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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
The objective of the study was to assess the irritant and/or corrosive effects of “Sodium Salicylate (CAS: 54-21-7)” on eye, when exposed by the ocular route in rabbits.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Sodium salicylate
- Molecular Formula: C7H6O3.Na
- Molecular Weight: 160.105 g/mol
- Smiles Notation: c1(c(cccc1)O)C(=O)[O-].[Na+]
- InChI: 1S/C7H6O3.Na/c8-6-4-2-1-3-5(6)7(9)10;/h1-4,8H,(H,9,10);/q;+1/p-1
- Substance Type: Organic
- Physical State: Solid
Specific details on test material used for the study:
- Name of test material (as cited in study report): Sodium salicylate
- Molecular Formula: C7H6O3.Na
- Molecular Weight: 160.105 g/mol
- Smiles Notation: c1(c(cccc1)O)C(=O)[O-].[Na+]
- InChI: 1S/C7H6O3.Na/c8-6-4-2-1-3-5(6)7(9)10;/h1-4,8H,(H,9,10);/q;+1/p-1
- Substance Type: Organic
- Physical State: Solid

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: LIVEON BIOLABS PVT. LTD
- Age at study initiation: 3 to 4 Months (Approximately)
- Weight at study initiation: Minimum: 2.120 kg and Maximum: 2.448 kg (Prior to Treatment)
- Housing: The animals were housed individually in stainless steel cages.
- Room Sanitation : The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle : All the cages and water bottles were changed minimum twice a week
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rabbit diet
- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum
- Acclimation period: Rabbits were acclimatised to the test conditions for a period of 7 days (Animal No.-1) and 9 days (Animal No. 2 and 3) prior to the application of the test item

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 20.30 °C Maximum: 22.70 °C
- Humidity (%): Minimum: 48.30 % Maximum: 68.30 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg



Duration of treatment / exposure:
24 hours
Observation period (in vivo):
All the animals were observed at 1, 24, 48 and 72 hours and on day 7 after instillation of test item.
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
Three female rabbits
Details on study design:
Details on study design
REMOVAL OF TEST SUBSTANCE
- Washing (if done):The treated eye of rabbit was washed with normal saline.
- Time after start of exposure:24 hours
SCORING SYSTEM:Grading of irritation lesions was carried out as per Draize Method
TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescent strips.REMOVAL OF TEST SUBSTANCE

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
Animal 1,2 and 3
Time point:
24/48/72 h
Score:
> 0.67 - <= 1
Max. score:
3
Reversibility:
fully reversible within: 7 days observation period
Remarks on result:
probability of mild irritation
Irritation parameter:
iris score
Basis:
mean
Remarks:
Animal 1,2 and3
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
probability of mild irritation
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal 1,2 and 3
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 7 days observation period
Remarks on result:
probability of mild irritation
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
Animal 1, 2 and 3
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days observation period
Remarks on result:
probability of mild irritation
Irritant / corrosive response data:
The following were observed in treated rabbits.
Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals.
Observation at 24 hours after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in animal no. 1 whereas animal no. 2 and 3 showed no ulceration or opacity; Area of Opacity- One quarter (or less) but not zero in animal no. 1 and Zero in animal no. 2 and 3; Iris: Normal in all the animals. Conjunctivae – Diffuse, crimson color; individual vessels not easily discernible was observed in all animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all animals.
At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 40%, 20%, 20% damage in animal no. 1, 2 and 3 respectively.
Observation at 48 hour after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all animals; Area of Opacity- One quarter (or less) but not zero in all 3 animals; Iris: Normal in all the animals. Conjunctivae – Diffuse, crimson color; individual vessels not easily discernible was observed in all animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all animals.
Observation at 72 hour after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all animals; Area of Opacity- One quarter (or less) but not zero in all 3 animals; Iris: Normal in all the animals. Conjunctivae – Diffuse, crimson color; individual vessels not easily discernible was observed in all animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all animals.
Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all 3 animals; Iris: Normal in all the animals. Conjunctivae – Blood vessels were normal in all 3 animals; Chemosis: No swelling (Normal) was observed in all 3 animals.
The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 1.00, 0.00, 2.00, 1.00; 0.67, 0.00, 2.00, 1.00 and 0.67, 0.00, 2.00, 1.00 respectively
Other effects:
Clinical Observation
No systemic toxicity was observed in treated rabbits during the experimental period

Mortality
No mortality was observed during the observation period


Body weight
All rabbits were weighed on test day 0 (prior to application) and at termination



Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:100 mg (0.1gm) of test item                                              

 Sex:Female

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

Day 7

*

1

 24

48

72

Day 7

*

1

24

48

72

Day 7

Corneal Opacity

0

0

1

1

1

0

0

0

0

1

1

0

0

0

0

1

1

0

Area of Opacity

0

0

1

1

1

0

0

0

0

1

1

0

0

0

0

1

1

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

2

2

2

0

0

1

2

2

2

0

0

1

2

2

2

0

Chemosis

0

1

1

1

1

0

0

1

1

1

1

0

0

1

1

1

1

0

Corneal Damage%

40

20

20

 

Dose:Untreated (Control Eye)                                                                       Sex:Female

 

Animal Numbers

1

2

3

Application Side

Left

Left

Left

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

Day 7

*

1

 24

48

72

Day 7

*

1

24

48

72

Day 7

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

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

0

0

0

0

0

0

Conjunctiva

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

0

0

 

Key:*= Pre-exposure eye examination.

 

 

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Treated eye)

            Animal No.

 Eye Reaction

1

2

3

Cornea

1.00

0.67

0.67

Iris

0.00

0.00

0.00

Conjunctiva

2.00

2.00

2.00

Chemosis

1.00

1.00

1.00

 

 Formula :

 Mean Irritation Score= Sum of the Individual Animal Score for eye reactions at 24, 48 and 72 hours

Number of the Observations (3)

Table 2 : Individual Animal Clinical Signs

 

Sex:Female

Animal No.

Days (Post application observation)

0

1

2

3

4

5

6

7

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

Key:1 = Norma

Individual Animal Body Weight

Sex :Female

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.448

2.612

2

2.120

2.156

3

2.236

2.402

Key:kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 1.00, 0.00, 2.00, 1.00; 0.67, 0.00, 2.00, 1.00 and 0.67, 0.00, 2.00, 1.00 respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within day 7.
Hence, under the experimental test conditions, “Sodium salicylate (CAS No. – 54-21-7) is “Mildly Irritating to eyes” of New Zealand White female rabbit eyes and is being classified "Category 2"as per the CLP
regulation.
Executive summary:

Acute Eye Irritation/Corrosion Study of Sodium Salicylate (CAS: 54 -21 -7) in Rabbits, was performed in a OECD GLP laboratory (Sustainability Support Services (Europe) has letter of access).The study was performed according to OECD 405 Guidelines. 3 female young adult rabbits were used for the study.

Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 100 mgof test itemwas instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hour and on day 7 after test item instillation.Ophthalmoscope was used for scoring of eye lesions.

In the initial test,100 mg of test item (pulverized form)was instilled into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);100 mgof test itemwas instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Ocular lesions were seen in animal no. 2 and 3 at 1, 24, 48 and 72 hour observation which recovered at day 7.

 

Untreated eye of all the three rabbits was normal throughout the experiment.

 

The following grading scores were observed in treated eye of treated rabbits.

Observation at 1 hour after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was observed in all the animals;Chemosis:Some swelling above normal (includes nictitating membranes) was observed in all the animals.

Observation at 24 hours after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in animal no. 1 whereas animal no. 2 and 3 showed no ulceration or opacity; Area of Opacity-One quarter (or less) but not zero in animal no. 1 and Zero in animal no. 2 and 3;Iris:Normal in all the animals.Conjunctivae –Diffuse, crimson color; individual vessels not easily discernible was observed in all animals;Chemosis:Some swelling above normal (includes nictitating membranes) was observed in all animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 40%, 20%, 20% damage in animal no. 1, 2 and 3 respectively.

Observation at 48 hour after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all animals; Area of Opacity-One quarter (or less) but not zero in all 3 animals;Iris:Normal in all the animals.Conjunctivae –Diffuse, crimson color; individual vessels not easily discernible was observed in all animals;Chemosis:Some swelling above normal (includes nictitating membranes) was observed in all animals.

Observation at 72 hour after instillation of test item revealed: Cornea-Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible in all animals; Area of Opacity-One quarter (or less) but not zero in all 3 animals;Iris:Normal in all the animals.Conjunctivae –Diffuse, crimson color; individual vessels not easily discernible was observed in all animals;Chemosis:Some swelling above normal (includes nictitating membranes) was observed in all animals.

Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all 3 animals; Iris: Normal in all the animals. Conjunctivae – Blood vessels were normal in all 3 animals; Chemosis: No swelling (Normal) was observed in all 3 animals.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 1.00, 0.00, 2.00, 1.00; 0.67, 0.00, 2.00, 1.00 and 0.67, 0.00, 2.00, 1.00 respectively. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed within day 7.  

Hence, under the experimental test conditions, “Sodium salicylate (CAS No. – 54-21-7) is “Mildly Irritating to eyes” of New Zealand White female rabbit eyes and is being classified "Category 2"as per the CLP regulation.