Amides, tall-oil fatty, N,N'-(iminodi-2,1-ethanediyl)bis-, phosphates

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:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples were taken from the single loading rate and the control at the start and at the end of the first and last of the four 24-hour test medium renewal periods.
All samples were stored deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

A water accommodated fraction (WAF) with a loading rate of 5.0 mg/L was prepared. Prior to the start of the test and prior to the start of the test medium renewals, the test medium was prepared by weighing nominal 25 mg of the test item into 5000 mL test water (ISO 6341, half-concentrated). Ultrasonic treatment for 15 minutes was applied and thereafter, the dispersions were stirred for 48 hours at room temperature in the dark. The stirring period of 48 hours was considered reasonable to achieve maximum solubility for the different components of the test item.
Then, the dispersion was filtered through membrane filters (Whatman, Type NC45, pore size 0.45 µm). The filters were pre-conditioned with 300 mL filtrate. The undiluted filtrate was tested as WAF.
The method of preparation of the test media was based on the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, 2000.
- Controls: test water (ISO 6341, half-concentrated)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Breeding culture at the testing laboratory

- Length at study initiation(length definition, mean, range and SD): measured from the mouth to the end of the tail fin, 2.2 ± 0.07 cm
- Weight at study initiation (mean and range, SD): 0.13 ± 0.01 g
- Feeding during test: none

HOLDING AND ACCLIMATION
No medication was applied during holding and acclimatization.
- Acclimation period: ≥ 1 week
- Acclimation conditions (same as test or not): same
- Feeding during acclimation: until one day before the start of the test, the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, TETRA-Werke, 49324 Melle / Germany)
- Health during acclimation (any mortality observed): During the last four weeks prior to the test, no fish died in the test fish batch and all fish were healthy.

FEEDING DURING TEST
none

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

1.25 mM (125 mg CaCO3/L)

Test temperature:

21 - 22 °C

pH:

6.9 - 7.7

Dissolved oxygen:

≥ 7.5 mg/L (≥ 83 % oxygen saturation)

Nominal and measured concentrations:

nominal loading rate: 5.0 mg/L as WAF
measured test concentrations: below the limit of quantification (LOQ) of 1 mg/L in all samples.
The biological results are based on the loading rate of the test item.

Details on test conditions:

TEST SYSTEM
- Test vessel: full glass aquarium, containing five liters test medium
- Type (delete if not applicable): open
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): after 24 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.25 g fish wet weight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified water
- Alkalinity: 0.4 mM
- Ca/Mg ratio: 4:1
- Culture medium different from test medium: no
- Intervals of water quality measurement: water temperature, pH values and oxygen concentrations were measured for each treatment at the start of the test and once daily during the test in the freshly prepared and aged test medium. At the same dates the appearance of the test medium was recorded.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour light to 8-hour dark photoperiod, with a 30-minute transition period
- Light intensity 13 – 18 µE s-1 m-2

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable (limit test/threshold approach)
- Results used to determine the conditions for the definitive study: A limit test based on the threshold approach was performed (OECD Series on testing and assessment No. 126 (2010): Short Guidance on the Threshold Approach for Acute Fish Toxicity). The selected loading rate of 5.0 mg/L corresponded approximately to the lower EL50 value determined in an algae study (OECD 201, EC C.3) and an acute daphnia study (OECD 202, EC C.2). The lower EL50 value was determined in the algae test to be 4.8 mg/L, based on growth rate (EL50 for daphnia was 57 mg/L).

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

EL50

Effect conc.:

> 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

nominal loading rates

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

highest concentration tested in a threshold approach

Key result

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

nominal loading rates

Basis for effect:

other: mortality and sub-lethal effects

Remarks on result:

other:

Remarks:

highest concentration tested in a threshold approach

Details on results:

- Behavioural abnormalities:
- Other biological observations: none; all fish survived and no visible abnormalities were observed.
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Effect concentrations exceeding solubility of substance in test medium: Yes; the solubility of the test item in test water was below the LOQ of the analytical method (< 1 mg/L).

The test medium was a clear solution throughout the test medium renewal periods of 24 hours.

Reported statistics and error estimates:

The NOELR and LL0 were determined directly from the raw data. The LOELR, LL100 and LL50 at the observation times could not be quantified due to the absence of a toxic effect of the test item at the tested loading rate.

Validity criteria fulfilled:

yes

Remarks:

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

Description of key information

effect value for Danio rerio, based on loading rates (OECD 203, EU C.1; threshold approach):

96-h LL50: > 5.0 mg/L (solubility in test water < 1 mg/L)

Additional information

LL50 for freshwater fish: > 5.0 mg/L (threshold approach).

The results indicate that the test material is not toxic to fish up to the limits of its solubility (< 1 mg/L in test water) and a NOEC and thus a PNEC cannot be determined.