[No public or meaningful name is available]

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:

01 - 23 Aug 2018

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)

Version / remarks:

2004

Deviations:

yes

Remarks:

See section "Any other information on material and methods incl. tables"

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

yes

Remarks:

See section "Any other information on material and methods incl. tables"

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Mainz, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and test item concentration
- Sampling method: The content of phosphorus (total P and P from orthophosphate) of the test item in the test solutions was measured at the beginning and at the end of the test.
- Sample storage conditions before analysis: A sample was taken after 24 h and stored at -20 ± 5 °C.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution containing 177.6 mg/L (= 100 mg/L active ingredient due to 43.1% water content of test item) resp. 112 µL/L test item (based on a density of 1.59 g/mL stated in the SDS) in dilution water was prepared and used unfiltered for the test. This was done by filling 56 µL of the test item to 500 mL with dilution water. Homogenization was performed by shaking in the measuring cylinder. The resulting solution was clear.
- Controls: Yes, test medium only

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water-fleas
- Strain/clone: STRAUS
- Source: Umweltbundesamt Berlin
- Feeding during test: No feeding during the test
- Food type: Stock culture was feed with green algae (Desmodesmus subspicatus)
- Frequency: Stock culture was feed twice a week

ACCLIMATION
- Acclimation period: M4-Medium (recipe of ELENDT) was used for the culture and dilution water was used for the test (Composition: CaCl2*2H2O: 293.810 mg/L, MgSO4*7H2O: 123.3 mg/L, NaHCO3: 64.8 mg/L, KCl: 5.8 mg/L)
- Acclimation conditions: Same

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: 19 h before the start of the test, the adult animals were separated from the young. 35 min before test start, the adults were caught with the help of a glass tube, and the newborn Daphnia, aged between 0 and 18 h 25 min, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

2.502 mmol/L corresponding to 250 mg CaCO3/L

Test temperature:

22.7 – 23.8 °C

pH:

7.8 - 7.9 (Control)
8.5 - 8.0 (Highest test item concentration)

Dissolved oxygen:

8.9 - 8.4 (Control)
8.7 - 8.2 (Highest test item concentration)

Nominal and measured concentrations:

Control and 176 mg/L (corresponding to 100 mg/L active ingredient (nominal) due to 43.1% water content of test item)

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: open (covered with glass lids)
- Material, size, headspace, fill volume: Glass beakers, nominal volume 50 mL, tall shape, 25 - 30 mL headspace
- 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: Dilution water (Daphnia medium, composition see "Any other information on material and methods incl tables")
- Culture medium different from test medium: M4 Elendt medium was used for the culture and dilution water (Daphnia medium) for the test

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 hours, day/night
- Light intensity: Neon tubes at 1000 lux

EFFECT PARAMETERS MEASURED: Immobility after 24 and 48 h. Daphnia are considered as immobilised when they do not perform any movements or are only able to move their antennae when the beaker is gently agitated. Daphnia which are trapped at the surface of the test solution are also considered immobilised.

RANGE-FINDING STUDY: non-GLP study
- Test concentrations: 1, 10, 100, 176 mg/L (corresponding to 100 mg/L active ingredient (nominal) due to 43.1% water content of test item).
- Results used to determine the conditions for the definitive study: No immobilisation was observed after 48 h for all test item concentrations

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 110.82 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: constituents 1 and 2 and active ingredient due to 43.1% water content of test item)

Basis for effect:

mobility

Remarks on result:

other: see table 1 and 2 in section "Any other information onn results incl tables" for detailed information

Details on results:

- Behavioural abnormalities: no abnormalities were observed
- Mortality of control: no mortality was observed

Results with reference substance (positive control):

The 24h-EC50 value was determined as 1.9 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline.

Reported statistics and error estimates:

Since no immobilisation in the treatment was observed, no statistical evaluation of results was conducted but values are stated as a range.

Table 1: Biological Results of Test Item based on constituent 1-determination and on active ingredient

Parameter	Value based on constituent 1	Value based on constituent 1 and active ingredient (due to 43.1% water content of test item)
24h EC50	> 194.77 mg/L	> 110.82 mg/L
48h EC50	> 194.77 mg/L	> 110.82 mg/L
48h NOEC	≥ 194.77 mg/L	≥ 110.82 mg/L
48h LOEC	> 194.77 mg/L	> 110.82 mg/L

  

Table 2: Biological Results of Test Item based on constituent 3 -determination and on active ingredient

Parameter	Value based on constituent 3	Value based on constituent 3 and active ingredient (due to 43.1% water content of test item)
24h EC50	> 229.94 mg/L	> 130.84 mg/L
48h EC50	> 229.94 mg/L	> 130.84 mg/L
48h NOEC	≥ 229.94 mg/L	≥ 130.84 mg/L
48h LOEC	> 229.94 mg/L	> 130.84 mg/L

Table 3: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	No immobilization in the control	Yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	8.2 mg/L	Yes

Analytical Results:

At the beginning and at the end of the test the content of phosphorus (total P and P from orthophosphate) of the test item in the test solutions was measured via ICP-OES determination and via photometer, respectively. A sample was taken after 24 hours and stored at -20 ± 5 °C. The measured test item concentration based on measurement of P from ortho-phosphate via photometer at the beginning of the test was at 130% of the nominal concentration and after 48 hours the measured concentration was 132% of the nominal concentration. Since the measured concentration was above 120% of nominal tThe determination of the biological results was based on the geometric mean of the measured concentration. The measured test item concentration based on measurement of total P via ICP-OES at the beginning of the test was at 119% of the nominal concentration and after 48 hours the measured concentration was 103% of the nominal concentration.

Table 4: Calculated Test item Concentration based on constituent 3 (= 16.8% of test item) and % of nominal

Nominal Concentration Test Item	Calculated Concentration  Test Item  t = 0 h	Calculated Concentration  Test Item  t = 48 h	% of Nominal concentration t = 0 h	% of Nominal concentration t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	--	--	--	--
176	228.08	231.83	130	132

 

Table 5: Geometric Mean of calculated test item concentration based on constituent 3

Nominal Concentration Test Item	Geometric Mean of calculated test item concentration based on constituent 3	Geometric Mean calculated test item concentration based on constituent 3 and active ingredient (due to 43.1% water content of test item)
mg/L	mg/L	mg/L
Blank control	--	--
176	229.94	130.84

Table 6: Calculated Test item Concentration based on constituent 1 (= 22.4% of test item) and % of nominal

Nominal Concentration Test Item	Calculated Concentration  Test Item  t = 0 h	Calculated Concentration  Test Item  t = 48 h	% of Nominal concentration t = 0 h	% of Nominal concentration t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	--	--	--	--
176	209.27	181.27	119	103

 

Table 7: Geometric Mean

Nominal Concentration Test Item	Geometric Mean of calculated test item concentration based on constituent 1	Geometric Mean calculated test item concentration based on constituent 1 and active ingredient (due to 43.1% water content of test item)
mg/L	mg/L	mg/L
Blank control	--	--
176	194.77	110.82

Validity criteria fulfilled:

yes

Remarks:

See details in secation "Any other information on results incl. tables"

Conclusions:

No effects were observed up to the highest test item concentration.

Description of key information

No effects observed, NOEC is 110 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

110 mg/L