L-Arginine, N2-(2,3-dihydroxypropyl)-, monohydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

01 April 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Detail on test item is not clear

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: The Hen's Egg Test- Utilizing the Chorioallantonic Membrane (HET-CAM)

Version / remarks:

Kemper & Luepke

Deviations:

not specified

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

L-Arginine, N2-(2,3-dihydroxypropyl)-, monohydrochloride

EC Number:

843-143-1

Cas Number:

709647-81-4

Molecular formula:

C9H21ClN4O4

IUPAC Name:

L-Arginine, N2-(2,3-dihydroxypropyl)-, monohydrochloride

Test material form:

liquid

Remarks:

Test item 15 % in distilled water

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL: Not specified
- Source and lot/batch No. of test material:
- Expiration date of the lot/batch:
- Purity test date:

RADIOLABELLING INFORMATION (if applicable): N/A
- Radiochemical purity:
- Specific activity:
- Locations of the label:
- Expiration date of radiochemical substance:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL: Not specified
- Storage condition of test material:
- Stability under test conditions:
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING: Applied diluted
- Treatment of test material prior to testing: diluted to 15% in distilled water
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material):N/A

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): N/A

OTHER SPECIFICS: N/A
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is

Test animals / tissue source

Species:

chicken

Strain:

other: White Leghorn

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EGGS
- Source: Moyer’s Chicks, Inc. Quakertown Pennsylvania
- Number of eggs: Not specified
- Characteristics of donor animals (e.g. age, sex, weight): Not specified
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Fresh eggs stored in test facility up 7 days at 500 F, the eggs were incubated in a Kuhl incubator which rotated automatically once every hour. Incubation temperature was controlled at 990 F (+/- 10 F) with relative humidity of 60 – 70% for the 10 days of incubation. On day 8, the eggs were turned so that the acutely angled end faced down. On day 10 the eggs were removed from the incubator and placed in preheated and humidified Plexigals work encloser.

Test system

Vehicle:

water

Remarks:

Distilled water

Controls:

other: Nivea moisturing lotion 50% and vaseline dermatology formula lotion 50%

Amount / concentration applied:

0.3 ml

Duration of treatment / exposure:

20 seconds

Observation period (in vivo):

30 seconds, 2 and 5 minutes

Duration of post- treatment incubation (in vitro):

Not applicable

Number of animals or in vitro replicates:

4 CAMs

Details on study design:

Fresh eggs stored in test facility up 7 days at 500 F, the eggs were incubated in a Kuhl incubator which rotated automatically once every hour. Incubation temperature was controlled at 990 F (+/- 10 F) with relative humidity of 60 – 70 % for the 10 days of incubation. On day 8, the eggs were turned so that the acutely angled end faced down. On day 10 the eggs were removed from the incubator and placed in preheated and humidified Plexigals work encloser. Forceps were then used to remove the shell down to the shell-membrane junction. The inner egg membrane was then hydrated with a warm, physiological saline solution. The saline was removed after 2 – 5 minutes exposure. Utilizing point forceps, the inner egg membrane was then carefully removed to reveal the CAM.
The test or reference article, at a dosage of three-tenth of one millilitre (0.3ml) of liquid was administered to each of four CAM’s. Twenty second later, the test or control article was rinsed from each CAM with 5 ml of physiological saline. All CAMs were observed immediately prior to test article administration, at 30 seconds, 2 and 5 minutes after exposure to the test article. The reaction of the CAM, the blood vessels, including the capillaries, and the albumin were examined and score for initiation effects as detailed below.
Effects score
Time (min) 0.5 2 5
Hyperemia 5 3 1
Minimal Haemorrhage (‘Feathering) 7 5 3
Haemorrhage (Obvious Leakage) 9 7 5
Coagulation and/or Thrombosis 11 9 7
The numerical, time dependant scores were tolerated for each CAM. Each reaction type can be recorded only once for each CAM, therefore the maximum score per CAM is 32. The mean score was determined for all CAM’s similarly tested.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

1.75

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

not specified

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The reference substances have historically been categorised as being practically non-irritating, eliciting score approaching 0, at 24 hours, when dosed at 100% and tested using Draize occular irritation methodologies (Draize Scale: 0 - 110). At 50%, the score of 1.00 and 2.00 all within the acceptability range.

Any other information on results incl. tables

Table 3. Experimental scoring results for irritation

Test Group	CAM #	Score@
		0.5 min.	2 min.	5 min.	Total
N-Glyceryl Arginine HCL (15% in distilled water)	1	0	0	0	0
	2	0	0	1	1
	3	0	3	0	3
	4	0	3	0	3 Average: 1.75
Nivea Moisturizing Lotion (50%)	1	0	0	1	1
	2	0	0	1	1
	3	0	0	1	1
	4	0	0	1	1 Average: 1.00
Vaseline Dermatology Formula Lotion (50%)	1	0	0	1	1
	2	0	0	1	1
	3	5	0	0	5
	4	0	0	1	1 Average: 2.00

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the study, the test article was not considered an occular irritant

Executive summary:

The test or reference article, at a dosage of three-tenth of one millilitre (0.3ml) of liquid was administered to each of four CAM’s. Twenty seconds later, the test or control article was rinsed from each CAM with 5 ml of physiological saline.  All CAMs were observed immediately prior to test article administration, at 30 seconds, 2 and 5 minutes after exposure to the test article. The reaction of the CAM, the blood vessels, including the capillaries, and the albumin were examined and score for initiation effects.

 

The numerical, time dependant scores were tolerated for each CAM. Each reaction type can be recorded only once for each CAM, therefore the maximum score per CAM is 32. The mean score was determined for all CAM’s similarly tested.

The reference substances have historically been categorised as being practically non-irritating, eliciting score approaching 0, at 24 hours, when dosed at 100% and tested using Draized ocular irritation methodologies (Draize Scale: 0 - 110).  at 50%, the score of 1.00 and 2.00 all within the acceptability range.

 

The average score for the test article was 1.75, falling within the classification for no irritation. Under the condition of the study, the results indicate that N-Glyceryl Arginine HCl, at 30%, would have practically no occular irritation potential in vivo.