1H-Imidazolium, 3-ethyl-1-methyl-, C11-13-branched alkylbenzenesulfonates

Eye irritation

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: Study was conducted in a GLP laboratory using OECD Testing Guideline 405.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

see box below

Principles of method if other than guideline:

The relative humidity (38 - 76%) and temperature (16.6 – 23.4 oC) were out of the target range during the study.
These deviations are considered to have no impact on the outcome of the study and interpretation of the results.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1H-Imidazolium, 3-ethyl-1-methyl-, C9-14-branched alkylbenzenesulfonates

IUPAC Name:

1H-Imidazolium, 3-ethyl-1-methyl-, C9-14-branched alkylbenzenesulfonates

Constituent 2

Reference substance name:

EMI-DBS

IUPAC Name:

EMI-DBS

Constituent 3

Reference substance name:

1065642-34-3

Cas Number:

1065642-34-3

IUPAC Name:

1065642-34-3

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Details on test material:

Substance name: EMI-DBS
Description of test item: Yellow paste
Storage conditions: Room temperature (15 - 25°C), in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species and strain: New Zealand White rabbits
Source: S&K-LAP Kft. 2173 Kartal, Császár út 135, Hungary
Justification of strain: The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined approximately one hour before starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
Number of animals: 3
Age of animals at treatment: ~12 weeks old (adult)
Sex: Male
Body weight range at the
- beginning of the life phase: 3105-3202 g
- end of the life phase: 3780-3882 g
Date of receipt: 24 August 2011
Acclimatization time: 13 days
Animal identification: The individual identification was by engraved ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number.
Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 607
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 16.6 – 23.4°C
Relative humidity: 38-76%
Housing/Enrichment: 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
Ventilation: 15-20 air exchanges/hour

The environmental parameters were recorded twice daily during the study. Variations from the target humidity and temperature range were observed during the study. These deviations were considered to have no impact on the animal health, as certified by the Clinical Veterinarian, or on the outcome of the study and interpretation of the results.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A single dose of 0.1 ml of the paste test item EMI-DBS was administered to each animal.

Duration of treatment / exposure:

The test item was applied once and observations made up to 3 weeks after application.

Observation period (in vivo):

The eyes were examined at 1, 24, 48, 72 hours and 1, 2 and 3 weeks after treatment. 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.
At the end of the observation period, each animal was sacrificed by intramuscular injections of CP-Ketamin 10% and CP-Xylazin 2% followed by iv. Euthasol® 40% anaesthesia. Death was verified by checking pupil and corneal reflex and the absence of respiration.

Number of animals or in vitro replicates:

3

Details on study design:

Application of the Test Item

Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition and any with ocular lesions were rejected.
An initial test was performed using one animal. The test item was instilled into the conjunctival sac of the left eye. The eyelids were held closed for several seconds to prevent the loss of the test item. The contralateral eye served as the control. Immediately after the administration of the test item, an assessment of the initial pain reaction was made according a six point scale.
After consideration of the ocular responses produced in the first animal, two additional animals were treated.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: Considering readings at 24, 48 and 72 hours after treatment

Score:

1.33

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

1.67

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

1.67

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 4

Irritation parameter:

iris score

Basis:

other: animal number 1, 2 and 3

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

0

Remarks on result:

other: Animal's individual mean score

Results 5

Irritation parameter:

other: Conjunctivae redness

Basis:

animal #1

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.33

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 6

Irritation parameter:

other: Conjunctivae redness

Basis:

animal #2

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.67

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 7

Irritation parameter:

other: Conjunctivae redness

Basis:

animal #3

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.67

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 8

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

3

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 9

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.33

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 10

Irritation parameter:

other: Conjunctivae discharge

Basis:

animal #1

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

3

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 11

Irritation parameter:

other: Conjunctivae discharge

Basis:

animal #2

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.33

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 12

Irritation parameter:

other: Conjunctivae discharge

Basis:

animal #3

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.33

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Results 13

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: Considering readings at 24, 48 and 72 hours after the treatment

Score:

2.67

Reversibility:

not fully reversible within: 3 weeks

Remarks on result:

other: Animal's individual mean score

Other effects:

There was no mortality observed during the study. The body weight and body weight change were considered to be normal with no indication of treatment related effect. There were no clinical signs observed that could be related to treatment.

Any other information on results incl. tables

Examination of eye-irritancy

The eyes were examined at 1, 24, 48, 72 hours and 1, 2 and 3 weeks after the application.

Initial Pain Reaction (IPR) (score 2 or 3) was observed in all animals after test item administration.

One hour after the application: Conjunctival redness (score 2), conjunctival chemosis (score 3) and discharge (score 3) were observed in all animals. All animals showed corneal opacity (score 1, area 4).

At 24 hours after treatment: Conjunctival redness, chemosis and discharge (score 3) were seen in all rabbits. All animals showed corneal opacity (score 1, area 4).

At 48 hours after treatment: Conjunctival redness (score 2 or 3), chemosis and discharge (score 3 - 3) were seen in all rabbits. All animals showed corneal opacity (score 1 or 2 , areas 4, 3 or 2).

At 72 hours after treatment: Conjunctival redness (score 2), chemosis (score 1, 2 or 3) and discharge (score 1 or 3) were seen in all rabbits. All animals showed corneal opacity (score 2 , areas 1 or 3).

At 1 week after treatment: Conjunctival redness (score 1 or 2) and discharge (score 1 or 3) were seen in all rabbits. Conjuctival chemosis (score 1 or 2) was observed in two animals. Two animals showed corneal opacity (score 2 , area 3).

At 2 weeks after treatment: Conjunctival redness (score 1) was seen in all rabbits. Conjuctival chemosis (score 1) and discharge (score 1) were observed in two animals. Two animals showed corneal opacity (score 2 , areas 1 or 2). In two animals, vascularisation was observed on the surface of the cornea. the extension of the vascularisation is not greater than 1/4 of the surface of the cornea.

At 3 weeks after treatment: Conjunctival redness (score 1) was seen in all rabbits. Conjuctival chemosis (score 1) was observed in one animal and discharge (score 1) was observed in two animals. Two animals showed corneal opacity (score 2 , areas 1 or 2). In one animals, vascularisation was observed on the surface of the cornea. The extension of the vascularisation is greater than 1/4 of the surface of the cornea.

The study was terminated after the 3-week observation.

The animal’s individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

Animal Number	Cornea Opacity	Iris	Conjunctivae
			Redness	Chemosis	Discharge
00369	1.33	0.00	2.33	3.00	3.00
00445	1.67	0.00	2.67	2.33	2.33
00443	1.67	0.00	2.37	2.67	2.33

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item EMI-DBS, applied to rabbit eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced at 2 weeks after application. The effects were not fully reversible within 3 weeks. According to Regulation (EC) No 1272/2008, EMI-DBS requires classification as an eye irritant (Irreversible effects on the eye/serious damage to eyes (Category 1)).

Executive summary:

The eye irritation potential of the test substance was determined in accordance with the OECD Guideline for Testing of Chemicals 405. The study was conducted by applying 0.1 ml of the test item in a single dose into the conjunctival sac of the left eye of male New Zealand White rabbits. Individual body weight was recorded at the beginning and end of the study. Morbidity and clinical signs of toxicity were checked daily. The eyes were examined at 1, 24, 48, 72 hours and 1, 2 and 3 weeks after the application.

No adverse effects on body weight development were noted during the study period. The general state and behavior of animals were normal throughout the study period. During the study, no signs of eye irritation were observed in the control eye of all animals.

Initial Pain Reaction (IPR) (score 2 or 3) was observed in all animals after test item administration.

One hour after the application: Conjunctival redness (score 2), conjunctival chemosis (score 3) and discharge (score 3) were observed in all animals. All animals showed corneal opacity (score 1, area 4). At 24 hours after treatment: Conjunctival redness, chemosis and discharge (score 3) were seen in all rabbits. All animals showed corneal opacity (score 1, area 4). At 48 hours after treatment: Conjunctival redness (score 2 or 3), chemosis and discharge (score 3 - 3) were seen in all rabbits. All animals showed corneal opacity (score 1 or 2 , areas 4, 3 or 2). At 72 hours after treatment: Conjunctival redness (score 2), chemosis (score 1, 2 or 3) and discharge (score 1 or 3) were seen in all rabbits. All animals showed corneal opacity (score 2 , areas 1 or 3). At 1 week after treatment: Conjunctival redness (score 1 or 2) and discharge (score 1 or 3) were seen in all rabbits. Conjuctival chemosis (score 1 or 2) was observed in two animals. Two animals showed corneal opacity (score 2 , area 3). At 2 weeks after treatment: Conjunctival redness (score 1) was seen in all rabbits. Conjuctival chemosis (score 1) and discharge (score 1) were observed in two animals. Two animals showed corneal opacity (score 2 , areas 1 or 2). In two animals, vascularisation was observed on the surface of the cornea. the extension of the vascularisation is not greater than 1/4 of the surface of the cornea. At 3 weeks after treatment: Conjunctival redness (score 1) was seen in all rabbits. Conjuctival chemosis (score 1) was observed in one animal and discharge (score 1) was observed in two animals. Two animals showed corneal opacity (score 2 , areas 1 or 2). In one animals, vascularisation was observed on the surface of the cornea. The extension of the vascularisation is greater than 1/4 of the surface of the cornea. The study was terminated after the 3-week observation. The test item EMI-DBS, applied to rabbit eye mucosa, caused significant conjunctival irritant effects at one hour which were reduced at 2 weeks after application. The effects were not fully reversible within 3 weeks.

According to Regulation (EC) No 1272/2008, EMI-DBS requires classification as an eye irritant (Irreversible effects on the eye/serious damage to eyes (Category 1)).