Castor oil, ethoxylated

Eye irritation

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

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 the test chemical on eye, when exposed by the ocular route in rabbits.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Castor oil, ethoxylated

EC Number:

500-151-7

EC Name:

Castor oil, ethoxylated

Cas Number:

61791-12-6

Molecular formula:

C57H104O9(CH2CH2O)n

IUPAC Name:

Castor oil, ethoxylated

Test material form:

liquid

Details on test material:

- IUPAC Name: Castor Oil, ethoxylated
- Smiles: CCCCCCC(CC=CCCCCCCCC(=O)OCC(C(=O)OCCCCCCCC=CCC(O)CCCCCC)OC(=O)CCCCCCC=CCC(O)CCCCCC)OCCOCCOCCOCCOCCO
- Molecular formula:C65H120O14
- Substance type:Organic
- Physical state:Liquid

Specific details on test material used for the study:

- IUPAC Name: Castor Oil, ethoxylated
- Smiles: CCCCCCC(CC=CCCCCCCCC(=O)OCC(C(=O)OCCCCCCCC=CCC(O)CCCCCC)OC(=O)CCCCCCC=CCC(O)CCCCCC)OCCOCCOCCOCCOCCO
- Molecular formula: C65H120O14
- Substance type: Organic
- Physical state: Liquid
- Batch number: Lot 1/12
- AI Content: 99.12 %
- Manufactured date: November, 2013
- Expiry Date: November, 2014
Handling and Disposal
- Safety precautions: Aprons, caps, mask, gloves and goggles were used to ensure the health and safety of the Personnel.
- Disposal: The remaining unused test item was disposed as per internal SOPs
- The pH of the test chemical was confirmed prior to the test and it was 6.9

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Sex: Male
- Source:Procured from RABBI ROOF Animal Facility, A. P., India.
- Age at study initiation:4.5 to 5.5 Months (Approximately)
- Weight at study initiation:Minimum: 1.602 kg and Maximum: 2.038 kg (Prior to Treatment)
- Health Status :Healthy young adult.
- 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 (Nutrivet Life Sciences, Pune) ad libitum.
- 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 5 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: 19.20 °C and Maximum: 22.70 °C
- Humidity (%)::Minimum: Minimum: 42.00 % and Maximum: 62.40 %
- 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):0.1 ml

VEHICLE
- Amount(s) applied (volume or weight with unit):N/A
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A

Duration of treatment / exposure:

24 hrs

Observation period (in vivo):

The eye was observed at 1, 24, 48, 72 hours after test item instillation.

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

3 male rabbits

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:

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescein strips.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

no indication of 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) in all the animals; Chemosis: Some swelling above normal (includes nictating membranes) were observed in all the animals.

Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the 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: No swelling was observed, recovered to normal in all the Animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0 % damage in Animal Nos 1, 2 and 3, respectively.

Observation at 48 and 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels were recovered to normal in all the animals at 48 hours; Chemosis: No swelling was observed in all the animals.

The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found 0.00, 0.00, 0.33, 0.00 ; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00, respectively.

Other effects:

Clinical Observation
No systemic toxicity was observed at 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.
The body weights were marginally increased in all the animals when compared with pre-treatment body weight.

Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

 

In Treated area Dose:0.1 ml of test item                                                               Sex:Male

Animal Numbers	1					2					3
Application Side	Left					Left					Left
Eye Reactions	At hour					At hour					At hour
	*	1	24	48	72	*	1	24	48	72	*	1	24	48	72
Corneal Opacity	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
Iris	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Conjunctiva	0	1	1	0	0	0	1	1	0	0	0	1	1	0	0
Chemosis	0	1	0	0	0	0	1	0	0	0	0	1	0	0	0
Corneal Damage%	0					0					0

Dose:Untreated (Control Eye)                                                                       Sex:Male

Animal Numbers	1					2					3
Application Side	Left					Left					Left
Eye Reactions	At hour					At hour					At hour
	*	1	24	48	72	*	1	24	48	72	*	1	24	48	72
Corneal Opacity	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
Iris	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
Chemosis	0	0	0	0	0	0	0	0	0	0	0	1	0	0	0
Corneal Damage%	0					0					0

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

Animal No.  Eye Reaction	1	2	3
Corneal Opacity	0.00	0.00	0.00
Iris	0.00	0.00	0.00
Conjunctiva	0.33	0.33	0.33
Chemosis	0.00	0.00	0.00

 

 

Formula :

 

Mean Eye Irritation Score =

                          

Sum of the Individual Animal Score for eye reactionat24, 48 and 72 hours

Number of the Observations (3)

Table 2: Individual AnimalClinicalSigns

 

Sex:Male

Animal No.	Days (Post application observation)
	0	1	2	3
1	1	1	1	1
2	1	1	1	1
3	1	1	1	1

Key:1 = Normal

 

Table 3: Individual Animal Body Weight

Sex :Male

Animal No.	Animal Body Weight (kg)
	Prior to application	At termination
1	1.602	1.750
2	2.038	2.101
3	1.902	1.993

Key:kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:

other: not irritating

Conclusions:

The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found 0.00, 0.00, 0.33, 0.00 ; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00, respectively.
Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours.
Hence under the experimental test conditions, the test chemicalwas “Non Irritant” to New Zealand White Male rabbit eyes and is being classified as "Not Classified".

Executive summary:

Acute eye irritation/corrosion study was conducted in rabbits to evaluate the eye irrritant nature of the test chemical. The study was performed as per OECD 405 Guidelines using 3 male New Zealand White rabbits.

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;0.1 ml of test item (as such)was instilled in the other (treated) eye of rabbits.The eye was observed at 1, 24, 48, 72 hours after test item instillation.Ophthalmoscope was used for scoring of eye lesions.

In the initial test,0.1 ml of test itemwas applied into the conjunctival sac of the right eye of Animal No.1. The left eye of the rabbit served as the control. Animal No. 1 presented ocular lesions at 1 hour observation period. Hence the confirmatory test was conducted on additional two rabbits (Animal No. 2 and 3);0.1 ml of test itemwas instilled into the conjunctival sac of right eye and left eye served as the control. Ocular lesions were observed at 1, 24 and 48 hour in Animal Number 2 whereas in Animal Number 3 ocular lesions presented only at 1 hour observation period.

Untreated eye of the treated rabbits was normal throughout the experimental period of 72 hours.

The following grading scores were observed in treated eye of tested 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) in all the animals;Chemosis:Some swelling above normal (includes nictating membranes) were observed in all the animals.

Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all the 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:No swelling was observed, recovered to normal in all the Animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0 % damage in Animal Nos 1, 2 and 3, respectively.

Observation at 48 and 72 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Blood vessels were recovered to normal in all the animals at 48 hours;Chemosis:No swelling was observed in all the animals.

The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found  0.00, 0.00,  0.33, 0.00 ; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00,  respectively.

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours.  

Hence under the experimental test conditions, the test chemicalwas “Non Irritant” to New Zealand White Male rabbit eyes and is being classified as "Not Classified".