Ethanol, 2,2'-oxybis-, reaction products with ammonia, morpholine derivs. residues

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The in vivo eye irritation study was performed in 2010, according to EU method B.5 and OPPTS 870.2400. This type of data is intended to fulfill the testing requirements for the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) (7 U.S.C. 136, et seq.) and the Toxic Substances Control Act (TSCA) (15 U.S.C. 2601).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ethanol, 2,2'-oxybis-, reaction products with ammonia, morpholine derivs. residues

EC Number:

272-712-1

EC Name:

Ethanol, 2,2'-oxybis-, reaction products with ammonia, morpholine derivs. residues

Cas Number:

68909-77-3

Molecular formula:

Not applicable (UVCB)

IUPAC Name:

2-(2-hydroxyethoxy)ethan-1-ol; 2-[1-(morpholin-4-yl)ethoxy]ethan-1-amine; 2-{2-[bis(2-hydroxyethyl)amino]ethoxy}ethan-1-ol; 4-{2-[2-(morpholin-4-yl)ethoxy]ethyl}morpholine; morpholin-3-one

Test material form:

liquid

Details on test material:

Name: Ethanol, 2,2'-oxybis-, reaction products with ammonia, morpholine derivs. residues
Physical state: liquid
Appearance: brown liquid

Specific details on test material used for the study:

- Name of test material (as cited in study report): Amine C8
- Substance type: no data
- Physical state: viscous amber liquid
- Storage condition of test material: 15-30 °C, room temperature

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan or any other USDA approved vendor
- Age at study initiation: young adult (12-18 weeks) at start of dosing; records of dates of birth for animals used in this study will be retained in the Calvert archives.
- Weight at study initiation: 2.0-3.0 kilograms at the outset (Day 1) of the study.
- Housing: Animals are housed individually in compliance with USDA Guidelines. The room in which the animals will be kept will be documented in the study records. no other species will be kept in the same room.
- Diet (e.g. ad libitum): All animals will have access to PMI certified Hi-Fiber Rabbit Diet or equivalent as per Calvert SOP, unless otherwise specified. The lot number and specifications of each lot used are archived in the facility records at Calvert. No contaminants are known to be present in the certified diet at levels that would be expected to interfere with the results of this study. analysis of the diet was limited to that performed by the manufacturer, records of which will be maintained in the Calvert archives.
- Water (e.g. ad libitum): Tap water is available ad libitum, to each animal via an automatic watering device. The water is routinely analyzed for contaminants as per Calvert SOP's. No contaminants are known to be present in the water at levels that would be expected to interfere with the results of this study. Results of the water analysis will be maintained in the Calvert archives.
- Acclimation period: study animals were acclimated to their housing for a minimum of seven days prior to their first day of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 21°C
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Test system

Vehicle:

not specified

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:

Only once instilled

Observation period (in vivo):

mortality/morbidity: once daily
clinical observations: prior to dose administration and then daily
ocular observations: at 1 (± 15 minutes), 24, 48 and 72 hours (± 1 hour); if there is no evidence of irritation at 72 hours, the study will be complete. Extended observations may be necessary if there is persistent corneal involvement or other ocular irritation is observed to determine the progress of the lesions and their reversibility or irreversibility. If the cornea, iris or conjunctivae have not healed completely by Day 8, the unhealed animals will be retained and reexamined on Days 15 and 22, if injury persists. In addition to the observations of the cornea, iris and conjunctivae, any other lesions, which are noted, will be recorded and reported. The grades of ocular reaction according to Draize will be manually recorded at each examination.
Body weight: prior to dose administration

Number of animals or in vitro replicates:

Three

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eyes of the animals will remain un-rinsed for at least 24 hours after instillation of the test article. The treated eyes may be gently washed out (if necessary) using room temperature physiological saline.
- Time after start of exposure: see above

SCORING SYSTEM: An animal has exhibited a positive reaction if the test article has produced at any observation one or more of the following signs: opacity of the cornea (other than a slight dulling of the normal luster), inflammation of the iris (other than a slight deepening of the rugae or light hyperemia of the circum corneal blood vessels) or an obvious swelling in the conjunctivae (excluding the cornea and iris) with partial eversion of the eyelids or a diffuse crimson color with individual vessels not easily discernible.
EEC classification of ocular irritants: The total ocular irritation score for the 24, 48 and 72 hour invertvals are individually added for corneal opacity, iris lesion, conjunctival redness and conjunctival edema. For a group of the three animals, the calculated mean scores will remain expressed individually and compared to the EEC ocular evaluation criteria. Scores for two or more animals above the appropriate mean will classify the test article in that particular category. A test substance will be classified as a nonirritant, irritating to the eyes (R36) or risk of serious damage to eye (R41 Risk Phrase) based on the EEC Ocular Evaluation Criteria.

Globally harmonized system of classification evaluation criteria: for this classification system, the 24-, 48- and 72- hours scores will be added separately for each animal and each total divided by 3 (three time points) to yield the individual mean scores for each animal.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 22 days

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 15 days

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 4

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 15 days

Results 6

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 15 days

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

not reversible

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 22 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

not fully reversible within: 22 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

fully reversible within: 15 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Results 13

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24 hours post dose

Score:

26

Max. score:

110

Results 14

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 48 hours

Score:

31

Max. score:

110

Results 15

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 72 hours

Score:

32.67

Max. score:

110

Results 16

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: day 5

Score:

30.67

Max. score:

110

Results 17

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 8 days

Score:

28.67

Max. score:

110

Reversibility:

fully reversible

Irritant / corrosive response data:

No corneal, iris or conjunctival irritation was observed in the untreated eyes of the rabbit at any time point. In the first animal, scattered to diffuse corneal opacity covering 1/4 or less of the eye was observed at 24 hours post dose. Iritis (grade 1) was observed at 1 hour after dosing, at 24, 48 and 72 hours post dose, on Days 5 through 8. Conjunctival redness (grade 1) was noted at 1 hour after dosing up until study termination on Day 22. Chemosis (grades 3 to 4) was also seen starting at 1 hour post dose and throughout the study until it diminished to grade 1 on Day 22. Discharge (grade 3) was observed in the rabbit throughout the entire duration of the study.
In the second animal, scattered to diffuse corneal opacity covering 1/4 to 1/2 of the total surface area of the eye was observed from 24 hours post dose out to Day 8. Iritis (grade 1) was observed at 1 hour after dosing through to day 8. Conjunctival redness (grade 1) was noted at 1 hour after dosing up until Day 15. Chemosis (grades 3 to 4) was also seen from 1 hour post dose until Day 5. From Days 6-8, chemosis declined to grade 2 and was not apparent by Day 15. Discharge (grade 3) was observed from 1 hour post dose until Day 5. On Days 6-8, discharge decreased to grade 2 and by Day 15 further lowered to grade 1. At Day 22, no discharge was noted.
For the third animal, scattered or diffuse areas of corneal opacity with details of the iris clearly visible covering greater than 1/2 to 3/4 of the whole area of the eye was observed from 48 hrs post dose until Day 15. On Day 22, no ulceration or opacity was observed. Iritis (grade 1) was observed at 1 hour after dosing through to Day 22. Conjunctival redness (grade 1) was noted at 1 hour after dosing through to study termination on Day 22. Chemosis (grades 3 to 4) was also seen from 1 hour post dose through to Day 5. From Days 6 to 22, chemosis declined to grade 2. Discharge (grade 3) was observed from 1 hour post dose until study termination on Day 22.

Other effects:

No mortality or adverse clinical signs were observed in the animals during the study

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Based on the data of this EPA OPPTS 870.2400 and EC Method B.5 study and according to the criteria of the CLP Regulation, the test substance is classified as serious eye damage (category 1).