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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods
Qualifier:
according to guideline
Guideline:
other:
Version / remarks:
Testing method relating to the new chemical substances, etc. (Yakushokuhatsu No. 1121002, Heisei 15-11-13 Seikyoku No. 2, Kanpokihatsu No. 031121002 on November 21, 2003).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Principles of method if other than guideline:
To assess the toxicity of the test substance by conducting acute toxicity study to fish for notification of the new chemical substance under the Chemical Substance Control Law (CSCL) in Japan.
GLP compliance:
yes
Specific details on test material used for the study:
During the study period, the test substance was stored in desiccators (storage conditions: room temperature, under nitrogen purging).
Analytical monitoring:
yes
Details on sampling:
- Concentrations:
- Sampling method: 1.5 mL of test solution, at preparation and immediately before water renewal (or end of exposure)
- Sample storage conditions before analysis: samples analysed immediately, not stored
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution prepared (test substance and test water) stirred for 3 minutes, used to prepare test solutions. 5L of test water per concentration, various volume of stock solution added for each concentration to be tested
- Controls: test water used for control group
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): No vehicle used
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Not specified
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Killifish
- Source: In-house breeding
- Age at study initiation: less than 6 months after hatching
- Length at study initiation (mean and range): 1.96cm (1.78 to 2.12 cm)
- Weight at study initiation (mean and range): 0.054g (0.034 to 0.069g)

ACCLIMATION
- Acclimation period: January 7 2008 - February 4 2008
- Acclimation conditions (same as test or not): test water, flow-through system
- Type and amount of food during acclimation: Tetramin: Approx. 2% of fish weight/day
- Feeding frequency during acclimation: Daily (no food given from 24 hours before the start of exposure)
- Health during acclimation (any mortality observed): At 7 days before the start of exposure: Below 5%

FEEDING DURING TEST : None
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
24 +/- 1°C
pH:
6.0 - 8.5
Dissolved oxygen:
60% and above the saturated oxygen concentration
Salinity:
not applicable
Conductivity:
15 mS/m
Nominal and measured concentrations:
Nominal: 20, 36, 63, 110 and 200 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 5L glass tank
- Type - closed: Surface of the solution was covered by transparent Teflon sheet
- Material, size, headspace, fill volume: glass, 5L, No headspace
- Aeration: no
- Renewal rate of test solution (frequency): Every 24 hours
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water supplied by Yokohama public water system, which was dechlorinated by filtration with activated charcoal and addition of sodium thiosulfate.
- Total organic carbon: <0.5 mg/L
- Particulate matter: <1.0 mg/L
- Metals: - Pesticides: <0.001 mg/L (LoD)
- Chlorine: - Alkalinity: 41 CaCO3/L

- Intervals of water quality measurement: 6 months

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: 16-hour light and 8-hour dark cycle
- Light intensity: 1000 lux or below

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
1) Measurement of water quality and observation of the appearance of test water
Water temperature, dissolved oxygen concentration and pH of the test water were measured and the appearance was observed every day. Both test water immediately before water renewal and at preparation were measured and observed when water was renewed.
2) Measurement of fish weight and total length
At the start of exposure, 10 fish which were acclimatised in the same water tank as the test fish of the same lot were collected at random as the representative examples of the relevant lot and their weight and total length were measured
3) Observation of test fish
Fish were observed at 24, 48, 72 and 96 hours of exposure for mortality and abnormal behaviour. When dead fish were found, they was immediately removed to avoid deterioration of water quality.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study : Yes (results see table 1)
- Test concentrations: 20 and 200 mg/L
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
105 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
32.6 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
195 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: None noted
- Other biological observations: None noted
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: Yes (see table 2)
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: No
Results with reference substance (positive control):
Not applicable.
Reported statistics and error estimates:
Probit method, 95% confidence limits calculated
Sublethal observations / clinical signs:

Table 2: Results

Control

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

24

48

72

96

0.0

0.0

Death (cumulative)

0

0

0

0

Critical

0

0

0

0

Abnormal

0

0

0

0

Normal

10

10

10

10

TOTAL

10

10

10

10

 

Conc. 1

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

24

48

72

96

20.0

18.2

Death (cumulative)

0

0

 

0

0

Critical

0

0

0

0

Abnormal

0

0

0

0

Normal

10

10

10

10

TOTAL

10

10

10

10

 

Conc. 2

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

24

48

72

96

36.0

32.6

Death (cumulative)

0

0

0

0

Critical

0

0

0

0

Abnormal

0

0

0

3a

Normal

10

10

10

7

TOTAL

10

10

10

10

 

Conc. 3

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

24

48

72

96

63.0

58.5

Death (cumulative)

0

0

0

2

Critical

0

0

0

0

Abnormal

0

0

10a

8a

Normal

10

10

0

0

TOTAL

10

10

10

10

 

Conc. 4

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

24

48

72

96

110.0

107

Death (cumulative)

0

0

1

3

Critical

0

0

1b

0

Abnormal

0

3a

8a

7a

Normal

10

7

0

0

TOTAL

10

10

10

10

 

Conc. 5

Nominal Conc. (mg/L)

Measured conc. (mg/L)

Symptoms

Time (h)

Time (h)

Time (h)

Time (h)

24

48

72

96

200.0

195.0

Death (cumulative)

0

1

8

10

Critical

0

0

2b

0

Abnormal

0

9a

0

0

Normal

10

0

0

0

TOTAL

10

10

10

10

a - abnormal swimming

b - unable to swim

Validity criteria fulfilled:
yes
Remarks:
1) mortality in control group less than 10% at the end of exposure; 2) Disssolved oxygen concentration remained above 60% of the saturated oxygen concentration during the exposure period
Conclusions:
In the acute toxicity to fish test, the 96h LC50 was determined as 105 mg/L.

Description of key information

In an GLP-compliant study conducted with analytical monitoring in accordance with a standard method equivalent to the OECD guideline, the toxicity of the test item to fish was assessed. The 96h LC50 value was determined to be 105 mg/L indicating that the substance is of low toxicity to fish. The 96h LC50 value is used for the Chemical Safety Assessment.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
105 mg/L

Additional information