Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 Dec - 20 Dec 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

EC Number:

947-862-2

Molecular formula:

not applicable, substance is UVCB

IUPAC Name:

Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations incl. control
- Sampling method: Samples were taken at the beginning and end of exposure period.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test sample (0.150 g) was dissolved in dilution water (1500 mL) to prepare the stock solution of 100 mg/L. Required volumes of the stock solution and dilution water were mixed and stirred in a preparation container to prepare the test solution which was divided into each test vessel. The 100 mg/L test solution was prepared by direct addition.
- Eluate: no
- Differential loading: no (except for the highest test concentration)
- Controls: yes, test medium control
- Evidence of undissolved material: no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: The University of Sheffield, UK (obtained 9 July 1990)
- Age of daphnids: < 24 h
- Feeding during test: none

ACCLIMATION
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: Chlorella vulgaris of 0.1-0.2 mg C (Organic carbon content)/day.
- Feeding frequency: daily

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

32 mg/L

Test temperature:

19.9 - 20.2 °C

pH:

7.8 - 8.0

Dissolved oxygen:

8.7 - 9.0 mg O2/L

Conductivity:

13 mS/m

Nominal and measured concentrations:

nominal: control, 9.53, 17.1, 30.9, 55.6, 100 mg/L
Measured (geometric mean): < LOD, 8.65, 16.1, 29.6, 52.1, 97.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beaker covered with a transparent plastic lid (incubated in a water bath with automatic warming/cooling unit)
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water
- Culture medium different from test medium: no
- Intervals of water quality measurement: Water quality was measured in regular intervals (temperature, O2, pH).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h darkness
- Light intensity: room light

EFFECT PARAMETERS MEASURED
- Immobilisation was measured after 24 and 48 h.

RANGE-FINDING STUDY
- Test concentrations: 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: 100% immobilisation was recorded at 100 mg/L whereas no immobilisation was observed at 1 and 10 mg/L. The measured test concentrations during range finder were in the range of 93.2 - 105% of nominal.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate was periodically tested in a 48 hour test design.

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

74.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0%
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test solutions were clear and colorless.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: EC50 (48 h) 0.12 mg/L
- Other: Test was conducted from 18 - 20 Oct 2017.

Reported statistics and error estimates:

The EC50 value was calculated using Excel by binominal analysis.

Any other information on results incl. tables

The measured test concentrations were in the range of 80 - 120% of nominal (81.5 - 101%; see Table 2). Thus, it is justified that the effect concentrations were based on the nominal concentrations.

Table 1: Immobilisation of Daphnia magna after 24 and 48 h of exposure.

Nominal concentration [mg/L]	Vessel	24 h		48 h
		Number of immobilized Daphnids/total Daphnids	%	Number of immobilized Daphnids/total Daphnids	%
Control	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
9.53	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
17.1	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
30.9	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
55.6	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
100	A	3 / 5	80	5 / 5	100
	B	5 / 5		5 / 5
	C	4 / 5		5 / 5
	D	4 / 5		5 / 5

Table 2: Measured concentrations (mg/L + % of nominal) after 0 and 48 h of exposure)

Nominal concentration [mg/]	Measured concentration [mg/L] (%)
	0 h	48 h	Geometric mean
Control	Not detected	Not detected	-
9.53	9.63 (101)	7.77 (81.5)	8.65 (90.7)
17.1	17.1 (100)	15.1 (88.3)	16.1 (94.1)
30.9	30.8 (99.8)	28.4 (91.9)	29.6 (95.8)
55.6	56.3 (101)	48.3 (86.9)	52.1 (93.8)
100	97.9 (97.9)	96.1 (96.1)	97.0 (97.0)

Table 3: Validity criteria for OECD 202.

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	8.7 - 9.0 mg/L	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.