N-[2-[bis(carboxymethyl)amino]ethyl]-N-(1-oxononyl)Glycine

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:

2022-02-16 to 2022-10-20

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Patheon Austria GmbH & Co KG; lot/batch number: LFND3B1003
- Purity, including information on contaminants, isomers, etc.: 100 % (according to certificate of analysis)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature. Store in a dry place, keep away from moisture. Protect from heat and direct sunlight.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): The test solution was prepared as Water accommodated fraction (WAF) with a loading rate of 100 mg/L in Elendt M4 test medium and shaking for 24 hours in an overhead shaker at 24.4 - 25.1°C.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and 100 mg/L after 0 h and 48 h of exposure
- Sampling method: 3 mL samples were transferred from the test vessels directly into 8 mL glass vials.
- Sample storage conditions before analysis: To each vial, 3 mL acetonitrile was added and samples were stored at ≤ -18 °C in the dark.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test solution was prepared as Water accommodated fraction (WAF) with a loading rate of 100 mg/L in Elendt M4 test medium and shaking for 24 hours in an overhead shaker at 24.4 - 25.1°C.
- Controls: negative control with nutrition medium only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Test concentration separation factor: none since limit test

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Age at study initiation (mean and range, SD): 1 - 22 h old
- Source: from the clone breeding of the Federal Environment Agency, department IV 2.4, Berlin, and has been cultivated at Hydrotox GmbH since October 2012.
- Feeding during test: none

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS: The daphnids are cultured at 20 +/- 2 °C with 16 h light : 8 h dark and placed into fresh Elendt M4 medium twice a week. They are kept at approx. 10 animals / 200 mL and fed on workdays with Desmodesmus subspicatus algae corresponding to 0.1 mg C × daphnid-1 × day-1.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

18.9 – 20.1 °C

pH:

8.0 – 8.2 in the control and 6.4 – 7.4 in the test item treatments

Dissolved oxygen:

8.2 – 8.3 mg/L in the control and in the test item

Nominal and measured concentrations:

nominal: 100 mg/L
mean measured: 98.1 mg/L after 0 h and 99.6 mg/L after 48 h

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers 50 mL
- Material, size, headspace, fill volume: glass, 50 mL
- Volume of solution: 20 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 4 mL/daphnia

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium according to OECD 202 (2004)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The swimming capability of the daphnids was determined after 24 h and 48 h exposure.

RANGE-FINDING STUDY
- Test concentrations: 6.8, 16 and 100 mg/L test item
- Results used to determine the conditions for the definitive study: Nominal concentrations of 6.8, 16 and 100 mg/L test item were tested and resulted in in 0 %, 0 % and 5 % immobility (48 h), respectively.

Reference substance (positive control):

no

Remarks:

No positive control was used in this study. However, the recent quality testing was performed in November 2021 (GLP 1663) with EC50 (24h) = 2.02 mg/L, which is in the declared range of 0.6 – 2.1 mg/L (24 h) according to OECD 202.

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:

The measured test item concentrations in the test item treatments were 98.1 – 99.6 % of the nominal concentrations. As the measured test item concentration is within ± 20 % of the nominal concentration, according to OECD 202 (2004), the results are given in relation to the nominal test item concentration.
The immobilities of the test organisms in the control (NC) and the test item treatments are 0 % throughout the complete test duration of 48 h.

Results with reference substance (positive control):

Quality assurance takes place at regular intervals by testing the sensitivity of the test organisms to the reference item potassium dichromate (Honeywell Speciality Chemicals Seelze GmbH, Seelze, Germany; Lot no.: H1030). The recent quality testing was performed in November 2021 (GLP 1663) with EC50 (24h) = 2.02 mg/L, which is in the declared range of 0.6 – 2.1 mg/L (24 h) according to OECD 202.

Validity criteria:

Immobilisation of the test organisms in the control at the end of the test was 0 % and therefore ≤ 10 %.

Dissolved oxygen concentration in the control and the test item treatment at the end of the test was ≥ 8.2 mg/L and therefore ≥ 3 mg/L.

Validity criteria fulfilled:

yes

Remarks:

The test is valid according to OECD Test Guideline 202 (13 April 2004).

Conclusions:

The test item had no statistically significant effect on the immobilization of the daphnids at the nominal test concentration of 100 mg/L.

Executive summary:

The OECD 202 Guideline study was performed to GLP standards and met validity criteria. The 48 h acute toxicity of N-[2-[bis(carboxymethyl)amino]ethyl]-N-(1-oxononyl)Glycine to Daphnia magna was studied under static conditions. Test species were exposed to the test chemical at a nominal concentration of 100 mg/L for 48 h. Immobilisation was recorded daily. The 48 h EC50 was > 100 mg/l test item.
This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.
Results Synopsis
Test Organism: Daphnia magna (<24 h old)
Test Type: Static

EC50: > 100 mg/l (test item)
Endpoint(s) Effected: Mobility

Description of key information

The 48 hours acute EC50 was >100 mg/L, tested in a limit test following OECD 202.