N,N-diethyl-3-oxobutyramide

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

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:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

The aim of this study was to assess the dermal toxicity of Diethylacetoacetamide (DEAAA) (CAS No. 2235-46-3) after single dose application by dermal route in Wistar rats and 14 day observation period.

GLP compliance:

yes

Test type:

other: Acute Dermal Toxicity

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N,N-diethyl-3-oxobutyramide

EC Number:

218-792-3

EC Name:

N,N-diethyl-3-oxobutyramide

Cas Number:

2235-46-3

Molecular formula:

C8H15NO2

IUPAC Name:

N,N-diethyl-3-oxobutanamide

Test material form:

liquid

Details on test material:

- IUPAC Name: N,N-diethyl-3-oxobutyramide- InChI: 1S/C8H15NO2/c1-4-9(5-2)8(11)6-7(3)10/h4-6H2,1-3H3- Smiles: N(C(CC(C)=O)=O)(CC)CC- Molecular formula :C8H15NO2- Molecular weight :157.212 g/mol- Substance type:Organic- Physical state:Pale yellow clear liquid- Analytical purity: 98.15%- Lot/batch No.: 31- Expiration date of the lot/batch: May 03, 2016

Specific details on test material used for the study:

- IUPAC Name: N,N-diethyl-3-oxobutyramide- InChI: 1S/C8H15NO2/c1-4-9(5-2)8(11)6-7(3)10/h4-6H2,1-3H3- Smiles: N(C(CC(C)=O)=O)(CC)CC- Molecular formula :C8H15NO2- Molecular weight :157.212 g/mol- Substance type:Organic- Physical state:Pale yellow clear liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: In-House Bred- Females (if applicable) nulliparous and non-pregnant: yes- Age at study initiation: young adult animals - Weight at study initiation: Male:Minimum: 214 g and Maximum: 235 g; Female:Minimum: 229 g and Maximum: 259 g - Housing:The animals were housed individually in polycarbonate cages (size 37 [cm] x 21 [cm], height 20 [cm]).- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet, ad libitum. - Water (e.g. ad libitum):Aqua guard filtered tap water was provided via drinking bottles, ad libitum - Acclimation period:All animals were acclimatized to the test conditions for 6 days prior to application of the test item. ENVIRONMENTAL CONDITIONS - Temperature (°C): Minimum: 19.40 °C; Maximum: 23.60 °C - Humidity (%): Minimum: 43.30%; Maximum: 68.10% - Air changes (per hr): More than 12 changes per hour - Photoperiod (hrs dark / hrs light): 12 hour light and 12 hour dark IN-LIFE DATES: From: December 08, 2015 To:December 28, 2015

Administration / exposure

Type of coverage:

occlusive

Vehicle:

not specified

Details on dermal exposure:

TEST SITE - Area of exposure: clipped dorsal area of rat skin - % coverage: greater than 10% body surface area - Type of wrap if used: Test item was held in contact with the skin with a porous gauze dressing (Approx. 10% of body surface area of rat) and non-irritating tape REMOVAL OF TEST SUBSTANCE - Washing (if done): test item was removed by using distilled water - Time after start of exposure:24-hour

Duration of exposure:

24-hour

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Total = 10 (Five per sex)

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days - Frequency of observations and weighing:After test item administration, individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period for clinical signs. All animals were observed for Local signs/skin reactions (in common with clinical signs) once daily and for mortality and morbidity twice daily.All rats were weighed on days 0 (prior to dosing), 7 and 14. - Necropsy of survivors performed: yes, at the end of 14 day observation period, all the surviving rats were euthanized by overdose of CO2 and subjected to gross pathology examination, for external and internal observations.

Statistics:

No statistical analysis was performed for LD50 calculation since the study was terminated with limit test.

Results and discussion

Preliminary study:

not specified

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

No mortality was observed at limit dose of 2000 mg/kg body weight during the 14 day observation period.

Clinical signs:

other: At 2000 mg/kg, all the animals were observed normal throughout the experimental period.

Gross pathology:

The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Other findings:

not specified

Any other information on results incl. tables

Table 1: Dose Volume (ml), Individual Animal Body Weight (g) andBody Weight Changes(%)

 

Dose:2000 mg/ kg bodyweight                   Group:G1                           Density:0.98475

Animal No.	Sex	Dose Volume* (ml)	Body Weight (gram)			Body Weight Change (%)
			Day 0	Day 7	Day 14	Day 0-7	Day 0-14
01	Male	0.44	219	235	259	7.31	18.26
02		0.43	214	229	251	7.01	17.29
03		0.48	235	246	267	4.68	13.62
04		0.45	221	245	278	10.86	25.79
05		0.43	214	239	266	11.68	24.30
06	Female	0.51	252	256	263	1.59	4.37
07		0.52	258	270	281	4.65	8.91
08		0.51	249	256	264	2.81	6.02
09		0.53	259	276	285	6.56	10.04
10		0.47	229	238	249	3.93	8.73

Key: * = based on the test item density and day 0 body weight

Table 2:Summaryof Animal Body Weight (g) and Body Weight Changes (%)

Dose: 2000 mg/kg body weight                                                                                            Group: G1

Sex		Body Weight (gram)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	Day 0-7	Day 0-14
Male	Mean	220.60	238.80	264.20	8.31	19.85
	SD	8.62	7.09	10.03	2.90	5.07
	n	5	5	5	5	5
Female	Mean	249.40	259.20	268.40	3.91	7.62
	SD	12.14	14.74	14.66	1.88	2.34
	n	5	5	5	5	5

Keys: SD= Standard deviation, n = Number of animals

Table 3: Individual Animal Clinical Signs and Symptoms

 

Dose:2000 mg/kg body weight                                                                                            Group:G1

Animal No.	Sex	Hour(s) - Day 0				Day(s)
		1	2	3	4	1	2	3	4	5	6	7
01	Male	1	1	1	1	1	1	1	1	1	1	1
02		1	1	1	1	1	1	1	1	1	1	1
03		1	1	1	1	1	1	1	1	1	1	1
04		1	1	1	1	1	1	1	1	1	1	1
05		1	1	1	1	1	1	1	1	1	1	1
06	Female	1	1	1	1	1	1	1	1	1	1	1
07		1	1	1	1	1	1	1	1	1	1	1
08		1	1	1	1	1	1	1	1	1	1	1
09		1	1	1	1	1	1	1	1	1	1	1
10		1	1	1	1	1	1	1	1	1	1	1

 

Animal No.	Sex	Day(s)
		8	9	10	11	12	13	14
01	Male	1	1	1	1	1	1	1
02		1	1	1	1	1	1	1
03		1	1	1	1	1	1	1
04		1	1	1	1	1	1	1
05		1	1	1	1	1	1	1
06	Female	1	1	1	1	1	1	1
07		1	1	1	1	1	1	1
08		1	1	1	1	1	1	1
09		1	1	1	1	1	1	1
10		1	1	1	1	1	1	1

Key: 1 = Normal

Table 4: Individual Animal Mortality Record

 

Dose:2000 mg/kg body weight                                                                                           Group:G1

Animal No.	Sex	Days of Observation (0 to 14)
		Morning Observations	Evening Observations
01	Male	No mortality and morbidity	No mortality and morbidity
02		No mortality and morbidity	No mortality and morbidity
03		No mortality and morbidity	No mortality and morbidity
04		No mortality and morbidity	No mortality and morbidity
05		No mortality and morbidity	No mortality and morbidity
06	Female	No mortality and morbidity	No mortality and morbidity
07		No mortality and morbidity	No mortality and morbidity
08		No mortality and morbidity	No mortality and morbidity
09		No mortality and morbidity	No mortality and morbidity
10		No mortality and morbidity	No mortality and morbidity

Table 5: GrossNecropsyObservation

 

 Dose:2000 mg/kg body weight               Group:G1             Mode of Death:Terminal Sacrifice

Animal No.	Sex	Gross Observation
		External	Internal
01	Male	No abnormality detected	No abnormality detected
02		No abnormality detected	No abnormality detected
03		No abnormality detected	No abnormality detected
04		No abnormality detected	No abnormality detected
05		No abnormality detected	No abnormality detected
06	Female	No abnormality detected	No abnormality detected
07		No abnormality detected	No abnormality detected
08		No abnormality detected	No abnormality detected
09		No abnormality detected	No abnormality detected
10		No abnormality detected	No abnormality detected

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

The LD50 was considered to be >2000 mg/kg bw, when male and female Wistar rats were treated with N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) by dermal application.

Executive summary:

Acute Dermal Toxicity Study was conducted using N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) as per OECD No.402 in 10 male and female healthy young adult Wistar rats which were randomly selected and used for conducting acute dermal toxicity study at the concentration of 2000 mg/kg bw. Rats free from injury and irritation of skin were selected for the study. 24 hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight based on the test item density (0.98475) and latest body weight was applied by single dermal application and observed for 14 days after treatment. On test day 0, test item was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed.The animals were observed daily for mortality and clinical signs, during the acclimatization period and post dosing till the termination. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1, 14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14. Body weights were recorded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.No mortality was observed in any animal till the end of the experimental period.At 2000 mg/kg, all the animals were observed normal throughout the experimental period. Mean body weight was observed with gain on day 7 and 14 of male and female animals, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Hence, LD50 was considered to be >2000 mg/kg bw, when male and female Wistar rats were treated with N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) by dermal application.