3-methyl-1-vinyl-1H-imidazolium methyl sulphate

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other: GLP compliant guideline study, available as unpublished report, fully adequate for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No 440/2008

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nousan No. 8147

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-methyl-1-vinyl-1H-imidazolium methyl sulphate

EC Number:

247-832-2

EC Name:

3-methyl-1-vinyl-1H-imidazolium methyl sulphate

Cas Number:

26591-72-0

Molecular formula:

C6H9N2.CH3O4S

IUPAC Name:

1-ethenyl-3-methyl-1H-imidazol-3-ium methyl sulfate

Details on test material:

- Name of test item: 3-Methyl-1-vinyl-1H-imidazolium methyl sulfate
- Test-item No.: 11/0584-1
- Batch identification: 77761968E0
- Purity: Main component: 96.2 g/100g; w (C6H9N2, cation in the main component): 47.3 g/100 g; w (CH3O4S, anion in the main component): 48.9 g/100 g; water: 1.9 g/100 g. Determined by 1H-NMR-spectroscopy
- Homogeneity: The test item was homogeneous by visual inspection.
- Storage stability: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Expiry date: August 19, 2013
- Storage conditions: Room temperature
- Physical state/ color: Solid / yellowish
- pH-value: approx. 5 (undiluted test item, determined by Bioassay)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: approx. 3 months
- Weight at study initiation: 2.36 kg – 2.60 kg
- Housing: single housing in stainless steel wire mesh cages with grating with shallow cage body; floor area: 4225 cm²
- Enrichment: Wooden gnawing blocks (Typ KNH E-041); Abedd ® Lab. and Vet. Service GmbH Vienna, Austria
- Diet (e.g. ad libitum): STANRAB (P) SQC; SDS Special Diets Services, 67122 Altrip, Germany
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: at least 5 days before application

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: negative control: untreated left eye

Amount / concentration applied:

Test item preparation: For better handling the test item was ground with mortar and pestle until the test item was liquid.

Application volume: 0.1 mL (undiluted)

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3

Details on study design:

EXPERIMENTAL PROCEDURE
- Body weight determination: Just before application of the test item and after the last reading.
- Route of application: The test item was applied in a single dose to the conjunctival sac of the right eyelid.
- Rinsing of the eye: About 24 hours after application of the test item the treated eye of the animal(s) was rinsed with 3 to 6 mL of hand warm tap water for 1 to 2 minutes using a syringe with a blunt probe.
- Readings: Approx. 1, 24, 48 and 72 h after application and after seven days.
- Examination of the eye(s): An additional eye examination was performed 24 h after application with the instillation of a fluorescein solution.
- Illumination used for reading: At the 24 hours reading reactions on cornea or iris were observed using a slit lamp and otoscope lamp.
- Mortality: A check for any dead or moribund animal was made at least once each workday.

SCORING SYSTEM:
The evaluation of eye irritation was performed according to the quoted guidelines. In addition to specific observations recommended by the regulatory authorities, evaluations were made of discharge from the eye and the area of cornea affected by the lesions.

Cornea opacity (op): Degree of density (the most dense area is taken for reading)
Grading:
0 No ulceration or opacity
1 slight: Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 moderate: easily discernible translucent area, details of iris slightly obscured
3 marked: nacreous area, no details of iris visible, size of pupil barely discernible
4 severe: opaque cornea, iris not discernible through the opacity

Area of cornea involved (ar) (The assessment of these ocular reactions is performed independent of the quoted guidelines):
Grading
1 > 0 ≤ ¼
2 > ¼ < ½
3 > ½ < ¾
4 > ¾

Area with fluorescein:
Grading
1 > 0 ≤ ¼
2 > ¼ < ½
3 > ½ < ¾
4 > ¾

Iris:
Grading
0 Normal
1 moderate: markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these observations or combination of
any thereof, iris still reacting to light (sluggish reaction is positive)
2 severe: no reaction to light, haemorrhage, gross destruction (any or all of these)

Conjunctivae redness (red) (Refers to palpebral and bulbar conjunctivae, not to cornea and iris):
Grading
0 Blood vessels normal
1 slight: some blood vessels definitely hyperemic (injected)
2 obvious: diffuse, crimson color, individual vessels not easily discernible
3 severe: diffuse beefy red

Chemosis (sw) (Lids and/or nictitating membrane):
Grading
0 No swelling
1 slight: any swelling above normal (includes nictitating membranes)
2 moderate: obvious swelling with partial eversion of lids
3 marked: swelling with lids about half closed
4 severe: swelling with lids more than half closed

Discharge (di)(The assessment of these ocular reactions is performed independent of the quoted guidelines):
Grading
0 No discharge
1 slight: any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 obvious: discharge with moistening of the lids and hairs just adjacent to lids
3 severe: discharge with moistening of the lids and hairs, and considerable area around the eye

Descriptions of any ocular findings not covered by this scale were recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1, 24, 48, 72 hours and 7 days

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 24 hours

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48, 72 h and 7 days

Score:

0

Max. score:

2

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 - 72 hours

Score:

1

Max. score:

3

Reversibility:

fully reversible

Results 5

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 7 days

Score:

0

Max. score:

3

Results 6

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 1 hour

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 7

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 8

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 48, 72 h and 7 days

Score:

0

Max. score:

4

Irritant / corrosive response data:

No corneal opacity was observed in any animal at any examination time point.

Moderate iritis was observed in all animals 1 hour after application only.
Slight conjunctival redness (grade 1) was noted in all animals from 1 hour up to 72 hours after application.
Slight conjunctival chemosis (grade 1) was noted in two animals 1 hour after application, and persisted in one animal until hour 24. In one animal moderate
conjunctival chemosis (grade 2) was noted 1 hour after administration. Furthermore slight or obvious discharge (grade 1 or 2) was observed in all animals 1
hour after application.
Additional findings like injected scleral vessels in a circumscribed or circular area were noted in all animals from hour 1 up to hour 72 at the latest.
The ocular reactions were reversible in all animals within 7 days after application.

Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0, 0.0 and 0.0 for corneal opacity, 0.0, 0.0 and 0.0 for iris lesions, 1.0, 1.0 and 1.0 for redness of the conjunctiva and 0.3, 0.0 and 0.0 for chemosis.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU