Ethylene dinitrate

Long-term toxicity to fish

Administrative data

Endpoint:

fish, juvenile growth test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Test conducted according to Draft 10 of the proposed ASTM guideline for conducting early life-stage toxicity tests with fish.

Deviations:

not specified

GLP compliance:

no

Remarks:

Study is based on a standard national protocol with no variations or quality control issues and is very well-documented. Note that it is unlikely that the laboratory was operating according to all GLP principles at the time of the testing.

Test material

Test material information

Constituent 1

Reference substance name:

Glycerol trinitrate

EC Number:

200-240-8

EC Name:

Glycerol trinitrate

Cas Number:

55-63-0

Molecular formula:

C3H5N3O9

IUPAC Name:

propane-1,2,3-triyl trinitrate

Specific details on test material used for the study:

NG dissolved in JHU/APL well water at 1g/L was obtained from the US Naval Ordnance Section, Indian Head, MD.
Reverse phase HPLC determined that no impurities were present during testing.
Stability of stock solution was checked periodically during the study period.

Test solutions

Vehicle:

no

Details on test solutions:

NG dissolved in JHU/APL well water at 1 g/L was obtained from the US Naval Ordnance Section, Indian Head, MD.
Reverse phase HPLC determined that no impurities were present during testing.
Stability of stock solution was checked periodically during the study period.

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

Test species was fathead minnow (Pimephales promelas) juveniles. They were obtained from the JHU/APL culture maintained at 25 °C.
Embryos were collected on spawning substrates placed in the culture tanks.
No embryos used if they exhibited signs of disease within 10 days preceding test start or if >3% died preceding start of test.
40 fish were randomly assigned to each control or treatment replicate.
Post-hatch fish were fed brine shrimp nauplii 3x/2x per day (weekday/weekend) starting on day 2 post-hatch

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

28 d

Post exposure observation period:

Raw data are presented for each replicate.
Embryos were observed 3 times daily and those exhibiting fungus were removed and not included in endpoint estimates.
Post-hatch fish were observed daily for developmental abnormalities and mortality.
Length, blotted wet weight and dry weight were determined for surviving fish at end of test.

Test conditions

Details on test conditions:

Flow through system.
Chronic
Negative control with 2 replicates was run concurrently.
2 replicates per treatment
Exposure period 4 days in embryonic stage followed by 28 days post-hatch exposure. Exposure pathway was aqueous
Geometric dilution series of 5 concentrations was tested.

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

4 d

Dose descriptor:

LOEC

Effect conc.:

0.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: Hatching success

Effect concentrations 2

Duration:

4 d

Dose descriptor:

NOEC

Effect conc.:

0.12 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: Hatching success

Effect concentrations 3

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

0.33 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 4

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Endpoints: hatching success, growth, development and survival
Hatch success LOEC = 0.2 mg/L NOEC = 0.12 mg/L
Growth LOEC = 0.33 mg/L; NOEC = 0.2 mg/L
Survival LOEC = 0.33 mg/L; NOEC = 0.2 mg/L.
Malformations observed >0.2 mg/L and in one control fish.

Reported statistics and error estimates:

Statistical analysis of concentration data and organism response was based on mean measured concentrations of NG.
Survival and hatch success data were arc-sin sqrt transformed.
Hatch success, growth and survival were tested for normality (Chi-square) and equal variance (Bartlett’s test) and then analyzed with ANOVA or a non-parametric Kruskal-Wallis test. Where a significant difference was detected, Dunnet’s test was used to compare treatments.