N,N-diethyl-3-oxobutyramide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

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

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Principles of method if other than guideline:

The aim of this study was to assess the toxicity potential of Diethylacetoacetamide (DEAAA) (CAS No. 2235-46-3) after single oral administration in Wistar Rats and an observation period of 14 days

GLP compliance:

yes

Test type:

acute toxic class method

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

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- 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- Lot/batch No.: 31- Expiration date of the lot/batch: May 03, 2016

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: National Institute of Biosciences Pune- Females (if applicable) nulliparous and non-pregnant:yes- Age at study initiation: 9 - 12 weeks- Weight at study initiation: Minimum: 120 gm ;Maximum: 150 gm (Individual body weights were within ± 20 % prior to treatment after overnight fasting)- Fasting period before study: Rat were foasted overnight - Housing:Group of three animals were housed 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 ad libitum via drinking bottles.- Acclimation period:Animal nos. 01-03 was acclimatized for 8 days, 04-06 for 11 days, 07-09 for 18 days and 10-12 for 8 days prior to administration of the test item. ENVIRONMENTAL CONDITIONS - Temperature (°C): Minimum: 19.30 °C; Maximum: 23.90 °C - Humidity (%): Minimum: 50.80%; Maximum: 66.50% - 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: November 09, 2015 To:December 22, 2015

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE - Concentration in vehicle: 300 mg/kg and 2000 mg/kg - Amount of vehicle (if gavage):10 ml/kg - Justification for choice of vehicle:Corn oil was selected as a vehicle based on solubility testing - Lot/batch no. (if required): MR301015 DOSAGE PREPARATION (if unusual):Vehicle (Corn oil) was added to the weighed quantity of test item (300 mg and 2000 mg) slowly and mixed well. Transferred the formulation to the measuring cylinder and made the final volume up to 10 ml. Hence, stock dose concentration was achieved as 30 mg/ml and 200 mg/ml. The test item was miscible in corn oil. The dosing solution was prepared freshly, shortly prior to dose administration. CLASS METHOD (if applicable) - Rationale for the selection of the starting dose:The starting dose level was that, which most likely to produce mortality in some of the dosed animals. The starting dose level was selected as 300 mg/kg body weight as no information was available to conduct the study.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

Total = 12 Females (Group of 3 animals/dose)

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 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the animals were observed once a day during the 14 day observation period.All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.All the rats were weighed on days 0 (prior to dosing), 7 and 14. - Necropsy of survivors performed: yes - Other examinations performed: All the animals were observed for external and internal gross pathology.

Statistics:

not specified

Results and discussion

Preliminary study:

not specified

Effect levels

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in animals treated with 300 and 2000 mg/kg body weight throughout the 14 days observation period

Clinical signs:

other: At 300 and 2000 mg/kg body weight, all the animals were observed normal throughout the experimental period

Gross pathology:

During external and internal gross necropsy examination, terminally sacrificed animals treated with 300 and 2000 mg/kg body weight showed no abnormalities

Other findings:

not specified

Any other information on results incl. tables

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.	Group/ Dose (mg/kg body weight)		Body Weight (gram)			Body Weight Change (%)
		Dose Volume* (ml)	Day 0	Day 7	Day 14	Day 0-7	Day 0-14
01	G1/ 300	1.5	145	163	198	12.41	36.55
02		1.5	150	168	200	12.00	33.33
03		1.4	144	159	186	10.42	29.17
04		1.3	134	159	182	18.66	35.82
05		1.4	140	167	173	19.29	23.57
06		1.3	132	155	181	17.42	37.12
07	G2/ 2000	1.2	120	143	192	19.17	60.00
08		1.4	142	177	210	24.65	47.89
09		1.4	139	168	200	20.86	43.88
10		1.4	136	156	178	14.71	30.88
11		1.4	142	162	194	14.08	36.62
12		1.4	139	155	171	11.51	23.02

*= Dose volume calculated based on day 0 body weight

 

 

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

 

Sex:Female

Group/ Dose (mg/kg body weight)		Rats Body Weight (g)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	0-7	0-14
G1/ 300	Mean	140.83	161.83	186.67	15.03	32.59
	SD	6.88	5.08	10.46	3.85	5.30
	n	6	6	6	6	6
G2/ 2000	Mean	136.33	160.17	190.83	17.50	40.38
	SD	8.31	11.72	14.29	4.91	13.11
	n	6	6	6	6	6

Keys:SD = Standard Deviation, n = Number of Animals

Table 3: Individual Animal Clinical Signs and Symptoms

Sex:Female

Animal No.	Group/ Dose (mg/kg body weight)	Hours (Day 0)
		1/2	1	2	3	4
01	G1/ 300	1	1	1	1	1
02		1	1	1	1	1
03		1	1	1	1	1
04		1	1	1	1	1
05		1	1	1	1	1
06		1	1	1	1	1
07	G2/ 2000	1	1	1	1	1
08		1	1	1	1	1
09		1	1	1	1	1
10		1	1	1	1	1
11		1	1	1	1	1
12		1	1	1	1	1

Key: 1 = Normal

Table 3: Individual Animal Clinical Signs and Symptoms (Continued)

 

Sex:Female

 

Animal No.	Group/ Dose (mg/kg body weight)	Days post dosing
		1	2	3	4	5	6	7	8	9	10	11	12	13	14
01	G1/ 300	1	1	1	1	1	1	1	1	1	1	1	1	1	1
02		1	1	1	1	1	1	1	1	1	1	1	1	1	1
03		1	1	1	1	1	1	1	1	1	1	1	1	1	1
04		1	1	1	1	1	1	1	1	1	1	1	1	1	1
05		1	1	1	1	1	1	1	1	1	1	1	1	1	1
06		1	1	1	1	1	1	1	1	1	1	1	1	1	1
07	G2/ 2000	1	1	1	1	1	1	1	1	1	1	1	1	1	1
08		1	1	1	1	1	1	1	1	1	1	1	1	1	1
09		1	1	1	1	1	1	1	1	1	1	1	1	1	1
10		1	1	1	1	1	1	1	1	1	1	1	1	1	1
11		1	1	1	1	1	1	1	1	1	1	1	1	1	1
12		1	1	1	1	1	1	1	1	1	1	1	1	1	1

Key: 1 = Normal

Table 4: Individual Animal Mortality Record

 

Sex:Female

Animal No.	Group/ Dose (mg/kg body weight)	Day of Observation (Day 0 to 14)
		Morning Observation	Evening Observation
01	G1/ 300	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		No mortality and morbidity	No mortality and morbidity
07	G2/ 2000	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
11		No mortality and morbidity	No mortality and morbidity
12		No mortality and morbidity	No mortality and morbidity

Key:TS = Terminal Sacrifice

Table 5: Gross Necropsy Observation

 

Sex:Female                                                                                                                                        

Animal No.	Group/ Dose (mg/kg body weight)	Mode of Death	Gross Observation
			External	Internal
01	G1/ 300	TS	No abnormality detected	No abnormality detected
02		TS	No abnormality detected	No abnormality detected
03		TS	No abnormality detected	No abnormality detected
04		TS	No abnormality detected	No abnormality detected
05		TS	No abnormality detected	No abnormality detected
06		TS	No abnormality detected	No abnormality detected
07	G2/ 2000	TS	No abnormality detected	No abnormality detected
08		TS	No abnormality detected	No abnormality detected
09		TS	No abnormality detected	No abnormality detected
10		TS	No abnormality detected	No abnormality detected
11		TS	No abnormality detected	No abnormality detected
12		TS	No abnormality detected	No abnormality detected

Key : TS = Terminal Sacrifice

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

The LD50 cut-off value was considered to be 5000 mg/kg bw, when female Wistar rats were treated with N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) via oral gavage route.

Executive summary:

Acute Oral Toxicity Study of N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) was conducted as per OECD No. 423 on 12 female Wistar rats at the dose concentration of 300 and 2000 mg/kg bw. The animals were fasted for minimum 16-17.5 hours prior to dosing and for 4 hours post dosing, feed was withheld but drinking water provided ad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.3 rats of the first group G1 were dosed with starting dose of 300 mg/kg body weight and the animals did not show any mortality. So another 3 animals of the same group G1 were dosed with 300 mg/kg body weight and no mortality was observed. Next, 3 animals of group G2 were dosed with 2000 mg/kg body weight and no mortality was observed. So, another 3 animals of the same group G2 were dosed with 2000 mg/kg body weight and no mortality was observed. Hence, further dosing was stopped.After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the animals were observed once a day during the 14 day observation period. All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period. All the rats were weighed on days 0 (prior to dosing), 7 and 14.All the animals were observed for external and internal gross pathology.No mortality was observed in animals treated with 300 and 2000 mg/kg body weight throughout the 14 days observation period. At 300 and 2000 mg/kg body weight, all the animals were observed normal throughout the experimental period.Mean body weight of all the animals treated with 300 and 2000 mg/kg body weight were observed with gain on day 7 and 14, as compared to day 0. During external and internal gross necropsy examination, terminally sacrificed animals treated with 300 and 2000 mg/kg body weight showed no abnormalities. Hence, The LD50 cut-off value was considered to be 5000 mg/kg bw, when female Wistar rats were treated with N,N-diethyl-3-oxobutyramide (CAS no: 2235-46-3) via oral gavage route.