3-methyl-1,1-diphenylurea

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:

13.04.-24.04.2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

Principal investigator for analytical determination of the test substance concentrations was not Pavla Vlasáková, Dipl.Eng. In this study principal investigator was Jana Netušilová, Dipl.Eng., CETA, GLP Test Facility, Analytical Group I.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100, 50, 10, 5, 1 mg/L (measured at 0, 24, 48 and 96h)

Vehicle:

no

Details on test solutions:

The stock solution of the test substance was prepared in the dilution water. 100 mg of the test substance was weighed into 1000 mL of the dilution water for the preliminary test. Because the test substance was poorly soluble for dilution was used ultrasound for 1 hours and stirring for 2 hours. The concentrations of solutions used in the preliminary test were obtained by dilution of the stock solution with dilution water.

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM- Common name: Poecilia reticulata - Source: certified supplier EMPLA s.r.o., Hradec Králové (Czech Republic) - Age at study initiation: 3-4 months- Length at study initiation: 2.0-3.0 cm- Method of breeding: in all-glass aquaria in aerated drinking water.- Feeding during test: without feedingACCLIMATION- Acclimation period: 7 days in dilution water- Acclimation conditions: same as test- Feeding during test: without feeding- Food type: “Tropica” - feed for aquarium fish (manufacturer: DajanaPET, s.r.o., Bohuňovice- Frequency: once a day- Health during acclimation (any mortality observed): mortality control, The sensitivity of test species is periodically monitored by testing with the reference substance - potassium dichromate.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Ca + Mg ions 2.5 mmol.L-1

Test temperature:

22±2 °C

pH:

7.8±0.2

Dissolved oxygen:

air saturation value at 22 °C is 8.74

Nominal and measured concentrations:

100, 50, 10, 5, 1 mg/L

Details on test conditions:

TEST SYSTEM- Test vessel: glass beakers, fill volume 1000 ml- Type: open- Aeration: no- Renewal rate of test solution (frequency/flow rate): 24h- No. of organisms per vessel: 7 fishes for each concentration- No. of vessels per concentration (replicates): 1- No. of vessels per control (replicates): 1- Biomass loading rate: 1 g/lDILUTION WATERStandard dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D in 1000 mL of deionized water.A.117.6 g CaCl2.2H20 in 1L deionized waterB.49.3 g MgSO4.7H2O in 1L deionized waterC.25.9 g NaHCO3 in 1L deionized waterD.2.3 g KCl in 1L deionized waterThe dilution water should be of pH 7.8 ± 0.2, value measured in this study was 7.8. It is saturated by oxygen by the aeration at temperature of 20 – 24 °C. The content of sum Ca + Mg ions should have the value of 2.5 mmol.L-1, value measured in this study was 2.5 mmol.L-1OTHER TEST CONDITIONS- Photoperiod: natural variation of daily light and darkEFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The fish mortality was observed each 24 hours. The pH, temperature and dissolved oxygen were measured at the same intervals.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

See table No.: 1

Results with reference substance (positive control):

The sensitivity of the test species and correctness of test performance is periodically verified in six-month period by testing the reference substance, potassium dichromate.The results of verification test with K2Cr2O7 carried out in period from 13.04. to 17.04. 2015. are the following:48 hour – LC50 = 180 mg.L-1 (95% confidence limit: 153 – 206 mg.L-1)96 hour – LC50 = 165 mg.L-1 (95% confidence limit: 137 – 192 mg.L-1)

Sublethal observations / clinical signs:

Table No.: 1          Fish mortality – preliminary test

Nominal concentration	mg·L^-1	100	50	10	5	1	C
Number of live fish	2 hours	7	7	7	7	7	7
Mortality %	2 hours	0	0	0	0	0	0
Number of live fish	24 hours	7	7	7	7	7	7
Mortality %	24 hours	0	0	0	0	0	0
Number of live fish	48 hours	7	7	7	7	7	7
Mortality %	48 hours	0	0	0	0	0	0
Number of live fish	72 hours	7	7	7	7	7	7
Mortality %	72 hours	0	0	0	0	0	0
Number of live fish	96 hours	7	7	7	7	7	7
Mortality %	96 hours	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

No mortality was observed in the preliminary test. The exact values of LC50 could not be calculated and the values of LC are given in the form of a range. The LC100 and LC0 values were identified by direct observation.Based on no toxicity of the test substance found in preliminary test, the study was terminated. The experiment with the concentration of 100 mg•l-1 included in the preliminary test has been performed using 7 fish, therethrough the conditions of the limit test have been fullfiled. It was demonstrated that the LC50 is higher than this concentration.The analytical results from the preliminary test showed, that the test substance Akardit was stable in dilution water at conditions of the test, therefore the nominal concentrations were used for all evaluations and results.

Executive summary:

The test substance, Akardit, was tested for acute toxicity on fish Poecilia reticulata.

The test was performed according to method C.1 – Acute toxicity for fish, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

The preliminary test was performed in range of test substance nominal concentrations
1 – 100 mg·L^-1. Based on no toxicity of the test substance found in preliminary test,
the study was terminated.The experiment with the concentration of 100 mg·l^-1included
in the preliminary test has been performed using 7 fish, therethrough the conditions
of the limit test have been fullfiled. It was demonstrated that the LC₅₀is higher than this concentration.

 

Samples for analytical determination were taken at the beginning and at the end of the test. The analytical results from the preliminary test showed, that the test substance
Akarditwas stable in dilution water at conditions of the test, therefore the nominal concentrations were used for all evaluations and results.

The test was performed as static.

 

 Test results:                                     

24 hour – LC₅₀>100 mg·L^-1         (nominal concentration)

48 hour – LC₅₀>100 mg·L^-1         (nominal concentration)

72 hour – LC₅₀>100 mg·L^-1         (nominal concentration)

96 hour – LC₅₀>100 mg·L^-1         (nominal concentration)

 

24 hour – LC₀=100 mg·L^-1          (nominal concentration)

48 hour – LC₀=100 mg·L^-1          (nominal concentration)

72 hour – LC₀=100mg·L^-1          (nominal concentration)

96 hour – LC₀=100mg·L^-1          (nominal concentration)

 

24 hour – LC₁₀₀>100 mg·L^-1       (nominal concentration)

48 hour – LC₁₀₀>100 mg·L^-1       (nominal concentration)

72 hour – LC₁₀₀>100 mg·L^-1       (nominal concentration)

96 hour – LC₁₀₀>100 mg·L^-1        (nominal concentration

Description of key information

Akardit was tested for acute toxicity on fish Poecilia reticulata. The test was performed according to method C.1 – Acute toxicity for fish, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

Only one study is available.

GLP study.

Klimish score 1.

The analytical results from the preliminary test showed, that the test substance Akardit was stable in dilution water at conditions of the test, therefore the nominal concentrations were used for all evaluations and results.

96 hour – LC₅₀>100 mg·L^-1

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L