Oxydiethylene dinitrate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1.5. - 5.6.1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study report which meets basic scientific principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Acute exposure, oral toxicity. In: Health effects test guidelines. Washington, DC: EPA, Aug 1982, EPA 560/6-82-001

Deviations:

yes

Remarks:

See Overall remarks

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Acute oral toxicity study (ADL and LD50). LAIR Standard Operating Procedure OP-STX-36, Letterman Army Institute of Research, Presidio of San Francisco, CA. 15 Jun 1984

Deviations:

yes

Remarks:

See Overall remarks

GLP compliance:

yes

Test type:

other: modified up-and-down procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Oxydiethylene dinitrate

EC Number:

211-745-8

EC Name:

Oxydiethylene dinitrate

Cas Number:

693-21-0

Molecular formula:

C4H8N2O7

IUPAC Name:

2-[2-(nitrooxy)ethoxy]ethyl nitrate

Details on test material:

- Name of test material (as cited in study report): Diethyleneglycol Dinitrate (DEGDN)
- Physical state: pale yellow liquid
- Analytical purity: 96.6%
- Impurities (identity and concentrations): not identified
- Lot/batch No.: RAD84MOO1S214
- Stability under test conditions: stable

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bantin and Kingman, Inc, Fremont, CA
- Weight at study initiation: 168 - 279 g
- Fasting period before study: overnight
- Housing:individually in stainless steel wire mesh cages in racks equipped with automatic flushing dumptanks, no bedding was used
- Diet (e.g. ad libitum): Certified Purina Rodent Chow Diet 5002 (Ralston Purina Company, St Louis, MO)
- Water (e.g. ad libitum): from central line
- Acclimation period: 12-19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.3 - 25.6 °C
- Humidity (%): 41 - 55%
- Photoperiod (hrs dark / hrs light): 12 hours of light per day

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 ml/ kg b.w. (1.98-2.79 ml/males, 1.68-2.4 ml/females)

DOSAGE PREPARATION (if unusual): DEGDN was received as a solution containing 18% acetone. The acetone was removed with a rotary evaporator. DEGDN was then suspended in corn oil to form a viscous yellow oily suspension.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The results of an approximate lethal dose (ALD) determination suggested that the median lethal dose (MLD) was between 500 and 1000 mg/kg. Based on these data, test doses were selected.

Doses:

Males: 794, 891, 1000, 1120, 1260 mg/kg
Females: 631, 794, 891, 1000, 1260 mg/kg

No. of animals per sex per dose:

10 males and 10 females per dose

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations:
- mortality and signs of acute toxicity: on the day of dosing 1, 2, 4, 6 hours after dosing and then daily
- weighting: weekly
- Necropsy of survivors performed: yes

Statistics:

The LD10, LD50 and LD90 were derived by probit analysis using the maximum likelihood method. The program, PROBIT, Model MV8000 was used to plot the probit curve and lethal dose values.

Results and discussion

Effect levels

Effect levels 1

Sex:

male

Dose descriptor:

LD50

Effect level:

990 mg/kg bw

95% CL:

924 - 1 060

Effect levels 2

Sex:

female

Dose descriptor:

LD50

Effect level:

753 mg/kg bw

95% CL:

654 - 826

Mortality:

Forty-eight animals died as a result of the dosing. Twenty-five deaths (52.1%) occured within 24 hours of dosing. Fifteen deaths (31.2%) occured between 24 and 48 hours after dosing. The remaining 8 deaths (16.7%) occured between 18 and 120 hours after dosing.

Clinical signs:

other: Behavioral disturbances (73 of 79 animals) - inactivity, twitching, tremors, hypertonia, irritability, jumping and ataxia. They were first observed 1 to 3 hours after dosing and, with exception of inactivity and irritability, were no longer observed 72 ho

Gross pathology:

The mortalities which occured after dosing appear to have been caused by the test compound.
A dose-response effect on deaths was apparent in both male and female rats.
Multifocal necrohemorrhagic gastritis - probably treatment related.
Liver vacuolization - maybe related to treatment or maybe not
Renal autolysis, renal autolysis and renal necrosis

Applicant's summary and conclusion

Conclusions:

Diethyleneglycol dinitrate produces signs of neurotoxicity in addition to standard symptoms of nitrate ester poisoning. Calculated MLD values
were 990.4 ± 30.0 mg/kg in male Sprague-Dawley rats and 753.1 ± 35.9 mg/kg in female Sprague-Dawley rats.

Executive summary:

The acute oral toxicity of the nitrate ester, diethyleneglycol dinitrate (DEGDN), was determined in male and female Sprague-Dawley rats by using the oral gavage single-dose method. The median lethal dose (MDL) was 990.4 +/- 30 mg/kg for male rats and 753.1 +/- 35.9 mg/kg for female rats. Clinical signs produced by DEGDN included twitching, tremors, hypertonia, squinting, lacrimation, depression of grasping and righting reflexes, jumping, increased startle reflex, ataxia, cyamosis, inactivity, and prostration. The extent of the neurotoxic component of this clinical signs profile suggests that DEGDN possesses additional pharmacological properties to those routinely associated with the nitrate esters. The duration of clinical signs was acute. Most animals were exhibiting signs by 2 hours after dosing and had either died or cleared by 72 hours after dosing.