Tetrabutylammonium bromide

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 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 “Tetrabutylammonium bromide (CAS No. - 1643-19-2)” on eye, when exposed by the ocular route in rabbits.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Tetrabutylammonium bromide

EC Number:

216-699-2

EC Name:

Tetrabutylammonium bromide

Cas Number:

1643-19-2

Molecular formula:

C16H36N.Br

IUPAC Name:

tetrabutylazanium bromide

Test material form:

solid

Details on test material:

- Name of test material (as cited in study report):Tetrabutylammonium bromide
- Molecular formula :C16H36N.Br
- Molecular weight :322.37 g/mol
- Substance type:Organic
- Physical state:White Solid (Crystals)
- Lot/batch No.:Lot 1/02
- AI Content:99.9 %
- Storage condition of test material:The test item was stored in a cool, dry place and kept in a closed.
- 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 test item was disposed during the conduct of the study and the same was documented in the raw data.

Specific details on test material used for the study:

- Name of test material (as cited in study report):Tetrabutylammonium bromide
- Molecular formula :C16H36N.Br
- Molecular weight :322.37 g/mol
- Substance type:Organic
- Physical state:White Solid (Crystals)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source:Procured from Rabbiroof, Hyderabad, India.
- Age at study initiation:4.5 to 5.5 Months (Approximately)
- Weight at study initiation:Minimum: 1.932 kg and Maximum: 2.438 kg (Prior to Treatment).
- Health Status :Healthy young adult, Females were nulliparous and non pregnant.
- 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 6 days (Animal No.-1) and 11 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.50 °C and Maximum: 22.70 °C
- Humidity (%):Minimum: 41.20 % and Maximum: 67.30 %
- 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

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):100 mg (0.1 gm)
- 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 hrs

Observation period (in vivo):

All the animals were observed at 1, 24, 48, 72 hours and at day7 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 hours

SCORING SYSTEM:Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescent strips.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1,#2 and #3

Time point:

other: 24,48,72 hrs and 7 days

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Untreated (Control Eye)

Results 2

Irritation parameter:

iris score

Basis:

animal: #1,#2 and #3

Time point:

other: 24,48,72 hrs and 7 days

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Untreated (Control Eye)

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1,#2 and #3

Time point:

other: 24,48,72 hrs and 7 days

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Untreated (Control Eye)

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1,#2 and #3

Time point:

other: 24,48,72 hrs and 7 days

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Untreated (Control Eye)

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: After 24, 48, 72 Hours

Score:

1.33

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 6

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: After 24, 48, 72 Hours

Score:

1.67

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 7

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: After 24, 48, 72 Hours

Score:

1

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 8

Irritation parameter:

cornea opacity score

Basis:

animal: #1,#2 and #3

Time point:

other: Day 7

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 9

Irritation parameter:

iris score

Basis:

animal: #1,#2 and #3

Time point:

other: after 24, 48, 72 Hours and day 7

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 10

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: After 24, 48, 72 Hours

Score:

1.67

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: After 24, 48, 72 Hours

Score:

2

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 12

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: After 24, 48, 72 Hours

Score:

2

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 13

Irritation parameter:

conjunctivae score

Basis:

animal: #1,#2 and #3

Time point:

other: Day 7

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 14

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: After 24, 48, 72 Hours

Score:

1.33

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 15

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: After 24, 48, 72 Hours

Score:

2

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 16

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: After 24, 48, 72 Hours

Score:

1.67

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

Results 17

Irritation parameter:

chemosis score

Basis:

animal: #1,#2 and #3

Time point:

other: Day 7

Score:

0

Max. score:

3

Reversibility:

not specified

Remarks on result:

other: Treated eye

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 was seen in animal no. 1 and 3 whereas scattered or diffuse areas of opacity (other than slight dulling of normal lustre) details of iris clearly visible was seen in animal no. 2; Area of Opacity- Zero in animal no. 1 and 3 whereas one quarter (or less) but not zero was seen in animal no.2; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible in animal no. 2 and some blood vessels definitely hyperaemic (injected) animal no. 1 and 3; Chemosis: Some swelling above normal (includes nictitating membranes) in animal no. 1 and obvious swelling with partial eversion of lids was observed in animal no. 2 and 3.

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 all 3 animals; Area of Opacity- Greater than one quarter, but less than half was seen in animal no. 1 and 2 whereas one quarter (or less) but not zero was seen in animal no.3; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in animal no.1 whereas obvious swelling with partial eversion of lids was seen in animal no.2 and 3.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 50%, 60% and 45% damage in animal no. 1, 2 and 3 respectively.

Observation at 48 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 all 3 animals; Area of Opacity- One quarter (or less) but not zero was seen in animal no.1 and 3 whereas greater than one quarter, but less than half was seen in animal no. 2; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal no. 1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3; Chemosis: Obvious swelling with partial eversion of lids was seen in all 3 animals.

Observation at 72 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 all 3 animals; Area of Opacity- One quarter (or less) but not zero was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in animal no. 1 and 3 whereas obvious swelling with partial eversion of lids was seen in animal no. 2.

Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero percent was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in all the 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 1.33, 0.00, 1.67, 1.33; 1.67, 0.00, 2.00, 2.00; 1.00, and 1.00 0.00, 2.00, 1.67, respectively.

Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity was seen in animal no. 1 and 3 whereas scattered or diffuse areas of opacity (other than slight dulling of normal lustre) details of iris clearly visible was seen in animal no. 2; Area of Opacity- Zero in animal no. 1 and 3 whereas one quarter (or less) but not zero was seen in animal no.2; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible in animal no. 2 and some blood vessels definitely hyperaemic (injected) animal no. 1 and 3; Chemosis: Some swelling above normal (includes nictitating membranes) in animal no. 1 and obvious swelling with partial eversion of lids was observed in animal no. 2 and 3.

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 all 3 animals; Area of Opacity- Greater than one quarter, but less than half was seen in animal no. 1 and 2 whereas one quarter (or less) but not zero was seen in animal no.3; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in animal no.1 whereas obvious swelling with partial eversion of lids was seen in animal no.2 and 3.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 50%, 60% and 45% damage in animal no. 1, 2 and 3 respectively.

Observation at 48 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 all 3 animals; Area of Opacity- One quarter (or less) but not zero was seen in animal no.1 and 3 whereas greater than one quarter, but less than half was seen in animal no. 2; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was seen in animal no. 1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3; Chemosis: Obvious swelling with partial eversion of lids was seen in all 3 animals.

Observation at 72 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 all 3 animals; Area of Opacity- One quarter (or less) but not zero was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in animal no. 1 and 3 whereas obvious swelling with partial eversion of lids was seen in animal no. 2.
Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero percent was seen in all 3 animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal was seen in all 3 animals; Chemosis: No swelling (Normal) was seen in all the 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, 1.00, 1.00; 1.33, 1.67, 1.00; 1.67, 2.00, 2.00 and 1.33, 2.00, 1.67, 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 of test item                                                            Sex:Female

Animal Numbers		1							2								3
Application Side		Left							Right								Right
Eye Reactions		At hour						Day		At hour						Day			At hour						Day
		*		1		24		48		72		7		*		1		24		48		72			7		*		1		24		48		72			7
Corneal Opacity		0		0		1		1		1		0		0		1		1		1		1			0		0		0		1		1		1			0
Area of Opacity		0		0		2		1		1		0		0		1		2		2		1			0		0		0		1		1		1			0
Iris		0		0		0		0		0		0		0		0		0		0		0			0		0		0		0		0		0			0
Conjunctiva		0		2		2		1		2		0		0		1		2		2		2			0		0		1		2		2		2			0
Chemosis		0		1		1		2		1		0		0		2		2		2		2			0		0		2		2		2		1			0
Corneal Damage%		50%							60%								45%

 

Dose:Untreated (Control Eye)                                                                       Sex:Female

Animal Numbers	1							2							3
Application Side	Right							Left							Left
Eye Reactions	At hour					Day		At hour					Day		At hour					Day
	*	1	24	48	72		7	*	1	24	48	72		7	*	1	24	48	72		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.

 

Table 1 Continued…

 

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

Animal No.  Eye Reaction	1	2	3
Corneal Opacity	1.33	1.67	1.00
Iris	0.00	0.00	0.00
Conjunctiva	1.67	2.00	2.00
Chemosis	1.33	2.00	1.67

 

 

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: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 = Normal

 

Table 3: Individual Animal Body Weight

Sex :Female

Animal No.	Animal Body Weight (kg)
	Prior to application	At termination
1	2.208	2.387
2	1.932	2.214
3	2.438	2.646

Key:kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:

Category II

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 1, 2 and 3 was observed till 72 hours which were recovered on day 7.
Hence under the experimental test conditions, “Tetrabutylammonium bromide (CAS No. - 1643-19-2) solid is “Mildly Irritating to Eyes” to New Zealand White female rabbit eyes and is being classified as an eye irritant in 'category 2' as per the CLP regulation.

Executive summary:

Acute Eye Irritation/Corrosion Study of test chemical in Rabbits, sponsored bySustainability Support Services (Europe) AB was conducted at sa-FORD (Sanctuary for Research and Development), Maharashtra, India. 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; 100 mgof test item (pulverised)was instilled in the other (treated) eye of each rabbit.The eye was observed at 1, 24, 48, 72 hour and at day 7 after test item instillation.Ophthalmoscope was used for scoring of eye lesions.

 

In the initial test,100 mg (0.1 gm) of test item (pulverized)was applied into the conjunctival sac of the left eye of animal no.1 whereas the right eye of the rabbit served as the control. Ocular lesions were seen in animal no.1 at till 72 hours observation which recovered on day 7 hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 100 mgof test item(pulverised)was 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 till 72 hours observation which was recovered on day 7 observation.

 

Untreated eye of all the three 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 was seen in animal no. 1 and 3 whereas scattered or diffuse areas of opacity (other than slight dulling of normal lustre) details of iris clearly visible was seen in animal no. 2;Area of Opacity-Zero in animal no. 1 and 3 whereas one quarter (or less) but not zero was seen in animal no.2;Iris:Normal in all the animals.Conjunctivae -Diffuse, crimson color; individual vessels not easily discerniblein animal no. 2 and some blood vessels definitely hyperaemic (injected) animal no. 1 and 3;Chemosis:Some swelling above normal (includes nictitating membranes) in animal no. 1 and obvious swelling with partial eversion of lids was observed in animal no. 2 and 3.

 

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 all 3 animals; Area of Opacity-Greater than one quarter, but less than half was seen in animal no. 1 and 2 whereas one quarter (or less) but not zero was seen in animal no.3;Iris:Normal in all the animals.Conjunctivae -Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in animal no.1 whereas obvious swelling with partial eversion of lids was seen in animal no.2 and 3.

 

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 50%, 60% and 45% damage in animal no. 1, 2 and 3 respectively.

 

Observation at 48 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 all 3 animals; Area of Opacity-One quarter (or less) but not zero was seen in animal no.1 and 3 whereas greater than one quarter, but less than half was seen in animal no. 2;Iris:Normal in all the animals.Conjunctivae -Some blood vessels definitely hyperaemic (injected) was seen in animal no. 1 whereas diffuse, crimson color; individual vessels not easily discernible was seen in animal no. 2 and 3;Chemosis:Obvious swelling with partial eversion of lids was seen in all 3 animals.

 

Observation at 72 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 all 3 animals; Area of Opacity-One quarter (or less) but not zero was seen in all 3 animals;Iris:Normal in all the animals.Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all 3 animals;Chemosis:Some swelling above normal (includes nictitating membranes) was seen in animal no. 1 and 3 whereas obvious swelling with partial eversion of lids was seen in animal no. 2.

 

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

 

The individual mean score for animal nos. 1, 2 and 3at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 1.33, 0.00, 1.67, 1.33; 1.67, 0.00, 2.00, 2.00; 1.00, and 1.00 0.00, 2.00, 1.67, respectively.

 

Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbit no. 1, 2 and 3 was observed till 72 hours which were recovered on day 7.

Hence under the experimental test conditions, the test chemical solid is “Mildly Irritating to Eyes” to New Zealand White female rabbit eyes and is being classified as an eye irritant in 'category 2' as per the CLP regulation.