Registration Dossier

Ecotoxicological information

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:
2016-07-27 - 2016-10-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
13-Apr-2004
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Other: Storage conditions: Ambient temperature

Sampling and analysis

Analytical monitoring:
yes
Remarks:
TOC
Details on sampling:
- Concentrations (nominal with measured in brackets): 0.5 (no net TOC), 1.3 (no net TOC), 3.6 (0.27), 10.2 (2.23), 28.6 (12.68), 80 (32.29) mg/L
- Sampling method: The supporting analyses were performed on the same day of collection of the specimens. Additionally, the remaining amounts of specimens were stored deep-frozen and re-analyzed for control purpose later-on.

Test solutions

Vehicle:
no
Remarks:
medium was used
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was weighed in ELENDT M4 dilution water followed by ultrasonic treatment for 33 to 55 minutes and stirring on a magnetic stirrer for 1 hr (screening test) to 24hrs (main test) being necessary to dissolve the test item sufficiently in the test medium.
Before weighing the substance into the vessels for treatment preparation it was milled in a mortar made of porcelain. Four parallels were prepared for the test treatments with the test item and for the control without any test item, respectively, using 5 animals each parallel (main test). In the screening test (range-finder), two parallels were prepared for each concentration and the control without any test item, respectively, using 5 animals each parallel.
- Controls: medium without any test item
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): none stated

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna STRAUS (clone 5)
- Source: obtained from Dr. M. Bergtold, BASF, D-67117 Limburgerhof; the strain originally derived from the Institut National de Recherche Chimique Appliquee, France in 1978
- Feeding during test: no

ACCLIMATION
- Acclimation period: The cultivation of the daphnia is performed in a way that the animals are transferred in new test medium at an interval of 1 to 3 days followed by feeding.
- Type and amount of food: Desmodesmus subspicatus- and/or Chlorella-cells
- Health during acclimation (any mortality observed): none stated

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
Daphnias used for the test are not older than 24 hr. Therefore, in every case, daphnia are transferred into fresh medium one day before starting of the daphnia test in order to get daphnia for the test younger than 24 hr.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Test temperature:
20 ± 1°C
pH:
7.76 - 9.21 during main test (screening: 7.73 - 8.78)
Dissolved oxygen:
8.0 - 8.5 mg O2/L during main test (screening: 7.5 - 8.5 mg O2/L)
Nominal and measured concentrations:
Nominal: 0.5, 1.3, 3.6, 10.2, 28.6, 80 mg/L
Found (t0): n.n., n.n., 0.27, 2.23, 12.68, 32.29 mg/L
Found (t48h): n.n., n.n., 0.28, 1.62, 11.53, 31.29 mg/L

n.n. = no net TOC
Details on test conditions:
TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 50mL, glass
- 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:
Ultrapure water generating systems: Purelab Classic D1l; Ser.-No. 1470984-10 01
Purelab Ultra Analytic Mk2, Seral; Ser.-No. UAJ268941 (“CA 03”), ELGA Labwater

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hr of light and 8 hr of darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility, daily

VEHICLE CONTROL PERFORMED: yes, medium only

RANGE-FINDING STUDY
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: the results of the screening test were indifferent at 10 mg/L. That's why there was one more treatment than indicated in the study plan (6 instead of 5 treatments).
Reference substance (positive control):
no
Remarks:
A reference item was not used within the biological part of this study. Within the analytical part of the study, the test item served as the reference.

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.206 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: nominal: 3.0 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
< 0.2 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: nominal: 0.5 mg/L
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
<= 0.2 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: nominal: 1.3 mg/L
Duration:
48 h
Dose descriptor:
other: EC20
Effect conc.:
0.063 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: nominal: 1.3 mg/L
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
0.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: measured: not determined for methematical reasons

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The study was performed according to OECD TG 202 without deviations, the results were so obtained via a scientifically reasonable method. Hence, there is no doubt that the obtained results are not reliable:
Under the conditions used for the test, there was a significant immobilisation of the daphnias at concentrations >0.5 mg/L (main test).
As a conclusion of the analytical part of this study, it can be stated that the concentrations of the test item at concentration steps around the EC50 remained sufficiently stable during incubation of 48hr. At lower concentrations it is assumed that a certain amount of the test item may have been adsorbed to surfaces as minor concentrations were found. As a result of the supporting analyse it can be stated that the results of the biological part should be based on the measured concentrations applied because the recovery of the nominal concentrations was <80% though the necessary stability of ≥ 80% could be shown. The following EC-values (48h; measured concentrations) were determined accordingly:
LOEC ≤ 0.2 mg/L
NOEC < 0.2 mg/L
EC50 = 0.206 mg/L
Based on these results, the test item has to be classified as aquatic acute Cat. 1 (EC50 <1mg/L), M factor 1. Further, with regard to chronic toxicity and taking into account the facts that the test item is soluble in water and not readily biodegradable, the test item should be classified as aquatic chronic Cat. 1, M factor 1.
Executive summary:

Octadecanoic acid, reaction products with tetraethylenepentamine was tested for acute toxicity towards daphnia according to OECD-Test Guideline 202 (version dated 13-Apr-2004; GLP). In order to investigate the influence of the test item towards the daphnias, the swimming behaviour of the animals was recorded. Under the conditions used for the test, there was a significant immobilisation of the daphnia at concentrations >0.5 mg/L (main test). As a conclusion of the analytical part of this study, it can be stated that the concentrations of Octadecanoic acid, reaction products with tetraethylenepentamine at concentration steps around the EC50 remained sufficiently stable during incubation of 48hr. At lower concentrations it is assumed that a certain amount of the test item may have been adsorbed to surfaces as minor concentrations were found. As a result of the supporting analyse it can be stated that the results of the biological part should be based on the measured concentrations applied because the recovery of the nominal concentrations was <80% though the necessary stability of ≥ 80% could be shown. The following EC-values (effective concentrations) were determined accordingly:

      Basis: Nominal Concentrations [mg/L]   
 t24hr  t48hr
 NOEC  0.5  0.5
 LOEC  1.3  1.3
 EC10  1.3  0.9
 EC20  2.3  1.3
 EC50  7.6  3.0

      Basis: Measured Concentrations [mg/L]   
 t24hr  t48hr
 NOEC  0.2  < 0.20
 LOEC  1.37  ≤ 0.20
 EC10  0.21  n.d.
 EC20  0.41  0.063
 EC50  1.39  0.206

n.d. = not determined for mathematical reasons.

The test is considered valid as all conditions for validity were met.