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 aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

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:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted April 13, 2004

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Remarks:

HPLC analysis and UV/VIS-detection

Details on sampling:

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:

Pre-treatment of test item and preparation of test item concentrations
To produce the only test item concentration, 100.2 mg of the test item was added to 1 litre 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 7.8.
50 mL of the solution were taken and diluted with 0.5 mL of dilution water containing 5 daphnids resulting in a final concentration of 100 mg/L. For each test item concentration and the control 4 replicates were prepared.

Test organisms (species):

Daphnia sp.

Details on test organisms:

Name : Daphnia magna STRAUS, parthenogenetic females
Source : Strain of Bundesgesundheitsamt Berlin
Maintenance and Acclimatisation : A population of parthenogenetic females of synchronized age structure has been maintained for more than 20 years in the test facility under constant temperature conditions (20 +/- 2 °C) at a 16 : 8 hour light-dark photoperiod (light intensity: < 20 μE x m-2 x s-1). The culture water (so-called 'M4 medium') was partly renewed once a week. The Daphnia were exclusively fed unicellular green algae (Desmodesmus subspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period were recorded daily. The neonates were separated from their parent Daphnia by filtration prior to the acute test.

Test type:

static

Water media type:

freshwater

Remarks:

Reconstituted water (so-called 'M4 medium' according to OECD 202 and EC Method C.2, annex 1) was used for the maintenance of the test animals and the preparation of stock and test solutions of the test item.

Limit test:

yes

Total exposure duration:

48 h

Hardness:

14.2 °dH (= 253 mg/L CaCO3)

Test temperature:

18 - 22 °C (with a maximum temperature fluctuation of +/- 1 °C in each individual test)

pH:

7.8 - 8.1

Dissolved oxygen:

8.7 - 10.0 mg/L (95 - 101 %)

Nominal and measured concentrations:

100 mg/L

Details on test conditions:

Preparation of 10 L M4-Medium
The medium is prepared weekly and is aerated until usage (stability: 1 week). To reduce bacteria growth the prepared Medium is cleaned by filtration and continuous ultraviolet radiation.
40 mL of solution 1.1 (CaCl2 solution)
10 mL of solution 1.2 (MgSO4 x 7H2O solution)
10 mL of solution 1.3 (KCl solution)
10 mL of solution 1.4 (NaHCO3 solution)
1 mL of solution 2.1 (cationic solution)
5 mL of solution 2.2 (anionic solution)
2 mL of solution 2.3 (silicate solution)
50 mL of solution 2.4 (ion/ EDTA solution)
5 mL of solution 2.5 (phosphate solution)
1 mL of solution 2.6 (vitamin solution)
Make up with deionized water to 10 L.
After aeration the pH value of the medium will be ca. pH 8.3 – 8.4.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobilisation rates were recorded at 24-hour intervals. Additionally any abnormal behaviour or appearance of the Daphnia was reported every 24 hours.

Reported statistics and error estimates:

At a concentration of 100 mg/L no toxic effects against Daphnia were observed at the end of the 48 hour study period, thus no statistical analysis was required to determine the EC 50

Analysis

Nominal concentration	HPLC value [mg/L]
[mg/L]	at 0 hours	at 48 hours
control	---	< 0.858
100	100.701	97.707

Validity criteria fulfilled:

yes

Remarks:

The immobilisation and other abnormalities in the controls did not exceed 10 % by the end of the test. The dissolved oxygen concentration remained above 3 mg/L throughout the exposure period.

Conclusions:

An EC50 value of > 100 mg/L was determined after 48 h for EGGE 2806-1. No toxic effects against Daphnia were observed at a limit test concentration of 100 mg/L under exposure conditions.

Executive summary:

A study was performed to assess the acute toxicity of EGGE 2806-1 to Daphnia magna STRAUS under static conditions. The study was following OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test'. The Daphnia were exposed to a limit test concentration of nominally 100 mg/L of EGGE 2806-1 dissolved in dilution water. Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. An EC50 value of > 100 mg/L was determined after 48 h. No toxic effects against Daphnia were observed at a limit test concentration of 100 mg/L under exposure conditions.

The results are expressed in terms of nominal concentrations. .

Effective concentrations correspond to to 97.7% of nominal values at 48 hours.

Description of key information

A study was performed to assess the acute toxicity of EGGE 2806-1 to Daphnia magna STRAUS under static conditions. The study was following OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test'. The Daphnia were exposed to a limit test concentration of nominally 100 mg/L of EGGE 2806-1 dissolved in dilution water. Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. An EC50 value of > 100 mg/L was determined after 48 h. No toxic effects against Daphnia were observed at a limit test concentration of 100 mg/L under exposure conditions.

The results are expressed in terms of nominal concentrations. .

Effective concentrations correspond to to 97.7% of nominal values at 48 hours.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Should read >100 mg/L.