ethyl N2-dodecanoyl-.sc.l.sc.-argininate hydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From November 11, 1996 to February 18, 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Guideline study with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

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

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

434-630-6

EC Name:

-

Cas Number:

60372-77-2

Molecular formula:

Hill formula: C20H41N4O3Cl

IUPAC Name:

ethyl N2-dodecanoyl-l-argininate hydrochloride

Test material form:

solid: particulate/powder

Details on test material:

Nature of substance: powder
- Name of test material: Nα-Lauroyl-L-arginine ethyl ester monohydrochloride
- Analytical purity: 90%
- Lot/batch No.: 5159
- Storage condition of test material: Room temperature, darkness

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: B&K Universal GJ, S. L.
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 2.4 Kg
- Housing: Individually in steel cages (48x59x37 cm)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22 ºC
- Humidity (%): 45-80 %
- Photoperiod (hrs dark / hrs light): 12 hours each 24 hours.

IN-LIFE DATES: From: November 11, 1996; To: December 2, 1996

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1 mL (84 mg)

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3

Details on study design:

SCORING SYSTEM:

Cornea Opacity: Degree of density (area most dense taken for reading)
- 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

Iris:
- Normal: 0
- 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): 1
- No reaction to light, haemorrhage, gross destruction (any or all of these): 2

Conjunctivae: Redness (refers to palpebral and bulbar conjunctivae)
- Blood vessels normal: 0
- Some blood vessels definitely hyperaemic (injected): 1
- Diffuse, crimson colour, individual vessels not easily discernible: 2
- Diffuse beefy red: 3

Edema: Lids and/or nictitating membranes
- No swelling: 0
- Slight swelling (includes nictitating membranes): 1
- Obvious swelling with partial eversion of lids: 2
- Swelling with lids about half-closed: 3
- Swelling with lids more than half-closed: 4

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

7 d

Score:

3

Remarks on result:

other: At 24h: not possible (white corneal opacity and suppuration)

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Reversibility:

not fully reversible within: 21 d

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Reversibility:

not fully reversible within: 21 d

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 9

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 21 d

Results 10

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Remarks on result:

not determinable

Results 11

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Remarks on result:

not determinable

Results 12

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

not determinable

Irritant / corrosive response data:

Reversibility of any observed effect: Changes not fully reversible within 21 days

Other effects:

It was not possible to assess the effects of the test substance on the iris due to the opacity of the cornea.

Any other information on results incl. tables

Table 1: Acute eye irritation: Results.

Animal Nº.	REGION OF EYE		1 h	24 h	48 h	72 h	7d	14 d	21 d
1	Cornea	Density	3	4	4	4	4	3	4
	Iris		0	-	-	-	-	-	-
	Conjunctiva	Hyperaemia	2	*	*	*	3	2	2
		Edema	4	4	4	4	4	3	3
2	Cornea	Density	3	4	4	4	4	4	4
	Iris		0	-	-	-	-	-	-
	Conjunctiva	Hyperaemia	2	*	3	3	3	2	2
		Edema	4	4	4	4	4	4	3
3	Cornea	Density	3	4	4	4	4	4	4
	Iris		0	-	-	-	-	-	-
	Conjunctiva	Hyperaemia	2	*	*	3	3	2	2
		Edema	4	4	4	3	3	2	2

-: Not possible (corneal opacity)

*: Not possible (white corneal opacity and suppuration)

Applicant's summary and conclusion

Interpretation of results:

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

Remarks:

CLP Implementation.

Conclusions:

The scope of this experimental study was to evaluate the degree of eye irritation or corrosion caused by Nα-Lauroyl-L-arginine ethyl ester monohydrochloride (LAE) after a single application to the surface of 3 rabbit's eye. Observations to obtain current results were recorded until 21 days after administration. Results showed that the substance is causing severe ocular damage and is classified as eye irritating.

Executive summary:

The aim of this Study was to determine the degree of eye irritation or corrosion caused by N^α-Lauroyl-L-arginine ethyl ester monohydrochloride (LAE) after a single application to the surface of the rabbit's eye.

A total of 3 albino rabbits were tested by a single application of 0.1 ml (90% purity of the test substance) to one of the eye, the untreated eye serves as the control. The degree of eye irritation/corrosion was evaluated by scoring lesions of conjunctiva, cornea and iris at specific intervals.

The animal behavior was observed immediately after instillation of the product and then were returned to their cages and checked at regular intervals.The degree of ocular irritation was evaluated in each rabbit at 1, 24, 48 and 72 hours after treatment, following the criteria set out below. Additional observations were made at 7, 14 and 21 days after administration to assess the reversibility or irreversibility of the same.

After assessment and evaluation of the alterations according to the OECD GUIDELINE FOR TESTING OF CHEMICALS, protocol no. 405, of 24^thFebruary 1987 and the EEC Directive of the 31^stMy 1992 (92/69/EEC) Annex, PartB, Method B.5, it is considered to cause irreversible effects on the eye (Category I) according to CLP Regulation.