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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:
2012-03-21 to 2012-03-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
(2004)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
(2008)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed 2010-10-20

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Physical state: solid, whitish to white crystals
- Storage condition of test material: ambient temperature, dark, dry and in original container
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
SAMPLING
- Samples for determination of the test item concentration were taken of the fresh media (after 0 hours) and of the old media (after 48 hours).
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until sample preparation, if necessary.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION OF TEST SOLUTION
- Preparation of the saturated solution acc. to OECD Series, No. 23 (2000)6:
The saturated test item solution (8 mg/L test item was weighed out) was prepared with dilution water two days before the start of the exposure (0 h). The test item solution was shaken with approximately 20 rpm for 48 hours at room temperature. Thereafter the test item solution was filtrated (0.45 µm).

APPLICATION
20 g of the test solution each was weighed out into the test vessels. The daphnids were inserted with a small amount of dilution water by a pipette.

CONTROL
Four replicates with dilution water without test item, tested under the same conditions as the test groups

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Strain: Daphnia magna STRAUS (Clone 5)
- Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
- Breeder: DR.U.NOACK-LABORATORIEN, 31157 Sarstedt, Germany
- Culture/Method of breeding: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 +-2°C, in an incubator, 16 h illumination; light intensity of max. 20 μE*m-2 * s-1
- Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Culture feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata, Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 10E6 cells/mL.
- Age at study initiation: 2 to 24 h
- Feeding during test: no

ACCLIMATION
- Acclimation period: At least 2 hours in dilution water

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
no data

Test conditions

Hardness:
nominal hardness (dilution water): 272 mg CaCO3/L
Test temperature:
19 - 20 °C
pH:
At test start: 7.85 - 7.86
At test end: 7.5 - 7.74
Dissolved oxygen:
At test start: 9.13 - 9.23 (control / treatment)
At test end: 8.81 - 9.09 (control / treatment)
Salinity:
not applicable
Nominal and measured concentrations:
Nominal loading of the test item 8.00 mg/L (Saturated solution)
Geometric mean measured moncentration of 0.367 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 mL capacity
- Test volume per vessel: 20 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: Water quality parameters (pH values, dissolved oxygen concentration, conductivity, total hardness and temperature) of the dilution water were measured at the beginning of the exposure (0 hours) and after 48 hours in the old media (saturated solution and the control).
- Dilution water: ISO Test water, according to OECD 202, Annex 3

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity max. 20 µE*m-2 * s-1

EFFECT PARAMETERS MEASURED
- Biological Parameters: Immobilisation was determined in all groups after 24 and 48 h.

PRELIMINARY RANGE FINDING TEST
- Two non-GLP preliminary range finding tests were conducted under static conditions.
- The 2nd preliminary range finding test was carried out with the saturated solution with a nominal loading of 8.00 mg test item/L. The test conditions of the 2nd preliminary test were the same as specified for the definitive test. No immobilisation of Dahnia magna occurred at the saturated solution and at the control. The recovery rate was 104 %.
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate p.a.

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.37 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
> 0.37 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 0.37 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 0.37 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mobility
Details on results:
BIOLOGICAL DATA
No effect observed, either in the limit concentration or in the control group.

DETERMINATION OF THE TEST ITEM
The initial measured concentration of the test item in the saturated solution was 0.389 mg/L (fresh medium, 0 hours). In the old medium (48 hours) of the saturated test item solution, a test item concentration of 0.346 mg/L was determined, which corresponds to 89 % of the initial concentration.

WATER QUALITY PARAMETERS
The environmental conditions (dissolved oxygen concentration, temperature, conductivity and total hardness) were determined to be within the acceptable limits.
Results with reference substance (positive control):
The percentage immobility for the reference item was determined after 24 hours. The EC50-value with 95 % confidence interval (CI) was calculated by sigmoidal dose-response regression to be:
EC50: 1.64 mg/L (CI 1.59 - 1.69 mg/L)
The EC50-value of the reference item potassium dichromate after 24 hours is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according to AQS P 9/2 (05/1996) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.
Reported statistics and error estimates:
Since an immobilisation of daphnia magna was not observed at the tested limit concentration, the EC50 (48 h) was estimated with > limit concentration.

Any other information on results incl. tables

Immobilization Rates of the Definitive Test after 24 and 48 hours of Exposure

(n = 20, divided into 4 replicates with 5 daphnids each)

Test item

IMMOBILIZATION [%]

Geometric Mean

Measured Concentration

 

[mg/L]

24 hours

48 hours

Replicates

Replicates

1.

2.

3.

4.

MV

1.

2.

3.

4.

MV

0.367

0

0

0

0

0

0

0

0

0

0

Control

0

0

0

0

0

0

0

0

0

0

MV = Mean value

Measured Concentrationsin the Definitive Test

Sampling date

2012-03-21

(Fresh media, 0 hours)

2012-03-23

(Old media, 48 hours)

 

Start of analysis

2012-03-21

2012-03-23

 

Nominal loading

of the test item

[mg/L]

Test item

Geometric mean

measured concentration

[mg/L]

Measured

concentration*
[mg/L]

Measured

concentration*

[mg/L]

%

8.00

(Saturated solution)

0.389

0.346

89

0.367

Control

<LOQM

<LOQM

 

* = Mean value of 2 injections

% = Percentage of the initially measured concentration of the test item

LOQM= Limit of quantification of the analytical method (0.1 mg/L of the test item)

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
At the saturated solution of the substance with a mean measured concentration of 0.37 mg/L, effects on the mobility of Daphnia magna were not observed over 48 hours. The 48-h EC50 is above the water solubility limit and estimated with > 0.37 mg/L.