3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not reported

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:

1978

Report date:

1978

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Method according to the Federal Register 38 No. 187 § 1500.42 (eye)

Version / remarks:

1973

Deviations:

not specified

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Remarks:

Study pre-dates GLP

Test material

Test material information

Constituent 1

Reference substance name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

EC Number:

221-326-1

EC Name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Cas Number:

3068-39-1

Molecular formula:

C27H29N2O3.Cl

IUPAC Name:

3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

Test animals / tissue source

Species:

rabbit

Strain:

other: Albino

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: Approximately 3 to 4 kg
- Housing: Animals were singly housed in a stainless steel cage without litter
- Diet: Standard pellet feed for rabbits provided ad libitum
- Water: ad libitum
- Acclimation period: >3 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1 °C
- Humidity: 55 ± 10 % (relative)
- Photoperiod: 12 hour light/12 hour dark cycle (07:00 to 19:00)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg

Observation period (in vivo):

Animals were observed for signs of irritation and toxicity 1, 24, 48 and 72 hours after test material instillation and observations were continued for 7 days.

Number of animals or in vitro replicates:

6 animals

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No rinsing occurred

SCORING SYSTEM: Draize scale

CORNEA
Opacity: degree of density (area most dense is taken for reading)
0: No opacity
1: Scattered or diffuse areas of opacity, details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Opalescent area, no details of iris visible, size of pupil barely discernible
4: Opaque, iris invisible

IRIS
0: Normal
1: Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) still reacting to light (sluggish reaction is positive)
2: No reaction to light, haemorrhage, gross destruction (any of all these or all together)

CONJUNCTIVAE
Redness: (refers to the most severe effect of palpebral and bulbar conjunctivae excluding cornea and iris)
0: Vessels normal
1: Vessels definitely injected above normal
2: More diffuse, deeper crimson red, individual vessels not easily discernible
3: Diffuse beefy red
Chemosis
0: No swelling
1: Any swelling above normal (includes nictating membranes)
2: Obvious swelling with partial eversion of lids.
3: Swelling with lids about half closed
4: Swelling with lids about half closed to completely closed
Discharge
0: No discharge
1: Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs a considerable area around the eye

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of all animals that could be scored

Time point:

24/48/72 h

Score:

3.9

Max. score:

4

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

of all animals that could be scored

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

not determinable

Remarks:

Not possible to evaluate due to very strong swelling of cornea and iris

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of all animals

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Results 5

Irritation parameter:

other:

Remarks:

discharge

Basis:

mean

Remarks:

of all animals

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

1 hour post application, strong swelling of the conjunctivae with mild to severe oozing/ weeping of the eye was observed. Four rabbits were observed to have slight opacification of the cornea. From 24 hours post application until end of the observation period, the edges of the eyelids were strongly hardened with discharge for all rabbits. Eyes were initially covered in mucous, which later on became purulent and glued together. Any redness caused by the test material could not be determined due to extreme swelling of the eyes. After seven days post application, the observations were essentially the same as the observations after 72 hours.

Any other information on results incl. tables

Table 1: Individual Scores

Time Point (h)	Rabbit number	Evaluation
		Cornea	Iris	Conjunctival redness	Chemosis	Discharge
24	1	*	*	**	4	3
	2	*	*	**	4	3
	3	*	*	**	4	3
	4	*	*	**	4	3
	5	4	2	**	4	3
	6	3	2	**	4	3
48	1	*	*	**	4	3
	2	*	*	**	4	3
	3	4	2	**	4	3
	4	4	2	**	4	2
	5	4	2	**	4	2
	6	4	2	**	4	2
72	1	*	*	**	4	3
	2	*	*	**	4	3
	3	4	2	**	4	3
	4	4	2	**	4	3
	5	4	2	**	4	2
	6	4	2	**	4	1

*Assesment not possible due to staining of the eye as the test material is a red colour

**Not determined due to strong swelling of the cornea and iris

Applicant's summary and conclusion

Interpretation of results:

other: Requires classification as Category 1 in accordance with EU classification criteria

Conclusions:

Under the conditions of the study, the test material caused irreversible damage to the eyes and requires classification as Category 1.

Executive summary:

The potential of the test material to cause eye irritation was investigated in the albino rabbit using the German method according to the Federal Register 38 No. 187 § 1500.42 (eye) which is equivalent to the standardised guideline OECD 405.

A single 100 mg quantity of the test material was applied to one eye in each of 6 rabbits and the untreated eye was used as a control. Animals were observed for signs of irritation and toxicity 1, 24, 48 and 72 hours after test material instillation.

1 hour post application, strong swelling of the conjunctivae with mild to severe oozing/ weeping of the eye was observed. Four rabbits were observed to have slight opacification of the cornea. From 24 hours post application until end of the observation period, the edges of the eyelids were strongly hardened with discharge for all rabbits. Eyes were initially covered in mucous, which later on became purulent and glued together. Any redness caused by the test material could not be determined due to extreme swelling of the eyes. After seven days post application, the observations were essentially the same as the observations after 72 hours.

Under the conditions of the study, the test material caused irreversible damage to the eyes and requires classification as Category 1.