Reaction mass of 3-(difluoromethyl)-1-methyl-N-[(1RS,4SR,9RS)-1,2,3,4-tetrahydro-9-isopropyl-1,4-methanonaphthalen-5-yl]pyrazole-4-carboxamide and of 3-(difluoromethyl)-1-methyl-N-[(1RS,4SR,9SR)-1,2,3,4-tetrahydro-9-isopropyl-1,4-methanonaphthalen-5-yl]pyrazole-4-carboxamide; isopyrazam

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Jan 2014 to 05 Feb 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2012

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

632-619-2

EC Number:

632-619-2

Cas Number:

881685-58-1

Molecular formula:

C20 H23 F2 N3 O

IUPAC Name:

632-619-2

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Sex: Male
- Age at study initiation: 12-13 weeks
- Weight at study initiation: 3146 – 3169 g
- Housing: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
- Diet: UNI diet for rabbits. Ad libitum
- Water: Tap water. Ad libitum
- Acclimatisation period: at least 14 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17.3 – 21.7 °C
- Humidity: 24 – 58 %
- Air changes: 15-20 air exchanges/hour
- Photoperiod: 12 hours daily from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 29 Jan 2014 To: 05 Feb 2014

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g of the test material

Duration of treatment / exposure:

1 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

PRE-STUDY EXAMINATION
- The pH of the test material was measured as pH 4.0, permitting the test material to be used in the animal studies.
- The test was performed on 3 male rabbits. Before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect.
- Sixty minutes prior to test material application, a systemic opiate analgesic was administered by subcutaneous injection under direct Veterinary supervision. Repeat injections were given on the first day as appropriate to maintain an adequate level of analgesia. Five minutes prior to test material application, a topical ocular anaesthetic was applied to each eye (including the control eye) to ensure direct comparison of any ocular observations. Eight hours after test material application, a systemic opiate analgesic and a nonsteroidal anti-inflammatory drug (NSAID) were administered by subcutaneous injection under direct Veterinary supervision. The systemic opiate analgesic was again injected ~12 hours after the post-treatment analgesic and then every 12 hours, and NSAID injected every 24 hours, until eye scores were zero.

APPLICATION
The test material was placed in the conjunctival sac of the left eye of animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for at least one second in order to prevent loss of the material.

REMOVAL OF TEST SUBSTANCE
- Washing: The treated eye was rinsed with physiological saline solution at the first observation time point at 1 hour after the application of test item on the all animals as the irritation scores were greater than 1.
- Time after start of exposure: 1 hour

OBSERVATION TIME POINTS
Approximately 1, 24, 48 72 hours after the end of exposure. Observations with fluorescein staining were made approximately 24 hours before treatment and then 24, 48, 72 hours after treatment in all animals. All rabbits were examined for distress at least twice daily, with observations at least 6 hours apart.

SCORING SYSTEM:
The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (02 October 2012). For criteria, see 'Any other information on materials and methods incl. tables'.

TOOL USED TO ASSESS SCORE: fluorescein

OTHER OBSERVATIONS
- Individual body weight was recorded on the day of treatment and before euthanasia.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritant / corrosive response data:

- Initial Pain Reaction/ Pain Reaction (IPR/PR) was not observed.
- Conjunctival redness (score 2), chemosis (score 1) and discharge (score 1 or 2) were seen in all rabbits at 1 hour after treatment. Conjunctival redness (score 1) was seen in all rabbits at 24 hours after treatment.
- All symptoms had fully reversed in all animals after 48 hours.
- Fluorescein staining was negative in all animals at 24, 48 and 72 hours after instillation.
- The control eyes were symptom-free during the study.

Other effects:

OTHER EFFECTS
- The body weights of all rabbits were considered to be within the normal range of variability.
- No clinical signs of systemic toxicity were observed in any animals in this study.
- No mortality occurred in this study.

Any other information on results incl. tables

Table 1. Individual Draize Scores and Individual Total Scores* for Ocular Irritation.

Rabbit Number	#1				#2				#3
IPR	0				0				0
PR	0	0	0	0	0	0	0	0	0	0	0	0
Time after treatment (hour)	1	24	48	72	1	24	48	72	1	24	48	72
Cornea
Degree of Opacity (E)	0	0	0	0	0	0	0	0	0	0	0	0
Area involved (F)	0	0	0	0	0	0	0	0	0	0	0	0
*Score (E x F) x 5	0	0	0	0	0	0	0	0	0	0	0	0
Iris (D)	0	0	0	0	0	0	0	0	0	0	0	0
*Score (D x 5)	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctivae
Redness (A)	2	1	0	0	2	1	0	0	2	1	0	0
Chemosis (B)	1	0	0	0	1	0	0	0	1	0	0	0
Discharge (C)	1	0	0	0	2	0	0	0	1	0	0	0
*Score (A+B+C) x 2	8	2	0	0	10	2	0	0	8	2	0	0
*Total Score	8	2	0	0	10	2	0	0	8	2	0	0

* Kay J H and Calandra J C (1962)

Table 2. Individual body weights and body weight changes.

Rabbit Number	Body weight before treatment (g)	Body weight after treatment (g)	Body weight change (g)
#1	3169	3197	28
#2	3161	3209	48
#3	3146	3210	64

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The application of the test material did not result in signs of eye irritation.

Executive summary:

The test material was administered as an installation of a single dose of 0.1 g into the conjunctival sac of the left eye of 3 adult male New Zealand White rabbits in a study according to OECD TG 405 and GLP principles. The untreated right eye served as the control. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours after test material installation in all animals. Observations with fluorescein staining were made approximately 24 hours before treatment and then 24, 48 and 72 hours after the treatment in all animals. Results obtained from these three animals were used to classify the test item for irritation potential.

Initial Pain Reaction/Pain reaction (IPR/PR) was not observed. The average cornea opacity, iritis and chemosis score of all three animals over 24, 48 and 72 hours was 0. The average conjunctivae score of all three animals over 24, 48 and 72 hours was 0.33. All symptoms had fully reversed in all animals after 48 hours. Fluorescein staining was negative in all animals at 24, 48 and 72 hours after instillation. The control eye of animals was symptom-free during the study. No mortality occurred during the study. The body weights of all rabbits were considered to be within the normal range of variability.

The application of the test material did not result in signs of eye irritation.