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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP and valid methods and is considered relevant and reliable for classification.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Test animals / tissue source

Species:
rabbit
Strain:
Himalayan
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: LPT Laboratory of Pharmacology and Toxicology GmbH & Co. KG, branch Löhndorf, 24601 Löhndorf/¬Post Wankendorf Germany
- Age at dosing: Approx. 8 months
- Weight at dosing: Animal no.1: 3.11 kg; Animal no. 2: 2.83 kg; Animal no. 3: 2.76 kg
- Fasting period before study: Not provided
- Housing: For 8 hours following test item application, the animals were kept singly in restrainers which allowed free movement of the head but prevented a complete body turn, wiping of the eyes with the paws and excluded irritation of the eye by excrements and urine.
During the acclimatisation period and after the 8-hour period in restrainers, the animals were kept singly in cages with dimensions of 380 mm x 425 mm x 600 mm (manufacturer: Dipl. Ing. W. EHRET GmbH, 16352 Schönwalde, Germany)
- Diet (e.g. ad libitum): Commercial diet, ssniff® K-H V2333 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany), served as food. The food was available ad libitum before and after the exposure period.
- Water (e.g. ad libitum): Tap water was offered ad libitum before and after the exposure period.
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C ± 3°C (maximum range)
- Humidity (%): 30% - 70% (maximum range)
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: March 22, 2013 To: April 2, 2013

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The left eye, which remained untreated, served as a control.
Amount / concentration applied:
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test item were administered into one eye each of three animals.
Duration of treatment / exposure:
Single instillation into the conjunctival sac

Observation period (in vivo):
5 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No.
The test item was placed into the conjunctival sac of the right 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 in order to prevent loss of the material.

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 areas, 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, iris reactive to light (a sluggish reac¬tion is considered to be an effect): 1
haemorrhage, gross destruction, or no reaction to light: 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
CHEMOSIS
Swelling/ refers to lids and/or nictitating membranes
normal: 1
some swelling above normal: 1
obvious swelling with partial eversion of lid: 2
swelling with lids about half-closed: 3
swelling with lids more than half-closed: 4

TOOL USED TO ASSESS SCORE: hand-slit lamp/ fluorescein
The eyes were examined ophthalmoscopically with a slit lamp prior to the administration and 1, 24, 48, 72 hours, 4 to 5 days after the administration. The eye reactions were observed and registered.
24 hours after administration, fluorescein was applied to the eyes before being examined to aid evaluation of the cornea for possible lesions.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24. 48. 72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24. 48. 72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
other: 24. 48. 72 h
Score:
1.11
Max. score:
3
Reversibility:
fully reversible within: 5 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: 24. 48. 72 h
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Time point:
other: 24. 48. 72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 4 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: 24. 48. 72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 5 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24. 48. 72 h
Score:
0.45
Max. score:
4
Reversibility:
fully reversible within: 4 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24. 48. 72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 24. 48. 72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 24. 48. 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 4 days
Irritant / corrosive response data:
Cornea opacity (grade 1) was observed in all animals 60 minutes after instillation.
Conjunctival redness was observed in all animals:
- animal no. 1: 60 minutes to 48 hours (grade 1) after instillation;
- animal no. 2: 60 minutes and 24 hours (grade 2), 48 hours and 72 hours (grade 1) after instillation;
- animal no. 3: 60 minutes and 24 hours (grade 2), 48 hours to 4 days (grade 1) after instillation.
Chemosis was observed in all animals 1 hour, in animal no. 2 until 24 hours and in animal no. 3 until 72 hours after instillation.
The fluorescein tests performed 24 hours after instillation revealed corneal staining in animal no. 2 (1/2 to 3/4 of the surface) and in animal no. 3 (up to 1/4 of the surface).
The irises were not affected by instillation of the test item.

Any other information on results incl. tables

Table 1. Examination of the treated eye (Animal no. 1/2/3)

Time after administration

Cornea

opacity

Iris

Conjunctivae

Redness

Chemosis

Before dosing

0/0/0

0/0/0

0/0/0

0/0/0

60 minutes

1/1/1

0/0/0

1/2/2

1/1/1

24 hours

0/0/0

0/0/0

1/2/2

0/1/1

48 hours

0/0/0

0/0/0

1/1/1

0/0/1

72 hours

0/0/0

0/0/0

0/1/1

0/0/1

4 days

-/-/0

-/-/0

-/-/1

-/-/0

5 days

-/-/0

-/-/0

-/-/0

-/-/0

24 hours fluorescein test:

animal no. 1: no pathological findings
animal no. 2: corneal staining (1/2 to 3/4 of the surface)
animal no. 3: corneal staining (up to 1/4 of the surface)

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the EC-Commission directive 67/548/EEC and its subsequent amendments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions, Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts is non-irritating to eyes, hence, no labelling is required.
Also, according to the EC-Regulation 1272/2008 and subsequent regulations, the test item is non-irritating to eyes; classification and labelling of the substance is not necessary.
Executive summary:

Under the present test conditions, a single instillation of 0.1 mL Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts per animal into the conjunctival sac of the right eye of three rabbits caused the following changes:

Cornea opacity (grade 1) was observed in all animals 60 minutes after instillation.

Conjunctival redness was observed in all animals:

- animal no. 1: 60 minutes to 48 hours (grade 1) after instillation;

- animal no. 2: 60 minutes and 24 hours (grade 2), 48 hours and 72 hours (grade 1) after instillation;

- animal no. 3: 60 minutes and 24 hours (grade 2), 48 hours to 4 days (grade 1) after instillation.

Chemosis was observed in all animals 1 hour, in animal no. 2 until 24 hours and in animal no. 3 until 72 hours after instillation.

The fluorescein tests performed 24 hours after instillation revealed corneal staining in animal no. 2 (1/2 to 3/4 of the surface) and in animal no. 3 (up to 1/4 of the surface).

The irises were not affected by instillation of the test item.

All lesions observed had disappeared at 72 hours or 5 days after instillation.

There were no systemic intolerance reactions concerning behaviour, body weight and food consumption.