Diethyl acetylsuccinate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 April 1998 to 18 September 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline Study according to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EC-Guideline 92/69, Method B.1.

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Diethyl acetylsuccinate

EC Number:

214-223-8

EC Name:

Diethyl acetylsuccinate

Cas Number:

1115-30-6

Molecular formula:

C10H16O5

IUPAC Name:

1,4-diethyl 2-acetylbutanedioate

Constituent 2

Reference substance name:

Diethylacetylsuccinat (DEAS)

IUPAC Name:

Diethylacetylsuccinat (DEAS)

Details on test material:

- Name: "ACETYLBERNSTEINSÄUREDIETHYLESTER".- Chemical name: Acetonyl succinic acid diethylester.-Trade name: Diethylacetylsuccinat (DEAS).- CAS No.: 1115-30-6.- EINECS No.: 2142238.- Batch No.: SLBA 004.- Supplier: Sponsor.- Purity: 98 % (min.).- Solubility in water: 15 g/l (at 20 °C).- Density: 1.081 g/cm3 (at 20 °C).- Melting point: -8 °C.- Boiling point: 180-183 °C (at 67 mbar).- Flash-point: > 110 °C.- Ignition temperature: 385 °C.- Appearance: Light yellow liquid.- Odour: Perceptible.- Conditions of storage: In the refrigerator, in the dark.- Stability at conditions of storage: 12 months.- Stability at ambient temperature: 6 months.- Date of expiry: 31 December 1998.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Forschungsinstitut für Versuchstierzucht, A-2325 Himberg.- Age at study initiation: Approximately 8 weeks (males) and 12 weeks (females) at the time of administration.- Weight at study initiation: female mean: 164g and male 211g, for further information please see: "Any other information on results incl. tables"- Fasting period before study: Feed was withdrawn the evening before administration of the test substance and was offered again about three hours afterwards.- Housing: Single caging in Makrolon cages type III (39 cm x 23 cm x 15 cm). Wire mesh lids. Sanitation of cages once a week. Bedding material: Aspen wood chips, type "4 HV" (Finn Tapvei Oy, SF-73620 Kortteinen), autoclaved. Bedding material was changed weekly.- Diet: Altromin 1314 forte, gamma irradiated with 25 kGy 60Co, adlibitum (Producer: Altromin GmbH, D-32791 Lage). - Water: Tap water, offered in Makrolon bottles with stainless steel canules, ad libitum.- Acclimation period: 6 days.ENVIRONMENTAL CONDITIONS- Temperature (°C): Average of 22 °C.- Humidity (%): Average of 55 %.- Air changes (per hr): 12 per hour.- Photoperiod: Artificial light from 6 a.m. to 6 p.m.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLECorn oil was used for the suspension of the test substance. The suspension was prepared freshly before administration and was given within 20 minutes afterwards. A peroral administration was performed once in the morning by stomach intubation using a metal gavage. The dose volume was 10 ml per kg body weight. The individual dose volumes were calculated using the body weights determined at the day of the administration.CLASS METHOD (if applicable)- Rationale for the selection of the starting dose: In a range finding study three groups of one male and one female each were dosed with 200, 1000 or 2000 mg per kg body weight. All animals survived for 14 days p.a. Therefore it was decided to perform a limit-test with one dose of 2000 mg per kg body weight. Both animals, dosed with 2000 mg/kg in the preliminary test, were included into the main study.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

7 males and 7 females (including the animals for the rangefinding study.

Control animals:

no

Details on study design:

Observations were performed within the periods 0 - 0.5, 0.5 - 1, 1 - 2, 2 - 4 and 4 - 6 hours after administration (p.a.) of the test substance and then at least once a day for a total of 2 weeks. Observations included but were not limited to changes in skin, fur, eyes, the occurrence of secretions and excretions, autonomic activity, changes in gait, posture and the presence of convulsions.Body weight was determined before administration, 7 days p.a. and 14 days p.a.All animals were killed by CO2 14 days p.a. and subjected to a necropsy including a gross pathological examination.

Results and discussion

Preliminary study:

In a range finding study three groups of one male and one female each were dosed with 200, 1000 or 2000 mg per kg body weight. All animals survived for 14 days p.a. Therefore it was decided to perform a limit-test with one dose of 2000 mg per kg body weight. Both animals, dosed with 2000 mg/kg in the preliminary test, were included into the main study.

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived until the scheduled termination of the study.

Clinical signs:

other: All animals were affected in any way. The findings were:- Central nervous effects:Motor excitation or sedation, catalepsy, tremor and/or hyperalgesia in 3 males and onefemale immediately after administration of the test substance and lasting to a maximum

Gross pathology:

Pale glandular stomach mucosa in one male and in one female animal were noted. All other animals were normal at the post mortem examination.For further details please see: "Any other information on results incl. tables "

Any other information on results incl. tables

 Table 1: Synopsis of the results

Animal	Sex	Dose	Number of animals
No.		(mg/kg)	exposed	affected	dead
1 - 5	m	2000	5	5	0
6 - 10	f	2000	5	5	0

 

Table 2: Body weights and body weight gains Individual data, means and standard deviations SD.

Dose	Animal	Body weight (g)				Body weight gain (g)
(mg/kg) Sex	No.	before administr.	7 days p.a.	14 days p.a.	death	0-7 days p.a.	7-14 days p.a.
2000, m	1	171	244	297	-	73	53
	2	222	292	339	-	70	47
	3	238	305	339	-	67	34
	4	215	283	321	-	68	38
	5	210	272	303	-	62	31
	mean	211	279	320	-	68	41
	SD	25	23	20	-	4	9
2000, f	6	170	205	220	-	35	15
	7	167	199	221	-	32	22
	8	161	200	209	-	39	9
	9	159	202	223	-	43	21
	10	164	195	197	-	31	2
	mean	164	200	214	-	36	14
	SD	4	4	11	-	5	8

 

Table 3: Observations in life.

A grade of severity was recorded where applicable (low - mid - high). 

Findings	Dose (mg/kg), sex	No. of the affected animal	Observation time (p.a.)    first             last	Maximum grade of severity
normal at any time	2000, m	none	-  / -	-
	2000, f	none	-  / -	-
chromodacryorrhoea	2000, m	3	0.5 h / 3 d	low
		4	1 h / 1 d	low
	2000, f	10	2 h / 1 d	low
hyperalgesia	2000, f	6	2 h / 6 h	-
motor excitation	2000, m	1	0.5 h / 6 h	-
	2000, f	6	0.5 h / 1 d	-
piloerection	2000, m	1	0.5 h / 2 d	low
		2	0.5 h / 1 d	medium
		3	0.5 h / 2 d	low
		4	0.5 h / 2 d	low
		5	0.5 h / 1 d	low
	2000, f	6	0.5 h / 10 d	medium
		7	0.5 h / 1 d	low
		8	0.5 h / 1 d	low
		9	0.5 h / 1 d	low
		10	0.5 h / 1 d	low
righting reflex catalepsy	2000, m	2	1 h / 2 h	-
sedation	2000, m	2	0.5 h / 1 d	medium
		3	0.5 h / 1 d	low
tremor	2000, f	6	6 h / 6 h	-

 

Table 4: Necropsy findings

 

SYSTEM Organ, finding	Dose (mg/kg)	Sex	Nos. of the affected animals
Normal	2000	m	2, 3, 4, 5
	2000	f	7, 8, 9, 10
ALIMENTARY SYSTEM
Glandular stomach, mucosa, pale	2000	m	1
	2000	f	6

 

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated informationCriteria used for interpretation of results: EU

Conclusions:

The LD50,oral of "diethyl acetylsuccinate" is higher than 2000 mg/kg body weight in rats. Therefor no classificatio is required according to DSD and CLP.

Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance after a single peroral administration. The guidelines EC 92/69, method B.1., and OECD 401, 1987, were applied.

"Diethyl acetylsuccinate", freshly suspended in corn oil, was administered once orally by stomach intubation to Him:OFA Sprague Dawley rats. The dose was 2000 mg per kg body weight. The dose volume was 10 ml per kg body weight.

The test substance caused transient central nervous effects and signs of reduced well-being at the dose of 2000 mg/kg body weight. No mortality occurred.

The LD50,oral of "diethyl acetylsuccinate" is higher than 2000 mg/kg body weight in rats.