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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Published in peer reviewed literature, minor restrictions in design and/or reporting but otherwise adequate for assessment

Data source

Reference
Reference Type:
publication
Title:
Quantitative assessment of the embryotoxic potential of NSO-heterocyclic compounds using zebrafish (Danio rerio)
Author:
Peddinghaus et al.
Year:
2014
Bibliographic source:
Reproductive Toxicology 33 (2012) 224– 232

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: DIN 38415-6
Qualifier:
equivalent or similar to guideline
Guideline:
other: OECD. Fish embryo toxicity (FET) test. Draft OECD guideline for the testing of chemicals; 2006.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
2-methylpyridine
EC Number:
203-643-7
EC Name:
2-methylpyridine
Cas Number:
109-06-8
Molecular formula:
C6H7N
IUPAC Name:
2-methylpyridine
Details on test material:
- Name of test material (as cited in study report): 2-Methylpyridine

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Chemical name of vehicle: 100% dimethyl sulfoxide (DMSO; Sigma–Aldrich)
- Concentration of vehicle in test medium: The maximum content of DMSO during the test was below 0.25%.
- Solvent control: In order to exclude adverse effects caused by DMSO a solvent control of 0.25% was used.
- Negative control: Artificial water in the absence of the chemical.
- Positive control: 3.7 mg/L 3,4-dichloroaniline dissolved in millipore water.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Age at study initiation: The FET is initiated as soon as possible after fertilisation of the eggs and no later than 3 hpf (128-cell stage)
- Selection of eggs: To identify fertilised eggs, a binocular with a minimum magnification of 25×should be used. Freshly spawned eggs exhibit the following structures: The chorion surrounds the perivitelline space, which contains the yolk. The blastodisc is located at the animal pole of the yolk.

BREEDING:
- Method of breeding: Groups of 20, 3-month old zebrafish with a ratio of 3:2 (males:females) were used for egg production. Mating, spawning and fertilisation take place within 30 min after the onset of light in the morning.
- Housing: Spawning trays consisting of a flat basin and a metal net covering to which artificial plants are attached are placed into the aquaria. The artificial plants serve as a breeding stimulant and substrate while the metal net covering prevents the fish from feeding on their own spawn.
- Food type: artificial diets (e.g., TetraMinTM flakes; Tetra, Melle, Germany) and live nauplius larvae of Artemia sp. once daily ad libitum.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Nominal and measured concentrations:
- Nominal concentration: 5 concentrations ranging from ca. 20 mg/L to 200 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 24-well plates
- Type: covered with a gas-permeable foil (Renner, Darmstadt, Germany)
- Material, size, headspace, fill volume: 2 mL freshly prepared test solutions and controls per well.
- No. of organisms per vessel: ten eggs that had reached the 8-cell stadium
- No. of vessels per concentration: two independent replicates with ten embryos per test concentration
- No. of vessels per control: two independent replicates with ten embryos per control

EFFECT PARAMETERS MEASURED:
Evaluation of the test was carried out with an inverted microscope at magnifications of 40× and 100×. After an exposure time of 48 h, lethal endpoints (coagulation of the embryo, non-detachment of the tail, and non-detection of the heartbeat) were recorded


Reference substance (positive control):
yes
Remarks:
3.7 mg/L 3,4-dichloroaniline

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
103.51 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Reported statistics and error estimates:
In order to calculate LC50 values, the results of the FET were used to create concentration-response curves.

Any other information on results incl. tables

Sublethal observations / clinical signs:

The experimental, nominal LC50 (103.51 mg/L) was compared with predicted value (266.89 mg/L) obtained from the ECOSAR program. This was done using the toxicity ratio (TR): LC50predicted/LC50 experimental. The TR value was 2.58.

Applicant's summary and conclusion