Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 April 2015 to 14 May 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant, guideline study, available as an unpublished report.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.
Cross-reference
Reason / purpose for cross-reference:
read-across: supporting information
Reference
Endpoint:
eye irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
30 April 2015 to 14 May 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant, guideline study, available as an unpublished report.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF Guidelines (2000)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L'Arbresle Cedex, France
- Age at study initiation: 23 - 24 weeks old
- Weight at study initiation: 3812 - 4172 g
- Housing: Individually housed in labeled cages with perforated floors (dimensions: 67 x 62 x 55 cm, Ebeco, Germany) and shelters (dimensions: 40 x 32 x 23 cm, Ebeco, Germany).
- Diet: Pelleted diet for rabbits (Global Diet 2030, Harlan Tekland, Mucedola, Milanese, Italy), approximately 100 g per day. Hay (Tecnilab-BMI BV, Someren, The Netherlands) and wooden sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study period.
- Water: Tap water, ad libitum.
- Acclimation period: At least 5 days before test start, under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 24 °C
- Humidity: 40 - 70 %
- Air changes: at least 10 air changes/hour
- Photoperiod: 12 hours dark: 12 hours light

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: Animals were treated by instillation of 18.5 mg (range 18.1 - 18.8 mg) of the test substance (a volume of approximately 0.1 mL) in the conjunctival sac of one eye after gently pulling the lower lid away from the eyeball. The second eye remianed untreated and served as the control.

Duration of treatment / exposure:
Single application.
Observation period (in vivo):
Observations of eyes: Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.
Observations were made twice daily for mortality/viability and at least once daily for toxicity. Body weight was measured prior instillation of test item and after the final observation.
Number of animals or in vitro replicates:
3 females. The study was performed in a stepwise manner and was started by treatment of a single rabbit. Two other animals were treated in a similar manner 11 days later, after considering the degree of eye irritation observed in the first animal.
Details on study design:
PREEMPTIVE PAIN MANAGEMENT
One hour prior to instillation of the test substance, buprenorphine (Buprenodale®, Dechra Ltd., Stoke-on-
Trent, United Kingdom) 0.01 mg/kg was administered by subcutaneous injection in order to provide
a therapeutic level of systemic analgesia.

Five minutes prior to instillation of the test substance, two drops of the topical anesthetic alcaine 0.5%
(SA Alcon-Couvreur NV, Puurs, Belgium) were applied to both eyes.

Immediately after fluorescein examination on Day 2, in order to provide a continued level of systemic
analgesia, buprenorphine 0.01 mg/kg and meloxicam (Metacam®, Boehringer Vetmed GmbH,
Ingelheim/Rhein, Germany) 0.5 mg/kg were administered by subcutaneous injection.

REMOVAL OF TEST SUBSTANCE: None.

SCORING SYSTEM: The irritation was assessed according to the numerical scoring system. At each observation, the highest scores given were recorded.

TOOL USED TO ASSESS SCORE: 2 % fluorescein (Merck, Germany) in water (adjusted to pH 7.0)
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritant / corrosive response data:
- Corneal effects: None were noted during the study
- Iridial effects: No iridial irritation observed during the study.
- Conjunctival effects: Irritation of the conjunctivae was observed in each of three rabbits at 1 hour after treatment (redness, chemosis and discharge), with minimal irritation at 48 hours (redness). The irritation had completely resolved within 72 hours in all animals.
Other effects:
No signs of systemic toxicity observed in the animals during the test period and no mortality occured.

Table 1. Individual eye irritation scores

Animal

Time after dosing (h)

Cornea

Iris

Conjunctivae

Comments

Opacity (0-4)

Area (0-4)

Fluor area (%)2

(0-2)

Redness (0-3)

Chemosis (0-4)

Discharge (0-3)

53

1

0

0

0

0

2

1

1

c

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

72

0

0

0

0

0

0

-

2A

1

0

0

0

0

2

1

1

-

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

 

0

0

0

0

0

0

-

61

1

0

0

0

0

2

1

1

-

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

72

0

0

0

0

0

0

-

C – Remnants of the test item on the outside of the eyelids

Table 2. Animal specification

Animal

Sex

Age at start (weeks)

Body weights (g)

 

Prior to application

At termination

53

Female

24

4172

4160

2A

Female

23

3864

3903

61

Female

23

3812

3850

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Dilithium azelate does not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.
Executive summary:

The eye irritation of dilithium azelate was assessed in a GLP-compliant, in vivo eye irritation study following OECD guidelines 405 (WIL Research 2015). A single treatment of dilithium azelate was applied to the non-irrigated eye of three rabbits and observations made at 1, 24, 48 and 72 hours for effects on conjunctivae, iris and cornea and for reversibility of effects.

Instillation of the dilithium azelate resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours in all animals. Based on the results, dilithium azelate does not have to be classified for eye irritation according to GHS and Regulation EC No. 1272/2008.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF Guidelines (2000)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Dilithium nonanedioate
IUPAC Name:
Dilithium nonanedioate
Constituent 2
Chemical structure
Reference substance name:
Dilithium azelate
EC Number:
254-184-4
EC Name:
Dilithium azelate
Cas Number:
38900-29-7
Molecular formula:
C9H14O4.2Li
IUPAC Name:
dilithium azelate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material: Dilithium azelate
- Molecular formula: C9H14O4.2Li
- Molecular weight: 200.1 g/mol
- Physical state: Powder
- pH: 6.3 - 6.4 (in 1 % water, indicative value)
- Analytical purity: 98.33 %
- Lot/batch No.: A058/99
- Expiration date of the lot/batch: 06 March 2018
- Stability under test conditions: No data
- Storage condition of test material: At room temperature protected from light

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L'Arbresle Cedex, France
- Age at study initiation: 23 - 24 weeks old
- Weight at study initiation: 3812 - 4172 g
- Housing: Individually housed in labeled cages with perforated floors (dimensions: 67 x 62 x 55 cm, Ebeco, Germany) and shelters (dimensions: 40 x 32 x 23 cm, Ebeco, Germany).
- Diet: Pelleted diet for rabbits (Global Diet 2030, Harlan Tekland, Mucedola, Milanese, Italy), approximately 100 g per day. Hay (Tecnilab-BMI BV, Someren, The Netherlands) and wooden sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study period.
- Water: Tap water, ad libitum.
- Acclimation period: At least 5 days before test start, under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 24 °C
- Humidity: 40 - 70 %
- Air changes: at least 10 air changes/hour
- Photoperiod: 12 hours dark: 12 hours light

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: Animals were treated by instillation of 18.5 mg (range 18.1 - 18.8 mg) of the test substance (a volume of approximately 0.1 mL) in the conjunctival sac of one eye after gently pulling the lower lid away from the eyeball. The second eye remianed untreated and served as the control.

Duration of treatment / exposure:
Single application.
Observation period (in vivo):
Observations of eyes: Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.
Observations were made twice daily for mortality/viability and at least once daily for toxicity. Body weight was measured prior instillation of test item and after the final observation.
Number of animals or in vitro replicates:
3 females. The study was performed in a stepwise manner and was started by treatment of a single rabbit. Two other animals were treated in a similar manner 11 days later, after considering the degree of eye irritation observed in the first animal.
Details on study design:
PREEMPTIVE PAIN MANAGEMENT
One hour prior to instillation of the test substance, buprenorphine (Buprenodale®, Dechra Ltd., Stoke-on-
Trent, United Kingdom) 0.01 mg/kg was administered by subcutaneous injection in order to provide
a therapeutic level of systemic analgesia.

Five minutes prior to instillation of the test substance, two drops of the topical anesthetic alcaine 0.5%
(SA Alcon-Couvreur NV, Puurs, Belgium) were applied to both eyes.

Immediately after fluorescein examination on Day 2, in order to provide a continued level of systemic
analgesia, buprenorphine 0.01 mg/kg and meloxicam (Metacam®, Boehringer Vetmed GmbH,
Ingelheim/Rhein, Germany) 0.5 mg/kg were administered by subcutaneous injection.

REMOVAL OF TEST SUBSTANCE: None.

SCORING SYSTEM: The irritation was assessed according to the numerical scoring system. At each observation, the highest scores given were recorded.

TOOL USED TO ASSESS SCORE: 2 % fluorescein (Merck, Germany) in water (adjusted to pH 7.0)

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 h
Irritant / corrosive response data:
- Corneal effects: None were noted during the study
- Iridial effects: No iridial irritation observed during the study.
- Conjunctival effects: Irritation of the conjunctivae was observed in each of three rabbits at 1 hour after treatment (redness, chemosis and discharge), with minimal irritation at 48 hours (redness). The irritation had completely resolved within 72 hours in all animals.
Other effects:
No signs of systemic toxicity observed in the animals during the test period and no mortality occured.

Any other information on results incl. tables

Table 1. Individual eye irritation scores

Animal

Time after dosing (h)

Cornea

Iris

Conjunctivae

Comments

Opacity (0-4)

Area (0-4)

Fluor area (%)2

(0-2)

Redness (0-3)

Chemosis (0-4)

Discharge (0-3)

53

1

0

0

0

0

2

1

1

c

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

72

0

0

0

0

0

0

-

2A

1

0

0

0

0

2

1

1

-

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

 

0

0

0

0

0

0

-

61

1

0

0

0

0

2

1

1

-

24

0

0

0

1

0

0

-

48

0

0

0

1

0

0

-

72

0

0

0

0

0

0

-

C – Remnants of the test item on the outside of the eyelids

Table 2. Animal specification

Animal

Sex

Age at start (weeks)

Body weights (g)

 

Prior to application

At termination

53

Female

24

4172

4160

2A

Female

23

3864

3903

61

Female

23

3812

3850

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Dilithium azelate does not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.
Executive summary:

The eye irritation of dilithium azelate was assessed in a GLP-compliant, in vivo eye irritation study following OECD guidelines 405 (WIL Research 2015). A single treatment of dilithium azelate was applied to the non-irrigated eye of three rabbits and observations made at 1, 24, 48 and 72 hours for effects on conjunctivae, iris and cornea and for reversibility of effects.

Instillation of the dilithium azelate resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 72 hours in all animals. Based on the results, dilithium azelate does not have to be classified for eye irritation according to GHS and Regulation EC No. 1272/2008.