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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
Study period:
From 23 September 2015 to 20 July 2017
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD series on testing and assessment number 23, 2000.
Deviations:
no
GLP compliance:
yes
Remarks:
The certificate copy was not included in the study report.
Test material information:
Composition 1
Specific details on test material used for the study:
APPEARANCE
Reddish solid below 25°C / liquid above 25°C

SOURCE OF TEST MATERIAL
- Source and batch/lot No.of test material: Solvay / MR92143651 / 206887/A
- Expiration date of the batch: 01 January 2017
- Purity/Composition: 86.4%
- Purity/composition correction factor: No correction factor required
- Purity test date: 13 April 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature.
- Stability under test conditions: until 01 January 2017 (expiry date)
- Solubility of the test substance in the vehicle (20°C): Insoluble in Water
- Stability of the test substance in the vehicle (20°C): Not indicated
- Stability at higher temperatures: Yes, maximum temperature: 50°C

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was melted at approximately 40°C before weighing.
- No correction was made for the purity/composition of the test item.
Analytical monitoring:
yes
Remarks:
UPLC-MS/MS
Details on sampling:
- Concentrations: WAFs prepared at loading rates of 4.6, 10, 22, 46 and 100 mg/L.
- Sampling method: During the final test single samples for possible analysis were taken from all test concentrations and the control.
- At the start of the test and after 72 hours from the freshly prepared solutions.
- At the first renewal (t=24h) and the end of the test from the 24-hour old solutions.
- Sample storage conditions before analysis: The test samples were stored in the freezer (≤ -15°C).
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was melted at approximately 40°C before weighing. Preparation of test solutions started with Water Accommodated Fractions (WAFs) individually prepared at loading rates of 1.0 mg/L and higher. A magnetic stirring period of three days followed by filtration was applied to ensure reaching maximum solubility in test medium.
- Evidence of undissolved material: The resulting dispersion was subsequently filtered through a 0.45 μm membrane filter (Whatman; RC55) to remove undissolved test item. The final test solutions were all clear and colourless except for the WAF prepared at 100 mg/L in the combined limit/range-finding test that was hazy.
- Controls: Test medium without test item or other additives.
Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Carp
- Strain: Cyprinus carpio, Teleostei, Cyprinidae [Linnaeus, 1758]
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Mean length at study initiation: 2.4 ± 0.2 cm
- Mean weight at study initiation: 0.18 ± 0.05 g
- Characteristics: F1 from a single parent-pair bred in UV-treated water.

HOLDING
- Quarantine/Acclimation period: at least 12 days after delivery.
- Medium: Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
* CaCl2.2H2O: 211.5 mg/L
* MgSO4.7H2O: 88.8 mg/L
* NaHCO3: 46.7 mg/L
* KCl: 4.2 mg/L
- Type and amount of food during acclimation: pelleted fish food.
- Feeding frequency during acclimation: daily
- Health during Quarantine/Acclimation: In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

FEEDING DURING TEST
- No feeding from 24 hours prior to the test and during the total test period.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
Adjusted ISO medium with a hardness of 180 mg CaCO3 per litre.
Test temperature:
The temperatures measured during the study in the various test vessels are presented in Table I (See field "Any other information on materials and methods incl. tables").
pH:
The results of measurement of pH are presented in Table II (See field "Any other information on materials and methods incl. tables").
Dissolved oxygen:
The results of measurement of oxygen concentrations are presented in Table III (See field "Any other information on materials and methods incl. tables").
Salinity:
Not applicable
Conductivity:
Unspecified
Nominal and measured concentrations:
WAFs prepared at 1-Phenyldecane-1,3-dione loading rates of 4.6, 10, 22, 46 and 100 mg/L ; equivalent to measured concentrations of 0.16, 0.45, 0.59 1.2 and 10.5 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: 3.5 litres, all-glass, containing 3 litres of test solution.
- Aeration: the test media were not aerated during the test.
- Renewal rate of test solution: Daily renewal of test solutions.
- No. of organisms per vessel: 7 per concentration (Loading: 0.42 g fish/Litre, i.e. 7 fish per 3 litres of test medium).
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: RO-water (tap-water purified by reverse osmosis)
- Culture medium different from test medium: No.
- Intervals of water quality measurement (Dissolved oxygen content, pH and temperature): Daily in all vessels with surviving fish, beginning at the start of the test (day 0).

OTHER TEST CONDITIONS
- Photoperiod: Photoperiod of 16h light and 8h darkness (LD, 16:8)
- Light intensity: Unspecified

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Range finding study: Yes
- Results used to determine the conditions for the definitive study: The final test was performed based on the results obtained in this preceding static combined limit/range-finding test (Control + WAFs prepared at 1, 10 and 100 mg/L).
- Test concentrations: Analysis of the samples taken at the start of the final test showed measured concentrations of 0.16, 0.45, 0.59 1.2 and 10.5 mg/L for the WAFs prepared at 4.6, 10, 22, 46 and 100 mg/L, respectively.
- Spacing factor for test concentrations: 2.2
Reference substance (positive control):
yes
Remarks:
The reference item was Pentachlorophenol (PCP)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.39 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: See field: "Any other information on results incl. tables"
- Mortality of control: No, See field: "Any other information on results incl. tables"
- Other adverse effects control: No, See field: "Any other information on results incl. tables"
- Any observations that might cause a difference between measured and nominal values: Analysis of the samples taken at the start of the final test showed measured concentrations of 0.16, 0.45, 0.59 1.2 and 10.5 mg/L for the WAFs prepared at 4.6, 10, 22, 46 and 100 mg/L, respectively. Measured concentrations decreased by 30 to 50% during the 1st 24-hour test period. Analyses of the two freshly prepared WAFs at 72 hours showed somewhat higher concentrations compared to the analysis at the start of the test and a slightly lower decrease of 18-37% over the 24-hour period (See Table VI in field "Any other information on results incl. tables")
Results with reference substance (positive control):
- Results with reference substance valid? The range of the 96h-LC50 for carp is generally between 0.10 and 0.46 mg/l based on historical data of reference tests performed approximately every 3 months from April 1988
until the end of 2000, and annually since then. Hence, the sensitivity of carp originating from the present batch for PCP falls within the range of sensitivities generally observed during the past years.
- See Table VII in field "Any other information on results incl. Tables"
- LC50: The 96h-LC50 for carp exposed to Pentachlorophenol was 0.24 mg/l.. This effect was already reached within 48 hours of exposure.
- Other: During the test the pH, oxygen concentration and the temperature of the medium were within the optimal ranges for fish.
Reported statistics and error estimates:
96h-LC50 should be considered estimates due to the fact that the analyses showed no increasing range of average measured concentrations that related to the prepared loading rates. Nonetheless, it can be safely concluded that all LC-values are definitely below 1 mg/L.
Table IV: Incidence of mortality and total mortality during the final test
1-Phenyldecane-1,3-dione
Average conc. (mg/L)
Initial Number of fish Cumulative mortality Total
3¼h 24h 48h 72h 96h Mortality (%)
Control 7 0 0 0 0 0 0
0.26 7 0 0 0 0 0 0
0.61 7 0 0 0 1 5 71
0.43 7 0 7 7 7 7 100
1.0 7 0 7 7 7 7 100
10.5 7 5 7 7 7 7 100

Table V: Clinical effects observed during the final test.
1-Phenyldecane-1,3-dione
Average conc. (mg/L)
Time of recording (hours) Specification of effects Relative
number
Control, 0.26 0-96 No effects 7/7
0.61 0-48 No effects 7/7
72 Loss of equilibrium 1/7
96 Hypoactive swimming 2/2
0.43 0-3¼ No effects 7/7
1.0 Immobilised 2/7
10.5 Immobilised 2/2

Table VI: Measured concentrations versus loading rates
1-Phenyldecane-1,3-dione
(WAF; mg/L)
Measured concentration (mg/L) Average
t=0h t=24h t=72h t=96h (mg/L)
4.6 0.162 0.0834 0.503 0.316 0.26
10 0.451 0.231 0.991 0.812 0.61
22 0.593 0.311 nm nm 0.43
46 1.21 0.88 nm nm 1.0
100 10.5 nm nm nm 10.5*
* Initial measured concentration instead of average.
nm; not measured (test groups not prepared, all fish dead) 

Table VII: Incidence of mortality observed in the reference study:

Concentration
PCP (mg/L)
Nominal
Initial
Number
Of fish
Cumulative number of dead fish recorded at various time points after start of exposure Total Mortality (%)
4h 24h 48h 72h 96h
0.1 5 0 0 0 0 0 0
0.22 5 0 0 1 1 1 20
0.46 5 0 5 5 5 5 100
Validity criteria fulfilled:
yes
Remarks:
OECD validity criteria, see field "overall remarks"
Conclusions:
The 96h-LC50 was estimated to correspond with 0.39 mg/L based on average measured exposure concentrations. Note that the 96h-LC50 should be considered as an estimate due to the fact that the analyses showed no increasing range of average measured concentrations that related to the prepared loading rates. Nonetheless, it can be safely concluded that the 96h-LC50 is below an actual exposure concentration of 1 mg/L.
Executive summary:

The acute toxicity of 1 -Phenyldecane-1,3 -dione, a reddish solid or reddish liquid (temperature dependent) with a purity of 86.4%, to Cyprinus carpio was investigated in a study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test item was not completely soluble in test medium at the concentrations tested. Note that the test item was melted at approximately 40°C before weighing.

A semi-static final test with daily renewals was performed based on the results obtained in a preceding static combined limit/range-finding test. Seven carp per test group were exposed to an untreated control and to WAFs prepared at 1-Phenyldecane-1,3-dione loading rates of 4.6, 10, 22, 46 and 100 mg/L. Samples for analytical confirmation of actual exposure concentrations were taken from fresh and 24-hour spent solutions during the first and last renewal interval.

Analysis of the samples taken at the start of the final test showed measured concentrations of 0.16, 0.45, 0.59 1.2 and 10.5 mg/L for the WAFs prepared at 4.6, 10, 22, 46 and 100 mg/L, respectively. Measured concentrations decreased by 30 to 50% during the 1st 24-hour test period. Analyses of the two freshly prepared WAFs at 72 hours showed somewhat higher concentrations compared to the analysis at the start of the test and a slightly lower decrease of 18-37% over the 24-hour period. The range tested based on average measured concentrations was 0.26, 0.61, 0.43, 1.0 and 10.5 mg/L for the WAFs prepared at 4.6, 10, 22, 46 and 100 mg/L, respectively.

The study met the acceptability criteria prescribed by the study plan and was considered valid.

The 96h-LC50 was estimated to correspond with 0.39 mg/L based on average measured exposure concentrations.

Note that the 96h-LC50 should be considered as an estimate due to the fact that the analyses showed no increasing range of average measured concentrations that related to the prepared loading rates. Nonetheless, it can be safely concluded that the 96h-LC50 is below an actual exposure concentration of 1 mg/L.

Description of key information

One study is available to cover this endpoint.

This key study carried out on Cyprinus carpio was realized in semi-static conditions according to the OECD Guideline 203 (Fish, Acute Toxicity Test) and GLP statement.

Seven carp per test group were exposed to an untreated control and to WAFs prepared at 1 -Phenyldecane-1,3 -dione loading rates of 4.6, 10, 22, 46 and 100 mg/L. Analysis of the samples taken at the start of the final test showed measured concentrations of 0.16, 0.45, 0.59 1.2 and 10.5 mg/L for the above WAFs. Measured concentrations decreased by 30 to 50% during the 1st 24-hour test period. Analyses of the two freshly prepared WAFs at 72 hours showed somewhat higher concentrations compared to the analysis at the start of the test and a slightly lower decrease of 18-37% over the 24-hour period. The range tested based on average measured concentrations was 0.26, 0.61, 0.43, 1.0 and 10.5 mg/L for the WAFs mentioned below.

The study met the acceptability criteria prescribed by the study plan and was considered valid.

The number of dead fish and the observations of fish behavior and toxic signs was followed. The 96h-LC50 was estimated to correspond with 0.39 mg/L based on average measured exposure concentrations. Note that the 96h-LC50 should be considered as an estimate due to the fact that the analyses showed no increasing range of average measured concentrations that related to the prepared loading rates. Nonetheless, it can be safely concluded that the 96h-LC50 is below an actual exposure concentration of 1 mg/L.

Key value for chemical safety assessment

LC50 for freshwater fish:
0.39 mg/L

Additional information