[D-gluco-heptonato-κO1,κO2,κO3,κO6,κO7]calcium(II) nitrate

Eye irritation

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

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

Results

Results 1

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 1 hour

Results 5

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 N°	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 N°	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