Reaction mass of 2-ethyl-2-[[3-(2-methylaziridin-1-yl)propionyl]methyl]propane-1,3-diyl bis(2-methylaziridine-1-propionate) and 2,2-bis({[3-(2-methylaziridin-1-yl)propanoyl]oxy}methyl)butyl 3-[2,2-bis({[3-(2-methylaziridin-1-yl)propanoyl]oxy}methyl)butoxy]propanoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Jun 2000- Nov 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The experiment was conducted according to current EC/OECD guidelines in compliance with GLP standards.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

IONAC XAMA®220

IUPAC Name:

IONAC XAMA®220

Details on test material:

- Substance type: Clear colorless liquid
- Expiration date of the lot/batch: not described
- Storage condition of test material: room temperature
- Receipt date of batch: June 22, 2000

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Myrtle's rabbitry, Thompson Station, TN
- Age at study initiation: appr. 15 weeks
- Weight at study initiation: 2.9-3.5kg (m)/ 2.8-3.5kg (f)
- Housing: individually, in suspended stainless steel cages
- Diet: ad libitum, PMI Certified Rabbit Chow #5322 (Purina Mills, Inc.)
- Water: ad libitum, purified tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-21
- Humidity (%): 42-80
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: July 10, 2000 To: July 31, 2000

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the contralateral eye remained untreated

Amount / concentration applied:

0.1ml

Duration of treatment / exposure:

Single instillation

Observation period (in vivo):

1, 24, 48, 72 hours and up to 7 days

Number of animals or in vitro replicates:

3 males, 3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no. In addition to the 6 rabbits whose eyes were not rinsed, 3 females were treated with 0.1ml of the test substance and subsequently the eyes were rinsed (30s after instillation) with sterile water. These animals were observed up to 21 days.

SCORING SYSTEM:
Ocular grading system according to Draize; details described in OECD guideline

OBSERVATIONS:
- General health and mortality was monitored twice daily.

TOOL USED TO ASSESS SCORE: lamp / fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

ca. 3.4

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: 2/6 rabbits had severe swelling that hampered ocular inspection at 24 hours.

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

ca. 1.1

Max. score:

2

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: For none of the rabbits the iris score could be assessed at 24 hours due to severe swelling.

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

2.8

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

2.2

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Irritant / corrosive response data:

The eyes of animals with rinsed eyes were only exposed for 30s. The following scores were obtained (where possible, based on observations at 24, 48 and 72 hours):
Cornea: ca. 2.6 (one animal's eye too swollen at t=48 hours to be assessed)
Iris: ca. 1.2 (one animal's eye too swollen to be assessed)
Conjunctivae- redness: ca. 2.8
Chemosis: ca. 4
Conjunctivae- discharge: ca. 2.3
After 21 days, effects were not reversed.

Other effects:

In the group of animals without rinsing of the eyes slight hairloss around the treated eye was observed in 2 males and 2 females. One male showed head tilt on days 0-2.
The three females whose eyes were rinsed also showed hairloss around the treated eye (slight to moderate). Two females showed head tilt on days 0-2.

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:

An eye irritation study according to current OECD/EC guidelines was conducted in compliance with GLP principles. Based on the results of this study, the substance has to be classified category 1 causing "irreversible effects on the eye/ serious damage to eyes" according to CLP Regulation (EC) No. 1272/2008.

Executive summary:

An eye irritation study according to current OECD/EC guidelines was conducted in compliance with GLP principles. In this study, 6 rabbits (3 males and 3 females) showed severe effects on the eyes the first 72 hours after treatment, which did not reverse after one week. In parallel, three females were treated with the substance, which was removed from the eye after 30s by rinsing. The effects to the eyes of these animals were comparable to the eyes of the unrinsed group. The effects did not reverse within 21 days after treatment. Additionally, three treated animals (two with rinsed and one with unrinsed eyes) showed tilting of the head, Seven rabbits showed slight hairloss around the treated eye. Based on the results of this study, the substance has to be classified with category 1, H318, causing "irreversible effects on the eye/ causes serious eye damage" according to CLP Regulation (EC) No. 1272/2008.