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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:
2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well performed OECD study with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity 99.0 % (w/w)

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
The test concentrations of Polysynthren-Braun R were analytically verified via HPLC-DAD in the saturated solution and the control only at the start of
the exposure (0 hours).

Sampling for the analytical monitoring: At the start of the exposure (0 hours), sampling was carried out after preparation of the saturated solution.


The measured concentrations were below the SysQL at the start of the exposure (0 hours) in the saturated solution and in the control.
Therefore, analytical monitoring at the end of the exposure was waived and the test item concentrations were considered to be below the SysQL in theaged media, too.

Test solutions

Vehicle:
no
Details on test solutions:
Preparation of the The saturated solution (1.00 mg/L test item were weighed out) was prepared with dilution water two days prior to the
saturated solution start of the exposure (at -48 hours).

Treatment The test item was dispersed in the dilution water by stirring with approximately 1100 rpm for 48 hours at room
temperature. After completion of stirring, the test item dispersion was filtered with a membrane filter (0.45 µm, RC,
MACHEREY-NAGEL) to achieve the saturated solution. Centrifugation was found to be unsuitable.

Test concentration The effects of the saturated solution of the test item were tested in a limit test.

Control 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
- Common name: Daphnia magna
- Strain: Clone 5
- Source:
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder: DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
- Age at study initiation (mean and range, SD): 2 to 24 h old daphnids from a healthy stock were used for the study
- Method of breeding:
Culture: 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 x m-2 x s-1
Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
Cultur feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and
Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: At least 2 h in dilution water.
- Acclimation conditions (same as test or not): Same as test conditions.
- Health during acclimation (any mortality observed): No mortality observed.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
262 mgCaCO3/L
Test temperature:
18 - 22°C, constant within ± 1°C
At start of exposure: 20 °C
pH:
pH at the Start of the Exposure (0 hours)
(measured in one additional replicate of the limit concentration and the control)

Nominal loading
of the test item
[mg/L] pH-value

1.00 7.75
(Saturated solution)
Control 7.93


pH at the End of the Exposure (48 hours)
(measured in all replicates)

Nominal loading
of the test item
[mg/L] pH-values
Replicates
1 2 3 4
1.00 7.70 7.70 7.72 7.71
(Saturated solution)
Control 7.65 7.70 7.71 7.70

Dissolved oxygen:
O2-concentration at the Start of the Exposure (0 hours)
(measured in one additional replicate of the limit concentration and the control)

Nomonal loading
of the test item
[mg/L]
Dissolved
O2-concentration
[mg/L]
1.00 6.20
(Saturated solution)
Control 8.97



O2-concentration at the End of the Exposure (48 hours)
(measured in all replicates)

Nominaltest loading
of the test item
[mg/L] Dissolved O2 -concentration [mg/L]
Replicates
1 2 3 4
1.00 8.76 8.63 8.66 8.70
(Saturated solution
Control 9.03 9.01 9.01 8.91
Nominal and measured concentrations:
Nominal loading: 1.00 mg/L

Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 mL capacity, loosely covered with watch glasses
- Test volume: 20 mL
- Aeration: None
- 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: ISO Test water, according to OECD 202, Annex 3
Component Concentration [mg/L]
KCl 5.75
NaHCO3 64.8
CaCl2 x 2 H2O 294
MgSO4 x 7 H2O 123

- Water Quality Parameters of the Dilution Water prior to the Start of the Exposure (0 h):
pH-value Dissolved O2 concentration [mg/L] Temperature [°C] Conductivity [µS/cm] Total hardness [mg CaCO3/L]
7.93 8.97 20.3 656 262

- Culture medium different from test medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: test start (0 h) and test end (48 h)

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m-2 x s-1 (<1340 lx)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Immobilisation, 24 h interval

TEST CONCENTRATIONS
-The effects of the saturated solution of the test item were tested in a limit test.




Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
other: EL10,50,100
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Duration:
48 h
Dose descriptor:
other: EL10,50,100
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Duration:
48 h
Dose descriptor:
other: NOEL
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Duration:
48 h
Dose descriptor:
other: LOEL
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Details on results:
See below
Results with reference substance (positive control):
The percentage immobility for the reference item was determined after 24 hours. The EC50-value with 95 % confidence limits (CI) was calculated by
sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50: 2.00 mg/L (CI 1.83 - 2.19 mg/L)
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 0.6 - 2.4 mg/L of quality criteria according to AQS P 9/2 (02/2000) 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.

Any other information on results incl. tables

Biological Data

The percentage immobility, determined in the saturated solution with a nominal loading of 1.00 mg/Lof the test item and

the control after 24 and 48 h under static conditions, is presented in Table3. There was no effect observed in the saturated solution and the

control.

 

Table 3:   Immobilisation Rates of the definitive Test after 24 and 48 hours of Exposure

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

Test item

Nominal loading

of the test item

[mg/L]

IMMOBILISATION [%]

24 hours

48 hours

Replicates

Replicates

1

2

3

4

MV

1

2

3

4

MV

1.00

(Saturated solution)

0

0

0

0

0

0

0

0

0

0

Control

0

0

0

0

0

0

0

0

0

0

 

All replicates of the saturated solution of the test item and the control were visually clear throughout the exposure period.

 

According to OECD Guidance Document No 23 (2000) if an effect cannot be detected in a test with a saturated solution the result has to be reported as no toxic effects at saturation. Exposures are expressed in terms of the original concentration of the test item Polysynthren-Braun R in water at the start of the mixing period (loading rate).

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
At the saturated solution with a nominal loading of 1.00 mg/L of the test item Polysynthren-Braun R, no effects were observed on Daphnia magna.




Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the saturated solution with a nominal loading of 1.00 mg/L of the test item was determined according to OECD 202 (2004). The limit test was conducted under static conditions over a period of 48 .

The test item is a brown powder and has a water solubility of < 0.02 mg/L at 20 °C. Therefore, a saturated solution with a nominal loading of

1.00 mg/L was prepared in ISO test water by stirring with approximately 1100 rpm for 48 hours at room temperature followed by filtration with a membrane filter (0.45 µm, RC, Macherey-Nagel) to remove undissolved test item to achieve the saturated solution. The saturated solution was visually clear throughout the exposure phase of 48 hours.

Twenty daphnids were exposed to the saturated solution and the control.

The test concentrations of the test item were analytically verified via HPLC-DAD in the saturated solution and the control only at the start of the exposure (0 hours).

The measured concentrations were below the SysQL at the start of the exposure (0 hours) in the saturated solution and in the control.

Therefore, analytical monitoring at the end of the exposure was waived and the test item concentrations were considered to be below the SysQL in the aged media, too.

According to OECD Guidance Document No 23 (2000), if an effect cannot be detected in a test with a saturated solution, the result has to be reported as no toxic effects at saturation. Exposures are expressed in terms of the original concentration of the test item in water at the start of the mixing period (loading rate).

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 hours) and at the end of the exposure

(48 hours), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

 

 

At the saturated solution with a nominal loading of 1.00 mg/L of the test item ,

no effects were observed on Daphnia magna.

 

 

 

Table1:   EL10-, EL50-EL100-Values, NOEL and LOEL

                  (based on the nominal loading levels of the test item)

Effect levels

Test

duration

[h]

Test item

Nominal loading level of the test item

[mg/L]

EL10 / 50 / 100

24

> 1.00

48

> 1.00

NOEL

48

  1.00

LOEL

48

> 1.00