the test substance and served as a control. The rabbits were then returned to their designated cages.

-Ocular Scoring
Ocular irritation was evaluated using a high-intensity white light (Mag Lite) in accordance with Draize at al. (see below) at 1, 24, 48, and 72 hours, and at 4, 7, 10, and 14 days post-instillation. The fluorescein dye evaluation procedure described above was used at 24 hours and as needed at subsequent scoring intervals to evaluate the extent of corneal damage or to verify reversal of effects. Individual scores were recorded for each animal. In addition to observations of the cornea, iris, and conjunctivae, any other observed lesions were noted. The average score for all rabbits at each scoring period was calculated to aid in data interpretation.

-Cage-Side Observations
The animals were observed for signs of gross toxicity and behavioural changes at least once daily during the test period. Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, and coma.

SCALE FOR SCORING OCULAR LESIONS

1. Cornea
A. Opacity-degree of density (area most dense taken for reading)

No Opacity 0
Scattered or diffuse area, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris invisible 4

B. Area of cornea involved

One quarter (or less) but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4
A x B x 5 Total Maximum = 80

2. Iris
A. Values

Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all 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
A x 5 Total Maximum = 10

3. Conjunctivae
A. Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)

Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

B. Chemosis

No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half-closed 3
Swelling with lids about half-closed to completely closed 4

C. Discharge

No discharge 0
Any amount different from normal (does not include small amounts observed in inner
canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3
Score (A + B + C) x 2 Total Maximum = 20

Total Maximum Score: 110 represents the sum of all scores obtained for the cornea, iris and conjunctivae.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #1

Time point:

other: mean score for 24, 48 and 72 hours

Score:

1

Max. score:

80

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

other: mean score for 24, 48 and 72 hours

Score:

0.33

Max. score:

80

Reversibility:

fully reversible within: 48 hours

Results 3

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #3

Time point:

other: mean score for 24, 48 and 72 hours

Score:

0.33

Max. score:

80

Reversibility:

not fully reversible within: 48 hours

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: mean score for 24, 48 and 72 hours

Score:

1

Max. score:

10

Reversibility:

fully reversible within: 4 days

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: mean score for 24, 48 and 72 hours

Score:

0.33

Max. score:

10

Reversibility:

fully reversible within: 48 hours

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: mean score for 24, 48 and 72 hours

Score:

1

Max. score:

10

Reversibility:

fully reversible within: 4 days

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

other: mean score for 24, 48 and 72 hours

Score:

2

Max. score:

6

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

other: mean score for 24, 48 and 72 hours

Score:

1.33

Max. score:

6

Reversibility:

fully reversible within: 7 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

other: mean score for 24, 48 and 72 hours

Score:

1.67

Max. score:

6

Reversibility:

fully reversible within: 7 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean score for 24, 48 and 72 hours

Score:

2

Max. score:

8

Reversibility:

fully reversible within: 14 days

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean score for 24, 48 and 72 hours

Score:

0.33

Max. score:

8

Reversibility:

fully reversible within: 48 hours

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean score for 24, 48 and 72 hours

Score:

0.33

Max. score:

8

Reversibility:

fully reversible within: 48 hours

Any other information on results incl. tables

Apart from the eye irritation, there were no other signs of gross toxicity, adverse clinical signs or abnormal behaviour.

Within 24 hours of test substance instillation, all three treated eyes exhibited corneal opacity, iritis, and conjunctivitis. The overall incidence and severity of irritation decreased gradually thereafter. All animals were free of ocular irritation by Day 14 (study termination).

 

Individual scores for ocular irritation:

 

Animal 1:

	Rabbit number 13619 (male)
	Hours				Days
	1	24	48	72	4	7	10	14
1. Cornea
A. Opacity	0	1*	1*	1	1*	1*	1*	0*
B. Area	4	4	4	4	3	2	1	4
(A x B) x 5	0	20	20	20	15	10	5	0
2. Iris
A. Values	0	1	1	1	0	0	0	0
A x 5	0	5	5	5	0	0	0	0
3. Conjunctivae
A. Redness	2	2	2	2	2	2	1	0
B. Chemosis	2	2	2	2	1	1	1	0
C. Discharge	2	2*	2*	2*	2*	1	0	0
(A + B + C) x 2	12	12	12	12	10	8	4	0
Total	12	37	37	37	25	18	9	0

*2% ophthalmic fluorescein sodium used to evaluate the extent or verify the absence of corneal opacity.

**Thick white discharge was observed.

Animal 2:

	Rabbit number 13620 (male)
	Hours				Days
	1	24	48	72	4	7	10	14
1. Cornea
A. Opacity	0	1*	0*	0	0	0	0	0
B. Area	4	1	4	4	4	4	4	4
(A x B) x 5	0	5	0	0	0	0	0	0
2. Iris
A. Values	0	1	0	0	0	0	0	0
A x 5	0	5	0	0	0	0	0	0
3. Conjunctivae
A. Redness	2	2	1	1	1	0	0	0
B. Chemosis	1	1	0	0	0	0	0	0
C. Discharge	2	2	1	1	0	0	0	0
(A + B + C) x 2	10	10	4	4	2	0	0	0
Total	10	20	4	4	2	0	0	0

*2% ophthalmic fluorescein sodium used to evaluate the extent or verify the absence of corneal opacity.

 

Animal 3:

	Rabbit number 13621 (female)
	Hours				Days
	1	24	48	72	4	7	10	14
1. Cornea
A. Opacity	0	1*	0*	0	0	0	0	0
B. Area	4	1	4	4	4	4	4	4
(A x B) x 5	0	5	0	0	0	0	0	0
2. Iris
A. Values	0	1	1	1	0	0	0	0
A x 5	0	5	5	5	0	0	0	0
3. Conjunctivae
A. Redness	2	2	2	1	1	0	0	0
B. Chemosis	2	1	0	0	0	0	0	0
C. Discharge	2	2	1	1	0	0	0	0
(A + B + C) x 2	12	10	6	4	2	0	0	0
Total	12	20	11	9	2	0	0	0

*2% ophthalmic fluorescein sodium used to evaluate the extent or verify the absence of corneal opacity.

 

 

Applicant's summary and conclusion

Interpretation of results:

other: Category 2

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the test substance caused corneal opacity, iritis, and conjunctival irritation which cleared by Day 14.

Executive summary:

Under the conditions of this study, the test substance caused corneal opacity, iritis, and conjunctival irritation which cleared by Day 14. The study was conducted in accordance with U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400 (1998).