Urea, polymer with ethanedial

Short-term toxicity to fish

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:

April 2, 2013 - April 6, 2013.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control), 260.3, 364.4, 510.2, 714.3 and 1000 mg a.i./L.- Sampling method: To avoid incorrect TOC data (because of the test system) there was used special sampling method: Organic material released by the animals increase the NPOC content of the solutions, therefore parallel with the flasks containing animals other flasks containing no animals were run during the test. The analytical samples were taken from the flasks containing no animals at the beginning and end of the renewal periods. Three replicate were analyzed from each diluted analytical samples.- Sample storage conditions before analysis: The samples were analysed immediately after sampling or immediately after dilution of the samples.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances): Active ingredient content of the test substance (46.3 % in water solution) was taken into account in order to prepare the appropriate test solutions, therefore an amount of 3 x 10800 mg test substance was diluted in 3 x 5000 mL aquarium water (stock solution concentration: 1000 mg a.i./L). The test solutions were prepared by appropriate dilution of this stock solution.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM- Common name: Zebrafish- Source: Ágnes Németh (self-employed), Veszprém, Hungary- Length at study initiation (length definition, mean, range and SD): 3.0 – 3.6 cm- Weight at study initiation (mean and range, SD): 1.66 - 2.24 g- Method of breeding: Fish were bred in a well-known fish farm, under disease- and parasite-controlled conditions.- Feeding during test: The fish were not fed during the testACCLIMATION- Acclimation period: >12 days- Feeding: three times per week or daily until 24 hours before the start of the test- Health during acclimation (any mortality observed): Fish were healthy and there was no mortality of the population for seven days before the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

54 mg/L (as CaCO3)

Test temperature:

20.7 - 21.4 ºC

pH:

7.04 – 7.41

Dissolved oxygen:

94 – 96 % of the air saturation.

Nominal and measured concentrations:

Nominal concentrations: 0 (control), 260.3, 364.4, 510.2, 714.3 and 1000 mg a.i./L.Measured concentrations: 0 (control), 266.5; 383.7; 524.8; 756.6 and 1067.0 mg a.i./L.

Details on test conditions:

TEST SYSTEM- Test vessel: One aquarium for test group and control group respectively.- Material, size, headspace, fill volume: 5 L- Renewal rate of test solution (frequency/flow rate): 24 hours- No. of organisms per vessel: 7 - No. of vessels per concentration (replicates): 1- No. of vessels per control (replicates): 1- Biomass loading rate: Less than 1.0 g fish/LTEST MEDIUM / WATER PARAMETERS- Source/preparation of dilution water: dilution water (circulated, filtered)OTHER TEST CONDITIONS- Adjustment of pH: No.- Photoperiod: 16 hours light and 8 hours darknessEFFECT PARAMETERS MEASURED (with observation intervals if applicable): The observations were carried out at 3, 6, 24, 48, 72 and 96 hours. Mortality and any sub-lethal effect were determined in each observation period. The test conditions (pH, temperature, oxygen saturation) were measured at the start and at the end of each renewal period during the test. Body weight was measured were recorded before the introduction of fish and the body length was measured at the end of the test (or when mortality occurred).TEST CONCENTRATIONS- Spacing factor for test concentrations: 1.4- Range finding study: Two concentration range-finding tests were conducted to determine the approximate toxicity of the test item so that appropriate test concentrations can be selected for use in the definitive test to get the LC50 value of the test item- Test concentrations:Test 1: 0, 0.1, 1, 10 and 100 mg a.i./L (4 fish per dose, semi-static, 96 hours)Test 2: 0, 125, 250, 500 and 1000 mg a.i./L (4 fish per dose, semi-static, 96 hours)- Results used to determine the conditions for the definitive study:Test 1: 0, 0, 0, 0 and 0% of mortality respectivelyTest 2: 0, 0, 0, 25 and 75% of mortality respectively

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

562.77 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 504.20–868.29 mg a.i./L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

364.4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

Decreased activity was observed at the concentration of 714.3 mg a.i./L (nominal) at the observation of 48 and 72 hours, while swimming problems, decreased activity and localisation at the bottom of the aquaria could be observed at the highest concentration of 1000 mg a.i./L (nominal) at the observation of 24 and 48 hours. Behaviour of fish was found as normal at any other concentrations tested. There was no considerable difference observed concerning body weights between the groups.

Reported statistics and error estimates:

The 24, 48, 72 and 96 hours LC50 values were calculated by Probit analysis with 95 % confidence limits using SPSS PC+ software (based on the nominal concentrations). The NOEC, LOEC and LC100 were determined directly from the raw data.

Sublethal observations / clinical signs:

The 24h LC50 value: > 1000.0 mg a.i,/L

The 48h LC50 value: 772.65 mg a.i./L (with 95 % conf. limits: 639.77 – 955.18 mg a.i./L)

The 72h LC50 value: 632.39 mg a.i./L (with 95 % conf. limits: 528.11 – 761.32 mg a.i./L)

The 96h LC50 value: 562.77 mg a.i./L (with 95 % conf. limits: 504.20 – 868.29 mg a.i./L)

The 96h LC100 value: 714.3 mg a.i./L

The 96h No-Observed Effect Concentration (NOEC): 364.4 mg a.i./L

The 96h Lowest Observed Effect Concentration (LOEC): 510.2 mg a.i./L

Cumulative mortality data:

Concentration	Number of fish (initial population = 7)
Nominal (mg a.i./L)	3h	6h	24h	48h	72h	96h
Control	0	0	0	0	0	0
260.3	0	0	0	0	0	0
364.4	0	0	0	0	0	0
510.2	0	0	0	0	1	1
714.3	0	0	0	3	5	7
1000	0	0	0	6	7	-

Remark: There were 8 fish in the 364.4 mg a.i./L test group.

As the analytically measured concentration deviated not more than 20 per cent from the nominal in all case, the biological results are based on the nominal concentrations.

Validity criteria fulfilled:

yes

Remarks:

(conditions were maintained, no mortality in control, dissolved oxygen throughout the test approx. 94%, concentration of the test item > 80% of the nominal)

Conclusions:

The LC50 of test item in Zebrafish (Brachydanio rerio) at 96 h exposure period was 562.77 mg a.i./L and the NOEC was determined to be 364.4 mg a.i./L.

Executive summary:

An acute toxicity test in Zebrafish (Brachydanio rerio) was performed in accordance with EU Method C1 and OECD 203. Based on the preliminary range-studies, 7 fish per group were exposed to 0 (control) 260.3, 364.4, 510.2, 714.3 and 1000 mg a.i./L test item (corresponding to geometric mean test measured concentrations of 266.5, 383.7, 524.8, 756.6 and 1067.0 mg a.i./L) over an exposure period of 96 hours in a semi-static system. All the validity criteria were fulfilled. Decreased activity was observed at the concentration of 714.3 mg a.i./L (nominal) at the observation of 48 and 72 hours, while swimming problems, decreased activity and localisation at the bottom of the aquaria could be observed at the highest concentration of 1000 mg a.i./L (nominal) at the observation of 24 and 48 hours. Behaviour of fish was found as normal at any other concentrations tested. The LC50 of test item in Zebrafish (Brachydanio rerio) at 96 h exposure period was 562.77 mg a.i./L and the NOEC was determined to be 364.4 mg a.i./L.

Description of key information

Key study: Test method OECD 203. GLP study. The LC50 at 96 h exposure period was 562.77 mg a.i./L and the NOEC was determined to be 364.4 mg a.i./L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

562.77 mg/L

Additional information

Key study: An acute toxicity test in Zebra fish (Brachydanio rerio) was performed in accordance with EU Method C1 and OECD 203. 7 fish per group were exposed up to 1000 mg a.i./L test item over an exposure period of 96 hours in a semi-static system. The LC50 at 96 h exposure period was 562.77 mg a.i./L and the NOEC was determined to be 364.4 mg a.i./L.