Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2013-05-27 to 2013-07-26
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
Adopted: 17 July 1992
GLP compliance:
Specific details on test material used for the study:
Lot No.: KNC110310
Purity: 100.0%
Analytical monitoring:
Details on sampling:
- Concentrations: Test substance concentrations in the test solutions were measured in all tested groups in the definitive test at initiation and termination of exposure.
- Sampling method: Test solutions for measurement were taken from mid-depth of the test vessels without agitation in each test group (n = 1).
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
A 0.5000 g aliquot of the test substance was weighed with the electronic balance. A stock solution was prepared by mixing the test substance with the dilution water to make volume to 100 mL. Test solution at 100 mg/L was prepared by diluting 100 mL of the stock solution with the dilution water to make volume to 5 L.
- Controls: yes, test medium control
Test organisms (species):
Oryzias latipes
Details on test organisms:
- Common name: Medaka
- Source: in-house culture
- Length at study initiation): Mean total length: 26 mm (22 - 29 mm), n=20
- Weight at study initiation: Mean body weight: 0.15 g (0.10 - 0.21 g), n=20

- Acclimation period: ≥ 12 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: commercial feed, Tetramin (Tetra Japan), amount: ca 5% of body weight / day
- Feeding frequency during acclimation: 5 times a week (not fed for 48 hours prior to exposure).
- Health during acclimation: Mortality during 7 days prior to exposure was less than 5% (practically 0.1 %).

Test type:
Water media type:
Limit test:
Total exposure duration:
96 h
42 mg CaCO3/L at the initiation of exposure, measured by Total hardness test kit HA-DT (HACH COMPANY Ltd.)
Test temperature:
23.1- 23.3℃, measured by Water temperature meter SK-250WP (Sato Keiryoki Mfg. Co., Ltd.)
7.6 - 8.1, measured by pH meter D-51 (Horiba Ltd.)
Dissolved oxygen:
6.2 - 8.4 mg/L, measured by DO meter Model 58 (YSI/Nanotech Inc.)
Nominal and measured concentrations:
Nominal: 100 mg/L
Measured: 98.2 mg/L
Details on test conditions:
- Test vessel: 5 L volume whole glass aquarium
- Type: closed
- Volume of test solution: 5 L 1 vessel
- Replication: 1 vessell test group
- Number of organisms tested: 10 individuals/ test group
- pH: No adjustment
- Lighting: Room lighting, 16-hour light / 8-hour dark condition

- Source/preparation of dilution water: Tap water (Takarazuka-shi, Hyogo) dechlorinated with charcoal filter was used for the dilution water after saturation with oxygen by aeration, and adjustment to the appropriate test temperature
- Total organic carbon: <0.5 mg/L
- Metals: Mercury: <0.0005 mg/L; Copper: <0.01 mg/L; Cadmium: <0.001 mg/L; Zinc: <0.01 mg/L; Lead: <0.005 mg/L; Aluminum: <0.02 mg/L; Nickel: < 0.02 mg/L; Chromium: <0.02 mg/L; Manganese( dissolved): <0.005 mg/L; Tin: <0.03 mg/L; lron(dissolved): <0.03 mg/L; Sodium: 38 mg/L; Potassium: 3.9 mg/L; Calcium: 12 mg/L; Magnesium: 3.9 mg/L
- Chlorine: . Residual chlorine in the dechlorinated tap water was checked by a residual chlorine meter to be less than the detection limit (<0.05 ppm).
- M-Alkalinity: 58 mg/L
- Culture medium different from test medium: no

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortalities and toxic symptoms as compared to the control were recorded at 3, 6, 24, 48, 72 and 96 hours after initiation of exposure. Dead fish were removed immediately after finding to prevent worsening of water quality. DO, water temperature and pH of the test solutions in all groups were measured before the initial of test and once daily. At the same time, appearance of the test solutions was recorded.

- Range finding study
- Test concentrations: 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: no adverse effect was observed even at the highest concentration (100 mg/L) when
compared with the control.
Reference substance (positive control):
Copper [cupric] sulfate pentahydrate
Key result
96 h
Dose descriptor:
Effect conc.:
> 98.2 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Cumulative Mortalities, Toxic Symptoms:
Mortalities in the dilution water control group were 0% during the exposure.
Cumulative mortality was 0% and no toxic symptoms were observed In the medaka exposed to the test substance at the mean measured concentration of 98.2 mg/L for 96 hours.
- Test Substance Concentrations and Appearance of the Test Solutions:
Measured concentrations of the test substance were stable and 97.9 and 98.5 mg/L at the initiation and termination of exposure, respectively (97.9 and 98.5% of the nominal concentration) and mean measured concentration was 98.2 mg/L (98.2% of the nominal concentration).
Recoveries of QC sample were 101 and 99.2% at the nominal concentration of 5.00 and 120 mg/L, respectively. No peak derived from the test substance was detected in the dilution water control and the measured concentration was found to be less than quantification limit (5.00 mg/L).
Appearance of the test solution was clear and no precipitation, suspension, floating material or coloration was observed in the test substance solution through the period from preparation to termination of exposure.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- LC50 (96hr): 1.40 mg/L
- Other: conducted at approximately 6-month intervals
Reported statistics and error estimates:
Since the test was conducted as a limit test at the nominal concentration of 100 mg/L, LC50 was determined to be greater than (>) the above concentration.
Validity criteria fulfilled:
96-hour LC50 value of the test substance for medaka after 96-hour exposure is >98.2 mg/L on the basis of the mean measured concentration.
Executive summary:

A 96-hour LC50 of test item for medaka resulting from an acute toxicity test was determined based on OECD 203. A limit static test at nominal concentration of 100 mg/L was conducted.

The 96-hour LC50 value was >98.2 mg/L on the basis of the mean measured concentration.

Description of key information

The acute toxicity to medaka was determined based on OECD 203.

The 96-hour LC50 value was >98.2 mg/L on the basis of the mean measured concentration.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
98.2 mg/L

Additional information