rac-(1R,4S,4aR,8R,8aS)-9-(dichloromethylidene)-8-hydroxyoctahydro-1,4-methanonaphthalen-5(1H)-one

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

27 May 2014 to 01 Jun 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:

Adopted: 2 October 2012

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

29 April 2004

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

August 1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

rac-(1R,4S,4aR,8R,8aS)-9-(dichloromethylidene)-8-hydroxyoctahydro-1,4-methanonaphthalen-5(1H)-one

EC Number:

941-628-3

Cas Number:

1263184-87-7

Molecular formula:

C12H14Cl2O2

IUPAC Name:

rac-(1R,4S,4aR,8R,8aS)-9-(dichloromethylidene)-8-hydroxyoctahydro-1,4-methanonaphthalen-5(1H)-one

Test material form:

solid

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: approximately 14 weeks
- Weight at study initiation: 3390 g – 3588 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 rabbit, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: At least 27 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 80
- Air changes (per hr): 15 – 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
27 May 2014 to 01 Jun 2014

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: A single dose of 0.1 g of test item was administered to the left eye of each animal (n=3).

Duration of treatment / exposure:

One hour

Observation period (in vivo):

Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours in all animals after test material installation. Observations with fluorescein staining were made approximately 24 hours before treatment and then 24, 48 and 72 hours after treatment in all animals.
The duration of the observation period was sufficient to identify reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill-health during the study were recorded. All rabbits were examined for distress at least twice daily, with observations at least 6 hours apart. Clinical observations or signs of ill-health were recorded.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: The treated eye was rinsed with physiological saline solution at the first observation time point at one hour after the application of test item on all animals as residual test item was in the eye.
- Time after start of exposure: One hour

SCORING SYSTEM:
The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (2nd October 2012). See ‘Any other information on materials and methods incl. tables’.

TOOL USED TO ASSESS SCORE: Observations with fluorescein staining were made approximately 24 hours before treatment and then 24, 48 and 72 hours after treatment in all animals.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

overall irritation score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: Weak irritation observed at the 1 hour reading (conjunctivae), fully reversible within 24 hours.

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: Weak irritation observed at the 1 hour reading, fully reversible within 24 hours.

Results 5

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

other: Weak irritation observed at the 1 hour reading, fully reversible within 24 hours.

Irritant / corrosive response data:

Initial Pain Reaction/Pain reaction (IPR/PR) was not observed. Conjunctival redness (score 1), chemosis (score 1) and discharge (score 1) were seen in all rabbits at 1 hour after treatment. Residual test item was noted on the eye in all animals at 1 hour after treatment. No other signs were observed and all animals were symptom free 24 hours after treatment. Fluorescein staining was negative in all animals at all-time points during the study.
The control eyes were symptom-free during the study.
Individual ocular reactions and individual total sores results are presented in Table 1 and 2 in 'Any other information on results incl. tables'.

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 and sex	#1 Male				#2 Male				#3 Male
	PR/PR = 0				IPR/PR = 0				IPR/PR = 0
Time after treatment	1 Hr	24 Hr	48 Hr	72 Hr	1 Hr	24 Hr	48 Hr	72 Hr	1 Hr	24 Hr	48 Hr	72 Hr
CORNEA
E = Degree of Opacity	0	0	0	0	0	0	0	0	0	0	0	0
F = Area of Cornea	0	0	0	0	0	0	0	0	0	0	0	0
involved
*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
A = Redness	1	0	0	0	1	0	0	0	1	0	0	0
B = Chemosis	1	0	0	0	1	0	0	0	1	0	0	0
C = Discharge	1	0	0	0	1	0	0	0	1	0	0	0
*Score (A+B+C) x 2	6	0	0	0	6	0	0	0	6	0	0	0
*Total Score	6	0	0	0	6	0	0	0	6	0	0	0

IPR: Initial pain reaction Hr: Hour(s)

PR: Pain reaction

Table 2. Individual Total Scores and Group Mean Scores for Ocular Irritation Calculated from the Draize Scores

Rabbit Number and Sex	*Individual Total Scores At:
	1 Hour	24 Hours	48 Hours	72 Hours
01021 Male	6	0	0	0
01038 Male	6	0	0	0
01043 Male	6	0	0	0
Group Total	18	0	0	0
Group Mean Score	6	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was graded as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to the modified Kay and Calandra classification system.

Executive summary:

In this GLP compliant OECD 405 study, the primary eye irritation effect of the test item was investigated using 3 young adult male New Zealand White rabbits. The test item was administered as an installation of a single dose of 0.1 g into the conjunctival sac of the left eye with the untreated right eyes serving as the control. The scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours in all animals after test material instillation. Observations with fluorescein staining were made approximately 24 hours before treatment and then 24, 48 and 72 hours after treatment in all animals. Rabbits were treated with analgesic and anaesthetic as per the regulatory guideline. Three animals were used to make the classification.

Initial Pain Reaction/Pain reaction (IPR/PR) was not observed. Conjunctival redness (score 1), chemosis (score 1) and discharge (score 1) were seen in all rabbits at 1 hour after treatment. Residual test item was noted on the eye in all animals at 1 hour after treatment. No other signs were observed and all animals were symptom free 24 hours after treatment. Fluorescein staining was negative in all animals at all-time points during the study. The control eyes were symptom-free during the study. No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred. The body weights of all rabbits were considered to be within the normal range of variability.

The test item was graded as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to the modified Kay and Calandra classification system.