N,N-diethylaniline

Endpoint summary

Description of key information

Skin irritation

The dermal irritation potential of test chemical was assessed in various experimental studies conducted on rabbits. Based on the available data for the key and supporting studies, it can be concluded that the test chemical is able to cause skin irritation and thus considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2”.

 Eye irritation

An ocular irritation potential of target chemical was assessed in various experimental studies conducted on rabbits for test and its structurally similar read across chemical. Based on the available data for the key and supporting study, it can be concluded that the test chemical is unable to cause eye damage and thus can be considered as not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

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

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

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

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

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

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.

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

 

 

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from authoritative databases

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To assess the ocular irritation potential of the test chemical

GLP compliance:

not specified

Species:

rabbit

Strain:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

100%

Duration of treatment / exposure:

no data

Observation period (in vivo):

no data

Duration of post- treatment incubation (in vitro):

no data

Number of animals or in vitro replicates:

no data

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: not specified

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No irritation was observed

Interpretation of results:

other: not irritating

Conclusions:

The test material did not induce any ocular effects. Hence the test chemical was considered to be not irritating to the eyes of treated rabbits.

Executive summary:

The ocular irritation potential of the test chemical was in rabbits according to OECD Guide–line 405 "Acute Eye Irritation/Corrosion"

 

When the undiluted test chemical was instilled into the eyes of each rabbit, no adverse effects were observed in treated eyes.

 

Since the test material did not induce any ocular effects, the test chemical was considered to be not irritating to the eyes of treated rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

Various studies has been investigated for the test chemical to observe the potential for dermal irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for target chemical that have been summarized as below;

 

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.

 

Another skin irritation study of test chemical was assessed in rabbits to determine its skin irritation potential.The undiluted test chemical was applied on back or ear of each rabbit for exposure period of 1 min., 5 min., 15 min. (back) and 20 hours (back or ear). The skin reactions were observed for 8 days.Since the test chemical caused moderate skin irritating effects, the test chemical can be considered as irritating to the rabbits’ skin.

 

The next dermal irritation study was examined on rabbit’s ear under semi-occlusive.About 500 µl of test chemical was applied on the ear of each rabbit for the exposure period of 24 hours and skin reactions were observed for 7 days.As the test chemical caused severe skin irritation and corrosive effects on treated rabbits’, it was considered as irritating to the skin of rabbits.

 

The overall results were further supported by the skin irritation study conducted on rabbits according to OECD Guide–line 404 "Acute Dermal Irritation/Corrosion.When the undiluted test chemical was applied onto each rabbit, slightly skin irritating effects were observed in treated eyes. Since the test material induced skin reactions, the test chemical was considered to be slightly irritating to the skin of treated rabbits.

 

Based on the available data for key and supporting studies, it can be concluded that test chemical is able to cause skin irritation and considered as irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Category 2”. 

 

Eye irritation:

In different studies, the test chemical has been investigated for potential for ocular irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for target chemical and its structurally similar read across chemical that have been summarized as below;

 

The ocular irritation potential of the test chemical was conducted in rabbits according to OECD Guide–line 405 "Acute Eye Irritation/Corrosion". When the undiluted test chemical was instilled into the eyes of each rabbit, no adverse effects were observed in treated eyes. Since the test material did not induce any ocular effects, the test chemical was considered to be not irritating to the eyes of treated rabbits.

 

Another eye irritation study was conducted to assess the adverse ocular effects caused by the test chemical on treated rabbits. The chemical did not induce any ocular lesions when 50 mg of undiluted test chemical was instilled into the eye of treated rabbits.Therefore the test chemical was considered to be not irritating to the rabbit’s eye.

 

In next ocular study, when the undiluted test chemical was applied on rabbit’s eye, little irritative effects (maximal effects 1 hour after application) were observed. These effects were completely reversible after 48–96 hours.As the observed effects were not persisted with the observation period the test chemical was considered to be not irritating to the rabbit’s eye.

 

An eye irritation study was performed on rabbits to determine the eye irritation potential of test chemical. The installation of test chemical into the rabbits’ eye caused slight irritation. Hence the test chemical was considered to be slightly irritating to the eye of treated rabbits’.

 

The overall negative results were further supported by the ocular irritation study conducted on rabbits to assess its irritation potential of test chemical. When the test chemical was applied into the eyes of each rabbit, no irritation was observed in treated rabbits. Hence the test chemical was considered to be not irritating to the eyes of treated rabbits.

 

Even though one study for the test chemical claims that there is a possibility of test chemical to cause slight eye irritation, but it is strongly negated by the results of the other studies. Also the potential of causing eye irritation was not sufficient to classify the test chemical as eye irritatnt. Henceon the basis of available data for the target as well as supporting study, it can be concluded that test chemical is unable to cause eye irritation and considered not irritating. Comparing the above annotations with the criteria of CLP regulation, it can be classified under the category “Not Classified”.

Justification for classification or non-classification

The skin and eye irritation potential of test chemical were observed in various studies. The results obtained from these studies indicates that the chemical is able to cause skin irritation but unlikely to cause eye damage. Hence the test chemical can be classified under the category “Category 2” for skin and “Not Classified” eye as per CLP.