Reaction mass of 2-(3-{6-[3-(2-Hydroxyethyl)-3-methylureido]hexyl}-1-methylureido)ethanol and 6-[3-(2-Hydroxyethyl)-3-methylureido]hexylamino 3-(methylamino)propionate

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:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Remarks:

HPLC analysis and UV/VIS-detection

Details on sampling:

Chemical analysis
Storage: Routinely, the samples were analysed immediately. Only in exceptional cases, they were stored overnight deep frozen and protected from light.

Vehicle:

no

Details on test solutions:

A stock solution was prepared to give the desired series of test concentrations. 1000.5 mg of the test item were added to 10 litres of dilution water, treated for 1 h in an ultrasonic bath and stirred for 24 h on a magnetic stirrer. The pH was measured to be 8.0. To produce the different test item concentrations appropriate amounts of the stock solution were diluted with dilution water to a volume of 5000 mL. Finally 7 fish were given to all test item concentrations and the control.
Auxiliaries used to prepare the test media were an ultrasonic bath and a magnetic stirrer.
Stability of test concentration/s during exposure : Examined by chemical analysis (HPLC) at 0, 24, 48, 72 and 96 hours.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Source : Interaquaristik.de Shop (Germany)
Date supplied : 2016-06-15
Acclimatisation : Stock held since 2016-06-15 and acclimatised to the test conditions since then.
Temperature : 20 - 24 °C
Dissolved oxygen : > 5 mg/L
Feeding : Commercial fish food, daily. Feeding discontinued 24 h prior to test start.
Mortalities during acclimatisation period : < 5 %
Medication : none
Mean standard length (n = 21) : 3.57 cm (S.D. = 0.14 cm)

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

13.3 °dH (= 237 mg/L CaCO3)

Test temperature:

21.8 to 22.1 °C (with a maximum temperature fluctuation of +/- 0.7 °C in the test)

pH:

7.6 - 8.0

Dissolved oxygen:

7.8 - 8.9 mg/L (Oxygen saturation 91 - 100 %)

Nominal and measured concentrations:

Test item concentration/s : 50 and 100 mg/L

Details on test conditions:

Exposure conditions
Test vessels : glass aquaria holding 5 L of test media covered by glass plates
Experimental design : 2 test concentrations plus 1 control, 7 fish per test concentration, no feeding during the exposure period, static system
Method of initiation : fish were placed in prepared media
Loading : 0.81 g body weight (wet weight) per litre
Photoperiod : 16 h light: 8 h dark
Temperature : 21.4 to 22.1 °C
Aeration : gentle aeration via narrow glass tubes
Method of administration : stock solution
Medium renewal : none

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

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)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The criterion of death used in this study was the absence of response to physical stimulation. In addition to observations on mortality at 2, 24, 48, 72 and 96 hours, type and incidence of sub-lethal effects compared with control fish were observed.
Effective concentrations ranged from 100.1 % to 100.5 % of nominal values at 96 hours.

Reported statistics and error estimates:

As no toxic effects against fish were observed, no statistical analysis was required to determine the LC 50.

Sublethal observations / clinical signs:

Some test fish had a somewhat larger size than the one recommended in the test guideline for the zebra fish. This deviation is not regarded to be relevant to the results.

Validity criteria fulfilled:

yes

Remarks:

The mortality in the controls did not exceed 10 % by the end of the test. The dissolved oxygen concentration remained above 60 % of the air-saturation value throughout the exposure period. The pH did not vary by more than 1 unit.

Conclusions:

No toxic effects against fish were observed up to a test item, EGGE 2806-1, concentration of 100 mg/L.

Executive summary:

A study was performed to assess the acute toxicity of EGGE 2806-1 to Danio rerio under static conditions. The study was conducted in accordance with GLP and OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992). Groups of seven fish of the recommended size were exposed to a range of concentrations, nominally 50 and 100 mg/L of EGGE 2806-1 dissolved in dilution water. Observations were made on the numbers of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. No toxic effects against fish were observed up to a test item concentration of 100 mg/L (LC50 > 100 mg/L after 96 h). All results are expressed in terms of nominal concentrations.

Description of key information

A study was performed to assess the acute toxicity of EGGE 2806-1 to Danio rerio under static conditions. The study was conducted in accordance with GLP and OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992). Groups of seven fish of the recommended size were exposed to a range of concentrations, nominally 50 and 100 mg/L of EGGE 2806-1 dissolved in dilution water. Observations were made on the numbers of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. No toxic effects against fish were observed up to a test item concentration of 100 mg/L (LC50 > 100 mg/L after 96 h). All results are expressed in terms of nominal concentrations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Should read >100 mg/L