Octadecanoic acid, reaction products with diethylenetriamine and urea, acetates

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

Study period:

2013

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: FAT #: 93580/A
Batch: 0022357000
Purity: 94.3 %
Expiry date: 15 December 2015
Description at Room Temperature: White-yellowish waxy solid
Storage Conditions: Room temperature, in the dark.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: limit test concentration based on the loading rate of 100 mg/L
- Sampling method: A semi-static test with test medium renewal after 24 hours was performed to keep the concentrations of the test item in the test medium as constant as possible during the test period of 48 hours. After 24 hours, the test organisms were transferred in clean test vessels with freshly prepared test medium by wide-bore pipette.
For the characterization of the water accommodated fraction (WAF), duplicate samples were taken from each treatment at the start and end of each test medium renewal period. For the 24 and 48 hour stability samples, additional flasks with adequate volumes of the freshly prepared test media of the single test concentration and the control were incubated during the test interval under the same conditions as in the actual test (but without daphnids).
- Sample storage conditions before analysis: All samples were analyzed immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the test organisms up to the loading rate of 100 mg/L. Thus, a single loading rate of 100 mg/L was tested. Additionally, a control was tested in parallel (test water without addition of the test item). The limit test was based on the results of a range-finding test and on results of pre-experiments to determine the solubility of the test item.
For preparation of the water accommodated fraction (WAF) with the loading rate of 100 mg/L, 300 mg of the test item (dosed amounts: 300.1 and 300.1 mg) was mixed into 3 litres of test water. First, the dispersion was treated ultrasonically for 15 minutes. Then, the dispersion was stirred for 96 hours to dissolve a maximum amount of the different compounds of the test item in the dispersion. The long stirring time was chosen in order to dissolve a maximum amount of the different components of the sparingly soluble test item and its degradation products in the dispersion. After stirring, the dispersion was left to settle for about 60 minutes to separate undissolved test material from the test water (as far as possible). Thereafter, the dispersion was filtered through a membrane filter (Schleicher & Schuell, Type Supor-200, pore size 0.20 µm) after preconditioning the filter material with about 200 mL of test medium to avoid test item loss in the filter. The undiluted filtrate was tested as a WAF.

The stirring period of 96 hours was chosen according to the results of pre-experiments which showed that the solution equilibrium was reached after this time. In these pre-experiments the same test item concentrations were analytically measured in the dispersion after stirring for 3, 24 and 96 hours.
The preparation of the test medium was based on the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures [OECD, 2000].
The test medium was prepared just before the start of exposure and prior to each test medium renewal period.
- Eluate: test water
- Differential loading: one loading rate at 100 mg/L
- Controls: test water
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Straus
- Source: originally supplied by the University of Sheffield / UK in 1992. Since that time, the clone has been bred at Harlan Laboratories.
- Age at study initiation: At the start of the test, the organisms used in the test were 6-24 hours old and were not first brood progeny.
- Method of breeding: the clone has been bred at Harlan Laboratories in reconstituted water of the quality identical to the water quality used in the tests (in respect to pH, main ions, and total hardness) and under temperature and light conditions identical to those of the test. During breeding, daphnids are generally fed three times a week with an algal suspension of the green algae Desmodesmus subspicatus CHODAT, Strain No. 86.81 SAG, supplied by the Collection of Algal Cultures (SAG, Institute for Plant Physiology, University of Göttingen, 37073 Göttingen / Germany) and cultivated at Harlan Laboratories under standardized conditions or a mixture of this algal suspension and a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, 49304 Melle / Germany).
- Feeding during test: no

ACCLIMATION
- Acclimation period: At the start of the test, the organisms used in the test were 6-24 hours old
- Acclimation conditions: same as in the test
- Type and amount of food: no feeding
- Health during acclimation: mortality observed

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Test terminated after 48 hours

Hardness:

250 mg/L as CaCO3

Test temperature:

20-21 °C

pH:

7.8-8.0 at 0 hours (new medium)
7.9 at 24 hours of exposure (old medium)
7.9 at 24 hours of exposure (new medium)
7.8 at 48 hours of exposure (old medium)

Dissolved oxygen:

8.6 at 0 hours (new medium)
8.6 at 24 hours of exposure (old medium)
8.6 at 24 hours of exposure (new medium)
8.8 at 48 hours of exposure (old medium)

Salinity:

not applicable

Nominal and measured concentrations:

Loading rate 100 mg/L
Mean measured concentration of 1.0 µg/L (calculated as arithmetic mean of the geometric means of the test item concentrations measured at the start and the end of each test medium renewal period)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100-mL glass beakers
- Type: open
- Material, size, headspace, fill volume: glass, 50 mL, 50 mL
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): once per 24 hours (semi-static)
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 5 individuals per 50 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water according to ISO 6341 (1996)
- Total organic carbon: not reported
- Particulate matter: not reported
- Metals:
CaCl2 × 2H2O: 2.0 mmol/L = 294 mg/L
MgSO4 × 7H2O: 0.5 mmlo/L = 123 mg/L
NaHCO3: 0.75 mmol/L = 65 mg/L
KCl: 0.075 mmol/L = 5.8 mg/L
- Pesticides: not reported
- Chlorine: not reported
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 1:4
- Conductivity: not reported but calculable from the concentrations of the salts
- Culture medium different from test medium: no
- Intervals of water quality measurement: 24 hours

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour light to 8-hour dark cycle with a 30-minute transition period
- Light intensity: 390-560 Lux

EFFECT PARAMETERS MEASURES: The daphnids were observed for immobility after 24 and 48 hours of exposure (daphnids not being able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile).

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable since a limit test was conducted
- Range finding study: no but two main tests were performed before and were not valid due to analytical problems with the difficult test item
- Test concentrations: not applicable
- Results used to determine the conditions for the definitive study: two main tests were performed before and were not valid due to analytical problems with the difficult test item.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: The results were based on the loading rate of 100 mg/L and on the mean measured concentration of 1 μg/L (calculated as arithmetic mean of the geometric means of the test item conc. measured at the start and the end of each test medium renewal period).

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 1 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no
- Mortality of control: 5 %
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: use of one loading rate of 100 mg/L
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

For evaluation of the quality of the daphnia clone and the experimental conditions, potassium dichromate is tested as a positive control twice a year. The result of the latest positive control test in October 2013 (48-hour EC50: 0.66 mg/L, Harlan Laboratories Study D74255) showed that the sensitivity of the test organisms was within the internal historical range (48-hour EC50 from 2000 to 2013: 0.43-1.1 mg/L).

Reported statistics and error estimates:

The NOEC/NOEL, EC0/EL0, EC50/EL50 and EC100/EL100 were directly determined from the raw data.
For each test medium renewal period, the mean concentration was calculated as geometric mean of the test item concentrations measured at the start and the end of the test medium renewal period [Sachs, 1984].

Validity criteria fulfilled:

yes

Conclusions:

FAT #: 93580/A had no acute toxic effects on Daphnia magna up to its solubility limit in water under the present test conditions.

Executive summary:

The acute toxicity of the test item FAT #: 93580/A to Daphnia magna was determined in a 48-hour semi-static test according to the EU Commission Directive 92/69/EEC, Part C.2, the Commission Regulation (EC) No. 440/2008, Part C.2 and the OECD Guideline for Testing of Chemicals, No. 202 (2004).

A limit test was performed in compliance with the test guidelines to demonstrate that water accommodated fractions (WAFs) of the test item have no toxic effect on Daphnia magna up to the loading rate of 100 mg/L. Thus, the only concentration tested was a WAF with the loading rate of 100 mg/L. Additionally, a control (test water without test item) was tested in parallel. The preparation of the test medium was based on the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, 2000.

For preparation of the test medium, a dispersion of the test item with the loading rate of 100 mg/L was prepared. The dispersion was stirred for 96 hours to dissolve a maximum amount of the different compounds of the test item in the dispersion. Then, the dispersion was filtered through a membrane filter (0.20 µm pore size) and the undiluted filtrate was tested as a WAF.

The analytically measured test item concentrations (based on the sum of six main components) in the test medium with the loading rate of 100 mg/L were 1.90 µg/L (day 0) and 0.865 µg/L (day 1) at the start of the test medium renewal periods. At the end of the test medium renewal periods the measured test item concentrations were 0.851 µg/L (day 1) and 0.767 µg/L (day 2).

The reported biological results were based on the loading rate of 100 mg/L and on the mean measured concentration of 1.0 µg/L (calculated as arithmetic mean of the geometric means of the test item concentrations measured at the start and the end of each test medium renewal period).

At the loading rate of 100 mg/L (mean measured concentration of 1.0 µg/L), no immobilized daphnids were determined during the test period of 48 hours. In the control, one test organism was immobile at the observation after 24 hours. However the test is considered to be valid, as in accordance with the test guidelines an immobilization rate of 10 % is tolerated in the control.

The summary of the biological results is as follows:

– 24-hour EL50 / EC50: >100 mg/L and >1.0 µg/L, respectively.

– 24-hour EL0 / EC0: >100 mg/L and >1.0 µg/L, respectively.

– 24-hour EL100 / EC100: >100 mg/L and >1.0 µg/L, respectively.

– 48-hour EL50 / EC50: >100 mg/L and >1.0 µg/L, respectively.

– 48-hour EL0 / EC0 and 48-hour NOELR / NOEC: ≥100 mg/L and ≥1.0 µg/L, respectively.

– 48-hour EL100 / EC100: >100 mg/L and >1.0 µg/L, respectively.

Based on the findings of this study, the test item FAT #: 93580/A had no acute toxic effects on Daphnia magna up to its solubility limit in water under the present test conditions.

Description of key information

The study describes acute toxicity of the test item FAT #: 93580/A to Daphnia magna was determined in a 48 hour semi-static test with daily test medium renewal according to OECD TG 202.

At the loading rate of 100 mg/L (mean measured concentration of 1.0 µg/L), no immobilized daphnids were determined during the test period of 48 hours. In the control, one test organism was immobile at the observation after 24 hours. However the test is considered to be valid, as in accordance with the test guidelines an immobilization rate of 10 % is tolerated in the control.

The summary of the biological results is as follows:

– 24-hour EL50 / EC50: >100 mg/L and >1.0 µg/L, respectively

– 24-hour EL0 / EC0: >100 mg/L and >1.0 µg/L, respectively

– 24-hour EL100 / EC100: >100 mg/L and >1.0 µg/L, respectively

– 48-hour EL50 / EC50: >100 mg/L and >1.0 µg/L, respectively

– 48-hour EL0 / EC0 and 48-hour NOELR / NOEC: ≥100 mg/L and ≥1.0 µg/L, respectively

– 48-hour EL100 / EC100: >100 mg/L and >1.0 µg/L, respectively

Based on the findings of the study, the test item FAT #: 93580/A had no acute toxic effects on Daphnia magna up to its solubility limit in water under the present test conditions.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L