Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

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:
22 June - 31 August 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study was conducted in accordance with International guidelines and GLP. All guideline validity criteria were met.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
April 2004
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark.
- Stability under test conditions: Biodegradation of test tem in the test system was monitored.
- Solubility and stability of the test substance in the solvent/vehicle: solvent vehicle not used.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: N/A

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No
- Preliminary purification step (if any): No
- Final dilution of a dissolved solid, stock liquid or gel: no
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Test item applied as a Water Accomodated Fraction (WAF) at a loading rate of 100 mg/L (nominal).

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): N/A

OTHER SPECIFICS: N/A
Analytical monitoring:
yes
Remarks:
One component of the UVCB substance was selected as a marker anlayte. The marker analyste was used to confirm the concentration of the the UVCB substance in the test system by extapolation.
Details on sampling:
- Concentrations: 100 mg/L loading rate (nominal)
- Sampling method: Samples were taken from the control and the 100 mg/L loading rate WAF test group from the bulk fresh test preparations at 0 and 24 hours and from the pooled replicates (R1 to R4) of the old media at 24 and 48 hours for quantitative analysis. Samples were taken for immediate analysis.
Duplicate sample sets of samples were taken at 0, 24 and 48 hours and stored frozen for further analysis if necessary.
- Sample storage conditions before analysis: See above.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A nominal amount of test item (500 mg) was added to the surface of 5 liters of test water (Elendt M7) to give the 100 mg/L loading rate. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 95 hours and the mixture allowed to stand for 1 hour. Microscopic observations made on the WAF indicated that a significant amount of dispersed test item was present in the water column and hence it was considered justifiable to remove the WAF by filtering through a glass wool plug (2 to 4 cm in length). A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. A glass wool plug was inserted into the opposite end of the tubing and the WAF removed by mid depth siphoning (the first 75 to 100 mL discarded) to give the 100 mg/L loading rate WAF. Microscopic observations of the WAFs were performed after filtering and showed no micro-dispersions of the test item to be present.
- Controls: Elendt M7 only (negative control). Positive control (potassium dichromate) test was conducted in a separate GLP study between 23-25 Jan 2018 using a concentration range of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): N/A
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea (Daphnia magna)
- Strain/clone: -
- Justification for species other than prescribed by test guideline: N/A
- Source: In-house laboratory culture.
- Age of parental stock (mean and range, SD): Not reported
- Feeding during test: No
- Food type: N/A
- Amount: N/A
- Frequency: N/A

ACCLIMATION
- Acclimation period: N/A
- Acclimation conditions (same as test or not): N/A
- Type and amount of food: N/A
- Feeding frequency: N/A
- Health during acclimation (any mortality observed): N/A

QUARANTINE (wild caught)
- Duration: N/A
- Health/mortality: N/A

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: Adult daphnids were maintained in 150 mL glass beakers containing 100 mL Elendt M7 medium in a temperature controlled room maintaining the water temperature at 18 to 22 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and GEMMA Micro 300 food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
Not reported
Test temperature:
21 ºC
pH:
7.8 - 8.2
Dissolved oxygen:
≥ 8.5 mg/L
Salinity:
N/A
Conductivity:
N/A
Nominal and measured concentrations:
100 mg/L nominal loading rate
Details on test conditions:
TEST SYSTEM
- Test vessel: Beaker
- Type (delete if not applicable): Open but covered to reduce evaporation.
- Material, size, headspace, fill volume: Glass, 150 mL capacity, filled to 100 mL.
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A
- Renewal rate of test solution (frequency/flow rate): Renewal at 24 h.
- No. of organisms per vessel: 5 per replicate/ beaker.
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): N/A
- Biomass loading rate: N/A

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: In-house preparation.
- Total organic carbon: Not reported
- Particulate matter: Not reported
- Metals: Not reported
- Pesticides: Not reported
- Chlorine: Not reported
- Alkalinity: Not reported
- Ca/mg ratio: Not reported
- Conductivity: Not reported
- Salinity: Not reported
- Culture medium different from test medium: No
- Intervals of water quality measurement: Daily (fresh and expired solutions)

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 808 - 848 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mobility and sub-lethal effects recorded daily.

VEHICLE CONTROL PERFORMED: N/A

RANGE-FINDING STUDY
- Test concentrations: Nominal loading rates of 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: Yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities:
- Observations on body length and weight: Not reported
- Other biological observations: No
- Mortality of control: 0 %
- Other adverse effects control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: Yes
- Limit test: No
- Dose-response test: Yes
- ECx: 48 h EC50 = 0.75 mg/L
- Other: 48 h NOEC = 0.56 mgL
Reported statistics and error estimates:
No immobilization was observed.

Table 1       Immobility

 

Nominal loading rate

(mg/L)

Replicate

Immobility

(%)

24 h

48 h

Rep.

Mean

Rep.

Mean

Control

R1

0

0

0

0

R2

0

0

R3

0

0

R4

0

0

100

R1

0

0

0

0

R2

0

0

R3

0

0

R4

0

0

Table 2       Chemical Analysis

 

Nominal Loading Rate

(mg/L)

Determined Concentration of Test Item

(mg/L)

0 h Fresh

24 h Expired

24 h Fresh

48 h Expired

Control

<LOQ

<LOQ

<LOQ

<LOQ

100

0.00248

<LOQ

0.0485

0.0270

LOQ: 0.0021 mg/L  

Validity criteria fulfilled:
yes
Conclusions:
The 48 h EL50, based on immobility, for Daphnia magna under the conditions of this test was >100 mg/L.
Executive summary:

OECD 202 (2018) - The 48 h acute toxicity of the test item to Daphnia magna was studied under semi-static conditions.  Test species were exposed to control and test chemical at a nominal loading rate of 100 mg/L.  The test solution was prepared from a water accommodated fraction which contained the test item suspended in dilution water at a nominal loading rate of 100 mg/L.  The solution was stirred for 95 h.  The solubilized fraction was decanted and filtered to prepare the treatment solution for this limit test.  Test substance concentration was confirmed by LC-MS analysis of a "marker analyte" of the UVCB using a validated method at 0 h, 24 h (fresh and expired solutions) and 48 h. The test substance concentration was calculated/extrapolated from the determined marker analyte concentration. Mortality/ immobilisation and sublethal effects were observed daily.

The 48 h EL50 was >100 mg/L.  The 48 h NOEL based on mortality/immobilization/sublethal adverse effects was 100 mg/L.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

Description of key information

48 h EL50 = >100 mg/L to Daphnia magna; OECD 202; Squance, D. (2019).

Key value for chemical safety assessment

Additional information

OECD 202 (2018) - The 48 h acute toxicity of the test item to Daphnia magna was studied under semi-static conditions.  Test species were exposed to control and test chemical at a nominal loading rate of 100 mg/L.  The test solution was prepared from a water accommodated fraction which contained the test item suspended in dilution water at a nominal loading rate of 100 mg/L.  The solution was stirred for 95 h.  The solubilized fraction was decanted and filtered to prepare the treatment solution for this limit test.  Test substance concentration was confirmed by LC-MS analysis of a "marker analyte" of the UVCB using a validated method at 0 h, 24 h (fresh and expired solutions) and 48 h. The test substance concentration was calculated/extrapolated from the determined marker analyte concentration. Mortality/ immobilisation and sublethal effects were observed daily.

The 48 h EL50 was >100 mg/L.  The 48 h NOEL based on mortality/immobilization/sublethal adverse effects was =>100 mg/L.

This study is acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.