[(±)-(4-amino-2-hydroxy-4-oxobutyl)trimethylammonium] chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 06 December 2012 to 04 February 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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:

[(±)-(4-amino-2-hydroxy-4-oxobutyl)trimethylammonium] chloride

EC Number:

226-073-0

EC Name:

[(±)-(4-amino-2-hydroxy-4-oxobutyl)trimethylammonium] chloride

Cas Number:

5261-99-4

Molecular formula:

C7H17N2O2.Cl

IUPAC Name:

4-amino-2-hydroxy-N,N,N-trimethyl-4-oxobutan-1-aminium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Physical state: white powder
- Analytical purity: 99,4%
- Lot/batch No.: WI 2274
- Storage condition of test material: at room temperature not exceeding 40°C.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Institute of Occupational Medicine, Łódź
- Age at study initiation: The experiment was conducted on three females which were 5.5 months old.
- Weight at study initiation: 4.2-4.7 kg
- Housing: The animals were kept individually in metal cages with the following dimensions: 60 x 35 x 40 cm (length x width x height).
- Diet (e.g. ad libitum): LSK standard granulated laboratory fodder, ad libitum
- Water (e.g. ad libitum): drinking tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21°C
- Humidity (%): 30 – 65%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light – 12 hours dark

IN-LIFE DATES: From: 12 December 2012 To: 24 December 2012

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The test item (grounded to a powder) in a volume of 0.1 mL (0.049 g) was administered to the conjunctival sac of one eye of the animal by gently pulling the lower eyelid away from the eyeball.

Observation period (in vivo):

1, 24, 48 and 72 hours as well as 7 days after the administration of the test item.

Number of animals or in vitro replicates:

Three females

Details on study design:

SCORING SYSTEM:

Cornea
Opacity: degree of density (readings should be taken from most dense area) *
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4
Maximum possible: 4 * The area of corneal opacity should be noted

Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, iris reactive to light (a sluggish reaction is considered to be an effect 1
Hemorrhage, gross destruction, or no reaction to light 2
Maximum possible: 2

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
Normal 0
Some blood vessels hyperaemic (injected) 1
Diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3
Maximum possible: 3

Chemosis
Swelling (refers to lids and/or nictitating membranes)
Normal 0
Some swelling above normal 1
Obvious swelling, with partial eversion of lids 2
Swelling, with lids about half closed 3
Swelling, with lids more than half closed 4
Maximum possible: 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Irritant / corrosive response data:

After the administration of the test item, some pathological changes were observed in the conjunctivas of the animals’ eyes. No pathological changes were stated in the cornea and iris.

During the reading which took place 1 hour after the administration of the test item, congestion of the nictitating membrane was observed in three animals. Moreover, erythema in the form of congestion (injection) of some blood vessels and circumcorneal injection were stated in animals no. 2 and 3. Swelling of the nictitating membrane was observed in animal no. 2.

During the reading which took place 24 hours after the administration of the test item, congestion of the nictitating membrane was observed in animals no. 2 and 3. Moreover, circumcorneal injection was stated in animal no. 2. No conjunctival edema was observed.

During the reading which took place 48 hours after the administration of the test item, congestion of the nictitating membrane was observed in animals no. 2 and 3.

During the reading which took place 72 hours after the administration of the test item, congestion of the nictitating membrane was observed in animal no. 2.

During the reading which took place 7 days after the administration of the test item, no pathological changes were observed in rabbit no. 2.

Any other information on results incl. tables

Table 1: Eye irritation scores

Animal	Region of eye	24 h	48 h	72 h	7 d
1	Cornea	0	0	0	-
	Iris	0	0	0	-
	Conjunctivae erythema	0	0	0	-
	Conjunctivae swelling	0	0	0	-
2	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae erythema	1	1	1	0
	Conjunctivae swelling	0	0	0	0
3	Cornea	0	0	0	-
	Iris	0	0	0	-
	Conjunctivae erythema	1	1	0	-
	Conjunctivae swelling	0	0	0	-

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the available results, the test substance is not classified as eye irritating.

Executive summary:

The acute eye irritation/corrosion study, according to the OECD Guideline No. 405 / EU Method B.5., was performed in order to obtain information on health hazards likely to arise from possible contact of the test item. The study started with a preliminary study on one animal. The test item was grounded to a powder and administered to the conjunctival sac of one eye of the animal (rabbit no. 1). Its volume was 0.1 mL. The second eye served as a control. After the evaluation of the treated eye, the test item was applied to the eyes of the next two animals (rabbit no. 2 and 3). General clinical observations of the animals for morbidity and mortality were performed daily during the entire experiment. Detailed clinical observations for changes in the cornea, iris and conjunctiva were performed 1, 24, 48 and 72 hours (all animals) as well as 7 days (rabbit no. 2) after the administration of the test item. The animals’ body weight was determined on the administration day (day 0) and on the last day of the experiment. After the administration of the test item, some pathological changes were observed in the conjunctivas of the animals’ eyes. No pathological changes were stated in the cornea and iris. Based on the available results, the test substance is not classified as eye irritating.