Fatty acids, tall-oil, reaction products with pentaethylenehexamine

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:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Analytical determinations of the test substance concentrations was based on dissolved organic carbon (DOC). DOC determinations of the test substance loading rate were conducted at the beginning of the exposure, after 24, 48, and 72 h, and at the end of the exposure. Each replicate was measured individually.

Vehicle:

no

Details on test solutions:

Prior to the definitive test, non-GLP screening tests were performed to evaluate the stirring duration for the preparation of the water soluble fraction (WSF), the mode of filtration and the method for the DOC determination.
A stock dispersion with a loading rate of 100 mg/l was prepared and stirred for 24, 48, 72 and 96 h. Two alternative methods of the initial filtration were compared for the dissolved organic carbon (DOC) determinations: 1) filtration with a MILLIPORE AP15 glass fiber filter, and 2) filtration with a 0.45 µm cellulose acetate membrane sterile syringe filter. Subsequently, two methods were compared for the measurement of all the samples: 1) measurement with acidification/sparging (NPOC mode), and 2) indirect measurement by using the TOC mode.
The nominal loading rate of 100 mg/l resulted in a turbid dispersion. Filtration with a glass fiber filter and an indirect measurement of the DOC using the indirect TOC mode proved to be the preferred option.
After filtration, DOC concentrations around 10 mg/l were measured after 24 h. This concentration remained essentially constant over the 96 h stirring time. Therefore, a 72 h stirring time for the WSF preparation was used.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test organism: Danio rerio (Zebrafish)
Culturing: Population of parental and young fish are held in either 200 l or 40 l glass aquaria. The bottom of the aquaria is covered with gravel (≤1.5 cm).
Illumination: 16 h a day
Temperature: 21-25 °C, controlled by a thermometer.
Medium: Continuously aerated natural water of the town of Schlieren.
Feed: The fish are fed commercially prepared diet Sera micropan and Sera vipan (Sera apanristic GmbH, D-52525 Heinsberg, Germany).

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Total Hardness
°f H: 16.1 - 20.9
mmol/l: 1.61 - 2.09
mg/l: 161 - 209

CO3 – Hardness
°f H: 14.6 - 18.8
mg/l: 146 - 188

Rest – Hardness
°f H: 1.5 - 2.1
mg/l: 15 - 21

Test temperature:

21 - 22 °C

pH:

8.4 - 8.8

The pH values of the blank control were in the range of 8.4-8.8 and not in the recommended preferred pH range of 6.0-8.5. It can be concluded, that the deviation had no influence as no adverse effects were observed in the blank control and in the treatments at the loading rates of 1.0, 1.5, 2.0 and 2.9 mg/l.

Dissolved oxygen:

6.9 - 7.7 mg/L

Nominal and measured concentrations:

In a 1st definitive test performed as a limit test, all seven fish died, and therefore, the study had to be repeated as a full study with nominal concentrations of 1.0, 1.5, 2.0, 2.9, 4.0 mg/L.

Details on test conditions:

Test vessel: Glass aquaria of 2 l (14.5 x 14.0 x 9.5 cm) filled with 1.0 liter of water.
Test medium: Continuously aerated natural water of the town of Schlieren
Oxygen concentration: The total dissolved oxygen content is kept >60% of the saturation value with aeration via Pasteur pipettes.
Number of fish: 7 fish per test concentration and control (total length of 1-3 cm)
Light: 16 h photoperiod a day, supplied by overhead white fluorescent tubes, no transition period.
Temperature (target): 21-25 °C, controlled by a thermometer
Test type: Static exposure conditions
Test duration: 96 h

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 2.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 2.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

DOC analysis of the test substance concentration was conducted at the beginning and after 24, 48, 72 and 96 h of exposure. All net DOC concentrations (after subtraction of the DOC of the blank control) were below the quantification limit of the analytical method (LOQ) of 0.5 mg C/l. Thus, the analysis was based on the loading rate.

Description of key information

Acutely toxic to fish. 

Additional information

Short-term toxicity to fish was assessed in an OECD 203 guideline study according to GLP. Due to the results of a range finding study the concentrations in the definite test were 1.0, 1.5, 2.0, 2.9 and 4.0 mg/L. The results are based on the loading rates due to the uvcb character of the test substance. After 96h of exposure, the lethal concentration (LC50) of the test substance was calculated to be 3.50 mg/L with respect to the loading rate. The LC0 and NOEC were calculated to be >= 2.9 mg/L, respectively.