Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: E.C. Directive No. 92/69/E.E.C.,B5
GLP compliance:
yes (incl. certificate)
Remarks:
OECD C(81)30(Final) Annex 2.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Phenylacetic Acid
- Molecular formula : C8H8O2
- Molecular weight : 136.1
- Substance type:Large cream flakes
- Physical state: solid
- Analytical purity: 99%
- Purity test date: 03/02/1997
- Lot/batch No.: S2969
- Expiration date of the lot/batch: February 1998
- Storage condition of test material:In dark and at room temperature
- Other: Physical and Chemical Properties
Odour : Persistent, unpleasant
Specific gravity: 1.1
Melting point:76.5°c
Boiling point: 265°c at 1013 mbar
Vapor pressure: 1.3 hPa at 97°C
Flash point:132°C
Solubility: Water: 1.66 g/100 ml @ 20°C (increasing temperature is likely to increase solubility)

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
- Age at study initiation: Not reported
- Weight at study initiation: On the day of treatment, the animals had a mean body weight ± standard deviation of 2.7±0.2 kg.
- Housing: The animals were housed individually in polystyrene cages (35 cm x 55 cm x 32 cm or 48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
- Diet : All the animals had free acess 112 C pelleted diet (U.A.R., 91360 Villemoisson - sur - Orge, France).
- Water : Drinking water filtered by a F.G. Millipore (0.22 micron) was provided ad libitum.
- 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): 12h/12h

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The right eye, which remained untreated, served as a control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): A single dose of 100 mg of the test substance was introduced into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball.

VEHICLE
No vehicle used. The test substance was used in its original form
Duration of treatment / exposure:
A single dose
Observation period (in vivo):
The eyes were examined approximately one hour, 24, 48, and 72 hours after administration of the test substance and then daily until the end of the study (day 22).
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing : The eye were not rinsed after administration of the test substance.
- Time after start of exposure: Not applicable

SCORING SYSTEM: Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation given in Table 1 (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24 h
Score:
3.3
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 48 h
Score:
3
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 72 h
Score:
3
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24 h
Score:
3
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 48 h
Score:
2.7
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 72 h
Score:
3
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24 h
Score:
1
Remarks on result:
other: Not calculable in one animal
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 48 h
Score:
1
Remarks on result:
other: Not calculable in one animal
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24 h
Score:
2
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 48 h
Score:
4
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 72 h
Score:
2.3
Irritant / corrosive response data:
Slight to severe conjunctive reactions were observed in all animals: very slight to severe chemosis, very slight to moderate redness of the conjunctivae and clear or whitish purulent discharge were noted, from Day 1 up to Day 17 at the latest.
A slight iritis was observed in two animals on Day 2: it persisted up to day 12 or 14. Reading was masked by corneal opacity in the remaining animal.
A moderate or severe corneal opacity was noted in all animals on Day 1: a slight to severe corneal opacity persisted up to the end of the observations period (Day 22) in two animals. The remaining animals was killed on Day 7 for ethical reasons, beacuse of the severity of ocular reactions.
A whitening of the conjunctivae was noted in all animals on Day 1; it persisted up to Day 17 at the lastest. Neovascularisation was noted in two animals from Day 5 up to Day 14 or from Day 7 up to the end of the observation period (Day 22) and alopecia around the eye was recorded in one animal, from Day 11 up to Day 20.

Any other information on results incl. tables

Table 2: Individual ocular examinations and mean values of the scores recorded at each reading (24, 48 and 72 hours) of each animals

 

 

Rabbit Number

 

Region of eye

 

Description of ocular reactions

 

            Scores

Mean irritation score (1)

Interpretation (+) (-)

 

 

 

 

01

 

Conjunctivae

Chemosis

3

4

3

3

3.3

(+)

Redness

2

3

3

3

3.0

(+)

Discharge

2

s

s

s

(2)

 

Iris

 

0

1

1

1

1.0

(+)

Corneal opacity

Intensity

3

2

2

2

2.0

(+)

Area

2

4

4

3

3.7

 

Other

 

B

B

B

B

 

 

Fluorescein

 

/

U

U

U

 

 

 

 

 

 

02

 

Conjunctivae

Chemosis

3

3

3

3

3.0

(+)

Redness

1

2

3

3

2.7

(+)

Discharge

0

S

S

S

(2)

 

Iris

 

OP

OP

OP

OP

(2)

(2)

Corneal opacity

Intensity

4

4

4

4

4.0

(+)

Area

1

2

2

3

2,3

 

Other

 

B

B

B

B

B

 

Fluorescein

 

/

U

U

U

U

 

 

 

 

03

 

Conjunctivae

Chemosis

2

3

3

3

3.0

(+)

Redness

2

3

3

3

 

(+)

Discharge

0

 

 

 

 

 

Iris

 

0

1

1

1

1

 

Corneal opacity

Intensity

4

3

2

2

2.3

(+)

Area

2

2

2

2

2.0

(+)

Other

 

B

B

B

B

 

 

Fluorescein

 

/

U

U

U

 

 

 

(I) mean of scores on days 2, 3 and 4

h = hour

D =day

(+) = irritant according to E.E.C criteria

(-) = non-irritant according to E.E.C criteria

(2) = not calculated

U =Fluorescein batches Nos. 5691 and 7239

/ = Fluorescein not used

S = Whitish purulent discharge

OP = Scoring masked by marked corneal opacity

B = Whitening of conjunctivae

 

Table 1 (Continued)

 

Rabbit number

Region of eye

Description of ocular reactions

Scores

D5

D6

D7

D8

D9

D10

D11

D12

D13

D14

 

 

 

 

 

01

Conjunctivae

Chemosis

3

3

2

2

2

2

2

2

2

2

Redness

3

3

3

3

2

2

2

2

2

1

Discharge

S

S

0

0

0

0

0

0

0

0

Iris

 

1

1

1

1

1

1

1

1

0

0

Corneal opacity

Intensity

2

2

2

2

2

2

2

2

2

2

Area

3

3

3

3

3

3

3

2

2

2

Other

 

B

B

B/N

B/N

B/N

B/N

B/N/A

B/N/A

B/N/A

N/A

Fluorescein

 

U

U

U

U

U

U

U

/

/

U

 

 

 

 

02

Conjunctivae

Chemosis

3

3

3

M

M

M

M

M

M

M

Redness

3

3

3

M

M

M

M

M

M

M

Discharge

S

S

S

M

M

M

M

M

M

M

Iris

 

OP

OP

OP

M

M

M

M

M

M

M

Corneal opacity

Intensity

4

4

4

M

M

M

M

M

M

M

Area

3

4

4

M

M

M

M

M

M

M

Other

 

B

B

B

M

M

M

M

M

M

M

Fluorescein

 

U

/

/

M

M

M

M

M

M

M

 

 

 

 

03

Conjunctivae

Chemosis

3

3

3

3

2

2

1

1

1

1

Redness

3

3

3

3

3

3

2

2

2

2

Discharge

S

S

S

S

S

S

0

0

0

0

Iris

 

1

1

1

1

1

1

1

1

1

1

Corneal opacity

Intensity

2

2

2

2

2

2

2

2

2

2

Area

2

2

2

2

2

2

2

3

3

3

Other

 

B/N

B/N

B/N

B/N

B/N

B/N

B/N

B/N

B/N

B/N

Fluorescein

 

U

U

U

U

U

U

U

/

/

/

 

D = Day

U = Fluorescein batches Nos. 5691 and 7239

/ = Fluorescein not used

S = Whitish purulent discharge

OP = Scoring masked by marked corneal opacity

B = Whitening of conjunctivae

N = Neovascularisation

A =Alopecia around the eye

M =Animal killed for humane reasons

 

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Remarks:
Migrated information
Conclusions:
Under the experimental conditions and according to the classification criteria laid down in Directive 93/21/E.E.C (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/E.E.C., the test substance phenylacetic acid was considered irritant when administered by ocular route in rabbits.