Reaction mass of 6-Chloro-2-{4[ethyl(2-hydroxyethyl)amino]phenyl}-1-methylbenzo[cd]indolium hydroxide and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Test start date: 16 June, 1980; Test end date: 23 June, 1980; Study completion date: 30 June 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1980

Report date:

1980

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: the United States Environmental Protection Agency (EPA) § 163.81-4 "Primary eye irritation study", Federal Register, Vol. 43, No. 163, August 22, 1978.

Deviations:

no

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-bromo-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium hydroxide

IUPAC Name:

Reaction mass of 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-bromo-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium acetate and 6-chloro-2-[4-[ethyl(2-hydroxyethyl)amino]phenyl]-1-methylbenz[cd]indolium hydroxide

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Code No.of test material: FAT 31064/C
- Purity: 42 %

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: In-house
- Age at study initiation: adult ones
- Weight at study initiation: 2 to 3 kgs
- Housing: individually in metal cages, numbered by ear tags
- Diet: ad libitum, standard rabbit food - NAFAG, No. 814, Gossau SG
- Water: ad libitum
- Acclimation period: minimum of 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Photoperiod: 10 hour light cycle day

IN-LIFE DATES: From: 4 February, 1980; To: 12 February, 1980

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Duration of treatment / exposure:

In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of physiological saline. No flushing was performed in remaining animals.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6 (3 males and 3 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: Yes, In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of physiological saline.

SCORING SYSTEM: the United States Environmental Protection Agency (EPA) § 163.81-4 "Primary eye irritation study", Federal Register, Vol. 43, No. 163, August 22, 1978.

TOOL USED TO ASSESS SCORE: Reading of reaction is facilitated by use of a binocular loupe, hand slit-lamp, or other expert means.

SCORING SYSTEM: 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

AxBx5 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
Ax5 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


TOOL USED TO ASSESS SCORE: Slit lamp
The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

Rinsed eyes

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Remarks:

Rised eyes

Basis:

mean

Time point:

24/48/72 h

Score:

0.78

Max. score:

2

Reversibility:

fully reversible within: 7 d

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Rinsed eyes

Basis:

mean

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 7 d

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

chemosis score

Remarks:

Rinsed eyes

Basis:

mean

Time point:

24/48/72 h

Score:

2.78

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Remarks on result:

positive indication of irritation

Results 5

Irritation parameter:

cornea opacity score

Remarks:

Eye not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

1.66

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Results 6

Irritation parameter:

iris score

Remarks:

Eye not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

0.66

Max. score:

2

Reversibility:

not fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Results 7

Irritation parameter:

conjunctivae score

Remarks:

Eye not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

2.44

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Results 8

Irritation parameter:

chemosis score

Remarks:

Eye not rinsed

Basis:

mean

Time point:

24/48/72 h

Score:

2.66

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

FAT 31064/C was found to cause a moderate irritation when applied to the rabbit eye mucosa. Rinsing the eyes following instillation was of little but assessable effect.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

FAT 31064/C can be classified as an irritant to the eye.

Executive summary:

The eye irritation potential of the test item was evaluated in a study conducted according to procedure described in the United States Environmental Protection Agency (EPA) § 163.81-4 "Primary eye irritation study", Federal Register, Vol. 43, No. 163, August 22, 1978. 6 New Zealand rabbits (3 males; 3 females) were used in this study. 0.1 mL of test material was inserted into the conjunctival sac of the left eye of the rabbits and the lids were gently closed for a few seconds. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of physiological saline. The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit. The mean reaction scores for cornea, iris and conjunctiva observed for the rabbits with rinsed eyes were 1.00, 0.78, 2.00 and 2.78, respectively. The instillation was found to cause moderate irritation when applied to the rabbit eye mucosa. Hence, the test item can be classified as an irritant to the eye, classified as eye irritant category 2 as per CLP (Regulation EC No. 1272/2008) criteria.