Pentanediamide, N1,N5-dibutyl-2-[(2-ethyl-1-oxohexyl)amino]-, (2S)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 March 2001 - 26 March 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(2S)-N,N'-dibutyl-2-(2-ethylhexanamido)pentanediamide

EC Number:

700-664-8

Cas Number:

486455-65-6

Molecular formula:

C21H41N3O3

IUPAC Name:

(2S)-N,N'-dibutyl-2-(2-ethylhexanamido)pentanediamide

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 Chalaronne / France
- Age at study initiation: 10-11 weeks
- Weight at study initiation: 2.4-2.5 kg
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with autoclaved wood (RCC Ltd, Fullinsdorf) and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 33/00) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Haysticks (QS no. 11/01) provided by Eberle Nafag AG, CH-9200 Gossau
- Water: Community tap water from Fullinsdorf, ad libitum.
- Acclimation: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Air-conditioned with target ranges for room temperature 20 ± 3 °C
- Humidity (%): relative humidity 30-70 %
- Air changes (per hr): approximately 10-14 air changes per hour
- Photoperiod (hrs dark / hrs light): The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the light period.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:

On the day of treatment, the test material was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test item.

Observation period (in vivo):

1, 24, 48 and 72 hours, as well as 7 days after application.

Number of animals or in vitro replicates:

3 (1 male and 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eyes were not rinsed after instillation.

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

TOOL USED TO ASSESS SCORE: Eye examinations were made with a Varta Cliptrix diagnostic-lamp (A. Riegger, Basel / Switzerland).

To evaluate the irritation potential of the test item (EEC Commission Directive 93/21/EEC, April 27, 1993), the mean values were calculated for each individual, using the scores between 24 and 72 hours.
The primary irritation score was calculated by totaling the mean cumulative scores at 24, 48 and 72 hours and then divided by the number of data points.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

24 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

48 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within:

Remarks:

7 days

Results 5

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within:

Remarks:

7 days

Results 6

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Application of the test item to healthy rabbit conjunctivae resulted in a primary irritation score of 0.56. The eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.56 redness of the conjunctivae and grade 0.00 chemosis of the conjunctivae.

No abnormal findings were observed in the cornea or the iris in any animal at any time point. Slight reddening of the conjunctivae was noted in two animals from 1 to 24 hours after treatment. Slight to moderate reddening was also observed in the other animal from 1 to 72 hours after treatment. Slight swelling of the conjunctivae was evident in all animals at the 1-hour reading. Slight reddening of the sclera was observed in all animals at the 1-hour reading and in one female this persisted up to 24 hours after treatment. A slight watery discharge was present in one female at the 1-hour reading.

All eye reactions had cleared within 7 days after treatment. No staining of the treated eyes by the test item was observed. White remnants of the test item were evident in the eye or conjunctival sac of all animals 1 hour after treatment. No corrosion was observed at any of the measuring intervals.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

Any other information on results incl. tables

BODY WEIGHTS

The body weights of all rabbits were considered to be within the normal range of variability.

Body weight in grams
Animal No.	Sex	First Day of Acclimatization	Day of Treatment	Last Day of Observation
88	male	2187	2468	2670
89	female	2063	2429	2648
90	female	2112	2423	2720

TABLE 1: EYE IRRITATION SCORES - INDIVIDUAL VALUES

Animal No.	Sex	Evaluation Interval	Corneal Opacity	Iris	Conjunctivae		Cumulative		Sclera
					Redness	Chemosis	Score	Mean
88	M	1 hour	0	0	1	1	2	2.33	1
89	F		0	0	1	1	2		1
90	F		0	0	2	1	3		1
88	M	24 hours	0	0	1	0	1	1	0
89	F		0	0	1	0	1		0
90	F		0	0	1	0	1		1
88	M	48 hours	0	0	0	0	0	0.33	0
89	F		0	0	0	0	0		0
90	F		0	0	1	0	1		0
88	M	72 hours	0	0	0	0	0	0.33	0
89	F		0	0	0	0	0		0
90	F		0	0	1	0	1		0
88	M	7 days	0	0	0	0	0	0	0
89	F		0	0	0	0	0		0
90	F		0	0	0	0	0		0

TABLE 2: EYE IRRITATION SCORES - MEAN VALUES AFTER 24, 48 AND 72 HOURS

Animal No.	Sex	Corneal Opacity	N	Iris	N	Conjunctivae				Primary Eye Irritation Score
						Redness	N	Chemosis	N
88	M	0	3	0	3	0.33	3	0	3	0.56
89	F	0	3	0	3	0.33	3	0	3
90	F	0	3	0	3	1	3	0	3
Mean score		0		0		0.56		0

N = number of available data points

TABLE 3: EYE IRRITATION SCORES - ASSESSMENT ACCORDING TO EEC GUIDELINES

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

Applicant's summary and conclusion

Interpretation of results:

other: Not classified according to EU criteria

Conclusions:

According to Regulation (EC) No 1272/2008, the test material does not require classification as irritating to the eye.

Executive summary:

The primary eye irritation potential of the test material was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7 days after test item application. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totaling the individual cumulative scores at 24, 48 and 72 hours and then dividing the resulting total by the number of data points. The primary irritation score was 0.56 (max. 13).

The eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.56 redness of the conjunctivae and grade 0.00 chemosis of the conjunctivae.

No abnormal findings were observed in the cornea or the iris in any animal at any reading.

Slight reddening of the conjunctivae was noted in two animals from 1 to 24 hours after treatment. Slight to moderate reddening was also observed in the other animal from 1 to 72 hours after treatment. Slight swelling of the conjunctivae was evident in all animals at the 1-hour reading. Slight reddening of the sclera was observed in all animals at the 1-hour reading and in one female this persisted up to 24 hours after treatment.

A slight watery discharge was present in one female at the 1-hour reading.

White remnants of the test item were evident in the eye or conjunctival sac of all animals 1 hour after treatment.

No staining of the treated eyes by the test item was observed. No corrosion was observed at any of the measuring intervals. All eye reactions had cleared within 7 days after treatment.

According to Regulation (EC) No 1272/2008, the test material does not require classification as irritating to the eye.