Ethyl diethoxyacetate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 23 September 1980 to 23 March 1981

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant, comparable to guideline study with deviations not affecting the validity of the study. Study Report very well detailed.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1981

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

yes

Remarks:

see "principles of method if other than guideline"

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

see "principles of method if other than guideline"

Principles of method if other than guideline:

Study perfomed prior to the adoption of guidelines. Overall, the study complies with the recommendations of the EU Method B.1 and OECD 401 guidelines except for the following: The minimum acclimatation period was 3 days instead of 5 days. The maximum volume of test material applied was 25 mL/kg instead of the recommended 10 mL/kg for rodents. Only hydric fasting was performed prior to dosing. In addition, according to the nowadays ethical considerations, the doses selected for the main study should not have included the dose causing 100 % mortality.

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl diethoxyacetate

EC Number:

227-999-8

EC Name:

Ethyl diethoxyacetate

Cas Number:

6065-82-3

Molecular formula:

C8H16O4

IUPAC Name:

ethyl 2,2-diethoxy-acetate

Test material form:

other: liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Centre d'élevage IFFA-CREDO, 69210 Saint Germain sur l'Arbresle
- Age at study initiation: no data
- Weight at study initiation: 120-150 g
- Fasting period before study: only hydric fasting was peformed 16 hours prior to the administration of the test material.
- Housing: animals (5/cage), were housed in polycarbonate cages with stainless steel lid sized 42 cm x 27 cm x 15 cm. Cages were cleaned and sterilized once a fortnight, according to the Standard Operating Procedures of the testing laboratory.
- Bedding: Sawdust (non resinous and non African wood), dusted and autoclaved. Litter was changed once or twice a week.
- Diet: standardised pellets (reference 890 from NAFAG CH., Switzerland, 9202 GOSSAU SG, with complete composition attached to the Study Report). The residue analysis was performed and communicated by the diet supplier.
- Water: ad libitum, in 500 or 800 mL bottles made of polypropylene with rubber stopper and stainless steel pipette. The bottles were washed and sterilized once a fortnight. The drinking water was sterilized by membrane filtration (FG, Millipore 0.2µ). The microbiological analysis was perfomed by the testing laboratory, and the chemical analysis by the municipal laboratory (76000 Rouen).
- Minimum acclimation period: 3 days

ENVIRONMENTAL CONDITIONS
- Cleaning: the animal unit rooms were cleaned according to the Standard Operating Procedures of the testing laboratory.
- Temperature (°C): 20-25
- Humidity (%): 30-80
- Air changes (per hr): 11
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 25 ml/kg

DOSAGE PREPARATION (if unusual): not applicable

RATIONALE FOR THE SELECTION OF DOSES:
A range finding test was performed: One male and one female per dose level were exposed to the following 3 doses: 5, 15 and 25 mL/kg bw (ca. 4950, 14850 and 24750 mg/kg bw). Clinical signs and mortalities were recorded up to 7 days post single exposure. No mortality was obserevd at the lowest dose level. 50% mortality was recorded at the medium dose (male died at Day2), and 100% mortality was observed for the highest dose level (male died at Day 1 and female at Day 2).

Doses:

5, 10, 13, 15 and 25 mL/kg bw corresponding to ca. 4950, 9900, 12870, 14850 and 24750 mg/kg bw .

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- A single oral administration was performed by gavage (Day 0).
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
1) Mortality was checked daily during the study and recorded at Day 0 (1 and 3 hours after administration of the test material), and then at Days 1, 4, 7 and 14.
2) Animals were weighted at Days 0, 4, 7 and 14 after oral administration of the test material, and weights were recorded.
3) Clinical signs: observations were performed during the first and the third hour following the test material administration, and then daily during the 14 days study period.
- Necropsy of survivors performed: yes. Macroscopic examinations were performed on main organs (liver, kidneys, heart, adrenals, spleen and stomach) of animals that died during the study, and animals that survived at the end of the study (Day 14), that were CO2-euthanised (Air Liquide, 27000 Evreux). Histopathological examinations were performed if required.
- Other examinations performed: no further examinations performed.

Statistics:

The oral LD50 and its confidence limits were determined according to the Litchfield and Wilcoxon method (J. Pharmacol. Exp. Therap., 1949, 96, 99-113).

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 14 850 mg/kg bw

Based on:

test mat.

95% CL:

> 12 573 - < 17 523

Remarks on result:

other: calculated based on the density of test material

Effect levels 2

Sex:

male/female

Dose descriptor:

LD50

Effect level:

15 mL/kg bw

Based on:

test mat.

95% CL:

> 12.7 - < 17.7

Mortality:

- 5 mL/kg bw: 0%
- 10 mL/kg bw: 20%
- 13 mL/kg bw: 10%
- 15 mL/kg bw: 60 %
- 25 mL/ kg bw: 100 %

Detailed recording of mortalities is presented in Table 1.

Clinical signs:

other: - 5 mL/kg bw: During the first hour post dosing, animals were sleepy. This effects was fully reversed 3 hours post dosing. - 10 mL/ kg bw: 15, 30 and 60 minutes post dosing, slight drowsiness was noted in all animals. At Day 1, piloerection was obsereved

Gross pathology:

Animals that died during the study showed the following macroscopic findings:
- Discoloration areas in liver, spleen and/or kidneys
- Food stasis

Other findings:

No further findings

Any other information on results incl. tables

Table 1 :Mortality

Doses (ml/kg)	Number of animals	Cumulative mortality
		J0 1h	J0 3h	J1	J4	J7	J14	% of mortality
5	5 males 5 females	0 0	0 0	0 0	0 0	0 0	0 0	0
10	5 males 5 females	0 0	0 0	1 0	2 0	2 0	2 0	20
13	5 males 5 females	0 0	0 0	0 0	1 0	1 0	1 0	10
15	5 males 5 females	0 0	0 0	2 1	4 2	4 2	4 2	60
25	5 males 5 females	0 0	3 2	4 4	5 5	5 5	5 5	100

Table 2 :Weight evolution of animals

Doses (ml/kg)	J0		J4		J7		J14
	Number of animals	Mean ± SD (g)	Number of animals	Mean ± SD (g)	Number of animals	Mean ± SD (g)	Number of animals	Mean ± SD (g)
5	10	145 ± 7	10	190 ± 13	10	201 ± 17	10	224 ± 17
10	10	140 ± 6	8	160 ± 10	8	174 ± 13	8	211 ± 19
13	10	124 ± 6	9	142 ± 5	9	159 ± 11	9	188 ± 16
15	10	134 ± 7	4	150 ± 9	4	174 ± 13	4	208 ± 23
25	10	129 ± 7	0		0		0

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions of this study, the oral LD50 of the test material was determined to be 15 mL/kg (ca. 14860 mg/kg). Thus, the test material is not classified as hazardous for acute oral toxicity according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Dangerous Substances Directive 67/548/EEC (DSD) criteria.

Executive summary:

This GLP-compliant study was performed to assess the acute oral toxicity of the test material, according to a method comparable to EU Method B.1 and OECD Guideline 401.

 

Material and methods

The test material was administered unchanged by gavage to 5 groups of 10 Wistar rats (5 animals/sex/ dose), at the following doses determined in a range finding test: 5, 10, 13, 15 and 25 mL/kg bw corresponding to ca. 4950, 9900, 12870, 14850 and 24750 mg/kg bw.

Animals were monitored during the 1^stand the 3^rdhour following the test material administration, and then daily during the 14 days study period. Mortality was checked daily and animals were weighted at Days 0, 4, 7 and 14. Necropsy of survivors and died animals was performed. Macroscopic examinations were carried out on main organs (liver, kidneys, heart, adrenals, spleen and stomach), and histopathological examinations were performed if required.

 

Results

The weight evolution of animals was typical of the species used in this study.

The mortalities (%) of the tested doses (5, 10, 13, 15 and 25 mL/kg bw) were as follows: 0, 20, 10, 60 and 100.

The clinical signs ranged from a transient drowsiness at the lowest dose, to drowsiness, strong lethargy, breathing decrease, slight loss of equilibrium and slight lachrymation, with all animals dying at the highest dose.

The macroscopical examination of animals that died during the study showed discoloration areas in liver, spleen and/or kidneys, and food stasis.

 

Conclusion

Under the experimental conditions of this study, the oral LD50 of the test material was determined to be 15 mL/kg (ca. 14860 mg/kg). Thus, the test material is not classified as hazardous for acute oral toxicity according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Dangerous Substances Directive 67/548/EEC (DSD) criteria.