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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:
January 15, 2016 - March, 10, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well documented GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
impurity
Type:
impurity
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Dabquel Complex CaL
- Substance type: metal complex
- Physical state: brown liquid
- Composition of test material, percentage of components:
Calcium complexed content: 6.10%
Total Calcium content: 6.11%
- Lot/batch No.: 4140H1112
- Expiration date of the lot/batch: September 25, 2018
- Storage condition of test material:Keep container tightly closed in a dry, cool and well ventilated place
Storage Temperature: Room temperature
Storage Container: Keep container tightly closed in a dry, cool and well ventilated place
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Dabeer S.A. (Batch/Lot No. 4140Z0925)
- Expiration date of the lot/batch: September, 25, 2018
- Purity test date: 2015-11-16


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in a cool well ventilated place
- Stability under test conditions: stable
- pH: 6.77

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation
- Age at study initiation: 16 to 17 weeks
- Weight at study initiation: Minimum: 2.561, Maximum: 2.741
- Housing: individually in stainless steel wire meshed cages
- Diet (e.g. ad libitum): Teklad certified Global High Fiber Rabbit pellet Feed manufactured by Harlan, USA ad libitum
- Water (e.g. ad libitum): UV sterilised drinking water filtered through Kent Reverse Osmosis water filtration system ad libitum
- Acclimation period: 8 - 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23 °C
- Humidity (%): 64 to 65%
- Air changes (per hr): Minimum 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 h artificial light and 12 h darkness, light hours being 06:00 - 18:00 h except light was kept ON at the time of the subcutaneous injection during night hours. Photoperiod was maintained through automatic timer.

Animal Identification
Each rabbit was serially numbered on the ear using a tattoo machine on day 1 of acclimatisation. Appropriate labels were attached to the cages indicating the study number, test item code, sex, dose, type of study, cage number and animal number.

Test system

Vehicle:
other: distilled water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 1 % solution in distilled water

VEHICLE
- Amount(s) applied (volume or weight with unit): distilled water, test item solved in it
- Concentration (if solution): 1 %
Duration of treatment / exposure:
8 hours
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 0.9 % normal saline
- Time after start of exposure: 24 after treatment

SCORING SYSTEM: according to OECD 405

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 1 hour
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 1 hour
Irritant / corrosive response data:
Conjunctival redness was evident at 1 h in all three rabbits which resolved by 24 h post test item application (TIA). Conjunctival chemosis was evident at 1 h in rabbit N° 1 which was resolved by 24 h post TIA. The individual animal mean eye irritation scores of the 24, 48 and 72 h post TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively.

Any other information on results incl. tables

Mean Eye Irritation Scores

The individual animal mean eye irritation scores observed for corneal opacity (0.00), iritis (0.00), conjunctival redness (0.00) and conjunctival chemosis (0.00) following grading at 24, 48 and 72 h post TIA.

 

Narrative Description of Eye Irritation

 

Treated Eye

At 1 h post TIA, the treated eyes revealed conjunctival redness [some blood vessels definitely hyperaemic (injected) in all three rabbits; score of 1] and conjunctival chemosis [some swelling above normal (includes nictitating membranes) in rabbit N° 1; score of 1].

At 24 h post TIA, the treated eyes of all three rabbits recovered completely and appeared normal throughout the experimental period.

Examination with fluorescein dye and cobalt blue filter [corneal epithelium damage showing as green fluorescein staining] revealed no (area) corneal epithelium damage in all three rabbits at 24 h post TIA.

Corneal opacity and Iritis were not observed in all three rabbits throughout the experimental period.

 

Control Eye

No abnormalities were detected in the control eye of each rabbit during the course of this study (Table 3). No corneal epithelium damage was observed during the examination with fluorescein dye and cobalt blue filter.

 

Clinical Observations other than Eye Irritation

Other than eye irritation, no signs of systemic toxicity including clinical observation and body weight were observed in the rabbits throughout the experimental period.

 

Interpretation of Results

Conjunctival redness was evident at 1 h in all three rabbits which resolved by 24 h post TIA. Conjunctival chemosis was evident at 1 h in rabbit N° 1 which was resolved by 24 h post TIA. The individual animal mean eye irritation scores of the 24, 48 and 72 h post TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively.

 

Conclusion

Based on the results of this study, an indication of the classification for DABQUEL COMPLEX CaL is as follows:

Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

TABLE1: Mean Eye Irritation Scores

Sex: Female

Rabbit N°

Mean Score at 24, 48 and 72 Hours

Opacity: Degree of Density

Iris Lesion

Conjunctivae

Redness

Chemosis

1

0.00

0.00

0.00

0.00

2

0.00

0.00

0.00

0.00

3

0.00

0.00

0.00

0.00

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

Table 2: Clinical Observations (Non Ocular) and Body Weight (kg) of Individual Rabbit 

A. Clinical Observation (Non Ocular)                                                                         

Sex:Female

Rabbit

Observations made on Day

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key:  0 = Day of treatment

          1 = Normal

B. Body Weight Record

Rabbit N°

Body Weight (kg)

Initial (Day 0)

Termination (72 h)

1

2.561

2.598

2

2.720

2.939

3

2.741

2.813

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits 

Table 3 :Individual Scores of Eye Reactions Post Application 

Control Eye                                                                                                                   

Sex: Female

Rabbit N°

1

2

3

Site of Application

Left

Left

Left

Reaction

Hour

Hour

Hour

1

24

48

72

1

24

48

72

1

24

48

72

Opacity:

Degree of Density

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctivae

(Redness)

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctivae

(Chemosis)

0

0

0

0

0

0

0

0

0

0

0

0

 Treated Eye

Rabbit N°

1

2

3

Site of Application

Right

Right

Right

Reaction

Hour

Hour

Hour

1

24

48

72

1

24

48

72

1

24

48

72

Opacity:

Degree of Density

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctivae

(Redness)

1

0

0

0

1

0

0

0

1

0

0

0

Conjunctivae

(Chemosis)

1

0

0

0

0

0

0

0

0

0

0

0

Acute Eye Irritation Study ofDABQUEL COMPLEX CaLin Rabbits

Table 4: Individual Corneal Observations 24 h Post Application using Fluorescein Dye Staining 

Control Eye                                                                                                                   

Sex: Female

Rabbit N°

Control Eye

Fluorescein Staining Response

Details of Corneal Damage

Observed in Control Eye

1

Left

Negative

No corneal epithelium damage was observed.

2

Left

Negative

No corneal epithelium damage was observed.

3

Left

Negative

No corneal epithelium damage was observed.

Treated Eye

Rabbit N°

Treated Eye

Fluorescein Staining Response

Details of Corneal Damage

Observed in Treated Eye

1

Right

Negative

No corneal epithelium damage was observed.

2

Right

Negative

No corneal epithelium damage was observed.

3

Right

Negative

No corneal epithelium damage was observed.

Acute Eye Irritation Study of DABQUEL COMPLEX CaL in Rabbits

 Table 5:Details of Injections and Applications 

Rabbit

On

Day

Administration of Systemic Analgesics

(Subcutaneous Injection)

Administration of Topical Anaesthetic

(1 to 2 drops)0.5% Proparacaine Hydrochloride

Time of Test Item

Application (0.1 mL)

Buprenorphine hydrochloride

(0.01 mg/kg body weight)

Meloxicam

(0.5 mg/kg body weight)

1

0

11:30 am and 8:50 pm

8:50 pm

12:25 pm

12:30 pm

1

8:42 am

-

-

-

2 and 3

0

11:47 am to 11:48 am and

8:49 pm to 8:50 pm

8:49 pm

to

8:50 pm

12:42 pm

to

12:43 pm

12:47 pm

to

12:48 pm

1

8:53 am to 8:54 am

-

-

-

Key : - = Not applicable

Note: Day 0 = Day of treatment

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Examination with fluorescein dye and cobalt blue filter at 24 h post test item application revealed no corneal epithelium damage in all rabbits. Based on the results of this study, an indication of the classification for DABQUEL COMPLEX CaL is as follows:
Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant
Executive summary:

In an acute eye irritation study, 3 adult male New Zealand White rabbits were given a single ocular application of 0.1 mL DABQUEL COMPLEX CaL in the right eye of the rabbit while the contralateral eye remained untreated and served as the control. Initially one rabbit was tested. Based on the results obtained at 24 h post Test Item Application (TIA), the irritation response was confirmed by testing two additional rabbits simultaneously. Observations were made at 1, 24, 48 and 72 h post TIA. General health status was also checked.

Conjunctival effects were evident at 1 h in all three rabbits which resolved by 24 h post TIA. The animal mean eye irritation scores of the 24, 48 and 72 h post TIA observation were 0.00, 0.00, 0.00 for corneal opacity, 0.00, 0.00, 0.00 for iris effects, 0.00, 0.00, 0.00 for conjunctival redness and 0.00, 0.00, 0.00 for conjunctival chemosis for rabbit N° 1, 2 and 3, respectively.

Examination with fluorescein dye and cobalt blue filter at 24 h post TIA revealed no corneal epithelium damage in all rabbits.

The control eye did not show any abnormal reaction during the study. Moreover, there were no signs of systemic toxicity in any animal observed.

Based on the results of this study, an indication of the classification for DABQUEL COMPLEX CaL is as follows:

Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2015): Not classified as an eye irritant