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

Skin irritation / corrosion

Administrative data

Endpoint:

skin 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 404 (Acute Dermal 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 after dermal application on the intact skin 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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species:Rabbit (Oryctolagus cuniculus)
Strain :New Zealand White
Age :4.0 to 5.5 Months (Approximately)
Sex :Female
Number of Animals: Three
Supplier/Source:Procured from GENTOX BIOSERVICES PVT. LTD., HYDERABAD, India.
CPCSEA Registration No.: 1242/bc/08/CPCSEA.
Health Status :Healthy young adult rabbits were used for the study.
Females were nulliparous and non-pregnant.
Body weight of animals:Minimum: 2.042 kg & Maximum: 2.146 kg (Prior to Treatment)
Acclimatisation :Rabbits were acclimatised to the test conditions for a period of 10 days (Animal No. 1) and 13 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 cards were labelled with study no., study type, test system, sex, dose, animal number, experimental start and completion date.

Diet:All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 200009.
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: 19.00 °C Maximum: 21.40 °C
Relative humidity: Minimum: 41.90% Maximum: 65.80%
Light-dark-rhythm: 12:12
Air Changes : More than 12 changes per hour

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 gm
VEHICLE (distilled water)
- Amount(s) applied (volume or weight with unit):0.5 ml of distilled water
- Lot/batch no. (if required):N/A
- Purity:N/A

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 female rabbits

Details on study design:

TEST SITE
- Area of exposure:The dorsal lumbar region at contralateral sites.
- % coverage:Approximately 6 X 6 cm.
- Type of wrap if used:A porous gauze dressing and non-irritating tape (Micropore 3”).
REMOVAL OF TEST SUBSTANCE
- Washing (if done):The residual test item was removed by using cotton soaked in distilled water.
- Time after start of exposure:4 hour
SCORING SYSTEM:Draize Method

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 hrs,48 hrs,72 hrs

Score:

0

Max. score:

4

Reversibility:

no data

Remarks on result:

other: not irritating

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 hrs,48 hrs,72 hrs

Score:

0

Max. score:

4

Reversibility:

no data

Remarks on result:

other: not irritating

Irritant / corrosive response data:

The following were observed in treated rabbits.
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.
After 4 hours of exposure in Animal No. 1,very slight erythema (barely perceptible) and no oedema was observed at 1 hour of observation.At 24 , 48 and
72 hour no erythema and no oedema was observed.
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1,24,48 and 72 hours after patch removal,evaluated and graded as per draize method.
At 1,24,48 and 72 hours,no erythema and oedema was observed in Animal No. 2.
Animal no. 3 at 1 hour observation revealed very slight erythema (barely perceptible) and no oedema.At 24 , 48 and 72 hour,Animal No. 3 revealed no erythema and no oedema and was found to be normal throughout the experimental period.
The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.

Other effects:

Body weight:
Increase in body weight at terminal sacrifice as compared to day 0 in all the three animals.
Clinical signs:
No systemic toxicity was observed at treated rabbits during the experimental period.
Mortality:
No mortality was observed during the observation period.

Any other information on results incl. tables

Table 1

Skin Reaction

 

In Treated area Dose:0.5 g of test item (Pulverized form)                       Sex:Female

 

Animal No.	Test	Treated  area	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Right	1	0	0	0	0	0	0	0
2	Confirmatory	Right	0	0	0	0	0	0	0	0
3		Left	1	0	0	0	0	0	0	0

 

 

 

 

In Control areaDose:0.5 ml of distilled water                                                        Sex:Female

 

Animal No.	Test	Treated area	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Left	0	0	0	0	0	0	0	0
2	Confirmatory	Left	0	0	0	0	0	0	0	0
3		Right	0	0	0	0	0	0	0	0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema

1 = Very slight erythema(barely perceptible)

Mean Individual Animal Score at 24, 48 and 72 hours (Treated Site)

 

Animal Number                   Observations	1	2	3
Erythema	0.00	0.00	0.00
Oedema	0.00	0.00	0.00

 

Applicant's summary and conclusion

Interpretation of results:

other: not irritating

Conclusions:

The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.Hence, the test chemical was re
garded as non-Irritating to the skin of Female New Zealand White rabbits under the experimental conditions tested and is thus not classified as a skin irritant.

Executive summary:

Acute Dermal Irritation/corrosion Study of test chemical was conducted in Rabbits.This study was performed as per OECD guideline No. 404.Three healthy young adult Female rabbits were used for conducting acute dermal irritation/corrosion study.Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment.A dose of 0.5 g (pulverized form)test item moistened with 0.5 ml distilled water wasappliedto the skin,over an area of approximately 6 x 6 cm clippedof haironone side of rabbits.The other untreated side was kept as control area and 0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch.After 4 hours of exposure in Animal No. 1,very slight erythema (barely perceptible) and no oedema was observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and no oedema was observed.Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3)to confirm the non irritant nature of the test item. The patch was removed after 4 hours and rabbits wereobserved for erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per draize method.At 1, 24, 48 and 72 hours, no erythema and oedema was observed in Animal No. 2. Animal No. 3 at 1 hour observation post patch removal revealed very slight erythema (barely perceptible) and no oedema. At 24, 48 and 72 hours, Animal No. 3 revealed no erythema and no oedema.The other untreated side revealed no erythema and no oedema and was found to be normal throughout the experimental period.The individual mean score at 24, 48 and 72 hoursfor animal nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.  No erythema and no oedema (skin irritation) were found at the end of 72 hour observation period after patch removal.Hence, it was concluded that test chemical was Non-Irritating to the skin of Female New Zealand White rabbits under the experimental conditions.