Reaction mass of l-Glutamic acid, N-coco acyl derivs., disodium salts and l-Glutamic acid, N-coco acyl derivs., monosodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 December 2011 to 03 January 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of l-Glutamic acid, N-coco acyl derivs., disodium salts and l-Glutamic acid, N-coco acyl derivs., monosodium salts

EC Number:

916-533-5

Molecular formula:

R-C6H6NO5Na2, R-C6H7NO5Na R: Coco fatty acid residue

IUPAC Name:

Reaction mass of l-Glutamic acid, N-coco acyl derivs., disodium salts and l-Glutamic acid, N-coco acyl derivs., monosodium salts

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Physical state: White to pale yellow powder with a slightly characteristic odour.
- Storage condition of test material: room temperature (15-25 ºC) in the dark.
- pH: 6.5, determined by measurement of the supernatant from a 1% w/v mixture in distilled water.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Sex: Males
- Age at study initiation: Approximately 16 weeks old.
- Weight at study initiation: Ranged from 3615 to 3904 g
- Housing: Individually in metal wire cages.
- Diet: Commercial rabbit feed, ad libitum.
- Water: Municipal tap water, as libitum.
- Acclimation period: 41 days.
- Health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined prior to starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 °C.
- Humidity (%): 28 to 67 %
- Air changes (per hr): 15 to 20 air charges per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light per day, from 06:00 to 18:00 hours.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Duration of treatment / exposure:

Single dose

Observation period (in vivo):

14 day

Number of animals or in vitro replicates:

Three

Details on study design:

APPLICATION OF THE TEST MATERIAL
- An initial test was performed using one animal. The test material was instilled into the conjunctival sac of the left eye. The eyelids were held closed for several seconds to prevent the loss of the test material. The contralateral eye served as the control. Immediately after the administration of the test material, an assessment of the initial pain reaction was made according to the six point scale shown in Table 1.
After consideration of the ocular responses produced in the first animal, one hour after the treatment of the first animal, two additional animals were treated.
All animals were sacrificed at the end of the observation period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Washing was performed on test animals.
- Time after start of exposure: 1 hour.

OBSERVATIONS
- The eyes were examined at 1, 24, 48, 72 hours and 1 and 2 weeks after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes.
- Any clinical signs of toxicity or signs of ill health during the study were recorded.
- Individual body weights were measured at the beginning and end of the observation period.

SCORING SYSTEM: The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (24 April 2002) shown in Table 2.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #1

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 3

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #3

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: Mean at 24, 48 and 72 hours

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: Mean at 24, 48 and 72 hours

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: Mean at 24, 48 and 72 hours

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

- Initial pain reaction: Slight initial pain was observed in all three animals, assigned a score of 2 (see Table 1).
- Control eye: The control eye of each animal was symptom-free.
- One hour after the application: conjunctival redness (score 2), discharge and chemosis (score 3) were seen in all rabbits. All animals showed corneal opacity (score 1, areas 2, 3 or 4).
- 24 hours after treatment: conjunctival redness (score 2), discharge (score 2 or 3) and chemosis (score 2) were seen in all rabbits. All animals showed corneal opacity (score 1 or 2, areas 1, 3 or 4).
- 48 hours after treatment, conjunctival redness (score 1 or 2), discharge (score 1 or 2) and chemosis (score 1) were seen in all rabbits. All animals showed corneal opacity (score 1, areas 1 or 3).
- 72 hours after treatment: conjunctival redness (score 1) was seen in all rabbits. Conjuctival discharge (score 1) and chemosis (score 1) were seen in two animals. All animals showed corneal opacity (score 1, areas 1 or 2).
- One week after treatment: conjunctival redness (score 1) was seen in two rabbits.
- Two weeks after treatment: no signs of eye irritation or other clinical signs were observed. The study was terminated after the 2-week observation as effects were fully reversible by this time.

Other effects:

- Mortality: None observed during the study.
- Bodyweights: The body weight and body weight change were considered to be normal with no indication of treatment related effect. Final weights ranged from 3799 to 4088 g, representing a weight gain of between 181 and 187 g.
- Clinical observations: There were no clinical signs observed that could be related to treatment. The general state and behaviour of animals were normal throughout the study period.

Any other information on results incl. tables

Table 3: Individual Scores for Ocular Irritation at 1 hour

Animal No.	Score of Irritation								IPR
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	2	3	3	1	4	0	0	-	2
2	2	3	3	1	3	0	0	-	2
3	2	3	3	1	2	0	0	-	2

 

Table 4: Individual Scores for Ocular Irritation at 24 hours

Animal No.	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	2	2	2	1	4	0	0	-
2	2	2	2	1	3	0	0	-
3	2	2	3	2	1	0	0	-

 

Table 5: Individual Scores for Ocular Irritation at 48 hours

Animal No.	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	2	1	1	1	3	0	0	-
2	1	1	2	1	3	0	0	-
3	1	1	2	1	1	0	0	-

 

Table 6: Individual Scores for Ocular Irritation at 72 hours

Animal No.	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	1	1	1	1	2	0	0	-
2	1	1	0	1	2	0	0	-
3	1	0	1	1	1	0	0	-

 

Table 7: Individual Scores for Ocular Irritation at 1 week

Animal No.	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	1	0	0	0	0	0	0	-
2	1	0	0	0	0	0	0	-
3	0	0	0	0	0	0	0	-

 

Table 8: Individual Scores for Ocular Irritation at 2 weeks

Animal No.	Score of Irritation
	Conjunctivae			Opacity of Cornea		Iris	Control Eye	Other Signs
	R	CH	D	OD	OE	R
1	0	0	0	0	0	0	0	-
2	0	0	0	0	0	0	0	-
3	0	0	0	0	0	0	0	-

R = redness, CH = chemosis; D = discharge; OD = opacity degree of density; OE = extent of opaque area; IPR = initial pain reaction.

Applicant's summary and conclusion

Interpretation of results:

other: Category 2

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the test material was determined to be irritating to the eye based on corneal opacity. Application of the test material to the rabbit’s eye caused significant conjunctival and corneal effects at one hour, which were reduced at 1 week after application. All effects were fully reversible within 14 days.

Executive summary:

The potential for the test material to cause eye irritation was assessed in an in vivo irritation study in rabbits. The study was performed under GLP conditions in line with standardised guidelines OECD 405, EU Method B.5 and EPA OPPTS 870.2400. The test material was administered to three male New Zealand white rabbits, in a single application which was washed after 1 hour. Rabbits were assessed over a 14 day observation period for signs of eye irritation, clinical sign of toxicity, mortality and bodyweight gain. Application of the test material to the rabbit’s eye caused significant conjunctival and corneal effects at one hour, which were reduced 1 week after application. The study was terminated after 14 day as all signs of irritation were fully reversed. Under the conditions of the study, the test material was determined to be an eye irritant (category 2).