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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 Mar 2013 - 17 December 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
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Concentrations of dissolved Nd were determined in at least one of the triplicate samples from each treatment per sampling time.
- Sampling method: Triplicate samples were taken from each treatment at the start and end of each test medium renewal period (after filtration through a membrane filter, Whatman, Type NC45, pore size 0.45 µm).
- Sample storage conditions before analysis: All samples are stored at room temperature in the dark after sampling until analysis.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low solubility of the test item in test water, a dispersion with the loading rate of 126 mg/L (corresponding to 100 mg/L anhydrous neodymium trinitrate) was prepared at the start of the test and before each test medium renewal by dispersing 126.2 mg of the test item (dosing range: 126.2–126.3 mg) in 1000 mL of test water. This preparation was supported by ultrasonic treatment for 15 minutes and intense stirring on a magnetic stirrer over 3 hours in the dark, to dissolve a maximum amount of the test item in the dispersion. No auxiliary solvent or emulsifier was used.

An optimal stirring period of 3 hours was chosen based on the results of a pre-experiment, which showed that the solution equilibrium was reached after this time. In this pre-experiment, similar concentrations of dissolved elemental neodymium were analytically measured in filtrates after stirring for 3, 24 and 96 hours.

After stirring and pH adjustment, the dispersion of the test item was filtered through a membrane filter (Whatman, Type NC45, pore size 0.45 µm). The undiluted filtrate was used as highest concentrated test medium and as a stock solution for preparation of the test media with lower test concentrations. For preparation of the latter, the filtrate was diluted with pH adjusted test medium.

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea (Daphnia magna)
- Source: University of Sheffield / UK in 1992
- Age at study initiation (mean and range, SD): 6-24 hours (not a first brood progeny)
- Feeding during test: no
- Method of breeding: clone has been bred at Harlan Laboratories in reconstituted water of the quality identical to the water quality used in the test

ACCLIMATION
- Type and amount of food: green algae Desmodesmus subspicatus CHODAT, Strain No. 86.81 SAG, supplied by the Collection of Algal Cultures or a mixture of this algal suspension and a commercial fish diet
- Feeding frequency: daphnids are generally fed three times a week
- Health during acclimation (any mortality observed): good. No mortalities observed
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
250 mg CaCO3/L
Test temperature:
21°C
pH:
New test media:
0 h = 6.0-6.1
24 h = 5.9-6.1
Old test media:
24 h = 6.9-7.1
48 h = 6.9-7.2
Dissolved oxygen:
8.0 - 8.5 mg/L
Salinity:
not applicable
Nominal and measured concentrations:
Analytically determined concentrations of dissolved neodymium in the test media (control, dilutions 1:100, 1:32, 1:10, 1:3.2, and the undiluted filtrate) were as follows:
Day 0, 0 h (start of experiment) = 0.00115/0.00215, 0.153, 0.489, 1.63, 5.31 and 16.8 mg Nd/L (i.e. 0.00263/0.00492, 0.35, 1.12, 3.74, 12.2, 38.5 mg Nd(NO3)3/L).
Day 1, 24 h (end of 1st renewal period) = < LOQ, 0.124, 0.430, 1.28, 4.73, 12.5 mg Nd/L (i.e. Day 2, 0 h (start of 2nd renewal period) = < LOQ, 1.98/1.92, 0.532, 1.87, 5.48 and 17.7 mg Nd/L (i.e. Day 2, 24 h (end of 2nd renewal period) = < LOQ, 1.93/1.87, 0.520, 1.80, 5.55 and 19.2 mg Nd/L (I.e.
The analytically determined levels of dissolved Nd in the 1:100-dilution at the start and at the end of the second 24-hour test period were in the same range as the levels determined in the 1:10 dilution; probably due to a mix-up during sampling. No biological effect was observed in this dilution. Therefore, the analyzed concentration of dissolved Nd in this dilution of the second test period was not taken further in mean measured calculations.

At the end of the test medium renewal periods of 24 hours the concentrations of dissolved Nd decreased (recovery 74 to 109% of initially measured concentrations, on average 87 to 95%). Further, the dissolved Nd levels were overall higher during the second renewal period. Biological results were based on mean measured concentrations calculated as the arithmetic mean of the two geometric means determined for dissolved Nd measured at the start and end of each test medium renewal period. Mean measured concentrations were as follows (not mentioning the control, for which dissolved Nd was < LOQ): 0.14, 0.49, 1.6, 5.3 and 16.0 mg Nd/L (corresponding to 0.32, 1.1, 3.8, 12, 38 mg Nd(NO3)3/L).
Details on test conditions:
TEST SYSTEM
- Test vessel: 100-mL glass beakers filled with 50 mL of test medium
- Type (delete if not applicable): open
- Aeration: The test water was aerated prior to the start of the study. No aeration during the study.
- 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: Reconstituted test water according to ISO 6341
- Ca/mg ratio: 4:1
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: 1) adjustment of dispersion to pH 6.0 after the 3-h stirring period using 1 M NaOH, 2) adjustment of test medium (for dilution of the filtrate) to pH 6.0 using 1 M HCl
- Photoperiod: A 16-hour light to 8-hour dark cycle with a 30-minute transition period
- Light intensity: Light intensity during the light period was approximately between 390 and 560 Lux.

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.2
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
3.6 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Remarks:
neodymium
Basis for effect:
mobility
Remarks on result:
other: 95% CL: 2.4-5.2
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
8.1 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
other: neodymium trinitrate, anhydrous
Basis for effect:
mobility
Remarks on result:
other: 95% CL: 5.5-12
Details on results:
48-hour EC0 and NOEC: 0.14 mg dissolved Nd/L (corresponding to 0.32 mg/L when expressed as anhydrous neodymium trinitrate)
48-hour EC100 was 16 mg dissolved Nd/L (corresponding to 38 mg/L when expressed as anhydrous neodymium trinitrate)
Results with reference substance (positive control):
- Results with reference substance valid? yes
- EC50/LC50: 48-hour EC50: 0.94 mg/L (internal historical range: 48-hour EC50 from 2000 to 2012: 0.43-1.1 mg/L).

Reported statistics and error estimates:
NOEC, EC0 and EC100 were determined directly from the raw data.
24-h EC50 could not be calculated due to absence of toxicity and was therefore also determined directly from the raw data.
48-h EC50 and 95% confidence limits were calculated using Weibull Analysis with linear weighted regression.
The two lowest treatments were excluded from the statistical evaluation, as they did not contribute to the dose-response relationship.
Validity criteria fulfilled:
yes
Conclusions:
The test item had an acute toxic effect on Daphnia magna. Based on mean measured concentrations, the 48-hour EC50 was determined to be 3.6 mg/L for dissolved neodymium with 95% confidence limits of 2.4 and 5.2 mg/L (corresponding to 8.1 mg/L of anhydrous neodymium trinitrate with 95% confidence limits of 5.5 and 12 mg/L).
Executive summary:

The acute toxicity of neodymium trinitrate to Daphnia magna was determined in a 48-hour semi-static test according to the OECD guideline 202 and in compliance with the principles of Good Laboratory Practice. Juvenile daphnids were exposed to a control treatment and the test chemical at various concentrations (a dispersion with the loading rate of 126 mg/L (corresponding to 100 mg/L of anhydrous neodymium trinitrate) was used as stock solution for test media preparation; the undiluted filtrate and dilutions 1:3.2, 1:10, 1:32 and 1:100 of this undiluted filtrate were used as test media). Organisms were observed after 24 and 48 hours for immobility.

After 48 hours of exposure, no immobilized test organisms were recorded in the control and up to and including the mean measured concentration of 0.14 mg/L of dissolved neodymium, corresponding to 0.32 mg/L of anhydrous neodymium trinitrate. At the mean measured concentration of 5.3 and 16 mg/L of dissolved neodymium (corresponding to 12 and 38 mg/L of anhydrous neodymium trinitrate), 50% and 100% of the daphnids were found to be immobile, respectively. The 48-h EC50 was determined to be 3.6 mg/L for dissolved neodymium and 8.1 mg/L for anhydrous neodymium trinitrate; meaning this substance is toxic to the aquatic invertebrates.

Description of key information

The test item had an acute toxic effect on Daphnia magna. Based on mean measured concentrations, the 48-hour EC50 was determined to be 3.6 mg/L for dissolved neodymium with 95% confidence limits of 2.4 and 5.2 mg/L (corresponding to 8.1 mg/L of anhydrous neodymium trinitrate with 95% confidence limits of 5.5 and 12 mg/L).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Dose descriptor:
EC50
Effect concentration:
3.6 mg/L

Additional information