Fatty acids, C18-unsatd., trimers, reaction products with triethylenetetramine

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

Two samples (50 mL) were taken from the freshly prepared control and test media at 0 and 72 hours and from the expired control and test media at 24 and 96 hours.
Samples were stored frozen until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: water accommodated fractions (WAFs)
8.54, 18.8, 41.4, 91 and 200 mg each was dispersed in dechlorinated water in glass aspirators (20 L). The contents of each vessel were stirred for approximately 24 hours iand then left to stand for approximately 24 hours. Aliquots (18 L) were then removed mid vessel to provide the Water Accomodated Fractions (WAF).

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source:commercial fish farm (UK)
- Length at study initiation: 5.01 cm (mean total length)
- Weight at study initiation: 1.19 g (mean wet weight)
- Feeding during test: none

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): same
- Type and amount of food: commercial fish food (TROUW (UK) Ltd. Nutra Fry 02); amount equivalent to 1.5-2% of the total wet-weight of fish in the holding tank; no food was given during the 20-hour period immediately before exposure.
- Health during acclimation: <1% mortality

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

154 mg/L as CaCO3

Test temperature:

15-16°C

pH:

8.1-8.5

Dissolved oxygen:

86-105% air saturation value

Nominal and measured concentrations:

nominal loading rates: 0.427, 0.939, 2.07, 4.55 and 10 mg/L as WAF
measured TOC levels:
- loading rates of 0.427 to 4.55 mg/L: below the limit of quantification (after background correction)
- loading rate of 10 mg: 2.04 mg C/L in fresh media and 2.95 mg C/L in expired (24-hour old) media (mean: 2.5 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria, containing 18 litres of medium to a dephth of 16.5 cm
- Type: open
- Aeration: yes
- Renewal rate of test solution: daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.46 g body weight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory tap water, dechlorinated and softened by passage through a water purification system consisting of activated carbon filter, water softener and reverse osmosis treatment.
- Total organic carbon: 0.6-1.9 mg C/L
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light : 8 hours dark with periods of subdued lighting at the beginning and end of each light phase

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and sub-lethal effects; recorded at approx. 2, 4, 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study
- Test concentrations: nominal loading rates 0.1, 1, 10 mg/L
- Results used to determine the conditions for the definitive study: 100% mortality after 24 hours at 10 mg/L; no mortality or sub-lethal effects after 96 hours at 1 mg/L

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence limits: 1.2-2.1 mg/L; results are expressed in terms of loading rates

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

0.43 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: results are expressed in terms of loading rates

Duration:

96 h

Dose descriptor:

LOELR

Effect conc.:

0.94 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

behaviour

Remarks:

and morphology

Remarks on result:

other: results are expressed in terms of loading rates

Duration:

96 h

Dose descriptor:

LL0

Effect conc.:

0.94 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: results are expressed in terms of loading rates

Duration:

96 h

Dose descriptor:

LL100

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: results are expressed in terms of loading rates

Details on results:

- Other biological observations: lethargy , hyperventilation and darkened pigmentation at nominal loading rates of ≥0.939 mg/L
- Mortality of control: zero
- Other adverse effects control: none

The WAFs at loading rates of 0.427 and 0.939 mg/L were colourless dispersions. Between the nominal loading rates of 2.07 to 10 mg/L, the WAFs ranged from slightly hazy to grey dispersions, which remained unchanged throughout the test.

Reported statistics and error estimates:

Statistical analysis was performed using the SAFEStat LD50 application (version 1.3), SAS 9.1.3 (SAS Institute, 2002) and the nominal loading rates.
The “no observed effect loading rate” (NOELR) was derived by direct inspection of the data for lethal and treatment-related-effects. An incidence rate of more than one affected fish out of seven is considered to be significant.

Validity criteria fulfilled:

yes

Remarks:

mortality of control animals <10%; concentration of dissolved oxygen in all test vessels >60% saturation

Description of key information

96-h LL50 (based on loading rates)  1.6 mg/L for Oncorhynchus mykiss (OECD 203, EU C.1)

Additional information

LL50 for freshwater fish: 1.6 mg/L