Bis(2-hydroxyethyl)ammonium acetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

1999-08-17 to 1999-08-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is the result of a structural analogue substance used as read-across substance. Study is conducted according to Guidelines in a GLP certified laboratory.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

RA1

IUPAC Name:

RA1

Test material form:

other: liquid

Details on test material:

- Physical state: Liquid
- Stability under test conditions: Maximum 365 days
- Storage condition of test material: At room temperature at about 20 ºC, away from direct sunlight

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes,
F-01400 Chatillon sur Charlaronne,
France.
- Age at study initiation: Males: 15 weeks
Females: 14 weeks
- Weight at study initiation: Male 1 = 2737 g, Female 1 = 2917 g, Female 2 = 2974 g
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): Community tap water ad libitum
- Acclimation period: Five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 50-80%
- Air changes (per hr): 10 -15
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light.


IN-LIFE DATES: From: 17th August 1999 To: 30th August 1999

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml/animal

Duration of treatment / exposure:

The test substance was applied once into one eye of each rabbit. The treated eyes were not rinsed after application. Scoring of irritation effects was performed up to 7 days after test substance application.

Observation period (in vivo):

There was a 7 day observation period after the application of the substance.

Number of animals or in vitro replicates:

Three animals: One male and two females.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye was not rinsed following application


SCORING SYSTEM:
The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 at approximately 1, 24, 48 and 72 hours, as well as 7 days after administration.

TOOL USED TO ASSESS SCORE: Varta Cliptrix diagnostic-lamp.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

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

Score:

0

Max. score:

4

Remarks on result:

other: Reversibility not applicable as no irritation seen

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 hour to 7 days

Score:

0

Max. score:

2

Remarks on result:

other: Reversibility not applicable as no irritation seen

Results 3

Irritation parameter:

conjunctivae score

Remarks:

chemosis

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Max. score:

13

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Irritation:
Application of the test article to healthy rabbit conjunctiva resulted in a primary irritation score of 0.33 (out of 13). Slight reddening and slight swelling of the conjunctiva was noted in all animals one hour after treatment. Slight reddening of the conjunctiva persisted 72 hours after the administration in one animal and disappeared on the 7-day reading. The sclera of one animal was slightly reddened at the 1-hour examination.

Corrosion:
No corrosion of the cornea was observed at any of the reading times.

Other effects:

Viability/Mortality and Clinical Signs:
No clinical signs of systemic toxicity were observed in the animlas during the test and observation period, and no mortality occurred.

Coloration:
No staining by the test article was observed.

Body Weights:
The body weight of all rabbits were considered to be within the normal range of variability for the species.

Any other information on results incl. tables

Table 1: Body Weights:

Body Weights in grams
Animal No.	Sex	First Day of Acclimatization	Day of Treatment	Last Day of Observation
1	Male	2737	2903	3118
2	Female	2917	3107	3291
3	Female	2974	2840	3095

Table 2: Eye Irritation Scores - Individual Values:

Animal Number	Sex	Evaluation Interval	Corneal Opacity	Iris	Conjunctivae		Cumulative
					Redness	Chemosis	Score	Mean
1 2 3	Male Female Female	1 hour	0 0 0	0 0 0	1 1 1	1 1 1	2.00 2.00 2.00	2.00
1 2 3	Male Female Female	24 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	48 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	72 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	7 days	0 0 0	0 0 0	0 0 0	0 0 0	0.00 0.00 0.00	0.00

Table 3: Eye Irritation Scores - Mean Values after 24, 48 and 72 hours:

Animal Number	Sex	Corneal    Opacity	Iris	Conjunctivae		Primary Eye Irritation Score
				Redness	Chemosis
1 2 3	Male Female Female	0.00          0.00          0.00	0.00    0.00 0.00	0.00       1.00      0.00	0.00               0.00               0.00	0.33

Table 4: Eye Irritation Scores - Mean Values over Time:

Evaluation Interval	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
1 hour	0.00	0.00	1.00	1.00
24 hours	0.00	0.00	0.33	0.00
48 hours	0.00	0.00	0.33	0.00
72 hours	0.00	0.00	0.33	0.00
7 days	0.00	0.00	0.00	0.00

Table 5: Eye Irritation Scores - Assessment according to EC Guidelines:

Evaluated Interval	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
24 hours 48 hours 72 hours	Not Irritating	Not Irritating	Not Irritating	Not Irritating

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In a GLP guideline study, RA1 was considered to be not irritating to the rabbit eye.

Executive summary:

RA1 was tested for its eye irritation potential. 0.1 ml of the undiluted test substance was placed in the conjunctival sac of the left eye of each animal. The lids were gently held together for about 1 second to prevent loss of the test article. The right eye remained untreated and served as the reference control.

The animals were observed for 72 hours for signs of eye irritation, clinical signs, mortality and changes in body weight.

Application of the test article to healthy rabbit conjunctiva resulted in a primary irritation score of 0.33 (out of 13). Slight reddening and slight swelling of the conjunctiva was noted in all animals one hour after treatment. Slight reddening of the conjunctiva persisted 72 hours after the administration in one animal and disappeared on the 7-day reading. The sclera of one animal was slightly reddened at the 1-hour examination.

No corrosion of the cornea was observed at any of the reading times. The test substance had no effect on viability, clinical signs or body weight.

Based on these results, RA1 is considered not to be an eye irritant.