Reaction mass of (2E)-Tridec-2-enenitrile and (2Z)-Tridec-2-enenitrile and (3E)-Tridec-3-enenitrile and (3Z)-Tridec-3-enenitrile

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2008-01-25 to 2008-03-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Preparation of samples: 10 mL of each sample and 0.1 mL Methanol were filled into a 20 mL headspace vial that was crimped and afterwards analyzed via headspace-SPME.
- Sample storage conditions before analysis: All samples were stored at 7 ± 2 °C until start of analysis.
- Sampling method: The test item concentrations were analytically verified via HPLC from freshly prepared media after 0 and 72 h and from 24 h old media after 24 and 96 h.
- Preparation of standard: A stock solution of the standard of 1000 mg/L in acetonitril was diluted to 8 concentrations with acetonitril and used for calibration. 0.1 mL of each standard dilution was pipetted into a 20 mL headspace vial filled with 10 mL dilution water.

Vehicle:

no

Details on test solutions:

- Test solution: Dilution water according to the guideline. Tap water of local origin was used for holding and testing. The water was filtered with an activated charcoal and aerated for at least 24 hours to remove chlorine.
- Nominal water parameters:
Total hardness: 10 - 250 mg CaCO3/L
pH-value: 6.0 - 8.5
The water was analyzed biannual according to German tap water Regulation.
- Controls: Dilution water according to OECD guideline
- Evidence of undissolved material: No

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Danio rerio (zebrafish)
- Source: AQUARIUM AM AEGI, Volgersweg 6, D-30175, Hannover, Germany. All fish used in the test were originated from the same delivery of the supplier.
- Length at study initiation: Average body Iength at the test start was 2.06 cm.
- Fish density in the tanks was less than 1 g fish per litre test solution.
- Weight at study initiation: Average body weight at the test start was 0.39 g.
- Feeding during test: The fish were not fed during the test.

ACCLIMATION
- Acclimation period: At least 12 days
- Acclimation conditions: Holding was performed at the testing facility at 23 ± 2 °C and diffuse light (0.1-10 µmol photons*m-2*s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value.
- Type and amount of food: The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started.
- Feeding frequency: Food was provided 3 times per week.
- Health during acclimation: The mortality was lower than 5 %. No disease treatments were administered throughout holding and testing.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

None

Hardness:

59 – 64 mg/L as CaCO3

Test temperature:

23 +/- 2 °C

pH:

- Test solutions: 7.18 – 7.25
- Holding water: 6.0 – 8.5

Dissolved oxygen:

The oxygen saturation in the old solution was determined to be in a range between 71 – 84 %. The oxygen saturation in the new solution was determined to in a range between 96 and 100 %: The dissolved oxygen concentration was more than 80 % of the air saturation value.

Nominal and measured concentrations:

- Nominal concentration: satuarted solution (limit test)
- Measured concentration: The mean measured concentration was determined to be 0.164 mg/L. The recovery rates of the test item in the old media were in the range of 27 to 30 %. Therefore all effect values are given based on the mean measured concentration of the test item. Response of blank values of test medium control samples was lower than 30 % of LOQ.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass-aquaria loosely covered by glass tops were used.
- Test volume: About 4 L per vessel.
- Renewal rate of test solution (frequency/flow rate): daily
- Aeration: No aeration was provided with regard of the high volatility of the test item.
- No. of organisms per vessel: 7
- Introduction of fish: Fish were introduced randomly to individual replicates.
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: Fish density in the tanks was less than 1 g fish per litre test solution.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water according to OECD guideline. Tap water of local origin was used for holding and testing. The water was filtered an activated charcoal and aerated for at least 24 hours to remove chlorine.
- Stock solution: A saturated solution was prepared with the slow stirring method. A Water Accommodated Fraction was used. 100 mg of the test substance was applied to 5 L test medium in a brown glass closed bottle and mixed with a magnetic stirrer for 24 h at 20 °C ± 2 °C. Care was taken to avoid turbulence during stirring. The saturated solution was filled into the test vessels. This procedure was carried out daily to provide a fresh saturated solution for each day of exposure.
- Culture medium different from test medium: No
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured in all vessels after beginning the test and every 24 h (from old and new media).
- Total hardness of the water was determined at the beginning out of the test control. During the test the room temperature was recorded continuously with a thermo-hygrograph.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 0.1 - 10 µmol photons*m-2*s-1
- Light intensity: A natural photoperiod occurred during the test.

EFFECT PARAMETERS MEASURED:
- Determination of the acute toxicity of the test substance to zebrafish with the saturated solution under semi-static conditions. The saturated solution will be tested as limit concentration.
- Criteria effects: Fish were considered dead if there was no visible movement (e.g. gilt covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.).
- Measurement of fish size: At test start body weight and total length of a representative number of 7 test fish was determined. These fish were not used for testing.

TEST CONCENTRATIONS
- Range finding study: No range finding study was performed

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.164 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % C.L. not determined.

Details on results:

The recovery rates of the test item in the old media were in the range of 27 to 30 %. Therefore all effect values are given based on the mean measured concentration of the test item. Response of blank values of test medium control samples was lower than 30 % of LOQ.The 96 h-LC50 was determined to be greater than 0.164 mg/L and the 96 h-NOEC was determined to be 0.164 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

In a semi-static acute toxicity test to fish, Danio rerio (zebrafish) were exposed for 96 hours according to OECD guideline 203 (1992). The LC50 was determined to be greater than 0.164 mg/L (measured concentration).

Executive summary:

The acute toxicity of the test substance to the Danio rerio (zebrafish) was investigated in a 96-hour semi-static test according to the OECD Guideline 203 (1992). The test substance was tested in a nominal limit concentration of 100 mg/L. The test was performed with an aqueous extract (eluate) of the test substance (WAF). The mean measured concentration was determined to be 0.164 mg/L. The recovery rates of the test item in the old media were in the range of 27 to 30 %. Therefore all effect values are given based on the mean measured concentration of the test item. No mortality or other adverse effects were observed up to the solubility limit of the test substance. The 96 h-LC50 was determined to be greater than 0.164 mg/L and the 96 h-NOEC was determined to be 0.164 mg/L. The validity criteria were fulfilled by the test.

Description of key information

 In a semi-static acute toxicity test to fish, Danio rerio (zebrafish) were exposed for 96 hours according to OECD guideline 203 (1992). The LC50 was determined to be greater than 0.164 mg/L (measured concentration).

Additional information

The acute toxicity of the test substance to the Danio rerio (zebrafish) was investigated in a 96-hour semi-static test according to the OECD 203 (1992). The test substance was tested in a nominal limit concentration of 100 mg/L. A WAF was performed. The mean measured concentration was determined to be 0.164 mg/L. The recovery rates of the test item in the old media were in the range of 27 to 30 %. Therefore all effect values are given based on the mean measured concentration of the test item. The 96 h-LC50 was determined to be greater than 0.164 mg/L. The validity criteria were fulfilled by the test.