Fatty acids, coco, tetraesters with bis[2,2-bis(hydroxymethyl)butyl] adipate

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:

2 April 2012 to 29 April 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in accordance with recognised guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1,6-bis({2,2-bis[(undecyloxy)methyl]butyl}) hexanedioate

EC Number:

700-772-5

Cas Number:

1190961-28-4

Molecular formula:

N/A - too complex

IUPAC Name:

1,6-bis({2,2-bis[(undecyloxy)methyl]butyl}) hexanedioate

Test material form:

other: liquid

Details on test material:

- Physical state: straw colored liquid
- Storage condition of test material: room temperature in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100mg/L
- Sample storage conditions before analysis: Water samples were taken from the control and loading rate WAF test group at 0 and 48 hours for TOC analysis.

Test solutions

Vehicle:

no

Details on test solutions:

Due to the low aqueous solubility and complex nature of the test item the test solutions were prepared as Water Accommodated Fractions. An amount of test item (250 mg) was added to the surface of 2.5 liters of reconstituted water to give the 100 mg/L loading rate. After the addition of the test item, the reconstitued water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 em from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 7 5-100 mL discarded) to give the 100 mg/L loading rate WAF.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water Flea
- Strain: Daphnia magna
- Source: In house laboratory cultures
- Age at study initiation (mean and range, SD): 1st instar, less than 24 h old
- Weight at study initiation (mean and range, SD): no data available.
- Length at study initiation (length definition, mean, range and SD): no data available.
- Valve height at study initiation, for shell deposition study (mean and range, SD): no data available.
- Peripheral shell growth removed prior to test initiation: no data available.
- Method of breeding: parthenogenesis
- Feeding during test : The daphnids received no food during exposure
- Food type: Not applicable.
- Amount: Not applicable.
- Frequency: Not applicable.

ACCLIMATION
- Acclimation period: Not applicable.
- Acclimation conditions (same as test or not): Not applicable.
- Type and amount of food: Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): Not applicable.

QUARANTINE (wild caught)
- Duration: Not applicable.
- Health/mortality: Not applicable.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

The number of immobilised Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours. The criterion of effect used was that Daphnia were considered to be immobilized if they were unable to swim for 15 seconds after gentle agitation.

Test conditions

Hardness:

The reconstituted water had an approximate theoretical total hardness of 250 mg/l as CaCO3.

Test temperature:

21 °C to 23 °C. Water temperature was recorded daily throughout the test.

pH:

There were no treatment related differences in pH (8.0). pH values were recorded at the start and termination of the test.

Dissolved oxygen:

There were no treatment related differences in oxygen concentration. Dissolved oxygen concentrations were recorded at the start and termination of the test (range: 8.2 - 8.4 mg O2/L).

Salinity:

Not applicable.

Nominal and measured concentrations:

Nominal loading rates of 100 mg/L

Details on test conditions:

TEST SYSTEM
As in the range finding tests 250 ml glass jars containing 200 ml of test preparation were used. At the start of the test 10 daphnids were placed in each test and control vessels at random in the test preparations. Duplicate test vessels were used for each test and control group. The test vessels were then covered to reduce evaporation and maintained in a temperature controlled room at approximately 21 degrees C with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified and received no food during exposure and the test vessels were not aerated. The control group was maintained under identical conditions but not exposed to the test item. The test preparations were not renewed during the exposure period.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The criterion of effect used was that Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation.

-Results used to determine the conditions for the definitive study: Based on the results of the range finding tests the loading rates for the definitive test were assigned. Results of the initial range finding test indicated that significant immobilization occurred at 1.0, 10 and 100 mg/L loading rate WAF so a second range finding test was conducted by exposing daphnids to nominal loading rates of 0.1, 1.0 and 10 mg/L.

TEST SYSTEM
- Test vessel: Each 250 mL test and control vessel contained 200 mL of test media
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 250 mL vessel contained 200 mL of test media
- No. of organisms per vessel: 10 in the range finding study, 5 in the main study
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water


OTHER TEST CONDITIONS
- Adjustment of pH: pH = 8.0
- Photoperiod: 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods.


TEST CONCENTRATIONS
- Range finding study:
Due to the low aqueous solubility and complex nature of the test item, for the purposes of the range-finding test the test item was prepared as a Water Accommodated Fraction (WAF). The loading rate to be used in the definitive test was determined by a preliminary range-finding test. In the range-finding test Daphnia magna were exposed to a series of nominal loading rates of 10 and 1 00 rng/L.

- Results used to determine the conditions for the definitive study: 100 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate (non-concurrent). A positive control used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L. Exposure conditions and data evaluation for the positive control were similar to those in the

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities:no
- Mortality of control: 0
- Other adverse effects control: None noted

Results with reference substance (positive control):

- Results with reference substance valid? Yes.
- EC50/LC50:
Exposure of Daphnia magna to the reference (potassium dichromate) item gave a 48 hour EC50 value of1.1 mg/L; The No Observed Effect Concentrations after 24 and 48 hours were 1.0 and 0.56 mg/L respectively. The No Observed Effect Concentration is based upon zero immobilization at this concentration.

Any other information on results incl. tables

Cumulative Immobilisation Data in the Definitive Test

NominalLoading Rate (mg/L)		CumulativeImmobilizedDaphnia (InitialPopulation:5PerReplicate)
		24Hours			48Hours
		No.Per Replicate	Total	%	No.Per Replicate	Total	%
Control R1	R1	0	0	0	0	0	0
	R2	0			0
	R3	0			0
		0			0
100	R1	0	0	0	0	0	0
	R2	0			0
	R3	0			0
		0			0

Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed measured concentrations of 12.90 and 10.66 mg C/L respectively were obtained.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48-Hour EL50 for the test item to Dahnia magna based on nominal loading rates was greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April2004) No 202, "Daphnia sp, Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

Methods

Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a Water Accommodated Fraction (WAF) of the test item at single nominal loading rate of 100 mg/L for 48 hours at a temperature of 21 °C to 23 °C under static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

Results

The 48-Hour EL50 for the test item to Dahnia magna based on nominal loading rates was greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF. It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L loading rate WAF. Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed measured concentrations of 12.90 and 10.66 mg C/L respectively were obtained. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole the results were based on nominal loading rates only.