N,N-diethylaniline

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 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:

N,N-diethylaniline

EC Number:

202-088-8

EC Name:

N,N-diethylaniline

Cas Number:

91-66-7

Molecular formula:

C10H15N

IUPAC Name:

N,N-diethylaniline

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report):N, N-diethylaniline
- Molecular formula :C8H11N
- Molecular weight :121.18 g/mol
- Substance type:Organic
- Physical state:clear light yellow oily liquid
- Lot/batch No.:Lot 1/07
- Storage condition of test material:Store in a cool place. Keep container tightly closed in a dry and well ventilated place.
- Other:
Handling and Disposal
Safety precautions : Avoid Contact with skin and eyes. Keep away from sources of ignition. Aprons, masks, caps, 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.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Procured from LIVEON BIOLABS PVT. LTD
- Age at study initiation:3.0 to 4.0 Months (Approximately)
- Weight (Prior to Treatment):Minimum: Minimum: 2.082 kg and Maximum: 2.250 kg
- Health Status : Healthy young adults rabbits were used for the study.
- 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 8 days (Animal No.-2 and 3) prior to the application of the test item.

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

Test system

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 ml
- 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:

4 hours

Observation period:

rabbits were observed for 1, 24, 48, 72 hr and 7th day after patch removal

Number of animals:

3 male 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:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: #1 and #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The following were observed in treated rabbits.
Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch. At 1 hour observation animal no. 1 revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible). At 24 hour observation well defined erythema and slight oedema (edges of area well defined by definite raising) was observed. After 24 hours no severe skin lesions were observed hence a confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the irritant nature of the test item. At 48 hour observation, well defined erythema and slight oedema (edges of area well defined by definite raising) was observed. At 72 hour observation, very slight erythema (barely perceptible) and very slight oedema (barely perceptible) was observed. Animal no.1 was recovered to normal on day 7.
Animal no. 2 and 3 revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible) at 1 hour observation. At 24 and 48 hour observation animal no. 2 and 3 revealed well defined erythema and slight oedema (edges of area well defined by definite raising). At 72 hour observation, animal no. 2 revealed well defined erythema and very slight oedema (barely perceptible). At 24, 48 and 72 hour observation, animal no. 3 revealed well defined erythema and slight oedema (edges of area well defined by definite raising). Animal no.2 and 3 recovered to normal on day 7.
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours and on day 7 post patch removal, evaluated and graded as per draize method.

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.

Any other information on results incl. tables

Table 1

Skin Reaction

In Treated area Dose:0.5 ml of test item                                                                          Sex:Male

 

Animal No.	Test	Treated  area*	Erythema score					Oedema score
			1h	24h	48h	72h	Day 7	1h	24h	48h	72h	Day 7
1	Initial	Left	1	2	2	2	0	1	2	2	1	0
2	Confirmatory	Right	1	2	2	2	0	1	2	2	1	0
3		Right	1	2	2	2	0	1	2	2	2	0

 

 In Control area  Dose:0.5 ml of distilled water                                                         Sex:Male

Animal No.	Test	Treated  area*	Erythema score					Oedema score
			1h	24h	48h	72h	Day 7	1h	24h	48h	72h	Day 7
1	Initial	Right	0	0	0	0	0	0	0	0	0	0
2	Confirmatory	Left	0	0	0	0	0	0	0	0	0	0
3		Left	0	0	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)                       1 =Very slight oedema (barely perceptible)

2 =Well defined erythema                                                   2 = Slight oedema (edges of defined by definite raising                            

Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours

 

Animal Number                   Observations	1	2	3
Erythema	2.00	2.00	2.00
Oedema	1.67	1.67	2.00

 

 

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 2.00, 2.00, 2.00 and 1.67, 1.67, 2.00, for erythema and oedema formation, respectively. Hence, it was concluded that the test chemical was Mild Skin Irritation to the skin of Male New Zealand White rabbits under the experimental conditions tested and classified as “Category 2 (irritant)” as per CLP regulation.

Executive summary:

Acute Dermal Irritation/corrosion Study of test chemical was performed as per OECD guideline No. 404 using three healthy young adult male.

 

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 ml of test item (as such) was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one 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. At 1 hour observation animal no. 1 revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible). At 24 hour observation well defined erythema and slight oedema (edges of area well defined by definite raising) was observed. After 24 hours no severe skin lesions were observed hence a confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the irritant nature of the test item. At 48 hour observation, well defined erythema and slight oedema (edges of area well defined by definite raising) was observed. At 72 hour observation, very slight erythema (barely perceptible) and very slight oedema (barely perceptible) was observed. Animal no.1 was recovered to normal on day 7.

Animal no. 2 and 3 revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible) at 1 hour observation. At 24 and 48 hour observation animal no. 2 and 3 revealed well defined erythema and slight oedema (edges of area well defined by definite raising). At 72 hour observation, animal no. 2 revealed well defined erythema and very slight oedema (barely perceptible). At 24, 48 and 72 hour observation, animal no. 3 revealed well defined erythema and slight oedema (edges of area well defined by definite raising). Animal no.2 and 3 recovered to normal on day 7.

The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours and on day 7 post patch removal, evaluated and graded as per draize method.

The individual mean score at24, 48 and 72 hours for animal nos. 1, 2 and 3 were 2.00, 2.00, 2.00 and The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 2.00, 2.00, 2.00 and 1.67, 1.67, 2.00, for erythema and oedema formation, respectively. 

Hence, it was concluded that the test chemical was Mild Skin Irritation to the skin of Male New Zealand White rabbits under the experimental conditions tested and classified as “Category 2 (irritant)” as per CLP regulation.