2-Pyridinecarboxylic acid, 4-(acetylamino)-3,6-dichloro-, methyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 January 2013 to 27 January 2013

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:

2013

Report date:

2013

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:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Appearance: Variable coloured powder
- Storage conditions of the test material: Ambient (18 to 36 °C)
- pH: The pH of the test material as a 10 % aqueous solution was 5.05

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7 to 8 months
- Weight at study initiation: 2.75 to 2.86 kg
- Fasting period before study: No
- Housing: The rabbits were housed individually in rabbit cages (size: approximately 65 cm long x 65 cm deep x 45 cm high) with a noryl shallow cage body and facilities for pelleted feed and drinking water. The litter collection trays (noryl waste tray) were changed daily (except on Sundays). A noryl perforated raised shelf was provided to the animals for enrichment.
- Diet (e.g. ad libitum): Rabbit feed was provided ad libitum in a stainless steel feed hopper.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in a water filter-cum-purifier was provided to animals ad libitum in 750 mL bottles with sipper tubes. The water in the bottles was replenished once daily and the water bottles were changed once a week.
- Acclimation period: Five to six days under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 23 °C
- Humidity (%): 65 to 66 % (relative)
- Air changes (per hr): 12 to 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours of light and 12 hours of dark

IN-LIFE DATES: From: 18 January 2013 To: 27 January 2013

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye of each rabbit remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:

The eye lids were gently held together for about one second, in order to minimise loss of the test material.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 male animals

Details on study design:

TEST MATERIAL INSTALLATION
- Eye examinations: Prior to instillation, both eyes of each rabbit were examined with the aid of a pen light for irritation and ocular defects. The eyes were then checked for opacity using ophthalmic fluorescein sodium strips. The eyes were then irrigated with enough physiological saline (0.9 % NaCl) to remove any excess fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes were checked and evaluated for pre-existing corneal damage. No corneal defects were observed in any of the animals.
The finely ground test material was placed in the everted lower lid (conjunctival sac) of the left eye of each of three rabbits in a stepwise manner.
- Stepwise exposure of animals: A single rabbit was initially exposed to the test material. Prior to instillation, a local anaesthetic (4 % Lidocaine) was used in both the test and control eye. No corrosive effect or irritation was observed; hence the test was completed using two additional animals.

SCORING SYSTEM: The eyes were evaluated with the illumination of a white light source at intervals of approximately 1, 24, 48 and 72 hours post instillation of the test material according to the "Scale for Scoring Ocular Lesions" (Draize, 1944).

TOOL USED TO ASSESS SCORE: At 24 hours post installation, the eyes were checked for opacity using ophthalmic fluorescein sodium strips. The eyes were then irrigated with enough physiological saline (0.9 % NaCl) to remove any excess fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes were evaluated to verify the extent or absence of corneal damage.

OBSERVATIONS
- Clinical Observations: The rabbits were observed for clinical signs of toxicity and mortality 4 times (at hourly intervals) on the day of test material instillation and once daily thereafter until the end of the observation period.
- Body Weights: Body weights were recorded at the start of acclimatisation, day 1 of treatment (just before test material instillation) and at termination of observation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: Mean score at 24, 48 and 72 hours

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Remarks on result:

other: The conjunctivae score for all animals at all time points was 0

Results 2

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: Mean score at 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: The chemosis score for all animals at all time points was 0

Results 3

Irritation parameter:

other: discharge

Basis:

mean

Remarks:

of 3 animals

Time point:

other: Mean score at 24, 48 and 72 hours

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Remarks on result:

other: The discharge score for all animals at all time points was 0

Results 4

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: Mean score at 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: The iris score for all animals at all time points was 0

Results 5

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: Mean score at 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: The cornea score for all animals at all time points was 0

Irritant / corrosive response data:

There was no eye irritation (iritis, conjunctival irritation or corneal opacity) observed in any treated eye during the study at 1, 24, 48 and 72 hours post instillation.

Other effects:

There were no clinical signs of toxicity observed, and none of the animals died, during the study.
The body weights of all rabbits increased slightly throughout the observation period. See Table 1.

Any other information on results incl. tables

Table 1: Body Weight

Rabbit Number	Body weight (kg)
	At start of acclimatisation	At treatment	At termination of observation
RB9718	2.65	2.75	2.82
RB9719	2.78	2.85	2.92
RB9720	2.80	2.86	2.94

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the test material caused no irritation to the eye of male New Zealand White rabbits and requires no classification in accordance with EU criteria.

Executive summary:

The potential of the test material to cause eye irritation in male New Zealand White rabbits was assessed in accordance with the standardised guidelines OECD 405 and US EPA OPPTS 870.2400 under GLP conditions.

The test was performed in a stepwise manner. A single rabbit was initially exposed to the test material. Since the test material was not corrosive or severely irritating the study was completed using two additional rabbits.

0.1 g of the finely ground test material was placed in the everted lower lid (conjunctival sac) of the left eye of each of three rabbits. The lids were gently held together for about one second, in order to minimise loss of the test material. The right eye of each rabbit remained untreated and served as the reference control. The eyes were evaluated at 1, 24, 48 and 72 hours post instillation according to the Draize scale for scoring ocular lesions.

There was no eye irritation (iritis, conjunctival irritation or corneal opacity) observed in any treated eye during the study at 1, 24, 48 and 72 hours post instillation.

There were no clinical signs of toxicity observed and none of the animals died during the study. All rabbits appeared healthy and gained body weight throughout the observation period.

Under the conditions of this study, the test material caused no irritation to the eye of male New Zealand White rabbits and requires no classification in accordance with EU criteria.