Bis(4-(1,1,3,3-tetramethylbutyl)phenyl)amine

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 study report

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 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:

Bis(4-(1,1,3,3-tetramethylbutyl)phenyl)amine

EC Number:

239-816-9

EC Name:

Bis(4-(1,1,3,3-tetramethylbutyl)phenyl)amine

Cas Number:

15721-78-5

Molecular formula:

C28-H43-N

IUPAC Name:

4-(2,4,4-trimethylpentan-2-yl)-N-[4-(2,4,4-trimethylpentan-2-yl)phenyl]aniline

Details on test material:

- Name of test material (as cited in study report):4-(1,1,3,3-tetramethylbutyl)-N-[4-(1,1,3,3-tetramethylbutyl)phenyl]aniline
- Molecular formula :C28H43N
- Molecular weight :393.655 g/mol
- Substance type:Organic
- Physical state:off white solid
- A.I Content:98.72%
- Lot/batch No.:Lot 03/31
- Storage condition of test material:Room temperature (20-30 0 °C)
- Other:
Handling and Disposal
Safety precautions : Aprons,masks, caps, gloves and goggles were used to ensure the health and safety of the personnel.
Disposal :No unused test item remained for disposal

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species: Rabbit (Oryctolagus cuniculus)
Strain : New Zealand White
Age : 4.0 to 5.5 Months (Approximately)
Sex : Male
Number of Animals: Three
Supplier/Source: Procured from LIVEON BIOLABS PVT. LTD., INDIA
(CPCSEA Reg. No. 1610/PO/RcBiBt/S/2012/CPCSEA).
Health Status : Healthy young adult animals were used for the study.
Body weight of animals: Minimum: 1.832 kg and Maximum: 2.530 kg
(Prior to Treatment)
Acclimatisation : Rabbits were acclimatised to the test conditions for a period of 5 days (Animal No.-1) and 8 days (Animal No. 2 and 3) prior to the application of the test item.
Identification: During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage card was labelled with at least study no., study type, test system, sex, dose, animal no. experiment start date and experiment completion date.

Diet: All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No. 200010 and 200011.
Water: Aqua guard filtered tap water was provided ad libitum.
Husbandry: 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.

Temperature : Minimum: 19.30 °C Maximum: 21.20 °C
Relative humidity: Minimum: 49.10 % Maximum: 61.50 %
Light-dark-rhythm: 12:12
Air Changes: More than 12 changes per hour

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):100 mg
- Concentration (if solution):N/A
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 hr

Observation period (in vivo):

All the animals were observed at 1, 24, 48 and 72 hours after instillation of test item.

Number of animals or in vitro replicates:

3 female 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 hrs

SCORING SYSTEM: Grading of irritation lesions was carried out as per Draize Method

TOOL USED TO ASSESS SCORE: ophthalmoscope

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:

4

Reversibility:

not specified

Remarks on result:

other: no indication of irritation for treated eye and untreated eye

Results 2

Irritation parameter:

iris score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

not specified

Remarks on result:

other: no indication of irritation For treated eye and untreated eye

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: no indication of irritation For treated eye and untreated eye

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1,#2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

other: no indication of irritation For treated eye and untreated eye

Irritant / corrosive response data:

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 was seen 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 (Normal) was observed in all the animals.
Observation at 24 hours after instillation of test item revealed: Cornea: No ulceration or opacity was seen in all the animals; Area of Opacity: Zero in all the animals; Iris: Normal in all the animals; Conjunctivae: Blood vessels normal in all the animals; Chemosis: No swelling (Normal) was observed in all the animals.
At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0%, 0% and 0% damage in animal no. 1, 2 and 3 respectively.
Observation at 48 and 72 hours after instillation of test item revealed: Cornea: No ulceration or opacity was seen in all the animals; Area of Opacity: Zero in all the animals; Iris: Normal in all the animals; Conjunctivae: Blood vessels normal in all the animals; Chemosis: No swelling (Normal) was observed in all the animals.The individual mean score (treated site) for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00 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:
Increase in body weights of all the animals at terminal sacrifice as compared to day 0 in all the three animals was observed

Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

 

Treated Dose:0.1 g of test item (in pulverized form)                                              Sex:Male

Animal Number	1
Side of Instillation	Right
Eye Reactions	*	Hour(s)
		1	24		48		72
Cornea	0	0	0		0		0
Area of Opacity	0	0	0		0		0
Iris	0	0	0		0		0
Conjunctiva	0	1	0		0		0
Chemosis	0	0	0		0		0
Corneal Damage (%)	0%	./.	0%	./.		./.

 

Animal Number	2
Side of Instillation	Right
Eye Reactions	*	Hour(s)
		1	24	48	72
Cornea	0	0	0	0	0
Area of Opacity	0	0	0	0	0
Iris	0	0	0	0	0
Conjunctiva	0	1	0	0	0
Chemosis	0	0	0	0	0
Corneal Damage (%)	0%	./.	0%	./.	./.

 

 

Animal Number	3
Side of Instillation	Right
Eye Reactions	*	Hour(s)
		1	24	48		72
Cornea	0	0	0	0		0
Area of Opacity	0	0	0	0		0
Iris	0	0	0	0		0
Conjunctiva	0	1	0	0		0
Chemosis	0	0	0	0		0
Corneal Damage (%)	0%	./.	0%		./.	./.

Key:*= Pre-treatment eye examination;  ./.= Not Applicable

 

 

 

 

 

 

Table 1 Continued…

Untreated Control                                                                                                                 Sex:Male

Animal Number	1
Side of Instillation	Left
Eye Reactions	*	Hour(s)
		1	24	48	72
Cornea	0	0	0	0	0
Area of Opacity	0	0	0	0	0
Iris	0	0	0	0	0
Conjunctiva	0	0	0	0	0
Chemosis	0	0	0	0	0
Corneal Damage (%)	0%	./.	0%	./.	./.

 

 

Animal Number	2
Side of Instillation	Left
Eye Reactions	*	Hour(s)
		1	24	48		72
Cornea	0	0	0	0		0
Area of Opacity	0	0	0	0		0
Iris	0	0	0	0		0
Conjunctiva	0	0	0	0		0
Chemosis	0	0	0	0		0
Corneal Damage (%)	0%	./.	0%		./.	./.

 

 

Animal Number	3
Side of Instillation	Left
Eye Reactions	*	Hour(s)
		1	24	48	72
Cornea	0	0	0	0	0
Area of Opacity	0	0	0	0	0
Iris	0	0	0	0	0
Conjunctiva	0	0	0	0	0
Chemosis	0	0	0	0	0
Corneal Damage (%)	0%	./.	0%	./.	./.

Key:*= Pre-treatment eye examination;  ./.= Not Applicable

Applicant's summary and conclusion

Interpretation of results:

other: not irritating

Conclusions:

The individual mean score (treated site) for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00 respectively.Hence, test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is thus not classified as a eye irritant.

Executive summary:

Acute Eye Irritation/Corrosion Study of test chemical was conducted in Rabbits. This study was performed as per OECD guideline no. 405.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 gof test item (pulverised form)was instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48 and 72 hour after test item instillation.Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 g 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 hence a confirmatory test was conducted on additional two rabbits (Animal No. 2 and 3); 0.1 gof test item (pulverized form)was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Untreated eye of all the three rabbits was normal throughout the experimental period. 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 was seen in all the animals;Area of Opacity:Zero inall the animals;Iris:Normal in all the animals;Conjunctivae:Some blood vessels definitely hyperaemic (injected) was observed in all the animals;Chemosis:No swelling (Normal) was observed in all the animals. Observation at 24 hours after instillation of test item revealed: Cornea:No ulceration or opacity was seen in all the animals;Area of Opacity:Zero inall the animals;Iris:Normal in all the animals;Conjunctivae:Blood vessels normal in all the animals;Chemosis:No swelling (Normal) was observed in all the animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0%, 0% and 0% damage in animal no. 1, 2 and 3 respectively. Observation at 48 and 72 hours after instillation of test item revealed:Cornea:No ulceration or opacity was seen in all the animals;Area of Opacity:Zero inall the animals;Iris

Normal in all the animals;Conjunctivae:Blood vessels normal in all the animals;Chemosis:No swelling (Normal) was observed in all the animals.The individual mean score (treated site) for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.00, 0.00; 0.00, 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 0.00 respectively.Hence, test chemical is “Non Irritant” to New Zealand White Female rabbit eyes and is thus not classified as a eye irritant.