Amidinourea phosphate

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:

18 Sept 2017 to 01 Dec 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992. OECD (2010): No.126 Short Guidance On The Threshold Approach for Acute Fish Toxicity, 2010.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 16VL8189
- Expiration date of the lot/batch: 04 Feb 2018
- Purity test date: 05 Aug 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: ambient (=< 30 deg C), dark, dry

Analytical monitoring:

yes

Details on sampling:

Analytical samples were taken from limit test concentration and control in 24-hours intervals. Analytical samples from control and the limit test concentration 35.0 mg/L were analysed at t = 0 h and t = 96 h

Vehicle:

no

Details on test solutions:

The necessary amount of test item for preparing the stock solution was weighed on a weighing scoop and transferred to a volumetric flask. Test medium was added up to the bench mark and the solution was homogenized by shaking and by ultrasonication for 10 minutes. Afterwards, the solution was clear and transparent. The solution was applied to the respective aquarium.
Nominal conc in test vessel: 35 mg/L
Test item required: 70mg
Final volume/application voume per test vessel: 2000 mL
Test solution per test vessel: 2 L

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Fish from the own stock breeding of the test facility were used. The parent animals were originally received from the University of Heidelberg (Heidelberg, Germany) in a healthy condition. According to the supplier they had been bred from wild strain (WA) zebrafish before. Since they have been purchased, the zebrafish were continuously reared in the laboratory of the testing facility. Only fish in good health and free from any apparent malformation were used. The last feeding was performed three days prior test start. The holding water was composed of reconstituted test water consisting of analytical grade salts dissolved in purified water. The ratio of Ca:Mg and Na:K was 4:1 and 10:1, respectively, based on molarity. The water parameters were controlled 3 times a week. The water temperature was between 23 -27 °C. The pH-value of the aerated water was between 6.0 - 8.5 and the total hardness was between 140 - 250 mg/L (as CaCO3), corresponding to 7.8 - 14°dH. For at least seven days prior to testing the dissolved oxygen was above 80 % saturation. The fish were exposed to light for 12 to 16 hours daily. The fish were fed each day with granular rearing food with approx. 2 % of their body-weight. The food was obtained from a commercial supplier.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

Fish were observed at 0, 4, 24, 48, 72 and 96 hours after test start.

Post exposure observation period:

None

Hardness:

13°dH corresponding to 232 mg CaCO3/L

Test temperature:

23.4 – 24.1 °C

pH:

7.19 – 7.97

Dissolved oxygen:

≥ 92 % of air saturation pH

Salinity:

Not applicable

Conductivity:

Not stated

Nominal and measured concentrations:

Nominal: 35 mg/L
Measured: 35.0 mg/L

Details on test conditions:

Test Medium
The test medium was reconstituted test water consisting of analytical grade salts dissolved in purified water with a ratio of Ca:Mg and Na:K of 4:1 and 10:1, respectively, based on molarity, and a conductivity of 710 µS/cm. The pH was between 6.0 and 8.5 and total hardness was 232 mg/L (as CaCO3), corresponding to 13°dH.
Test Units
For the test 2 L glass beakers, filled with 2 L of test solution were used.
Test Conditions
Temperature: 23.4 – 24.1 °C
Oxygen saturation: ≥ 92 % of air saturation
pH-value: 7.19 – 7.97
Hardness: 13°dH corresponding to 232 mg CaCO3/L
Exposure to light: 16 hours daily
Feeding: none
Loading: max. loading of 0.505 g fish/L
Aeration: continuous aeration of the test vessels with a membrane pump using a Pasteur pipette

Assessments
Mortality and Sub-lethal Effects
Fish were considered dead if there was no visible movement (e.g. gill movement), and if touching of the caudal peduncle produced no reaction. Records were made on visible abnormalities as: loss of equilibrium, swimming behaviour, respiratory function, pigmentation and all other observed events. Dead fish were removed if observed and mortality, length and weight was recorded. At termination of the test, all remaining fish were euthanized and all fish were weighed and measured.
Physico-Chemical Assessments
Measurements of temperature, pH-value and oxygen saturation were performed in 24 hour intervals in fresh and aged test solutions. Water hardness of the untreated medium was determined at the beginning of the test.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 35 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

35 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The LC50 (96 h) of the limit test was determined to be > 35.0 mg/L test item (nominal). The corresponding NOEC (mortality) (96 h) was 35.0 mg/L test item (nominal). No sublethal effects were observed in the control and in the limit test item concentration of 35.0 mg/L after 96 h.

Mortality
No mortality was observed in the control and in the limit test concentration of 35.0 mg/L.
Clinical signs
No sublethal effects were observed in the control and at the limit test concentration of 35.0 mg/L.
Length and weight
All fish were weighed and measured
The average weight of the test organisms was 0.1387 0.0299 g; the average length was 26 +/- 2 mm.
Analytical Results
The content of Guanyl urea phosphate in test solutions was analytically determined to confirm the correct application of the test item. The measured content of Guanyl urea phosphate in the fresh and aged test solution was 103 % of nominal. Since the content of Guanyl urea phosphate in the samples was between 80 and 120 % of nominal, the toxicological endpoints were evaluated using nominal test item concentrations.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not applicable. The measured content of Guanyl Urea Phosphate in the fresh and aged test solution was 103 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal, the toxicological endpoints were evaluated using nominal test item concentrations.

Validity criteria fulfilled:

yes

Conclusions:

The LC50 (96 h) of the test item was determined to be > 35.0 mg/L test item (nominal). The corresponding NOEC (mortality) (96 h) was 35.0 mg/L test item (nominal). No sublethal effects were observed in the control and the test item concentration of 35.0 mg/L after 96 h. The absence of mortality at the threshold concentration of 35.0 mg/L indicates that fish is not the most sensitive group of test organism and that the LC50 is greater than the limit test concentration.

Executive summary:

The short term toxicity to fish was determined in accordance with OECD Guideline 203 taking into consideration OECD (2010): No.126 Short Guidance On The Threshold Approach for Acute Fish Toxicity, 2010.

The test item was evaluated in a static limit test, 96 h exposure in treated test medium; 7 organisms ( zebrafish; Danio rerio) for the test concentration and the control were used. The test solution (35 mg/L) was prepared by dilution of the test item in test medium and application of a defined volume of the stock solution to the test vessel. Assessments on effects and mortality after 0, 4, 24, 48, 72 and 96 hours were conducted.

The content of Guanyl Urea Phosphate in test solutions was analytically determined to confirm the correct application of the test item. The measured content of Guanyl Urea Phosphate in the fresh and aged test solution was 103 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal, the toxicological endpoints were evaluated using nominal test item concentrations

The LC50 (96 h) of the test item was determined to be > 35.0 mg/L test item (nominal). The corresponding NOEC (mortality) (96 h) was 35.0 mg/L test item (nominal). No sublethal effects were observed in the control and the test item concentration of 35.0 mg/L after 96 h. The absence of mortality at the threshold concentration of 35.0 mg/L indicates that fish is not the most sensitive group of test organism and that the LC50 is greater than the limit test concentration.

Description of key information

The short term toxicity to fish was determined in accordance with OECD Guideline 203 taking into consideration OECD (2010): No.126 Short Guidance On The Threshold Approach for Acute Fish Toxicity, 2010.

The test item was evaluated in a static limit test, 96 h exposure in treated test medium; 7 organisms ( zebrafish; Danio rerio) for the test concentration and the control were used. The test solution (35 mg/L) was prepared by dilution of the test item in test medium and application of a defined volume of the stock solution to the test vessel. Assessments on effects and mortality after 0, 4, 24, 48, 72 and 96 hours were conducted.

The content of Guanyl Urea Phosphate in test solutions was analytically determined to confirm the correct application of the test item. The measured content of Guanyl Urea Phosphate in the fresh and aged test solution was 103 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal, the toxicological endpoints were evaluated using nominal test item concentrations

The LC50 (96 h) of the test item was determined to be > 35.0 mg/L test item (nominal). The corresponding NOEC (mortality) (96 h) was 35.0 mg/L test item (nominal). No sublethal effects were observed in the control and the test item concentration of 35.0 mg/L after 96 h. The absence of mortality at the threshold concentration of 35.0 mg/L indicates that fish is not the most sensitive group of test organism and that the LC50 is greater than the limit test concentration.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

35 mg/L