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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:
From September 15, 2017 to October 27, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 6K0001
- Purity test date: 97.5% (The test item was treated as 100% in purity.)
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: The test sample was stored in a dark storage place at room te
mperature.
- Stability under test conditions: IR spectrum of the test item after the completion of the experiment
was the same as that before the start of the experiment, and it indicated that the test item was stable
under the storage conditions.
- Solubility and stability of the test substance in the solvent/vehicle: Dimethylsulfoxide;>50 mg/mL,
Acetone;>50 mg/mL

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Concentrations: Stock solution content of 50.0, 27.8, 15.4, 8.57 and 4.76% (All test levels)
- Sampling method: Another solution sampled separately from the preparing container (at the prep
aration)
The mixed solution taken out with equal volume from the middle layer of the test solution in test v
essels in each test level (before the renewal and at the end of exposure)
- Sample storage conditions before analysis: immidiately pretreated

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:The test sample (100 mg) and dilution water (1 L) were mixed to prepare the nominal
concentration of 100 mg/L, and they were gently stirred with a magnetic stirrer for 48 hours at about
20°C. Then, they were settled for 1 hour at about 20°C, and the middle layer was collected as the
stock solution. Required volumes of the stock solution and dilution water processing similar to the
control without the test item were mixed and stirred in a preparing container to prepare the test solut
ion which was divided into each test vessel. This preparation was carried out for the start of exposure
and renewal of test solution.
- Controls:Dilution water without the test item, which was treated in the same manner as the test
solution except for collecting the middle layer

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Clone A
- Source: The University of Sheffield, UK
- Age study initiation: <24 hours
- Feeding during test: No
ACCLIMATION
- Acclimation period: 14-day
- Acclimation conditions (same as test or not):same as test
- Type and amount of food:Chlorella vulgaris of 0.1-0.2 mgC (Organic carbon content)/day per Daph
nia
- Feeding frequency: everyday
- Health during acclimation (any mortality observed): survival rate of parent animals was 100%
METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAG
ES:All young daphnid was removed from culture vessel within 24 hours from the start of exposure.

Study design

Test type:
semi-static
Water media type:
freshwater
Remarks:
Dechlorinated tap water
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
32 mg CaCO3/L
Test temperature:
20.0-20.1°C
pH:
7.8-7.9
Dissolved oxygen:
8.5-8.9
Nominal and measured concentrations:
Nominal concentrations: 50.0, 27.8, 15.4, 8.57 and 4.76% as the stock solution (saturated solution)
content.
Measured concentrations:1.37, 0.763, 0.421, 0.217 and 0.126 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 mL stoppered Erlenmeyer flask
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 50mL, fill volume without headspace
- Aeration: No
- Renewal rate of test solution (frequency/flow rate):renewal at 24 hours after
- No. of organisms per vessel:5 daphnids/test vessel
- No. of vessels per concentration (replicates):4 replicates/test level
- No. of vessels per control (replicates):4 replicates/test level
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:Dechlorinated tap water
Total hardness (Ca, Mg) 32mg/L
Suspended solid < 1mg/L
pH 7.8 (24.7°C)
Total organic carbon < 0.5mg/L
Chemical oxygen demand 1.0mg/L
Residual chlorine < 0.02mg/L
Ammonium ion < 0.1mg/L
Total cyanide < 0.05mg/L
Alkalinity 36mg/L
Electric conductivity 13mS/m
Total mercury < 0.0005mg/L
Cadmium < 0.001mg/L
Chromium (VI) < 0.01mg/L
Lead < 0.001mg/L
Arsenic < 0.005mg/L
Iron < 0.01mg/L
Copper < 0.001mg/L
Cobalt < 0.001mg/L
Manganese < 0.005mg/L
Aluminum 0.03mg/L
Zinc < 0.1mg/L
Nickel < 0.001mg/L
Silver < 0.0001mg/L
1,2-dichloropropane < 0.002mg/L
Chlorothalonil < 0.001mg/L
Propyzamide < 0.0008mg/L
Chlornitrofen < 0.0001mg/L
Simazine < 0.0003mg/L
Thiobencarb < 0.001mg/L
Diazinon < 0.0005mg/L
Isoxathion < 0.0008mg/L
Fenitrothion < 0.0003mg/L
EPN < 0.0006mg/L
Dichlorvos < 0.001mg/L
Iprobenfos < 0.0008mg/L
PCB < 0.0005mg/L
Boron < 0.1mg/L
Fluorine < 0.1mg/L
Sulfate ion 11mg/L
Chloride ion 9.3mg/L
Sodium 11mg/L
Potassium 3.1mg/L
Calcium 9.1mg/L
Magnesium 2.2mg/L
- Culture medium different from test medium: No differences
- Intervals of water quality measurement: 6 month
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16-hour light/8-hour dark
- Light intensity: Room light
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Observation about immobility and symptoms of test organism was conducted at 24 and 48 hours after
exposure. Daphnids were considered immobile when they were not able to swim within 15 seconds
after gentle agitation of the test vessel.
RANGE-FINDING STUDY
- Test concentrations: 100, 50.0, 10.0, 5.00% of the stock solution content
- Results used to determine the conditions for the definitive study: test range and renewal period
Reference substance (positive control):
not specified
Remarks:
There are periodically reference substance data.

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.209 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
0.126 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
other: neither immobilization nor other symptons
Details on results:
- Behavioural abnormalities: It was not observed.
- Observations on body length and weight: No
- Mortality of control:0%
- Other adverse effects control:No
- Abnormal responses: Immobilization, lethargic and reduced activity
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal
values: It was not observed
- Effect concentrations exceeding solubility of substance in test medium: No

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
48-hour EC50 was 0.209 mg/L and 48-hour NOEC was 0.126 mg/L.
Executive summary:

This study was conducted in order to confirm the effect of the test item on the test organisms below

the solubility of the test item in dilution water. As a result, 48-hour EC50was 0.209 mg/Land 48-hour

NOEC was 0.126 mg/L.

Since the test item concentration in the test solution at the end of exposure was not maintained within

±20% of that at the start in the preliminary study, the definitive study was conducted by semi-static

regime (renewal at 24 hours) to maintain the test item concentration. As a result, the maintenance rate

of test item concentrations in the test solution during exposure were 80.8-99.0% of the concentrations

at the preparation.

The environmental conditions were within the suitable range; therefore, it was concluded that this study

complied with the applied test guideline.