C16-18 (evennumbered, C18 unsaturated) alkyl bis(2-hydroxyethyl) amine oxide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 March 1990 -23 April 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to EC/OECD guidelines and under GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

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:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-(Z)-octadec-9-en-1-ylimino)bisethanol N-oxide

EC Number:

300-699-5

EC Name:

2,2'-(Z)-octadec-9-en-1-ylimino)bisethanol N-oxide

Cas Number:

93962-62-0

Molecular formula:

C22H45NO3

Details on test material:

Identification: Aromox 0/12
Chemical name: Oleylbis(2-hydroxyethyl)amineoxide
Description: Clear viscous liquid
Batch/Lot Number: RCD/HJ-173
Purity/Composition: Amine Oxide 58.2%
Instructions for test substance storage: In the original container at room temperature in the dark
Stability under storage conditions: stable
Expiry date: March 09, 1991

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute , Someren, The Netherlands
- Age at study initiation: 14 or 16 weeks
- Weight at study initiation: 2698-2851 gram
- Housing: Individually in cages with perforated floors and equipped with an automatic drinking system (ITL, Bergen, The Nether lands).
- Diet (e.g. ad libitum): 100g per day
- Water (e.g. ad libitum): ad linitum
- Acclimation period: At least five days under test conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 14-23
- Humidity (%): 46-75
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Temperature was approximately 4 hours below 18ºC , due to a technical failure of the heating system), this did not affect the outcoem of the study.

IN-LIFE DATES: From: 20 March 1990 -23 April 1990

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the left eye of each animals served as control

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

A single instillation of 0.1 ml in the right eye af the animals was observed for 21 days.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation o f the test substance.

Immediately after the 24 hours observation, a solution o f 2% fluorescein in water (pH adjusted to 7.0) was applied to both eyes of each animal to
quantitatively examine the potential for corneal injury. The bright green stained area indicating epithelial damage was estimated as a percentage of the total corneal area. This procedure was repeated in all three animals on day 4 and in animal 3068 also on days 8, 15 and 22.

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done

SCORING SYSTEM:
Scoring system eye irritation

Cornea: degree of density (area most dense taken for reading)
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 Easily discernible translucent areas, details of iris slightly obscured
3 Nacrous areas, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

Area of Cornea Involved
1 One quarter (or less) but not zero
2 Greater than one quarter but less than half
3 Greater than half but less than three quarters
4 Greater than three quarters, up to whole area

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

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae cornea and iris)

0 Blood vessels normal
1 Some blood vessels definitely hyperaemic
(injected)
2 Diffuse, crimson colour, individual vessels not easily discernible
3 Diffuse, beefy red

Chemosis (lids and/or nictitating membranes)
0 No swelling 0
1 Any swelling above normal (includes nictitating membranes)
2 Obvious swelling with partial eversion of lids
3 Swelling with lids about half-closed
4 Swelling with lids more than half-closed

Discharge
0 No discharge
1 Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 Discharge with moistening of the lids and hairs just adjacent to lids
3 Discharge with moistening of the lids and hairs a considerable area around the eye

TOOL USED TO ASSESS SCORE: fluorescein, In case standard lighting was considered inadequate for observing minor effects, eye examinations were performed using a pocket flash light.

Results and discussion

In vivo

Results

Irritation parameter:

overall irritation score

Remarks:

corneal opacity/iritis/conjunctival redness/oedema

Basis:

mean

Remarks:

all animals

Time point:

other: 24-48-72 hours

Score:

1.03

Max. score:

4

Reversibility:

not fully reversible within: 21 dyas

Remarks on result:

other: One animal still showed slight conjunctival redness after 21 days.

Irritant / corrosive response data:

Instillation o f AROMOX 0/12 into one of the eyes of each of three albino rabbits affected the cornea and the conjunctivae in all three animals and the
iris in animal 3068. The opacity of the cornea was reversible within 48 hours in animals 3070 and 3084 and within 21 days in animal 3068. The injec tion of the iris in animal 3068 was reversible within 7 days. The irritation of the conjunctivae was reversible within 14 days in animal 3070, within 21 days in animal 3084, while slight conjunctival redness was still observed in animal 3068 on day 22. No corrosion was observed.
Treatment o f the eyes with fluorescein 2%, 24 hours a f t e r t e s t substance
i n s t i l l a t i o n revealed corneal e p i t h e l i a l damage i n a l l three animals and i n
animal 3 0 6 8 on days 4, 8 and 15.
C

Other effects:

No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Rabbit No and sex	Region of the eye			Hours after instillation							Average 24-48 hours
				1	24	48	72	7(days)	14(days)	21(days
3068 f	Cornea	Degree of opacity		0	1	1	1	0	0	0	1
		Area of opacity		0	2	1	1	1	1	0
	Iris			0	0	1	1	0	0	0	0.7
	Conjunctivae		Redness	2	3	3	3	2	1	1	3
			Chemosis	2	2	1	1	0	1	0	1.7
			Discharge	0	2	1	1	0	0	0
3070 f	Cornea		Degree of opacity	0	0	0	0	0	0	0	0
			Area of opacity	0	1	0	0	0	0	0
	Iris			0	0	0	0	0	0	0	0
	Conjunctivae		Redness	1	2	2	2	1	0	0	2
			Chemosis	2	2	1	0	0	0	0	1
			Discharge	0	1	0	0	0	0	0
3084 f	Cornea		Degree of opacity	1	1	0	0	0	0	0	0.3
			Area of opacity	1	1	0	0	0	0	0
	Iris			0	0	0	0	0	0	0	0
	Conjunctivae		Redness	1	2	2	2	1	1	0	2
			Chemosis	2	1	1	0	0	0	0	0.7
			Discharge	0	1	0	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

Category 2A (irritating to eyes) based on GHS criteria

Remarks:

Migrated information

Conclusions:

In the study report a draize calculation is used and the classification follows the DSD (83/467/EEC, Annex VI of the EEC council Directive 67/548/EEC) which leads to a non-classification. However three animals have a ≥ 2 an average score 24-48-72 hours after exposure for conjuntival redness. The degree of opacity is ≥ 1 in in one animal. All effects execpt for slight redness in the eye of one animal are fully reversible within 21 days or sooner. No effects of corrosion are observed. Therefore based on these results, although some slight effects are still seen after 21 days, the substance is classified according GHS in category 2A (irritating to eyes).

Executive summary:

The Purpose of this study was to assess the possible irritation or corrosion potential when a single dose of the test item was placed in the conjunctival sac of a rabbit eye. This study was carried out in accordance with OECD Guideline No. 405 and EC guideline B.5. The test item was instilled in one o f the eyes of each o f the three animals, followed by seven observations at approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after test article administration. Under the conditions of this study, the test item resulted in adverse effects on the cornea, iris and conjunctivae. The opacity of the cornea was reversible within 48 hours in two animals and within 21 days in the third animal. The injection of the iris, observed in one animal, was reversible within 7 days. The irritation of the conjunctivae was reversible within 21 days in two animals, while the third animal still showed slight conjunctival redness at termination of the study. No corrosion and no signs o f systemic intoxication were observed. In the study report a draize calculation is used and the classification follows the DSD (83/467/EEC, Annex VI of the EEC council Directive 67/548/EEC) which leads to a non-classification. However three animals have a ≥ 2 an average score 24-48-72 hours after exposure for conjuntival redness. The degree of opacity is ≥ 1 in in one animal. All effects except for slight redness in the eye of one animal are fully reversible within 21 days or sooner. No effects of corrosion are observed. Therefore based on these results, although some slight effects are still seen after 21 days, the substance is classified according GHS in category 2A (irritating to eyes).