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Ecotoxicological information

Short-term toxicity to fish

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Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-11-02 to 2016-10-31
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental study performed according to the OECD 203 guideline and under GLP compliance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
OECD, 1993. OECD Guidelines for Testing of Chemicals No. 203 “Fish, Acute Toxicity Test (Adopted: July 17, 1992)”.
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Test concentrations were verified by chemical analysis. Please, see the study report attached for more details.
Vehicle:
no
Details on test solutions:
Preparation of the test solutions
The test was dispensed by elutriating with test water. The purity of test substance (content of Active ingredients) was calculated 98 % based on the test material information chart and COA (Annex 1, Annex 2). When dispensing test substance, which was 1000 mg/L of stock solution, the precipitation occurred, but the test solution was processed to fit the set concentration over stirring and dilution to verify the toxic values. To analyze toxic value and test concentration, definitive test was carried out after stock solution was dispensed
inevitably.

Preparation of stock solution
Stock solution concentration → 1000 mg/L
Test substance → 1.020 g (purity 98 %)
Vehicle → Water
Final volume → 1000 mL volumetric flask
Mix method → Stir 5 min
Test organisms (species):
Oryzias latipes
Details on test organisms:
CULTURING CONDITIONS
Vessel : A transparent rectangular vessel made of glass
Water : Dechlorinated water
Dissolve oxygen : 80 % or more of the saturation
Light : Fluorescent light, 16-hour light (300-600 lux) / 8-hour dark
Water temperature : (21 - 25) ℃
Feeding food : Top meal / Supply once per day

MONITORING OF BREEDING ENVIRONMENT
Temperature : Measure once a month with Orion 3 Star(Thermo Scientific Co., Lt, USA)
pH and DO : Measure once a month with Orion 3 Star(Thermo Scientific Co., Lt, USA)
Illumination and light period: Measure once per quarter with a illumination meter (ANA-F11, TOKYO Photoelectric Co., Ltd, Japan) and the recorder (KR100, Konics, Korea).
As a result of environmental monitoring of the fish breeding room (1), there were no changes that would affect the test.

ACCLIMATATION AND FOOD SUPPLY
The stock of fish, 80 fish in a 100 L glass container, was acclimated to test conditions 7 days before the definitive test. Food was not given during the 24 hours prior to exposure. The batch of fish was observed to be in good condition prior to use in the test. The test fish were maintained with water temperature of (21.0 - 25.0) ℃, and DO of 80 % or more of Air Saturation Value (ASV). No mortality was recorded during the acclimation period.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
24 mg/L
Test temperature:
22.6 ℃ (22.2 – 23.0)℃
pH:
7.28 (7.09 - 7.55)
Dissolved oxygen:
6.74 mg/L (5.33 - 8.32) mg/L 78.0 % (61.7 - 96.3)% of ASV
Nominal and measured concentrations:
- Nominal concentration, mg/L: Control, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 mg/L
- Mean measured concentration, mg/L: Control, 0.09, 0.16, 0.31, 0.73, 1.31, 2.51 mg/L
Details on test conditions:
TEST CONDITIONS
Room : Fish Test Room
Test species : Oryzias latipes
Size : Recommended total length (2.0 ± 1.0) cm
Duration of exposure : 96 hours
Exposure procedure : semi-static system
Volume of test solution : 10 L (glass vessel : 28 cm H. × 24 cm φ)
Number of vessels : 1 vessel / concentration
Biomass loading : 10 fish / concentration
Temperature : (21 - 25) ℃, ±1 ℃
D.O. : 60 % or more of the saturation
pH : 6.0 - 8.5 (Not adjusted)
Hardness : 10 mg/L - 250 mg/L
Light : 16-hour light (08:00 - 24:00), 8-hour dark (24:00 - 08:00)
Feeding : None

RANGE-FINDING TEST
The range-finding test of this test substance was conducted 0.1, 1, 10, 100 mg/L under static condition. Five fish were exposed to the each test vessel without replication. The number of dead fish was observed for 96 hours.
From preliminary experimental results, 100 % mortality in a setting density of 10 mg/L and 100 mg/L got shown and mortality at 0.1 mg/L wasn’t monitored. Therefore we would conduct definitive test in the concentration of settings; 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 mg/L in the difference between test groups and control.

OBSERVATION
The fish were inspected at test start 3, 24, 48, 72 and 96 hours. Fish were considered dead if there was the absence of respiratory movement and response to external stimulation. After completion of the experiment, 10 fish were taken from each of the control group and the treated group (concentration which all fish is live) and then measured body weight and total length of them.

ENVIRONMENTAL CONDITIONS
pH, dissolved oxygen and water temperature were recorded daily for all treatments. pH, dissolved oxygen (DO) and water temperature, during the exposure period were maintained as follows:
- pH : 7.28 (7.09 – 7.55, Table 3).
- Dissolved oxygen : 6.74 mg/L (5.33 – 8.32) mg/L 78.0 % (61.7 – 96.3) % of ASV (Table 4)
- Temperature : 22.6 ℃ (22.2℃ – 23.0℃, Table 5)
No food and air was provided during the test period.
Reference substance (positive control):
yes
Remarks:
Positive control : The test was performed in control, 0.10, 0.14, 0.20, 0.27, 0.38, 0.54 mg/L (geometric series 1.4, nominal concentration) as active ingredient of Pentachlorophenol sodium salt (Sigma-aldrich, Lot No. BCBJ8692V) as positive test
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.038 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.31 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
1.099 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
0.31 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
Please see the attached study report for more details on results

Validity of the test
- Mortality in the control was less than 10 % at the end of exposure.
- D.O. concentrations were 60 % or more of the saturation during the exposure period.
- pH of test solution (min. 7.09, max. 7.55), and it is suitable for validation criteria (6.0 – 8.5)
- Hardness of test water 24 mg/L, and it is suitable for validation criteria (10 – 250) mg/L (as CaCO3)]
- Temperature of test solution (min. 22.2℃, max.23.0℃), and it is suitable for validation criteria ±1 ℃
Reported statistics and error estimates:
- LC50: LC50 and 95 % confidence limit was calculated by Probit method (EPA600/4-85/13,1985) based on the result of the observation of mortality at the time of 48 and 96 hours after finishing test material processing.
- NOEC: NOEC does not have any toxic symptoms, and it indicated maximum test concentration (measured concentration) that mortality hasn’t occurred.
Sublethal observations / clinical signs:

Please, see the attached study report

Validity criteria fulfilled:
yes
Conclusions:
The result of an acute toxicity test in static system for the Fish (Oryzias latipes ) of Zinc cyanide is as follows, and LC50 and NOEC were designated by measured concentration.
- 96 hour-LC50 is 1.038 mg/L, and 95 % confidence interval is (0.819 – 1.315) mg/L
- 96 hour-NOEC is 0.31 mg/L
Executive summary:

The study was conducted to assess the acute toxicity of Zinc cyanide to ricefish (Oryzias latipes) under static system. This test was performed according to OECD guideline for the testing of chemicals 203 and under GLP conditions.

This test was conducted with a set concentrations of 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 mg/L (the average measured concentration: 0.09, 0.16, 0.31, 0.73, 1.31, 2.51 mg/L), and exposed 10 fish for each test concentration.

The result of this acute toxicity test in static system for the Fish (Oryzias latipes) of Zinc cyanide is as follows, and LC50 and NOEC were designated by measured concentration.

- 96 hour-LC50 is 1.038 mg/L, and 95 % confidence interval is (0.819 – 1.315) mg/L;

- 96 hour-NOEC is 0.31 mg/L.

The validity criteria were successful, therefore this study can be considered acceptable for that endpoint.

Description of key information

A reliable study was available to assess the acute toxicity of Zinc cyanide to ricefish (Oryzias latipes ) under static system. This test was performed according to OECD guideline for the testing of chemicals 203 and under GLP conditions.

This test was conducted with a set concentration of 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 mg/L (the average measured concentration: 0.09, 0.16, 0.31, 0.73, 1.31, 2.51 mg/L), and exposed 10 fish for each test concentration.

The result of the acute toxicity test in static system for the Fish (Oryzias latipes ) of Zinc cyanide is as follows, and LC50 and NOEC were designated by measured concentration.

- 96 hour-LC50 is 1.038 mg/L, and 95 % confidence interval is (0.819 – 1.315) mg/L

- 96 hour-NOEC is 0.31 mg/L.

The validity criteria were successful, therefore this study can be considered acceptable for that endpoint.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
1.038 mg/L

Additional information