Fatty acids, tall-oil, compds. with oleylamine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

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 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted Oct. 2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

adopted May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At the start of the test, two samples (50 mL) were taken from the freshly-prepared control and test media.
After 48 hours, the contents of the test vessels from each group were pooled and two samples (50 mL) were taken.
Samples were stored in a freezer until analysis. Two analyses were performed on each sample.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: water accommodated fractions (WAFs)
At each nominal loading rate, the test medium (WAF) was prepared by adding the required amount of test substance (21.4, 18.8, 20.7, 45.5 or 100 mg) to a volumetric flask (1, 2 or 5L) containing Elendt M4 media before being made up to volume. The mixtures were stirred for approximately 24 hours, after which the stirring was stopped and the contents then left to stand for approximately 17 hours in the dark. An aliquot (ca. 700 mL) was removed from the mid-vessel region at each level to produce the water accommodated fractions (WAF’s) used as the test medium at 4.27, 9.39, 20.7, 45.5 and 100 mg/L.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: in-house culture originating from a strain supplied by the National Institute for Applied Chemical Research (IRCHA), France
- Age at study initiation: < 24 h
- Method of breeding: Stock cultures of Daphnia magna were maintained in glass vessels containing approximately 0.5 to 0.8 litres of Elendt M4 culture medium in a temperature-controlled laboratory at nominally 20 ± 2°C. A photoperiod of 16 hours light : 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase. The culture medium was renewed three times each week.
Cultures were fed daily with a suspension of the unicellular green algae, Pseudokirchneriella subcapitata, to provide nominally 0.1 to 0.2 mg carbon per daphnid, per day, except during the initial three days when a slightly lower ration was given. Culture conditions ensure that the stock animals reproduce by parthenogenesis.
The day before the start of the study, all juvenile Daphnia were removed from the laboratory cultures. The following morning, juveniles produced by the gravid (egg-bearing) adult Daphnia were removed from the culture vessels and held in a separate holding vessel.
- Feeding during test: none

ACCLIMATION
- Acclimation period: not relevant, conditions were the same for maintenance and testing

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

270 mg/L as CaCO3

Test temperature:

19-20 °C

pH:

7.95-8.2

Dissolved oxygen:

91-101 % air saturation value

Nominal and measured concentrations:

nominal loading rates: 4.27, 9.39, 20.7, 45.5 and 100 mg/L as WAF
mean measured TOC levels (after background correction):
- loading rates of 4.27, 9.39, 20.7, 45.5 and 100 mg/L:
test start: 0.016, 0.043, 0.105, 0.182 and 0.336 mg C/L (control mean: 0.69 mg C/L)
after 48 hours: *, 0.198, 0.0870, 0.154 and 0.0525 mg C/L (control mean: 0.932 mg C/L)
geometric mean: not applicable, 0.121, 0.0957, 0.168 and 0.194 mg C/L.

*: At the nominal loading rate 4.27 mg/L measurements at 48 hours were below the limit of quantification of the analytical method after background correction.

Details on test conditions:

TEST SYSTEM
- Test vessel: covered glass dishes containing approx. 100 mL of medium
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 20 mL medium per organism

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised, reverse osmosis water
- Total organic carbon: 0.7 mg C/L
- Alkalinity: 44 mg /L as CaCO3
- Ca/Mg ratio: 3.7:1
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours 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) : numbers of mobile, immobile and floating Daphnia counted approx. 24 and 48 hours after the start of the study

RANGE FINDING STUDY
- Test concentrations: nominal loading rates of 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: 100% immobility after 48 hours at 100 mg/L WAF, no immobility at 1 and 10 mg/L WAF

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

15.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% confidence limits: 13.5-20.7 mg/L; results are expressed in terms of loading rates

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

9.39 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: results are expressed in terms of loading rates

Duration:

48 h

Dose descriptor:

LL100

Effect conc.:

45.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: results are expressed in terms of loading rates

Details on results:

- Immobilised Daphnia: see Table 1
- Behavioural abnormalities: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none

The test media were colourless at all nominal loading rates.

Results with reference substance (positive control):

- 48-hour EC50: 1.3 mg/L (within the range typically obtained in this laboratory)

Reported statistics and error estimates:

Statistical analysis was performed using the SAFEStat LD50 application (version 1.3), SAS 9.1.3 (SAS Institute, 2002).
The “no observed effect loading rate” (NOELR) was derived by direct inspection of the data on the immobility of the animals. An incidence rate of more than 10% was considered to be significant.

Table 1: Cumulative immobilisation

 

Nominal loading rate (mg/L)	Cumulative numbers of immobile Daphnia
	24 hours						48 hours
	R1	R2	R3	R4	total	%	R1	R2	R3	R4	total	%
Control	0	0	0	0	0	0	0	0	0	0	0	0
4.27	0	0	0	0	0	0	0	0	0	0	0	0
9.39	0	0	0	0	0	0	0	0	0	0	0	0
20.7	2	2	0	5	9	45	5	5	3	5	18	90
45.5	5	2	5	5	17	85	5	5	5	5	20	100
100	5	5	5	5	20	100	5	5	5	5	20	100

R: replicate number

Validity criteria fulfilled:

yes

Remarks:

In the control, no Daphnia have been immobilised or trapped at the surface of the water. The dissolved oxygen concentration at the end of the test was ≥60% of the air saturation value at the temperature used.

Description of key information

48-h EC50 (based on loading rates) 15 mg/L for Daphnia magna (OECD TG 202, EU C.2)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

15 mg/L

Additional information

The results indicate that the test material is not toxic to daphnia up to the limits of its water solubility and a NOEC and thus a PNEC cannot be determined.